lignans and honokiol

Magnolol and honokiol are natural lignans with good physiological effects. As the main active substances derived from Magnolia officinalis, their pharmacological activities have attracted extensive attention. It is reported that both of them can cross the blood-brain barrier (BBB) and exert neuroprotective effects through a variety of mechanisms. This suggests that these two ingredients can be used as effective therapeutic compounds to treat a wide range of neurological diseases. This article provides a review of the mechanisms involved in the therapeutic effects of magnolol and honokiol in combating diseases, such as cerebral ischemia, neuroinflammation, Alzheimer's disease, and brain tumors, as well as psychiatric disorders, such as anxiety and depression. Although magnolol and honokiol have the pharmacological effects described above, their clinical potential remains untapped. More research is needed to improve the bioavailability of magnolol and honokiol and perform experiments to examine the therapeutic potential of magnolol and honokiol.

Topics: Biphenyl Compounds; Humans; Lignans; Signal Transduction

Honokiol (3',5-di-(2-propenyl)-1,1'-biphenyl-2,2'-diol) is a biologically active natural product derived from

Topics: Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Lignans; Lung Neoplasms; Signal Transduction

Magnolol and Honokiol are the primary active components that have been identified and extracted from Magnolia officinalis, and several investigations have demonstrated that they have significant pharmacological effects. Despite their therapeutic benefits for a wide range of illnesses, research on and the implementation of these compounds have been hindered by their poor water solubility and low bioavailability. Researchers are continually using chemical methods to alter their structures to make them more effective in treating and preventing diseases. Researchers are also continuously developing derivative drugs with high efficacy and few adverse effects. This article summarizes and analyzes derivatives with significant biological activities reported in recent research obtained by structural modification. The modification sites have mainly focused on the phenolic hydroxy groups, benzene rings, and diene bonds. Changes to the allyl bisphenol structure will result in unexpected benefits, including high activity, low toxicity, and good bioavailability. Furthermore, alongside earlier experimental research in our laboratory, the structure-activity relationships of magnolol and honokiol were preliminarily summarized, providing experimental evidence for improving their development and utilization.

Topics: Biphenyl Compounds; Lignans; Magnolia; Structure-Activity Relationship

Lignans are plant-derived polyphenolic compounds with a plethora of biological applications. Also, regarded as phytoestrogens, the lignans offer a variety of health benefits of which the anti-cancer effects are the most attractive. Honokiol is a lignan isolated from various parts of trees belonging to the genus Magnolia. The bioactivity of honokiol is attributed to its characteristic physical properties, which include small size and the presence of two phenolic groups that may interact with proteins in cell membranes via hydrophobic interactions, aromatic pi orbital co-valency, and hydrogen bonding. The hydrophobicity of honokiol enables its rapid dissolution in lipids and the crossing of physiological barriers, including the blood-brain barrier and cerebrospinal fluid. These factors contribute towards the high bioavailability of honokiol which further support its candidature in medicinal research. Therefore, the anticancer properties of honokiol are of particular interest as many of the contemporary anticancer drugs suffer from bioavailability drawbacks, which necessitates the identification and development of novel candidate molecules directed as anticancer chemotherapeutics. The antioncogenic profile of honokiol also arises from the regulation of various signalling pathways associated with oncogenesis, arresting of the cell cycle by regulation of cyclic proteins, upregulation of epithelial markers and downregulation of mesenchymal markers leading to the inhibition of epithelial-mesenchymal transition, and preventing the metastasis by restricting cell migration and invasion due to the downregulation of matrix-metalloproteinases. In this review, we discuss the anticancer properties of honokiol.

Topics: Biphenyl Compounds; Cell Line, Tumor; Humans; Lignans; Neoplasms; Phenols; Structure-Activity Relationship

Honokiol (HNK) is a small-molecule polyphenol that has garnered considerable attention due to its diverse pharmacological properties, including antitumor, anti-inflammatory, anti-bacterial, and anti-obesity effects. However, its clinical application is restricted by challenges such as low solubility, poor bioavailability, and rapid metabolism. To overcome these limitations, researchers have developed a variety of nano-formulations for HNK delivery. These nano-formulations offer advantages such as enhanced solubility, improved bioavailability, extended circulation time, and targeted drug delivery. However, existing reviews of HNK primarily focus on its clinical and pharmacological features, leaving a gap in the comprehensive evaluation of HNK delivery systems based on nanotechnology. This paper aims to bridge this gap by comprehensively reviewing different types of nanomaterials used for HNK delivery over the past 15 years. These materials encompass vesicle delivery systems, nanoparticles, polymer micelles, nanogels, and various other nanocarriers. The paper details various HNK nano-delivery strategies and summarizes their latest applications, development prospects, and future challenges. To compile this review, we conducted an extensive search using keywords such as "honokiol", "nanotechnology", and "drug delivery system" on reputable databases, including PubMed, Scopus, and Web of Science, covering the period from 2008 to 2023. Through this search, we identified and selected approximately 90 articles that met our specific criteria.

Topics: Biphenyl Compounds; Drug Delivery Systems; Lignans; Micelles; Nanoparticles; Nanotechnology

Black and Hispanic cancer patients have a higher incidence of cancer mortality. Many factors (e.g., socioeconomic differences, insufficient access to healthcare) contribute to racial disparity. Emerging research implicates biological disparity in cancer outcomes. Studies show distinct differences in the tumor immune microenvironment (TIME) in Black cancer patients. Studies also have linked altered mitochondrial metabolism to changes in immune cell activation in TIME. Recent publications revealed a novel immunomodulatory role for triphenylphosphonium-based mitochondrial-targeted drugs (MTDs). These are synthetically modified, naturally occurring molecules (e.g., honokiol, magnolol, metformin) or FDA-approved small molecule drugs (e.g., atovaquone, hydroxyurea). Modifications involve conjugating the parent molecule via an alkyl linker chain to a triphenylphosphonium moiety. These modified molecules (e.g., Mito-honokiol, Mito-magnolol, Mito-metformin, Mito-atovaquone, Mito-hydroxyurea) accumulate in tumor cell mitochondria more effectively than in normal cells and inhibit mitochondrial respiration, induce reactive oxygen species, activate AMPK and redox transcription factors, and inhibit cancer cell proliferation. Besides these intrinsic effects of MTDs in redox signaling and proliferation in tumors, MTDs induced extrinsic effects in the TIME of mouse xenografts. MTD treatment inhibited tumor-suppressive immune cells, myeloid-derived suppressor cells, and regulatory T cells, and activated T cells and antitumor immune effects. One key biological disparity in Black cancer patients was related to altered mitochondrial oxidative metabolism; MTDs targeting vulnerabilities in tumor cells and the TIME may help us understand this biological disparity. Clinical trials should include an appropriate number of Black and Hispanic cancer patients and should validate the intratumoral, antihypoxic effects of MTDs with imaging.

Topics: Atovaquone; Biphenyl Compounds; Black People; Health Status Disparities; Hispanic or Latino; Humans; Immune Checkpoint Inhibitors; Lignans; Mitochondria; Neoplasms; Oxidative Phosphorylation; Tumor Microenvironment

Honokiol is one of the natural extracts of Magnolia officinalis. It is a small molecule, lipophilic compound with extensive biological effects. It has been used in the treatment of multisystem diseases, including digestive diseases, endocrine diseases, nervous system diseases, and various tumors. This paper reviews the biological effects of honokiol on the treatment of skin diseases in recent years, including anti-microbial, anti-oxidant, anti-inflammatory, anti-tumor, anti-fibrosis, anti-allergy, photo-protection, and immunomodulation. Most current researches are focused on the effects of anti-melanoma and photo-protection. Therefore, we summarized the specific mechanisms about these two effects. On the other side of treating skin diseases, the advantages of topical drugs cannot be replaced. As a small molecule fat-soluble compound, honokiol is suitable for external use. We reviewed the advantages and disadvantages of the topical mixed cream and various improved methods. These improvements include physical and chemical penetration enhancers, drug carriers, and chemical derivatives. In conclusion, honokiol has a wide range of effects, and its topical preparation provides a safe and effective way for treating skin diseases.

Topics: Allyl Compounds; Antioxidants; Biphenyl Compounds; Dermatology; Humans; Lignans; Phenols; Skin Diseases

Plant lignans constitute an important group of polyphenols, which have been demonstrated to significantly induce cancer cell death and suppress cancer cell proliferation with minimal toxicity against non-transformed cells. Numerous epidemiological studies have shown that the intake of lignans is associated with lower risk of several cancers. These natural compounds have the potential to inhibit carcinogenesis, tumor growth, and metastasis by targeting various signaling molecules and pathways. Growing evidence indicates that honokiol and magnolol as natural lignans possess potent anticancer activities against various types of human cancer. The aim of present review is to provide the reader with the newest findings in understanding the cellular and molecular mechanisms mediating anticancer effects of honokiol and magnolol. This review comprehensively elucidates the effects of honokiol and magnolol on the molecular targets and signal transduction pathways implicated in cancer cell proliferation and metastasis. The findings of current review indicate that honokiol and magnolol can be considered as promising carcinopreventive and anticancer agents.

Topics: Antineoplastic Agents; Biphenyl Compounds; Humans; Lignans

Alzheimer's disease (AD) is the most common neurodegenerative disease, and its incidence is increasing worldwide with increased lifespan. Currently, there is no effective treatment to cure or prevent the progression of AD, which indicates the need to develop novel therapeutic targets and agents. Sirtuins, especially SIRT3, a mitochondrial deacetylase, are NAD-dependent histone deacetylases involved in aging and longevity. Accumulating evidence indicates that SIRT3 dysfunction is strongly associated with pathologies of AD, hence, therapeutic modulation of SIRT3 activity may be a novel application to ameliorate the pathologies of AD. Natural products commonly used in traditional medicine have wide utility and appear to have therapeutic benefits for the treatment of neurodegenerative diseases such as AD. The present review summarizes the currently available natural SIRT3 activators and their potentially neuroprotective molecular mechanisms of action that make them a promising agent in the treatment and management of neurodegenerative diseases such as AD.

Topics: Aging; Alzheimer Disease; Animals; Biphenyl Compounds; Dietary Supplements; Drug Delivery Systems; Enzyme Inhibitors; Glucosides; Humans; Hydrazines; Indazoles; Lignans; Neuroprotection; Phenols; Sirtuin 3

Honokiol is a pleiotropic compound which been isolated from Magnolia species such as Magnolia grandiflora and Magnolia dealbata. Magnolia species Magnolia grandiflora is used in traditional medicine for the treatment of various diseases.. The objective of this review is to summarize the pharmacological potential and therapeutic insights of honokiol.. Honokiol has been specified as a novel alternative to treat various disorders such as liver cancer, neuroprotective, anti-spasmodic, antidepressant, anti-tumorigenic, antithrombotic, antimicrobial, analgesic properties and others. Therefore, this study designed to represent the in-depth therapeutic potential of honokiol.. Literature searches in electronic databases, such as Web of Science, Science Direct, PubMed, Google Scholar, and Scopus, were performed using the keywords 'Honokiol', 'Health Benefits' and 'Therapeutic Insights' as the keywords for primary searches and secondary search terms were used as follows: 'Anticancer', 'Oxidative Stress', 'Neuroprotective', 'Antimicrobial', 'Cardioprotection', 'Hepatoprotective', 'Anti-inflammatory', 'Arthritis', 'Reproductive Disorders'.. This promising bioactive compound presented an wide range of therapeutic and biological activities which include liver cancer, neuroprotective, anti-spasmodic, antidepressant, anti-tumorigenic, antithrombotic, antimicrobial, analgesic properties, and others. Its pharmacokinetics has been established in experimental animals, while in humans, this is still speculative. Some of its mechanism for exhibiting its pharmacological effects includes apoptosis of diseased cells, reduction in the expression of defective proteins like P-glycoproteins, inhibition of oxidative stress, suppression of pro-inflammatory cytokines (TNF-α, IL-10 and IL-6), amelioration of impaired hepatic enzymes and reversal of morphological alterations, among others.. All these actions displayed by this novel compound could make it serve as a lead in the formulation of drugs with higher efficacy and negligible side effects utilized in the treatment of several human diseases.

Topics: Animals; Biphenyl Compounds; Humans; Lignans; Magnolia; Plant Extracts

Honokiol and its isomer magnolol are poly-phenolic compounds isolated from the Magnolia officinalis that exert cardiovascular modulating effects via a variety of mechanisms. They are used as blood-quickening and stasis-dispelling agents in Traditional Chinese Medicine and confirmed to have therapeutic potential in atherosclerosis, thrombosis, hypertension, and cardiac hypertrophy. This comprehensive review summarizes the current data regarding the cardioprotective mechanisms of those compounds and identifies areas for further research.

Topics: Biphenyl Compounds; Humans; Lignans; Magnolia; Medicine, Chinese Traditional

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Humans; Lignans; Magnolia; Molecular Targeted Therapy; Neoplasms

The tumor suppressor LKB1 is an essential serine/threonine kinase, which regulates various cellular processes such as cell metabolism, cell proliferation, cell polarity, and cell migration. Germline mutations in the STK11 gene (encoding LKB1) are the cause of the Peutz-Jeghers syndrome, which is characterized by benign polyps in the intestine and a higher risk for the patients to develop intestinal and extraintestinal tumors. Moreover, mutations and misregulation of LKB1 have been reported to occur in most types of tumors and are among the most common aberrations in lung cancer. LKB1 activates several downstream kinases of the AMPK family by direct phosphorylation in the T-loop. In particular the activation of AMPK upon energetic stress has been intensively analyzed in various diseases, including cancer to induce a metabolic switch from anabolism towards catabolism to regulate energy homeostasis and cell survival. In contrast, the regulation of LKB1 itself has long been only poorly understood. Only in the last years, several proteins and posttranslational modifications of LKB1 have been analyzed to control its localization, activity and recognition of substrates. Here, we summarize the current knowledge about the upstream regulation of LKB1, which is important for the understanding of the pathogenesis of many types of tumors.

Topics: AMP-Activated Protein Kinase Kinases; AMP-Activated Protein Kinases; Biphenyl Compounds; Calcium-Binding Proteins; Gene Expression Regulation, Neoplastic; Genes, Tumor Suppressor; Humans; Lignans; Lung Neoplasms; Lysosomes; Mutation; Peutz-Jeghers Syndrome; Phosphorylation; Protein Serine-Threonine Kinases; Sumoylation; Ubiquitination

Topics: Animals; Biphenyl Compounds; Drug Interactions; Humans; Lignans; Magnolia; Mutagenicity Tests; Plant Extracts; Tissue Distribution

Honokiol ((3',5-di-(2-propenyl)-1,1'-biphenyl-2,2'-diol), a lignan, is a promising antitumor compound, having exerted activity against a number of human cancer cell lines. Honokiol has inhibitory role on the proliferation, invasion and survival of cancer cells in in vitro as well as in vivo studies. It interferes with signaling pathways components in order to elicit the anticancer effect.. In present review, the published data on the efficacy of honokiol against various cancer cell lines and tumor-bearing animal models has been presented and discussed.. Honokiol lowers the expression of pluripotency-factors, the formation of mammosphere, P-glycoprotein expression, receptor CXCR4 level, c-FLIP, steroid receptor coactivator-3 (SRC-3), Twist1, matrix metalloproteinases, class I histone deacetylases, H3K27 methyltransferase among numerous other anticancer functions. It increases bone morphogenetic protein 7 (BMP7), Bax protein, among others. It does so by interfering with the major checkpoints such as nuclear factor kappa B (NF-κB), and activator of transcription 3 (STAT3), mammalian target of rapamycin (m-TOR), epidermal growth factor receptor (EGFR), Sonic hedgehog (SHH). It promotes the efficacy of several anticancer drugs and radiation tolerance. The derivatization of honokiol results in compounds with interesting attributes in terms of cancer control. This review will shed light on the scopes and hurdles in the relevance of the bioactive lignan honokiol in cancer management.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Humans; Lignans; Treatment Outcome

Magnolia bark contains magnolol, metabolized to tetrahydromagnolol and honokiol, with both GABA-ergic/cannabimimetic activities, hence of possible attraction to vulnerable individuals/recreational misusers.. A literature review, assessment of related anecdotal online Magnolia misuse's reports and an overview of Magnolia products' online acquisition possibilities has been here described.. No peer-reviewed papers about Magnolia abuse/misuse/dependence/addiction were identified. Conversely, from a range of websites emerged potentially 3 groups of Magnolia misusers: (a) subjects with a psychiatric history already treated with benzodiazepines, being attracted to Magnolia bark as a "natural sedative"; (b) polydrug misusers, ingesting Magnolia with a range of other herbs/plants, attracted by the GABA-ergic/cannabimimetic activities; (c) subjects naive to the misusing drugs' scenario, perceiving Magnolia as a natural dietary supplement/weight-control compound.. To the best of our knowledge, this is the first paper commenting on the possible Magnolia derivatives' potential of misuse. Magnolia's recent increase in popularity, mainly as a sedative, may be arguably due to its peculiar pharmacological properties/acceptable affordability levels/virtually worldwide favorable legal status and customers' attraction to a product being perceived as "natural" and hence somehow "safe." Future/potent/synthetic magnolol and honokiol structural analogues could however contribute to increasing the number of synthetic GABA-ergic/cannabimimetic misusing compounds.

Topics: Biphenyl Compounds; Humans; Lignans; Magnolia; Plant Bark; Plant Extracts; Substance-Related Disorders

The incidence of neurological disorders is growing in developed countries together with increased lifespan. Nowadays, there are still no effective treatments for neurodegenerative pathologies, which make necessary to search for new therapeutic agents. Natural products, most of them used in traditional medicine, are considered promising alternatives for the treatment of neurodegenerative diseases. Honokiol is a natural bioactive phenylpropanoid compound, belonging to the class of neolignan, found in notable amounts in the bark of Magnolia tree, and has been reported to exert diverse pharmacological properties including neuroprotective activities. Honokiol can permeate the blood brain barrier and the blood-cerebrospinal fluid to increase its bioavailability in neurological tissues. Diverse studies have provided evidence on the neuroprotective effect of honokiol in the central nervous system, due to its potent antioxidant activity, and amelioration of the excitotoxicity mainly related to the blockade of glutamate receptors and reduction in neuroinflammation. In addition, recent studies suggest that honokiol can attenuate neurotoxicity exerted by abnormally aggregated Aβ in Alzheimer's disease. The present work summarizes what is currently known concerning the neuroprotective effects of honokiol and its potential molecular mechanisms of action, which make it considered as a promising agent in the treatment and management of neurodegenerative diseases. © 2017 BioFactors, 43(6):760-769, 2017.

Topics: Amyloid beta-Peptides; Antioxidants; Biological Products; Biological Transport; Biphenyl Compounds; Blood-Brain Barrier; Central Nervous System; Cytokines; Gene Expression Regulation; Humans; Lignans; Magnolia; Nerve Growth Factors; Neurodegenerative Diseases; Neuroprotective Agents; Plant Bark; Receptors, Glutamate

Honokiol(HNK), one of major biological active constituents of Mangnolia officinalis, exerts a wide range of biological functions, such as moderate anticancer effects. It inhibits the growth of lung cancer, gastrointestinal cancer, head and neck squamous cell carcinoma, breast cancer, prostate cancer, ovarian cancer, in vitro and in vivo through multiple potential molecular targets. It modulates apoptosis-associated signaling pathway, inhibits growth factor receptor-mediated signal transduction pathway, blocks nuclear factor-κB signaling pathway, decreases the expression level of androgen receptors, subsides m TOR and STAT3 signaling pathway, and so on. HNK enhances the inhibitory effects of traditional anticancer drugs or targeted antitumor drugs in vitro and in vivo. It reverses multidrug resistances of cancer cells to cisplatin, doxorubicin and paclitaxol. Therefore, HNK plays a role in the augmentation of antitumor effects of cancer drugs and the reversal of multidrug resistance of tumor cells. HNK is a promising biochemical modulator of anti-cancer medicines in the cancer therapy.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Drug Resistance, Multiple; Drug Synergism; Humans; Lignans; Neoplasms; Signal Transduction; Therapies, Investigational

Honokiol and magnolol (an isomer of honokiol) are small-molecule polyphenols isolated from the barks of Magnolia officinalis, which have been widely used in traditional Chinese and Japanese medicines. In the last decade, a variety of biological properties of honokiol and magnolol (e.g., anti-oxidativity, antitumor activity, anti-depressant activity, anti-inflammatory activity, neuroprotective activity, anti-diabetic activity, antiviral activity, and antimicrobial activity) have been reported. Meanwhile, certain mechanisms of action of some biological activities were also investigated. Moreover, many analogs of honokiol and magnolol were prepared by structural modification or total synthesis, and some exhibited very potent pharmacological activities with improved water solubility. Therefore, the present review will provide a systematic coverage on recent developments of honokiol and magnolol derivatives in regard to semisynthesis, total synthesis, and structure-activity relationships from 2000 up to now.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Line; Humans; Isomerism; Lignans; Magnolia; Plant Bark; Structure-Activity Relationship

Aberrant activation of the hypoxia inducible factor (HIF) pathway causing overexpression of angiogenic genes, like vascular endothelial growth factor (VEGF), is one of the underlying causes of ocular neovascularization (NV) and metastatic cancer. Consistently, along with surgical interventions, a number of anti-VEGF agents have been approved by FDA for the treatment of ocular neovascular diseases. These anti-VEGF agents, like ranibizumab/lucentis, have revolutionized the treatment in the past decade. However, substantial vision improvement is observed only in a subset of age-related macular degeneration patients receiving ranibizumab. Further, all current therapies are associated with limitations and side effects. For example, surgeries cause tissue destruction and inflammation while anti-VEGF therapies are expensive, require repeated administration, and offer temporary relief from vascular leakage. These factors impose significant cost and treatment burdens to both the patient and society. With an aging population in most western countries with a continually increasing number of patients on lifelong treatment for these retinal diseases, the focus of ocular drug development for neovascular diseases will be to improve efficacy while reducing treatment costs. Blocking the HIF pathway, a major regulator of ocular NV and cancer, offers an appealing therapeutic strategy. Therefore, this review summarizes HIF inhibitors that have been recently evaluated for the treatment of different cancers and ischemic retinopathies.

Topics: Angiogenesis Inducing Agents; Angiogenesis Inhibitors; Animals; Anthracyclines; Biphenyl Compounds; Cardiac Glycosides; Humans; Hypoxia-Inducible Factor 1; Indazoles; Lignans; Models, Biological; Neoplasms; Neovascularization, Pathologic; Retinal Neovascularization; Vascular Endothelial Growth Factors

Cancer continues to be the leading cause of death worldwide. Plants have a long history of use in the treatment of cancer. Honokiol (HNK) is an important bioactive compound found in the bark of Magnolia tree, and has been shown to inhibit cancer growth and metastasis in many cell types in vitro and in animal models. Resistance to chemotherapy and radiotherapy is the major obstacle for cure of cancer. Combination of HNK with many traditional chemotherapeutic drugs as well as radiation sensitizes cancer cells to apoptotic death, suggesting that HNK not only directly inhibits primary cancers and metastasis, but also has potential to overcome drug resistance. Ultimately, this may mean that HNK could be combined with traditional chemotherapies administered at lower doses to significantly reduce toxicity, meanwhile enhance efficacy. As a natural compound, HNK is composed of polyphenols and has been described in many studies targeting multiple key cell signaling molecules. Mitochondria are the main hub for cellular energy production and play an important role in cell survival, and are the key target identified for HNK to mediate cancer cell death, survival, and metastasis. In this review, we have summarized different aspects of HNK's anti-cancer effects from recent accumulated literature, as well as the underlying molecular mechanisms. This review is primarily focused on the effects of HNK on epidermal growth factor receptor (EGFR) and signal transduction and activator of transcription 3 (STAT3) signaling, as well as the broader regulation of mitochondrial function and cancer cell metabolism.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Line, Tumor; Disease Models, Animal; ErbB Receptors; Humans; Lignans; Magnolia; Mitochondria; Neoplasms; Plant Extracts; Polyphenols

SIRT3 belongs to a highly conserved protein family of histone deacetylases and it is rich in mitochondria. As acetyl-modification is one of the important post-translational modifications that prevail in the mitochondria, it is not surprising that SIRT3 plays a key regulatory role in this organelle. SIRT3 has a wide range of substrates that are involved in the physiological and pathological processes of oxidative stress, ischemia-reperfusion injury, mitochondrial metabolism homeostasis and cellular death. These pathophysiological processes are considered as the underlying mechanisms of diseases like cardiac hypertrophy, myocardial infarction and heart failure, indicating the potential roles of SIRT3 in cardiovascular diseases. In this review, we will summarize the emerging roles and therapeutic implications of SIRT3 in cardiovascular diseases by providing an update on the latest understanding of its functions.

Topics: Animals; Biphenyl Compounds; Cardiovascular Diseases; Enzyme Induction; Humans; Lignans; Oxidative Stress; Reactive Oxygen Species; Sirtuin 3

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Proliferation; Humans; Lignans; Neoplasms; Signal Transduction; Ultraviolet Rays

Despite the development of more advanced medical therapies, cancer management remains a problem. Head and neck squamous cell carcinoma (HNSCC) is a particularly challenging malignancy and requires more effective treatment strategies and a reduction in the debilitating morbidities associated with the therapies. Phytochemicals have long been used in ancient systems of medicine, and non-toxic phytochemicals are being considered as new options for the effective management of cancer. Here, we discuss the growth inhibitory and anti-cell migratory actions of proanthocyanidins from grape seeds (GSPs), polyphenols in green tea and honokiol, derived from the

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Carcinoma, Squamous Cell; Catechin; Cell Line, Tumor; Cell Movement; Cell Survival; Drug Evaluation, Preclinical; Head and Neck Neoplasms; Humans; Lignans; Magnolia; Polyphenols; Proanthocyanidins; Squamous Cell Carcinoma of Head and Neck; Tea; Vitis

Developed regions, including Japan, have become "aged societies," and the number of adults with senile dementias, such as Alzheimer's disease (AD), Parkinson's disease, and Huntington's disease, has also increased in such regions. Neurotrophins (NTs) may play a role in the treatment of AD because endogenous neurotrophic factors (NFs) prevent neuronal death. However, peptidyl compounds have been unable to cross the blood-brain barrier in clinical studies. Thus, small molecules, which can mimic the functions of NFs, might be promising alternatives for the treatment of neurodegenerative diseases. Natural products, such as or nutraceuticals or those used in traditional medicine, can potentially be used to develop new therapeutic agents against neurodegenerative diseases. In this review, we introduced the neurotrophic activities of polyphenols honokiol and magnolol, which are the main constituents of Magnolia obovata Thunb, and methanol extracts from Zingiber purpureum (BANGLE), which may have potential therapeutic applications in various neurodegenerative disorders.

Topics: Alzheimer Disease; Animals; Biphenyl Compounds; Cells, Cultured; Dietary Supplements; Hippocampus; Humans; Lignans; Magnolia; Mice; Molecular Weight; Nerve Growth Factors; Neurodegenerative Diseases; Neurogenesis; Phytotherapy; Polyphenols; Rats; Structure-Activity Relationship; Zingiberales

Honokiol (3,5-di-(2-propenyl)-1,1-biphenyl-2,2-diol) is a natural bioactive neolignan isolated from the genus Magnolia. In recent studies, honokiol has been observed to have anti-angiogenic, anticancer, anti-inflammatory, neuroprotective and GABA-modulating properties in vitro and in preclinical models. Honokiol and its analogs target multiple signaling pathways including NF-κB, STAT3, EGFR, mTOR and caspase-mediated common pathway, which regulate cancer initiation and progression. Honokiol and its targets of action may be helpful in the development of effective analogs and targeted cancer therapy. In this review, recent data describing the molecular targets of honokiol and its analogs with anticancer and some other bioactivities are discussed.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Caspases; Humans; Lignans; Magnolia; Neoplasms; NF-kappa B; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Phospholipase D; Signal Transduction; Structure-Activity Relationship; TOR Serine-Threonine Kinases

Honokiol (3',5-di-(2-propenyl)-1,1'-biphenyl-2,4'-diol) is a bioactive natural product derived from Magnolia spp. Recent studies have demonstrated anti-inflammatory, anti-angiogenic, anti-oxidative and anticancer properties of honokiol in vitro and in preclinical models. Honokiol targets multiple signaling pathways including nuclear factor kappa B (NF-κB), signal transducers and activator of transcription 3 (STAT3), epidermal growth factor receptor (EGFR) and mammalian target of rapamycin (m-TOR), which have great relevance during cancer initiation and progression. Furthermore, pharmacokinetic profile of honokiol has revealed a desirable spectrum of bioavailability after intravenous administration in animal models, thus making it a suitable agent for clinical trials. In this review, we discuss recent data describing the molecular targets of honokiol and its anti-cancer activities against various malignancies in pre-clinical models. Evaluation of honokiol in clinical trials will be the next step towards its possible human applications.

Topics: Animals; Anti-Inflammatory Agents; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; ErbB Receptors; Humans; Lignans; Magnolia; Mice; Neoplasms; NF-kappa B; Plant Preparations; Rats; Signal Transduction; STAT3 Transcription Factor; TOR Serine-Threonine Kinases

Honokiol is a small-molecule pharmacologically active component which has various medicinal applications. Increasing interest is paid on its multifunctional anti-tumor effects including inducing tumor cell death, anti-angiogenesis, anti-migration and anti-multiple drug resistance. We addressed a brief summary of the anti-tumor actions and potential applications of honokiol. This review is mainly focused on the multiple types of cell death induced by honokiol, and its potential role in overcoming multiple drug resistance.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Death; Cell Movement; Drug Resistance, Neoplasm; Humans; Lignans

Chinese herbs have been and still are widely used as important remedies in Oriental medicine. Over the recent years, a variety of biologically active constituents have been isolated from these sources and confirmed to have multifunctional activity in experimental studies. Honokiol is a small-molecule polyphenol isolated from the genus Magnolia. It is accompanied by other related polyphenols, including magnolol, with which it shares certain biological properties. Recently, honokiol and magnolol have been found to have anti-oxidative, anti-inflammatory, anti-tumor, and anti-microbial properties in preclinical models, without appreciable toxicity. These findings have increased interest in bringing honokiol and magnolol to the clinic as novel therapeutic agents in dermatology. In this review, the findings concerning the major mechanisms of action of honokiol and magnolol are described. Knowledge of the multiple activities of honokiol and magnolol can assist with the development of honokiol and magnolol derivatives and the design of clinical trials that will maximize the potential benefit of honokiol and magnolol in the patient setting for dermatologic disorders.

Topics: Antioxidants; Biphenyl Compounds; Dermatologic Agents; Drugs, Chinese Herbal; Humans; Lignans; Skin Diseases

Honokiol is a small-molecule polyphenol isolated from the genus Magnolia. It is accompanied by other related polyphenols, including magnolol, with which it shares certain biologic properties. Recently, honokiol has been found to have antiangiogenic, antiinflammatory, and antitumor properties in preclinical models, without appreciable toxicity. These findings have increased interest in bringing honokiol to the clinic as a novel chemotherapeutic agent. In addition, mechanistic studies have tried to find the mechanism(s) of action of honokiol, for two major reasons. First, knowledge of the mechanisms of action may assist development of novel synthetic analogues. Second, mechanistic actions of honokiol may lead to rational combinations with conventional chemotherapy or radiation for enhanced response to systemic cancers. In this review, we describe the findings that honokiol has two major mechanisms of action. First, it blocks signaling in tumors with defective p53 function and activated ras by directly blocking the activation of phospholipase D by activated ras. Second, honokiol induces cyclophilin D, thus potentiating the mitochondrial permeability transition pore, and causing death in cells with wild-type p53. Knowledge of the dual activities of honokiol can assist with the development of honokiol derivatives and the design of clinical trials that will maximize the potential benefit of honokiol in the patient setting.

Topics: Angiogenesis Inhibitors; Animals; Antineoplastic Agents; Antiviral Agents; Biphenyl Compounds; Cell Line, Tumor; Lignans; Mice; Reactive Oxygen Species

This review deals with the advances in the study of chemical constituents and analytical methods of the medicinal plant Houpo.

Topics: Alkaloids; Biphenyl Compounds; Chromatography; Drugs, Chinese Herbal; Lignans; Oils, Volatile

In men, the level of testosterone decreases with age. At the skin level, the result is observed as a decrease in density and in a lower elasticity. Identifying compounds that are able to increase the level of testosterone appears to be an attractive strategy to develop new antiaging bioactive ingredients for men. Reverse pharmacognosy was successfully applied to identify new natural compounds able to modulate testosterone levels. Among several in silico hits, honokiol was retained as a candidate as it has the greatest potential to become an active ingredient. This result was then validated in vitro on aromatase and 5-alpha-reductase type 1 and 2, which are two types of enzymes implicated in the degradation of free testosterone. Indeed, honokiol was identified as an inhibitor of aromatase, with a half-maximal inhibitory concentration (IC(50)) of about 50 μM. In addition, honokiol was shown to be an inhibitor of 5-alpha-reductase type 1, with an IC(50) of about 75 μM. Taken together, these data indicate that honokiol modulates testosterone levels, and its structure has the potential to serve as a lead for future designs of highly selective inhibitors of 5-alpha-reductase type 1.

Topics: 3-Oxo-5-alpha-Steroid 4-Dehydrogenase; Animals; Aromatase; Biphenyl Compounds; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Enzyme Inhibitors; Humans; Lignans; Magnolia; Male; Middle Aged; Skin Aging; Structure-Activity Relationship; Testosterone

Alcoholic liver disease is one of the leading causes of liver-related morbidity and mortality worldwide, but effective treatments are still lacking. Honokiol, a lignin-type natural compound isolated from the leaves and bark of Magnolia plants, has been widely studied for its beneficial effects on several chronic diseases. Accumulating studies have revealed that honokiol displays a potential therapeutic effect on alcoholic liver disease. In this study, the protective activity of honokiol on alcoholic liver disease was confirmed due to its significant inhibitory activity on the expression levels of inflammatory cytokines (such as tumor necrosis factor-alpha, interleukin-6, and interleukin-1

Topics: Animals; Cytokines; Interleukin-1beta; Interleukin-6; Lignans; Lipid Metabolism; Lipid Metabolism Disorders; Liver; Liver Diseases, Alcoholic; Mice; NF-kappa B; PPAR alpha; Sterols; Tumor Necrosis Factor-alpha

Plant-derived substances as antifungal agents have received considerable attention in recent years to reduce the use of chemical fungicides in food preservatives. In this study, honokiol, a type of phenolic compound in Magnolia officinalis, was found to inhibit spore germination and mycelia growth of Aspergillus flavus at 100 μg/mL. In addition, a pathogenicity assay showed that honokiol had potent antifungal activity against A. flavus in corn flour by suppressing conidia production. Fluorescence staining, transmission electron microscopy and biochemical assays were performed to explore its possible inhibition mechanisms against A. flavus. The results showed that the destructive effect of honokiol on A. flavus appeared to be related to increased plasma membrane permeability, the inhibition of ATPase activity, mitochondrial dysfunction and the accumulation of reactive oxygen species. Furthermore, a transcriptomic analysis showed that honokiol treatment resulted in 1578 different expressed genes. Gen Ontology and Kyoto Encyclopedia of Genes and Genomes analysis revealed that some genes of A. flavus related to spore development, integrity of the cell wall and membrane, oxidative stress and energy metabolism were significantly downregulated. In addition, RNA-seq results were validated by quantitative real-time polymerase chain reactions. Our finding enhanced the understanding of the antifungal activity of honokiol and underlying mechanisms of action at the molecular level, supporting honokiol as a potential agent in preventing contamination by A. flavus.

Topics: Antifungal Agents; Aspergillus flavus; Lignans; Transcriptome

This study aimed to investigate the antibiofilm and anticariogenic effects of honokiol, a traditional Chinese medicine, on the cariogenic bacterium Streptococcus mutans (S. mutans).. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of honokiol on S. mutans UA159 were measured. Then, S. mutans were treated with honokiol at concentrations of 1/2 MIC and 1/4 MIC. Extracellular polysaccharide (EPS) synthesis was assessed with confocal laser scanning microscopy (CLSM) and the anthrone-sulfuric method. Crystal violet staining and scanning electron microscopy (SEM) were used to demonstrate the characteristics and morphology of S. mutans biofilms. Colony-forming unit (CFU) assay was performed to observe the antibacterial effect of honokiol. Lactic acid production of 24-h biofilms was measured by the lactic acid assay. The expression level of caries-related genes (gtfB/C/D, comD/E and ldh) was identified by quantitative real-time PCR (qRTPCR) to explore the relevant mechanism. And the cytotoxic effect on human gingival fibroblasts (HGFs) was evaluated by the Cell Counting Kit-8 (CCK-8) assay.. The MIC and MBC of honokiol on S. mutans were 30 μg/mL and 60 μg/mL, respectively. Honokiol inhibited biofilm formation, EPS synthesis and lactic acid production. It also decreased the expression of glucosyltransferases (Gtfs) and quorum sensing (QS) system encoding genes. Moreover, honokiol showed favorable biocompatibility with HGFs.. Honokiol has an inhibitory effect on S. mutans and favorable biocompatibility, with application potential as a novel anticaries agent.

Topics: Biofilms; Dental Caries; Humans; Lactic Acid; Lignans; Streptococcus mutans

Honokiol is a natural bisphenol neolignan present in the bark of

Topics: Antioxidants; Biphenyl Compounds; Free Radical Scavengers; Free Radicals; Lignans; Phenols

The understanding of the use of Magnolia officinalis L. (Magnoliaceae) as a possible dietary supplement for supporting the treatment of airway pathologies might be of clinical interest. Two commercially available bark extracts (M. officinalis extract [MOE]) were characterized by quantitation in honokiol and magnolol content by means of high-performance liquid chromatography with UV detection. MOE effects, as well as those of the reference compounds per se, on some targets connected to airway pathologies (antibacterial- and lung and trachea relaxing- activities) were investigated. Results showed that MOE possessed interesting antibacterial activity against Staphylococcus aureus, Pseudomonas aeruginosa, and Streptococcus pneumoniae. This was accompanied by a spasmolytic and antispasmodic activity, possibly owing to its ability to concurrently modulate different targets such as H

Topics: Biphenyl Compounds; Humans; Lignans; Magnolia; Medicine, Chinese Traditional; Plant Bark; Plant Extracts; Respiratory Tract Diseases

Obesity is a complex disease defined as an excessive amount of body fat. It is considered a risk factor for several pathologies; therefore, there is an increasing interest in its treatment. Pancreatic lipase (PL) plays a key role in fat digestion, and its inhibition is a preliminary step in the search for anti-obesity agents. For this reason, many natural compounds and their derivatives are studied as new PL inhibitors. This study reports the synthesis of a library of new compounds inspired by two natural neolignans, honokiol (1) and magnolol (2) and bearing amino or nitro groups linked to a biphenyl core. The synthesis of unsymmetrically substituted biphenyls was achieved through an optimisation of the Suzuki-Miyaura cross-coupling reaction followed by the insertion of allyl chains, thus furnishing the O- and/or N-allyl derivatives, and finally, a sigmatropic rearrangement yielding in some cases, the C-allyl analogues. Magnolol, honokiol and the twenty-one synthesised biphenyls were evaluated for their in vitro inhibitory activity toward PL. Three compounds (15b, 16 and 17b) were more effective inhibitors than the natural neolignans (magnolol IC

Topics: Biphenyl Compounds; Lignans

Alzheimer's Dementia (AD) and Vascular Dementia (VaD) are two main types of dementias for which no specific treatment is available. Chronic Cerebral Hypoperfusion (CCH) is a pathogenesis underlying AD and VaD that promotes neuroinflammatory responses and oxidative stress. Honokiol (HNK) is a natural compound isolated from magnolia leaves that can easily cross blood brain barrier and has anti-inflammatory and antioxidant effects. In the present study, the effects of HNK on astrocyte polarization and neurological damage in in vivo and in vitro models of chronic cerebral hypoperfusion were explored. We found that HNK was able to inhibit the phosphorylation and nuclear translocation of STAT3, A1 polarization, and reduce conditioned medium's neuronal toxicity of astrocyte under chronic hypoxia induced by cobalt chloride; STAT3 phosphorylation inhibitor C188-9 was able to mimic the above effects of HNK, suggesting that HNK may inhibit chronic hypoxia-induced A1 polarization in astrocytes via STAT3. SIRT3 inhibitor 3-TYP reversed, while Sirt3 overexpression mimicked the inhibitory effects of HNK on oxidative stress, STAT3 phosphorylation and nuclear translocation, A1 polarization and neuronal toxicity of astrocyte under chronic hypoxic conditions. For in vivo research, continuous intraperitoneal injection of HNK (1 mg/kg) for 21 days ameliorated the decrease in SIRT3 activity and oxidative stress, inhibited astrocytic STAT3 nuclear translocation and A1 polarization, and prevented neuron and synaptic loss in the hippocampal of CCH rats. Besides, HNK application improved the spatial memory impairment of CCH rats, as assessed with Morris Water Maze. In conclusion, these results suggest that the phytochemical HNK can inhibit astrocyte A1 polarization via regulating SIRT3-STAT3 axis, thus improving CCH-induced neurological damage. These results highlight HNK as novel treatment for dementia with underlying vascular mechanisms.

Topics: Animals; Astrocytes; Brain Ischemia; Dementia; Lignans; Rats; Sirtuin 3

Topics: Adenocarcinoma; Biphenyl Compounds; Humans; Lignans

Topics: Choline; Deep Eutectic Solvents; Humans; Lignans; Male; Powders; Solvents

The weight coefficients of appearance traits, extract yield of standard decoction, and total content of honokiol and magnolol were determined by analytic hierarchy process(AHP), criteria importance though intercrieria correlation(CRITIC), and AHP-CRITIC weighting method, and the comprehensive scores were calculated. The effects of ginger juice dosage, moistening time, proces-sing temperature, and processing time on the quality of Magnoliae Officinalis Cortex(MOC) were investigated, and Box-Behnken design was employed to optimize the process parameters. To reveal the processing mechanism, MOC, ginger juice-processed Magnoliae Officinalis Cortex(GMOC), and water-processed Magnoliae Officinalis Cortex(WMOC) were compared. The results showed that the weight coefficients of the appearance traits, extract yield of standard decoction, and total content of honokiol and magnolol determined by AHP-CRITIC weighting method were 0.134, 0.287, and 0.579, respectively. The optimal processing parameters of GMOC were ginger juice dosage of 8%, moistening time of 120 min, and processing at 100 ℃ for 7 min. The content of syringoside and magnolflorine in MOC decreased after processing, and the content of honokiol and magnolol followed the trend of GMOC>MOC>WMOC, which suggested that the change in clinical efficacy of MOC after processing was associated with the changes of chemical composition. The optimized processing technology is stable and feasible and provides references for the modern production and processing of MOC.

Topics: Biphenyl Compounds; Drugs, Chinese Herbal; Lignans; Magnolia; Zingiber officinale

Topics: Biphenyl Compounds; Cell Membrane; Chromatography; Lignans; Vascular Endothelial Growth Factor A

Most current larynx cancer therapies are generally aimed at the global mass of tumor, targeting the non-tumorigenic cells, and unfortunately sparing the tumorigenic cancer stem cells (CSCs) that are responsible for sustained growth, metastasis, and chemo- and radioresistance. Phytochemicals and herbs have recently been introduced as therapeutic sources for eliminating CSCs. Therefore, we assessed the anti-tumor effects of two herbal ingredients, the green tea extract "Epigallocatechin-3-gallate (EGCG)" and Honokiol (HNK), on parental cells or CD44

Topics: Apoptosis; Biphenyl Compounds; Catechin; Cell Line, Tumor; Hedgehog Proteins; Humans; Hyaluronan Receptors; Laryngeal Neoplasms; Lignans; Neoplastic Stem Cells; Tumor Suppressor Protein p53

Development of cancer, including renal cancer, is a major problem in immunosuppressed patients. The mTOR inhibitor Rapamycin (RAPA) is used as an immunosuppressive agent in patients with organ transplants and other immunological disorders; and it also has antitumorigenic potential. However, long-term use of RAPA causes reactivation of Akt, and ultimately leads to enhanced tumor growth. Honokiol (HNK) is a natural compound, which possesses both anti-inflammatory and antitumorigenic properties. In this study, we investigated the effect of a novel combination therapy using RAPA + HNK on allograft survival and post-transplantation renal tumor growth. We observed that it effectively modulated the expression of some key regulatory molecules (like Carabin, an endogenous Ras inhibitor; and Rubicon, a negative regulator of autophagy) that play important roles in tumor cell growth and survival. This combination induced toxic autophagy and apoptosis to promote cancer cell death; and was associated with a reduced expression of the tumor-promoting receptor tyrosine kinase AXL. Finally, we utilized a novel murine model to examine the effect of RAPA + HNK on post-transplantation renal tumor growth. The combination treatment prolonged the allograft survival and significantly inhibited post-transplantation tumor growth. It was associated with reduced tumor expression of Rubicon and the cytoprotective/antioxidant heme oxygenase-1 to overcome therapeutic resistance. It also downregulated the coinhibitory programmed death-1 ligand, which plays major role(s) in the immune escape of tumor cells. Together, this combination treatment has a great potential to restrict renal tumor growth in transplant recipients as well as other immunosuppressed patients.

Topics: Animals; Apoptosis; Autophagy; Biphenyl Compounds; Cell Line, Tumor; Humans; Intracellular Signaling Peptides and Proteins; Kidney Neoplasms; Lignans; Mice; Organ Transplantation; Sirolimus; TOR Serine-Threonine Kinases

Honokiol (HO) exerts neuroprotective effects in several animal models of Alzheimer's disease (AD), but the poor dissolution hampers its bioavailability and therapeutic efficacy.. A novel honokiol nanoscale drug delivery system (Nano-HO) with smaller size and excellent stability was developed in this study to improve the solubility and bioavailability of HO. The anti-AD effects of Nano-HO was determined.. Male TgCRND8 mice were daily orally administered Nano-HO or HO at the same dosage (20 mg/kg) for 17 consecutive weeks, followed by assessment of the spatial learning and memory functions using the Morris Water Maze test (MWMT).. Our pharmacokinetic study indicated that the oral bioavailability was greatly improved by Nano-HO. In addition, Nano-HO significantly improved cognitive deficits and inhibited neuroinflammation via suppressing the levels of TNF-α, IL-6 and IL-1β in the brain, preventing the activation of microglia (IBA-1) and astrocyte (GFAP), and reducing β-amyloid (Aβ) deposition in the cortex and hippocampus of TgCRND8 mice. Moreover, Nano-HO was more effective than HO in modulating amyloid precursor protein (APP) processing via suppressing β-secretase, as well as enhancing Aβ-degrading enzymes like neprilysin (NEP). Furthermore, Nano-HO more markedly inhibited tau hyperphosphorylation via decreasing the ratio of p-Tau (Thr 205)/tau and regulating tau-related apoptosis proteins (caspase-3 and Bcl-2). In addition, Nano-HO more markedly attenuated the ratios of p-JNK/JNK and p-35/CDK5, while enhancing the ratio of p-GSK-3β (Ser9)/GSK-3β. Finally, Nano-HO prevented the gut microflora dysbiosis in TgCRND8 mice in a more potent manner than free HO.. Nano-HO was more potent than free HO in improving cognitive impairments in TgCRND8 mice via inhibiting Aβ deposition, tau hyperphosphorylation and neuroinflammation through suppressing the activation of JNK/CDK5/GSK-3β signaling pathway. Nano-HO also more potently modulated the gut microbiota community to protect its stability than free HO. These results suggest that Nano-HO has good potential for further development into therapeutic agent for AD treatment.

Topics: Alzheimer Disease; Animals; Biphenyl Compounds; Cognition; Cognitive Dysfunction; Gastrointestinal Microbiome; Glycogen Synthase Kinase 3 beta; Lignans; Male; Mice; Neuroinflammatory Diseases

Nanomedicine, constructed from therapeutics, presents an advantage in drug delivery for cancer therapies. However, nanocarrier-based treatment systems have problems such as interbatch variability, multicomponent complexity, poor drug delivery, and carrier-related toxicity. To solve these issues, the natural molecule honokiol (HK), an anticancer agent in a phase I clinical trial (CTR20170822), was used to form a self-assembly nanoparticle (SA) through hydrogen bonding and hydrophobicity. The preparation of SA needs no molecular precursors or excipients in aqueous solution, and 100% drug-loaded SA exhibited superior tumor-targeting ability due to the enhanced permeability and retention (EPR) effect. Moreover, SA significantly enhanced the antitumor immunity relative to free HK, and the mechanism has notable selectivity to the p53 pathway. Furthermore, SA exhibited excellent physiological stability and inappreciable toxicity. Taken together, this supramolecular self-assembly strategy provides a safe and "molecular economy" model for rational design of clinical therapies and is expected to promote targeted therapy of HK, especially in colorectal cancer patients with obvious p53 status.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biocompatible Materials; Biphenyl Compounds; Colorectal Neoplasms; Female; Humans; Immunotherapy; Lignans; Macromolecular Substances; Materials Testing; Mice; Mice, Inbred BALB C; Mice, Nude; Molecular Structure; Neoplasms, Experimental; Small Molecule Libraries; Tumor Cells, Cultured; Tumor Suppressor Protein p53

Magnolia officinalis constitutes a traditional Korean medicine used for the treatment of atopic dermatitis, and honokiol is an active diphenyl compound present in Magnolia officinalis.. The aim of the study was to investigate the therapeutic effects of honokiol on atopic dermatitis in vivo.. The therapeutic effects of honokiol were evaluated in a 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis model.. Administration of honokiol (10 mg/kg) significantly suppressed mast cell accumulation and inflammation induced by DNCB in skin tissues. Moreover, DNCB-induced increases in serum immunoglobulin E levels were reversed by honokiol treatment. In addition, DNCB-induced elevation of inflammatory cytokines (interleukin (IL)-4, IL-13, IL-17, and interferon-γ) in the skin and lymph nodes was significantly ameliorated by honokiol administration. Furthermore, the increase in lymph nodes sizes induced by DNCB treatment was reduced by honokiol administration.. DNCB-induced atopic responses in the ears and lymph nodes were significantly suppressed by honokiol treatment. These results suggested that honokiol is a potential therapeutic agent for atopic dermatitis.

Topics: Animals; Biphenyl Compounds; Cytokines; Dermatitis, Atopic; Dinitrochlorobenzene; Immunoglobulin E; Inflammation; Lignans; Magnolia; Male; Mast Cells; Mice; Mice, Inbred BALB C

Topics: Animals; Biphenyl Compounds; Breast Neoplasms; Drug Carriers; Female; Humans; Lignans; Mice; Nanoparticles; Particle Size; Sialic Acids; Tissue Distribution; Zein

Magnolia officinalis Cortex (M. officinalis) is a classical traditional Chinese medicine (TCM) widely used to treat digestive system diseases. It effectively regulates gastrointestinal motility to improve abdominal pain, abdominal distension and other symptoms. Magnolol (MAG) and honokiol (HON) are the main pharmacodynamic components responsible for the gastrointestinal activity of M. officinalis.. The transient receptor potential (TRP) family is highly expressed in the gastrointestinal tract and participates in the regulation of gastrointestinal motility, visceral hypersensitivity, visceral secretion and other physiological activities. In this study, the calcium-lowering mechanisms of MAG and HON contributing to the smooth muscle relaxation associated with TRP are discussed.. The relaxation smooth muscle effects of MAG and HON were tested by the isolated intestine tone tests. A synthetic MAG probe (MAG-P) was used to target fishing for their possible target. The distribution of MAG on the smooth muscle was identified by a molecular tracer based on chemical biology. Ca. After confirming the smooth muscle relaxation in the small intestine induced by MAG and HON, the relaxation effect was identified mainly due to the downregulation of intracellular calcium by controlling external calcium influx. Although MAG and HON inhibited both TRPV4 and TRPC4 channels to reduce calcium levels, the inhibitory effect on TRPC4 channels is an important mechanism of their smooth muscle relaxation effect, since TRPC4 is widely expressed in the small intestinal smooth muscle cells.. The inhibition of MAG and HON on TRPC4 channels contributes to the relaxation of intestinal smooth muscle.

Topics: Animals; Biphenyl Compounds; Calcium Signaling; HEK293 Cells; Humans; Intestines; Lignans; Male; Medicine, Chinese Traditional; Muscle Contraction; Muscle, Smooth; Myocytes, Smooth Muscle; Rats; Rats, Sprague-Dawley; TRPC Cation Channels; TRPV Cation Channels

Despite advantages of arsenic trioxide (ATO) in oncological practice, its clinical applications have been hampered by severe cardiotoxicity. The general mechanism of ATO-induced cardiotoxicity has been attributed to its damage to mitochondria, resulting in cardiac remodeling. Honokiol (HKL) is a naturally occurring compound derived from Magnolia bark. Previous studies have demonstrated that HKL exerts cardio-protective effects on ischemia/reperfusion (I/R) or chemical-induced cardiotoxicity by counteracting the toxic effects on mitochondria. The present study was conducted to investigate whether HKL pretreatment protects against ATO-induced cardiac oxidative damage and cell death. For the in vitro study, we evaluated the effects of ATO and/or Honokiol on reactive oxygen species (ROS) production and apoptosis induction in primary cultured cardiomyocytes; for the in vivo study, BALB/c mice were administrated with ATO and/or HKL for a period of 4 weeks, myocardial apoptosis, cardiac function, and cardiac remodeling (cardiac hypertrophy and cardiac fibrosis) were assessed at the end of administration. Our results demonstrated Honokiol pretreatment alleviated the ATO-induced boost in ROS concentration and the following apoptosis induction in primary cultured cardiomyocytes. In the mouse model, Honokiol pretreatment ameliorated ATO-induced myocardial apoptosis, cardiac dysfunction, and cardiac remodeling. Collectively, these results indicated that Honokiol provide a protection against ATO-induced cardiotoxicity by reducing mitochondrial damage. In addition, given that Honokiol has shown considerable suppressive effects on leukemia cells, our data also imply that ATO and Honokiol combination may possibly be a superior avenue in leukemia therapy.

Topics: Animals; Apoptosis; Arsenic Trioxide; Biphenyl Compounds; Cardiotoxicity; Lignans; Magnolia; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mitochondria; Myocytes, Cardiac; Oxidative Stress; Reactive Oxygen Species

We aimed to examine the effect of Honokiol (HKL) on endothelial dysfunction in type 2 diabetic rats and its possible mechanism. A high-fat diet and streptozotocin (STZ) were used to establish the type 2 diabetic model in rats. Part of these rats were intraperitoneally injected with HKL 10 mg/kg daily. Then the expression of Ser1177 phosphorylation of endothelial nitric oxide synthase (p-eNOS), eNOS, and CD31, vasodilation function, insulin signaling, indicators of oxidative stress and relative signaling pathway were measured. Human umbilical vein endothelial cells (HUVECs) were used to explore the underlying mechanism of the effect of HKL on high glucose-related endothelial injury in vitro. The data showed that HKL could reverse the decline of the expression of p-eNOS and CD31, endothelium-related vasodilation dysfunction, insulin resistance and activation of oxidative stress induced by type 2 diabetes in vivo. The similar results were obtained in vitro. In summary, our study demonstrates that HKL improves endothelial function and diminishes insulin resistance and oxidative stress, suggesting that HKL could be used as a treatment option for diabetes in the future.

Topics: Animals; Biphenyl Compounds; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Endothelium, Vascular; Human Umbilical Vein Endothelial Cells; Humans; Insulin Resistance; Lignans; Nitric Oxide; Nitric Oxide Synthase Type III; Oxidative Stress; Rats; Vascular Diseases; Vasodilation

Mitochondrial damage and oxidative stress are crucial contributors to the tubular cell injury and death in acute kidney injury. Novel therapeutic strategies targeting mitochondria protection and halting the progression of acute kidney injury are urgently needed. Honokiol is a small-molecule polyphenol that exhibits extraordinary cytoprotective effects, such as anti-inflammatory and anti-oxidative. Thus, we investigated whether honokiol could ameliorate cisplatin-induced acute kidney injury via preventing mitochondrial dysfunction.. Acute kidney injury was induced by cisplatin administration. Biochemical and histological analysis were used to determine kidney injury. The effect of honokiol on mitochondrial function and morphology were determined using immunohistochemistry, transmission electron microscopy, immunoblot and immunofluorescence. To investigate the mechanism by which honokiol alters mitochondrial dynamics, remodelling and resistance to apoptosis, we used transfection experiments, immunoblotting, immunoprecipitation and flow cytometry assay.. We demonstrated that the prominent mitochondrial fragmentation occurred in experimental models of cisplatin-induced nephrotoxicity, which was coupled to radical oxygen species (ROS) overproduction, deterioration of mitochondrial function, release of apoptogenic factors and the consequent apoptosis. Honokiol treatment caused notable reno-protection and attenuated of these cisplatin-induced changes. Mechanistically, honokiol treatment recovered the expression of SIRT3 and improved AMPK activity in tubular cells exposure to cisplatin, which preserved the Drp1 phosphorylation at Ser637 and blocked its translocation in mitochondria, consequently preventing mitochondrial fragmentation and subsequent cell injury and death.. Our results indicate that honokiol may protect against cisplatin-induced acute kidney injury by preserving mitochondrial integrity and function by SIRT3/AMPK-dependent mitochondrial dynamics remodelling.

Topics: Acute Kidney Injury; AMP-Activated Protein Kinases; Apoptosis; Biphenyl Compounds; Cisplatin; Humans; Lignans; Mitochondrial Dynamics; Sirtuin 3

Honokiol can reduce the MI/RI-induced cTnT and CK-MB levels, apoptosis index, and mitochondrial swelling in cardiomyocytes via activating the PI3K/AKT signaling pathway.. Honokiol provides cardiac protection from MI/RI by suppressing mitochondrial apoptosis through the PI3K/AKT signaling pathway.

Topics: Animals; Apoptosis; Biphenyl Compounds; Humans; Leukemia; Lignans; Lymphoma, B-Cell; Male; Mice; Mice, Inbred C57BL; Myocardial Reperfusion Injury; Myocytes, Cardiac; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Signal Transduction

Local administration of therapeutic agents provides a favorable approach to enhance drug accumulation at pathological sites. In this study, a novel honokiol nanosuspensions loaded thermosensitive injectable hydrogels (HK-NS-Gel) was designed as the local delivery system for the combination therapy with systemic paclitaxel (PTX). The formed HK-NS-Gel showed superior gelation time and temperature. In vitro release and in vivo drug retention assay showed that HK-NS-Gel can slowly and steadily release the HK during 12 days. Meanwhile, enhanced PTX accumulation in the tumor was observed after intratumoral injection of HK-NS-Gel. In vitro cytotoxicity and cell apoptosis tests against 4T1 cells proved the synergistic effects of free PTX combined with HK-NS-Gel. In vivo antitumor study was conducted on 4T1 bearing mice, indicating that co-administration HK-NS-Gel and PTX could effectively enhance tumor growth suppression, and the tumor inhibitory rate was as high as 72.51%. In conclusion, intravenous delivery of PTX combined with intratumoral delivery of HK-NS-Gel was a promising combination for breast cancer therapy with enhanced therapeutic response and safety.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Breast Neoplasms; Cell Line, Tumor; Female; Humans; Hydrogels; Lignans; Mice; Nanoparticles; Paclitaxel

Both PCAT19 and miR-25-3p have been reported in lung cancer studies, but whether there is a correlation between the two and whether they jointly regulate the progress of lung cancer have not been reported yet. Therefore, this study carried out a further in-depth research. The expression of PCAT19 was detected in lung cancer (LC) tissues and cells by quantitative real-time polymerase chain reaction (qRT-PCR). The effect of PCAT19 on tumor growth was detected in a tumor-bearing model of nude mice. PCAT19-transfected cells were treated with Honokiol and anisomycin. The effects of PCAT19 on proliferation, apoptosis, and cycle of LC cells were investigated by biomolecule experiments. The effects of PCAT19 on the expressions of mitogen-activated protein kinase- (MAPK-) related proteins were evaluated by western blotting. The expression of PCAT19 was decreased in LC tissues and related to patient survival, tumor size, and pathology. In addition, upregulation of PCAT19 hindered LC cell proliferation, miR-25-3p expression, and the activation of extracellular regulated protein kinases (ERK) 1/2, p38, and c-Jun N-terminal kinase (JNK), while facilitating LC cell apoptosis. Furthermore, upregulation of PCAT19 reversed the effects of Honokiol and anisomycin on promoting cell proliferation and inhibiting cell apoptosis. Collectively, our findings show that upregulated PCAT19 suppresses proliferation yet promotes the apoptosis of LC cells through modulating the miR-25-3p/MAP2K4 signaling axis.

Topics: Animals; Anisomycin; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Movement; Cell Proliferation; Gene Expression Regulation, Neoplastic; Humans; Lignans; Lung Neoplasms; MAP Kinase Kinase 4; Mice; Mice, Nude; MicroRNAs; RNA, Long Noncoding

Honokiol, isolated from a traditional Chinese medicine (TCM) Magnolia officinalis, is a biphenolic compound with several biological activities. To improve and broaden its biological activity, herein, two series of honokiol thioethers bearing 1,3,4-oxadiazole moieties were prepared and assessed for their α-glucosidase and SARS-CoV-2 entry inhibitory activities. Among all the honokiol thioethers, compound 7l exhibited the strongest α-glucosidase inhibitory effect with an IC

Topics: alpha-Glucosidases; Angiotensin-Converting Enzyme 2; Biphenyl Compounds; COVID-19 Drug Treatment; HEK293 Cells; Humans; Lignans; Molecular Docking Simulation; Oxadiazoles; Protein Binding; SARS-CoV-2; Spike Glycoprotein, Coronavirus; Sulfides

Cardiovascular diseases have consistently been the leading cause of death in the United States over the last two decades, with 30% of the adult American population having hypertension. The metabolites of arachidonic acid (AA) in the kidney play an important role in blood pressure regulation. The present study investigates the antihypertensive effect of honokiol (HON), a naturally occurring polyphenol, and examines its correlation to the modulation of AA metabolism.. Spontaneously hypertensive rats (SHR) were randomly divided into four groups. Treatment groups were administered HON intraperitoneally at concentrations of 5, 20, and 50 mg/kg. Blood pressure was monitored at seven-day intervals. After a total of 3 weeks of treatment, the rats were euthanized and the kidney tissues were collected to examine the activity of the two major enzymes involved in AA metabolism in the kidney, namely cytochrome P450 (CYP)4A and soluble epoxide hydrolase (sEH).. Rats treated with HON did not experience the rise in blood pressure observed in the untreated SHR. High-dose HON significantly reduced blood pressure and inhibited the activity and protein expression of the CYP4A enzyme in the rat kidney. The activity of the sEH enzyme in renal cytosol was significantly inhibited by medium and high doses of HON.. Our data demonstrate the antihypertensive effect of HON and provide a novel mechanism for its underlying cardioprotective properties.

Topics: Animals; Antihypertensive Agents; Arachidonic Acid; Biphenyl Compounds; Blood Pressure; Cytochrome P-450 CYP4A; Hypertension; Kidney; Lignans; Rats; Rats, Inbred SHR

Oxidative stress and mitochondrial dysfunction contribute greatly to fluoride-induced cognitive impairment and behavioural disorders. Honokiol, a natural biphenolic compound, possesses antioxidant and mitochondrial protective properties. The present study investigated the protective actions of honokiol on NaF-elicited cognitive deficits and elucidated the possible mechanism of honokiol-mediated protection. The results demonstrated that honokiol administration markedly attenuated fluoride-induced cognitive impairments and neural/synaptic injury in mice. Moreover, honokiol elevated the activity and expression of SOD2 and promoted mtROS scavenging through Sirt3 activation in NaF-treated mice and SH-SY5Y cell lines. Meanwhile, honokiol substantially lowered mtROS production by enhancing Sirt3-mediated mitochondrial DNA (mtDNA) transcription, thereby leading to significant increases in ATP synthesis and complex I activity. Further studies revealed that honokiol activated AMPK and upregulated the PGC-1α and Sirt3 protein expression in vivo and in vitro. Intriguingly, the protective actions of honokiol on oxidative stress and mitochondrial dysfunction were abolished by AMPK shRNA or Sirt3 shRNA. Notably, AMPK knockdown prevented the increase in PGC-1α and Sirt3 expression induced by honokiol, while Sirt3 shRNA suppressed Sirt3 signaling without significant effects on p-AMPK and PGC-1α expression. In conclusion, our findings indicate that honokiol mitigates NaF-induced oxidative stress and mitochondrial dysfunction by regulating mtROS homeostasis, partly via the AMPK/PGC-1α/Sirt3 pathway, which ultimately contributes to neuronal/synaptic injury and cognitive deficits.

Topics: AMP-Activated Protein Kinases; Animals; Biphenyl Compounds; Cognition; Cognitive Dysfunction; DNA, Mitochondrial; Fluorides; Humans; Lignans; Mice; Mitochondria; Neuroblastoma; Oxidative Stress; RNA, Small Interfering; Sirtuin 3

Infections caused by drug-resistant bacteria have emerged to be one of the greatest threats to global public health, and new antimicrobial agents with novel mechanisms of action hence are in an urgent need to combat bacterial resistance. Herein, we reported the design, synthesis, and antibacterial evaluation of novel honokiol derivatives as mimics of antimicrobial peptides (AMPs). These mimics showed potent antimicrobial properties against Gram-positive bacteria. Among them, the most promising compound 13b exhibited excellent antibacterial activity, rapid bactericidal properties, avoidance of antibiotic resistance, and weak hemolytic and cytotoxic activities. In addition, compound 13b not only inhibited the biofilm formation but also destroy the preformed biofilm. Mechanism studies further revealed that compound 13b killed bacteria rapidly by interrupting the bacterial membrane. More intriguingly, compound 13b exhibited potent in vivo antibacterial efficacy in a mouse septicemia model induced by Staphylococcus aureus ATCC43300. These results highlight the potential of 13b to be used as therapeutic agents.

Topics: Allyl Compounds; Animals; Anti-Bacterial Agents; Anti-Infective Agents; Bacteria; Biphenyl Compounds; Lignans; Mice; Microbial Sensitivity Tests; Phenols

The stem bark of Magnolia officinalis is a traditional Chinese medicine for the treatment of abdominal distention and functional dyspepsia. The pharmacokinetics of three glycosides (magnoloside A, magnoloside B, and syringin) and two lignans (honokiol and magnolol) in both normal and functional dyspepsia rats were firstly investigated by ultra-performance liquid chromatography-triple quadrupole mass spectrometry method and the influences of the coexisting compounds on the pharmacokinetic parameters of honokiol and magnolol were also studied. It was found that all of the five target compounds were quickly absorbed and eliminated in both normal and functional dyspepsia rats, while, their residence time was significantly decreased in pathological states except magnoloside A. The coexisting compounds in the stem bark of M. officinalis significantly reduced absorption and increased elimination of honokiol in vivo. It's worth noticing that the volume of distribution of lignan was quite lower than that of a glycoside. Moreover, the metabolic profiling of magnoloside A, honokiol, and magnolol in vivo was analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry method, from which three prototypes were identified and 35 metabolites were putatively characterized, and 18 unknown metabolites were reasonably characterized for the first time. The results indicated that sulfation and glucuronidation were the main metabolic pathways of honokiol and magnolol.

Topics: Animals; Biphenyl Compounds; Chromatography, High Pressure Liquid; Chromatography, Liquid; Dyspepsia; Glycosides; Lignans; Magnolia; Plant Bark; Rats; Tandem Mass Spectrometry

Among the sirtuin enzymes, Sirt3 is one of the most important deacetylases as it regulates acetylation levels in mitochondria, which are linked to the metabolism of multiple organs and therefore involved in many types of age-related human diseases such as cancer, heart diseases and metabolic diseases. Given the dearth of direct activators of Sirt3, the identification of new modulators could be a key step in the development of new therapeutics. Here we report the discovery of Sirt3 modulators, including activators, through the use of DNA encoded library technology (DEL) and computational high-throughput screening methodologies. Top hits from both screenings against Sirt3 were evaluated according to their activity and affinity. Our best activator is more potent than the previously reported activator Honokiol. Docking studies suggest that our activators identified from virtual screening interact with Sirt3 at a site similar to Honokiol, whereas the activators identified from DEL selection interact with Sirt3 at an atypical site. Our results establish the attractiveness of these high-throughput screening technologies in identifying novel and potent Sirt3 activators and, therefore, in associated therapeutic applications.

Topics: Acetylation; Allyl Compounds; Biphenyl Compounds; Humans; Lignans; Phenols; Sirtuin 3; Sirtuins

To investigate the effect of honokiol on demyelination after compressed spinal cord injury (CSCI) and it's possible mechanism.. Animal experiment study.. Institute of Neuroscience of Chongqing Medical University.. Total of 69 Sprague-Dawley (SD) rats were randomly divided into 3 groups: sham group (n=15), honokiol group (n=27) and vehicle group (n=27). After established CSCI model by a custom-made compressor successfully, the rats of sham group were subjected to the limited laminectomy without compression; the rats of honokiol group were subjected to CSCI surgery and intraperitoneal injection of 20 mg/kg honokiol; the rats of vehicle group were subjected to CSCI surgery and intraperitoneal injection of an equivalent volume of saline.. In the vehicle group, the rats became paralyzed and spastic after injury, and the myelin sheath became swollen and broken down along with decreased number of myelinated nerve fibers. Western blot analysis manifested that active caspase-3, caspase-12 and cytochrome C began to increase 1 d after injury while the expression of MBP decreased gradually. After intervened with honokiol for 6 days, compared with the vehicle group, the locomotor function and the pathomorphological changes of myelin sheath of the CSCD rats were improved with obviously decreased expression of active caspase-3, caspase-12 and cytochrome C.. Honokiol may improve locomotor function and protect neural myelin sheat from demyelination via prevention oligodendrocytes (OLs) apoptosis through mediate endoplasmic reticulum (ER)-mitochondria pathway after CSCI.

Topics: Animals; Apoptosis; Biphenyl Compounds; Caspase 12; Caspase 3; Cytochromes c; Demyelinating Diseases; Endoplasmic Reticulum; Humans; Lignans; Mitochondria; Myelin Sheath; Rats; Rats, Sprague-Dawley; Spinal Cord; Spinal Cord Injuries

Honokiol (HK) is a natural bioactive compound with proven antineoplastic properties against melanoma. However, it shows very low bioavailability when administered orally. Alternatively, topical administration may offer a promising route. The objective of the current study was to fabricate HK transfersomes (HKTs) for topical treatment of melanoma. As an ultradeformable carrier system, transfersomes can overcome the physiological barriers to topical treatment of melanoma: the stratum corneum and the anomalous tumor microenvironment. Moreover, the immunomodulatory and stemness-regulation roles of HKTs were the main interest of this study.. TFs were prepared using the modified scalable heating method. A three-factor, three-level Box-Behnken design was utilized for the optimization of the process and formulation variables. Intracellular uptake and cytotoxicity of HKTs were evaluated in nonactivated and stromal cell-activated B16F10 melanoma cells to investigate the influence of the complex tumor microenvironment on the efficacy of HK. Finally, ELISA and Western blot were performed to evaluate the expression levels of TGF-β and clusters of differentiation (CD47 and CD133, respectively).. The optimized formula exhibited a mean size of 190 nm, highly negative surface charge, high entrapment efficiency, and sustained release profile. HKTs showed potential to alleviate the immunosuppressive characteristics of B16F10 melanoma in vitro via downregulation of TGF-β signaling. In addition, HKTs reduced expression of the "do not eat me" signal - CD47. Moreover, HKTs possessed additional interesting potential to reduce the expression of the stem-like cell marker CD133. These outcomes were boosted upon combination with metformin, an antihyperglycemic drug recently reported to possess different functions in cancer, while combination with collagenase, an extracellular matrix-depleting enzyme, produced detrimental effects.. HKTs represent a promising scalable formulation for treatment of the aggressive B16F10 melanoma, which is jam-packed with immunosuppressive and stem-like cell markers.

Topics: Antineoplastic Agents; Biphenyl Compounds; Humans; Lignans; Melanoma; Tumor Microenvironment

Topics: Acetylation; Animals; Biphenyl Compounds; Down-Regulation; Epithelial Cells; Fibrosis; Gene Ontology; Humans; Kidney Tubules; Lignans; Male; Mice, Inbred C57BL; Mitochondrial Proteins; Peptide Fragments; Pyruvate Dehydrogenase Complex; Renal Insufficiency, Chronic; RNA, Messenger; Sirtuin 3; Transforming Growth Factor beta1; Up-Regulation; Ureteral Obstruction

Topics: Biphenyl Compounds; Cell Line, Tumor; Drug Carriers; Ethylene Glycols; Lignans; Nanomedicine; Nanoparticles; Particle Size; Polyesters; Polyethylene Glycols

Despite the advances in developing MMP-2/9 inhibitors, off-target side effects and pharmacokinetics problems remain major challenges hindering their clinical success in cancer therapy. However, recent targeting strategies have clearly revitalized MMP research. Herein, we introduce new s-triazine-based dendrimers endowed with intrinsic MMP-2/9 inhibitory potential and tetherable to hepatocellular carcinoma-specific targeting ligands and anticancer agents via biodegradable linkages for targeted therapy. The designed dendrimeric platform was built with potential zinc-binding branching linkers (hydrazides) and termini (carboxylic acids and hydrazides) to confer potency against MMP-2/9. Preliminary cytotoxicity screening and MMP-2/9 inhibition assay of the free dendrimers revealed promising potency (MMP-9; IC

Topics: Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Proliferation; Dendrimers; Dose-Response Relationship, Drug; Drug Screening Assays, Antitumor; Humans; Lignans; Liver Neoplasms; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Matrix Metalloproteinase Inhibitors; Molecular Structure; Structure-Activity Relationship; Triazines

Lung cancer, mainly non-small cell lung cancer (NSCLC), has been a global health problem, leading to maximum cancer death. Across adenocarcinoma patients, significant genetic and phenotypic heterogeneity was identified as responsible for individual cancer drug resistance, driving an urgent need for individualized treatment. High expectation has been set on individualized treatment for better responses and extended survival. There are pressing needs for and significant advantages of testing dosages and drugs directly on patient-specific cancer cells for preclinical drug testing and personalized drug selection. Monitoring the drug response based on patient-derived cells (PDCs) is a step toward effective drug development and individualized treatment. Despite the dependence on optical labels, optical equipment, and other complex manual operation, we here report a multidimensional biosensor system to guide adenocarcinoma individualized treatment by integrating 2D and 3D PDC models and cellular impedance biosensors. The cellular impedance biosensors were applied to quantitate drug response in 2D and 3D environments. Compared with 2D plate culture, 3D cultured cells were found to show higher resistance to anti-cancer drugs. Cell-cell, cell-ECM, and mechanical interactions in the 3D environment led to stronger drug resistance. The

Topics: Adenocarcinoma; Animals; Antineoplastic Agents; Biosensing Techniques; Biphenyl Compounds; Cell Culture Techniques; Cell Movement; Cell Proliferation; Drug Resistance, Neoplasm; Drug Screening Assays, Antitumor; Humans; Lignans; Lung Neoplasms; Male; Mice; Mice, Nude; Piperidines; Precision Medicine; Quinazolines; Tumor Cells, Cultured

To investigate the preventive effects of systemic honokiol and pentoxifylline treatments on epidural fibrosis (EF) in the experimental laminectomy model.. Thirty-two rats were divided into four equal groups. Laminectomy was performed in all rats except for the control group. One group was kept as the negative control group. Moreover, 10 mg/kg pentoxifylline and 10 mg/kg honokiol were administered intraperitoneally for 5 days, respectively, to the other two groups. The rats were sacrificed after 4 weeks. The samples were examined biochemically in terms of oxidative stress and inflammation induced by tissue damage. Histopathological and immunohistochemical investigations were also performed to detect EF severity.. In honokiol and pentoxifylline groups compared with the negative control group, tumor necrosis factor-beta and interleukin-10 levels (indicating inflammation); myeloperoxidase, malondialdehyde, and hydroxyproline levels (indicating oxidative stress); and intercellular adhesion molecule levels (indicating fibrosis) were decreased. Histopathologically and immunohistochemically, EF was significantly reduced in the pentoxifylline and honokiol groups. Biochemical findings were consistent with the histopathological and immunohistochemical findings.. Both pentoxifylline and honokiol prevent EF formation. However, this effect is more pronounced in honokiol.

Topics: Animals; Biphenyl Compounds; Epidural Space; Fibrosis; Laminectomy; Lignans; Pentoxifylline; Rats

Exploring and synthesizing the compounds with stronger antioxidant activity have always been the goal of researchers. Herein, the substitution effects of the amino (NH

Topics: Amines; Antioxidants; Biphenyl Compounds; Electrons; Hydrogen; Lignans; Molecular Conformation; Protons; Structure-Activity Relationship

Neolignans honokiol and 4'-

Topics: Animals; Anti-Inflammatory Agents; Biphenyl Compounds; Fluorine Radioisotopes; Hydrocarbons, Fluorinated; Inflammation; Lignans; Male; Mice; Mice, Inbred C57BL; Radiopharmaceuticals

Thrombus, bacterial infections, and severe inflammation are still serious problems that have to be faced with blood-contacting materials. However, it is a great challenge to simultaneously meet the above functional requirements in a simple, economical and efficient method. As such, we put forward a robust and versatile coating strategy by covalently modifying the multi-pharmacological drug honokiol (HK) with an amine-rich polydopamine/polyethyleneimine coating, through which anticoagulant, antibacterial and anti-inflammatory properties were obtained (DPHc) simultaneously. The amine content in the DPHc coating was lower than the detection limit, while it contained abundant phenolic hydroxyl groups (49 μmol cm

Topics: Animals; Anti-Bacterial Agents; Anti-Inflammatory Agents; Biphenyl Compounds; Cell Line; Coated Materials, Biocompatible; Escherichia coli; Fibrinolytic Agents; Lignans; Male; Mice; Microbial Sensitivity Tests; Rabbits; Rats, Sprague-Dawley; Staphylococcus aureus; Thrombosis

In the present study, the porous starch (PS) was used as an efficient carrier of honokiol (HK), and the HK-loaded PS (HPS) delivery system was prepared by melting method without using organic solvents. Its physical-chemical properties, solubility and oral bioavailability were also investigated. The obtained results proved that the HK in the HPS was mostly amorphous when it was loaded into the PSs with 87.54 ± 1.52% of encapsulation efficiency (EE) and 12.51 ± 0.22% of drug loading (DL) capacity. The water-solubility of the HPS was increased to 115.27 ± 2.92 μg/mL (pH = 1.2, artificial gastric juice (AGJ)), 161.58 ± 3.42 (pH = 6.8, artificial intestinal juice (AIJ)) and 148.5 ± 1.89 μg/mL (pH = 5.5, simulated tumor microenvironment), being 6.07, 4.38 and 4.87-folds higher than free HK.

Topics: Animals; Area Under Curve; Biphenyl Compounds; Drug Carriers; Drug Liberation; Half-Life; Hep G2 Cells; Humans; Lignans; Porosity; Random Allocation; Rats; Rats, Sprague-Dawley; Solubility; Starch; Surface Properties; Technology, Pharmaceutical; Tumor Microenvironment

Topics: Animals; ATP Binding Cassette Transporter, Subfamily G, Member 2; Biphenyl Compounds; Cell Survival; Dogs; Drug Resistance, Multiple; Drug Resistance, Neoplasm; ErbB Receptors; Lignans; Madin Darby Canine Kidney Cells; Magnolia; Neoplasm Proteins; Phosphatidylinositol 3-Kinases; Phosphorylation; Plant Extracts; Polyphenols; Signal Transduction

Topics: Allosteric Regulation; Antineoplastic Agents; Biphenyl Compounds; Cell Line; HSP90 Heat-Shock Proteins; Humans; Lignans; Mitochondria; Models, Molecular; Molecular Structure; Recombinant Proteins

Though honokiol, derived from the Magnolia tree, was known to suppress renal fibrosis, pulmonary fibrosis, non-alcoholic steatoheptitis, inflammation and cancers, the underlying antifibrotic mechanisms of honokiol are not fully understood in hepatic stellate cells until now. Thus, in the present study, inhibitory mechanism of honokiol on liver fibrosis was elucidated mainly in hepatic stellate cells (HSCs) by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell cycle analysis and western-blotting. Honokiol exerted cytotoxicity in LX-2, HSC-T6 and Hep-G2 cells. Honokiol increased sub G1 population and activated caspase 3 and cleaved poly (ADP-ribose) polymerase (PARP) in HSCs. Moreover, honokiol attenuated the expression of alpha smooth muscle actin (α-SMA), transforming growth factor beta 1 (TGF-β1), phospho-Smad3, phospho-AKT, cyclin D1, c-Myc, Wnt3a, β-catenin, and activated phosphorylation of glycogen synthase kinase 3 beta (GSK3β) in HSCs. Conversely, GSK3β inhibitor SB216763 reversed the effect of honokiol on PARP, α-SMA, phospho-GSK3β, β-catenin and sub G1 population in LX-2 cells. Overall, honokiol exerts apoptotic and antifibrotic effects via activation of GSK3β and inhibition of Wnt3a/β-catenin signalling pathway.

Topics: Actins; beta Catenin; Biphenyl Compounds; Caspase 3; Cell Line; Cyclin D1; Glycogen Synthase Kinase 3 beta; Hep G2 Cells; Hepatic Stellate Cells; Humans; Lignans; Liver Cirrhosis; Phosphorylation; Smad3 Protein; Transforming Growth Factor beta1; Wnt Signaling Pathway

Paclitaxel is an anticancer drug for the treatment of non-small cell lung cancer (NSCLC). However, drug-resistance remains a major problem. Honokiol is a natural component which has been found to exhibit anti-tumor activity. Paclitaxel and honokiol have been reported to be able to induce paraptosis. The aim of this study was to investigate whether honokiol can reverse paclitaxel resistance by inducing paraptosis in NSCLC cells.. NSCLC cell lines H1650 (paclitaxel-sensitive), H1299 and H1650/PTX (intrinsic and acquired paclitaxel-resistant, respectively) were used to assess the cytotoxic effects of paclitaxel and honokiol. Light and transmission electron microscopy were performed to detect cytoplasmic vacuolation. In vitro cell viability and clonogenic survival assays, as well as in vivo xenograft assays were conducted to test synergistic killing effects of paclitaxel and honokiol on NSCLC cells. Western blotting, flow cytometry and immunofluorescence were performed to evaluate paraptosis-regulating mechanisms.. Combination of honokiol and paclitaxel may represent a novel strategy for the treatment of paclitaxel-resistant NSCLC.

Topics: Animals; Apoptosis; Biphenyl Compounds; Calcium; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Drug Synergism; Endoplasmic Reticulum Stress; Humans; Lignans; Lung Neoplasms; Male; Mice, Inbred BALB C; Mice, Nude; Microtubule-Associated Proteins; Mitochondria; Paclitaxel; Proteasome Endopeptidase Complex; Ubiquitin-Activating Enzymes; Vacuoles

Honokiol and magnolol are natural components isolated from Magnolia bark that is used in traditional Chinese and Japanese herbal medicine. These two isomers are used as a component of dietary supplements and cosmetic products. In this study, we investigated the antimicrobial effect of honokiol and magnolol on pathogens causing oral diseases, their mechanism of action in biofilm formation and drug resistance of oral pathogens, and inflammatory regulation in mammalian cells.. We determined the minimum inhibitory concentration and minimum bactericidal concentration of honokiol and magnolol, and their stability at different temperatures and pH. We also evaluated their effect on biofilm formation, antibiotic-resistance gene expression in MRSA, and pro-inflammatory gene expression in mammalian cells.. Honokiol showed better antimicrobial activity than magnolol. Both honokiol and magnolol showed stable bacterial inhibitory activity over a wide range of temperature and pH, reduced biofilm formation, and antibiotic resistance in oral pathogens. The biofilm formation- and antibiotic resistance-related gene expression was consistent with the respective phenotypes. Furthermore, these two isomers repressed the expression of pro-inflammatory genes in RAW264.7 cells.. Our study provides evidence of the potential application of honokiol and magnolol in dental medicine to cure or prevent oral diseases.

Topics: Animals; Anti-Bacterial Agents; Biphenyl Compounds; Humans; Inflammation; Lignans; Macrophages

A novel surface plasmon resonance-based P-gp ligand screening system (SPR-PLSS) combined with lentiviral particle (LVP) stabilization strategy was constructed to screen out potential P-gp inhibitors from natural products. Firstly, we constructed LVPs with high and low expression levels of P-gp. The LVPs can ensure the natural conformation of P-gp based on the principle that LVPs germinated from packaging cells will contain cell membrane fragments and P-gp they carry. Then the LVPs with high P-gp expression for active channel and LVPs with low P-gp expression for reference channel were immobilized on CM5 chip respectively. The affinity detection was thus carried out with the signal reduction on the two channels. The P-gp inhibitors, Valspodar (Val) and cyclosporin (CsA), as positive compounds, were detected to characterize the chip's activity, and the K

Topics: Animals; ATP Binding Cassette Transporter, Subfamily B, Member 1; Biological Products; Biosensing Techniques; Biphenyl Compounds; Cell Line, Tumor; Cell Survival; Chemistry, Pharmaceutical; Cyclosporine; Cyclosporins; Dogs; Drug Evaluation, Preclinical; HEK293 Cells; Humans; In Vitro Techniques; Kinetics; Lentivirus; Ligands; Lignans; Madin Darby Canine Kidney Cells; MCF-7 Cells; Membrane Proteins; Resveratrol; Surface Plasmon Resonance

Type 2 diabetes mellitus is strongly associated with cardiac mitochondrial dysfunction, which is one of the main reasons for cardiovascular diseases. Among the mitochondrial metabolic changes, fatty acid metabolism is of great importance as cardiac tissues depend primarily on fatty acids. Honokiol, a constituent of Magnolia tree bark extract, is reported to strongly influence cardiac mitochondrial functions, via various mechanisms. The current study showed that honokiol decreased fatty acid-mediated complex I respiration and increased carbohydrate-mediated complex I and II respiration in diabetic C57BL/6 mice cardiac mitochondria. It was also found that honokiol treatment decreased expression of Cluster of Differentiation 36, AMP-activated kinases and nuclear transcription factors like, Peroxisome proliferator-activated receptor γ co-activator 1α/β and Peroxisome proliferator-activated receptor α, surrogating the evidence of decreased fatty acid-mediated complex I respiration. Honokiol treatment also reduced the levels of mitochondrial acetylated proteins, suggesting the possible action of honokiol via acetylation/deacetylation mechanism of regulation of protein functions in diabetic mitochondria. The antioxidant effect of honokiol is evidenced by the augmented expression of Manganese super oxide dismutase. In conclusion, honokiol imparts beneficial effect on diabetic cardiac mitochondria by decreasing the oxidant burden via regulating mitochondrial fatty acid respiration and expression of oxidant response factors.

Topics: Acetylation; AMP-Activated Protein Kinases; Animals; Antioxidants; Biphenyl Compounds; CD36 Antigens; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Energy Metabolism; Fatty Acids; Lignans; Male; Mice, Inbred C57BL; Mitochondria, Heart; Myocytes, Cardiac; NF-E2-Related Factor 2; Oxidative Stress; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; PPAR alpha; Streptozocin; Superoxide Dismutase

Honokiol, a natural phenolic compound derived from Magnolia plants, is a promising anti-tumor compound that exerts a wide range of anti-cancer effects. Herein, we investigated the effect of honokiol on doxorubicin resistance in breast cancer.. Doxorubicin-sensitive (MCF-7 and MDA-MB-231) and doxorubicin-resistant (MCF-7/ADR and MDA-MB-231/ADR) breast cancer cell lines were treated with doxorubicin in the absence or presence of honokiol; then, the following tests were performed: flow cytometry for cell apoptosis, WST-1 assay for cell viability, qPCR and western blot for the expression of miR-188-5p, FBXW7, and c-Myc. MiR-188-5p mimic, miR-188-5p inhibitor, siFBXW7, and c-Myc plasmids were transfected into cancer cells to evaluate whether miR-188-5p and FBXW7/c-Myc signaling are involved in the effect of honokiol on doxorubicin resistance in breast cancer. A dual luciferase reporter system was used to study the direct interaction between miR-188-5p and FBXW7.. Honokiol sensitized doxorubicin-resistant breast cancer cells to doxorubicin-induced apoptosis. Mechanically, upregulation of miR-188-5p was associated with doxorubicin resistance, and honokiol enhanced doxorubicin sensitivity by downregulating miR-188-5p. FBXW7 was confirmed to be a direct target gene of miR-188-5p. FBXW7/c-Myc signaling was involved in the chemosensitization effect of honokiol. Honokiol induced apoptosis in MCF-7/ADR and MDA-MB-231/ADR cells. However, FBXW7 silencing or c-Myc transfection resulted in resistance to the honokiol-induced apoptotic effect.. These findings suggest that downregulation of miR-188-5p by honokiol enhances doxorubicin sensitivity through FBXW7/c-Myc signaling in human breast cancer. Our study finds an important role of miR-188-5p in the development of doxorubicin resistance in breast cancer, and enriches our understanding of the mechanism of action of honokiol in cancer therapy.

Topics: Apoptosis; Biphenyl Compounds; Breast Neoplasms; Cell Line, Tumor; Doxorubicin; Drug Resistance, Neoplasm; F-Box-WD Repeat-Containing Protein 7; Female; Gene Expression Regulation, Neoplastic; Humans; Lignans; MicroRNAs; Proto-Oncogene Proteins c-myc; Signal Transduction

Acute promyelocytic leukemia (APL) is a blood system disease caused by the accumulation of a large number of immature blood cells in bone marrow. Although the introduction of all-trans retinoic acid (ATRA) and arsenic has reached a high level of complete remission rate and 5-year disease-free survival rate, the occurrence of various adverse reactions still severely affects the quality of life of patients. As a natural product, honokiol (HNK) has the advantages of low toxicity and high efficiency, and it is a potential drug for the treatment of cancer. Since cancer cells can escape apoptotic cell death through multiple adaptive mechanisms, HNK, a drug that induces cancer cell death in a nonapoptotic way, has attracted much interest. We found that HNK reduced the viability of human APL cell line (NB4 cells) by inducing paraptosis-like cell death. The process was accompanied by excessive reactive oxygen species (ROS), mitochondrial damage, endoplasmic reticulum stress, and increased microtubule-associated protein 1 light chain 3 (LC3) processing. The inactivation of proteasome activity was the main cause of misfolded and unfolded protein accumulation in endoplasmic reticulum, such as LC3II/I and p62. This phenomenon could be alleviated by adding cycloheximide (CHX), a protein synthesis inhibitor. We found that mTOR signaling pathway participated in paraptosis-like cell death induced by HNK in an autophagy-independent process. Moreover, the mitogen-activated protein kinase (MAPK) signaling pathway induced paraptosis of NB4 cells by promoting endoplasmic reticulum stress. In summary, these findings indicate that paraptosis may be a new way to treat APL, and provide novel insights into the potential mechanism of paraptosis-like cell death.

Topics: Apoptosis; Biological Products; Biphenyl Compounds; Cell Death; Cell Line, Tumor; Cell Proliferation; Endoplasmic Reticulum Stress; Humans; In Vitro Techniques; Leukemia, Promyelocytic, Acute; Lignans; MAP Kinase Signaling System; Proteasome Endopeptidase Complex; Reactive Oxygen Species; Signal Transduction; TOR Serine-Threonine Kinases

Hepatocellular carcinoma (HCC) is the fifth common types of cancer with poor prognosis in the world. Honokiol (HNK), a natural biphenyl compound derived from the magnolia plant, has been reported to exert anticancer effects, but its mechanism has not been elucidated exactly. In the present study, HNK treatment significantly suppressed the migration ability of HepG2 and Hep3B human hepatocellular carcinoma. The treatment reduced the expression levels of the genes associated with cell migration, such as S100A4, MMP-2, MMP-9 and Vimentin. Interestingly, treatment with HNK significantly reduced the expression level of Cyclophilin B (CypB) which stimulates cancer cell migration. However, overexpressed CypB abolished HNK-mediated suppression of cell migration, and reversed the apoptotic effects of HNK. Altogether, we concluded that the suppression of migration activities by HNK was through down-regulated CypB in HCC. These finding suggest that HNK may be a promising candidate for HCC treatment via regulation of CypB.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Movement; Cell Survival; Cyclophilins; Down-Regulation; Gene Expression Regulation, Neoplastic; Hep G2 Cells; Humans; Lignans; Liver Neoplasms; Signal Transduction

Traditional Chinese medicine (TCM) preparations are very complex mixtures, and the content of bioactive components is usually very low. Therefore, before final analysis, the preparation of an appropriate sample is necessary. Sample preparation is the most time-consuming and error-prone part of the analytical procedure, and the choice of purification technology greatly influences the reliability of the final analysis.. In the present study, we evaluated the feasibility of hollow fiber centrifugal ultrafiltrate (HFCF-UF) as a purification technology for the analysis of bioactive components in TCM preparations. The HFCF-UF technology was applied to analyze honokiol and magnolol in TCM preparations containing Cortex Magnoliae Officinalis (Hou Po in Chinese Pinyin). A mini centrifugal device based on hollow fiber was employed to remove the macromolecule components. A single step of simple centrifugation was required before the filtrate could be directly injected into an existing high performance liquid chromatography (HPLC) system without any further clean-up step or use of special columns. This greatly simplified the pretreatment steps, and improved the accuracy of analytic methods. The separation was achieved on a Diamonsil C18 column (i.d. 5 µm, 150 mm × 4.6 mm) with V (methanol):V (acetonitrile):V (0.5% acetic acid solution) =44:22:34 as the mobile phase at a flow rate of 1.0 mL/min.. It had good linear relationship between the peak areas of honokiol and magnolol and their concentrations at 6.40-205 and 3.15-101 µg/mL (r=0.9999), respectively. The method recovery was over 92.6% with a relative standard deviation (RSD) of less than 3.0%. The average recovery of honokiol was 97.7% with an RSD of 3.0%, and that of magnolol was 96.8% with RSD of 2.8%.. The application of HFCF-UF in TCM preparations could assist in making the quality control of TCM simple, rapid, and accurate. The HFCF-UF purification procedure can be used as an alternative means for analyzing bioactive components in TCM preparations.

Topics: Biphenyl Compounds; Humans; Lignans; Medicine, Chinese Traditional; Reproducibility of Results; Technology

The exposure of neurons to an excessive excitatory stimulation induces the alteration of the normal neuronal function. Mood disorders are among the first signs of alterations in the central nervous system function. Magnolia officinalis bark extract has been extensively used in the traditional medicine systems of several countries, showing several pharmacological activities. Honokiol, the main constituent of M. officinalis, is a GABA modulator and a CB1 agonist, which is deeply investigated for its role in modulating mood disorders.. Thus, we evaluated the possible neuroprotective effect of a standardized M. officinalis bark extract (MOE), enriched in honokiol, and its effect on animal mood behavioural tests and in an in vitro model of excitotoxicity.. MOE showed neuroprotective effect using SH-SY5Y cells, by normalizing brain-derived neurotrophic factor release. Then, we tested the effect of MOE in different behavioural tests evaluating anxiety and depression and we observed a selective anxiolytic-like effect. Finally, we confirmed the involvement of CB1 in the final effect of MOE by the co-administration of the CB1 antagonist, AM251.. These results suggest that MOE could be considered an effective and safe anxiolytic candidate with neuroprotective activity.

Topics: Animals; Anti-Anxiety Agents; Anxiety; Anxiety Disorders; Behavior, Animal; Biphenyl Compounds; Brain; Brain-Derived Neurotrophic Factor; GABA Modulators; Humans; Lignans; Magnolia; Male; Mice; Neuroprotective Agents; Phytotherapy; Plant Bark; Plant Extracts; Receptor, Cannabinoid, CB1

Honokiol plays an important role in anti-oxidation, but its role in diabetic vascular complications is unclear. In this study, the effects of honokiol in high glucose/high fat (HG/HF)-induced human umbilical vein endothelial cells (HUVECs) were explored. After pre-treatment with honokiol, the cells were transferred to an HG/HF medium, and cell viability and apoptosis were respectively measured by methyl tetrazolium and flow cytometry. Moreover, the contents of reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD) were measured. The expressions of C/EBP homologous protein (CHOP), glucose-regulated protein 78 (GRP78), phosphorylated-protein kinase RNA-like endoplasmic reticulum kinase (p-PERK), phosphorylated-inositol requiring enzyme-1α (p-IRE1α), cleaved caspase-3 and SIRT1 were determined by Western blot or quantitative reverse transcription PCR, respectively. Finally, the viability, apoptosis, and the contents of ROS, MDA, and SOD, as well as the expressions of CHOP, GRP78, p-PERK, p-IRE1α, cleaved caspase-3, Akt, p-Akt, and SIRT1 in the cells transfected with small interfering RNA SIRT1 (siSIRT1) were detected by the previously mentioned methods. Honokiol reversed the effect of HG/HF on promoting cell apoptosis, ROS and MDA contents, and the expressions of CHOP, GRP78, p-PERK, p-IRE1α and cleaved caspase-3, and also reversed the inhibitory effect of HG/HF on cell viability, SOD content and SIRT1 expression. However, siSIRT1 reversed the above effects caused by honokiol. Honokiol activated SIRT1 promoter. SIRT1 interacted with Akt, consequently promoting the activity of Akt. Therefore, honokiol activates the Akt pathway by regulating SIRT1 expression to regulate endoplasmic reticulum stress, promotes cell viability and inhibits the apoptosis of HG/HF-induced HUVECs.

Topics: Apoptosis; Biphenyl Compounds; Cell Survival; Endoplasmic Reticulum Stress; Human Umbilical Vein Endothelial Cells; Humans; Lignans; Malondialdehyde; Oxidative Stress; Proto-Oncogene Proteins c-akt; Reactive Oxygen Species; Signal Transduction; Sirtuin 1; Superoxide Dismutase

The habit of areca nut chewing has been regarded as an etiological factor of precancerous oral submucous fibrosis (OSF). In the present study, we aimed to evaluate the anti-fibrosis effect of honokiol, a polyphenolic component derived from Magnolia officinalis.. The cytotoxicity of honokiol was tested using normal and fibrotic buccal mucosal fibroblasts (fBMFs) derived from OSF tissues. Collagen gel contraction, Transwell migration, invasion, and wound healing capacities were examined. Besides, the expression of TGF-β/Smad2 signaling as well as α-SMA and type I collagen were measured as well.. Honokiol exerted higher cytotoxicity of fBMFs compared to normal cells. The arecoline-induced myofibroblast activities, including collagen gel contractility, cell motility and wound healing capacities were all suppressed by honokiol treatment. In addition, the expression of the TGF-β/Smad2 pathway was downregulated along with a lower expression of α-SMA and type I collagen in honokiol-receiving cells.. Our data suggest that honokiol may be a promising compound to alleviate the progression of oral fibrogenesis and prevent the transformation of OSF oral epithelium into cancer.

Topics: Areca; Arecoline; Biphenyl Compounds; Cell Transdifferentiation; Fibroblasts; Humans; Lignans; Mouth Mucosa; Oral Submucous Fibrosis; Smad2 Protein; Transforming Growth Factors

Dysfunction of microglia has been increasingly recognized as a causative factor in Alzheimer's disease (AD); thus, developing medicines capable of restoring microglial functions is critically important and constitutes a promising therapeutic strategy. Honokiol is a natural neuroprotective compound extracted from Magnolia officinalis, which may play roles in AD therapy.. This study aimed to evaluate the role and the underlying mechanisms of honokiol in microglial phagocytosis.. MTT and flow cytometry were used to assess the cell viability and apoptosis, respectively. Phagocytic capacity, mitochondrial reactive oxygen species production, and membrane potential were evaluated using fluorescence microscopy. Seahorse XF24 extracellular flux analyzer was for cell glycolysis and oxidative phosphorylation detection. Mass spectrometry was applied for metabolites measurement. Quantitative real-time polymerase chain reaction and western blotting were performed to detect the mRNA and protein level of PPARγ and PGC1α, respectively.. Honokiol alleviated Aβ42-induced BV2 neurotoxicity. Honokiol promoted phagocytic efficiency of BV2 cells through reversing a metabolic switch from oxidative phosphorylation to anaerobic glycolysis and enhancing ATP production. Meanwhile, honokiol reduced mitochondrial reactive oxygen species production and elevated mitochondrial membrane potential. Moreover, honokiol increased the expression of PPARγ and PGC1α, which might play positive roles in energy metabolism and microglial phagocytosis.. In this study, honokiol was identified as an effect natural product capable of enhancing mitochondrial function thus promoting microglial phagocytic function.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Apoptosis; Biphenyl Compounds; Cell Culture Techniques; Cell Survival; Humans; Lignans; Magnolia; Mice; Microglia; Phagocytosis; Reactive Oxygen Species

Existing agents to induce enamel self-repair and inhibit the progression of dental caries in the early stage have been proven to be inadequate and far from satisfactory. In this study, a honokiol-loaded poly(amido amine) (PAMAM) dendrimer (PAMH) was constructed to combat early caries lesions in enamel.. PAMH was prepared via a codissolution method. Computational simulation analysis was used to explore the mechanism of honokiol release. The cytotoxicity of PAMH was tested. The antibacterial effects of PAMH were tested by planktonic growth assays and biofilm formation inhibition assays. The remineralization effect of PAMH was examined via transverse microradiography and scanning electron microscopy after a pH cycling model. The in vivo anti-caries effect of PAMH was carried out in a rat model.. Honokiol released from PAMH was slower but more durable in a cariogenic pH environment than in a neutral pH environment, which could be explained through the computational simulation analysis results. Under electrostatic action, P3 beads with the same charge repelled each other and extended outwards, resulting in the rapid expansion of the PAMAM dendrimer and accelerating the release of the drug. At a low pH of 5.5, the protonated P3 beads were not charged and the protonated P1 beads were positively charged. However, the electrostatic repulsive interaction between protonated P1 beads was restricted by the P3 beads in the outermost layer of the PAMAM dendrimer, so the swelling rate was relatively slow, resulting in the slow release of drug molecules in the acidic environment. The cytotoxicity demonstration and the biocompatibility experiment in animal study showed that PAMH is biologically safe. PAMH showed excellent enamel remineralizing ability after pH cycling and showed a long-term antibacterial effect in vitro. Meanwhile, PAMH showed long-term anticaries efficacy in vivo.. Our findings indicated that PAMH had great potential to combat early caries lesions in enamel for future clinical application.

Topics: Amines; Animals; Anti-Bacterial Agents; Biphenyl Compounds; Cariostatic Agents; Dendrimers; Dental Caries; Dental Enamel; Dentin; Lignans; Rats; Tooth Remineralization

Ca. Patch-clamp techniques were performed to study the effect of honokiol on Ca. Honokiol blocked Ano1 currents in Ano1-overexpressing HEK293 cells and SW620 cells. Honokiol more potently inhibited Ano1 currents than Ano2 currents. Three amino acids (R429, K430 and N435) were critical for honokiol-induced Ano1 inhibition. The R429A/K430L/N435G mutation reduced the sensitivity of Ano1 to honokiol. Honokiol inhibited SW620 cell proliferation, and this effect was reduced by Ano1-shRNAs. Furthermore, Ano1 overexpression promoted proliferation in NCM460 cells with low Ano1 endogenous expression and resulted in an increased sensitivity to honokiol. Overexpression of the R429A/K430L/N435G mutation reduced WT Ano1-induced increase in the sensitivity of NCM460 cells to honokiol.. We identified a new anticancer mechanism of honokiol, through the inhibition of cell proliferation, by targeting Ano1 Ca

Topics: Anoctamin-1; Biphenyl Compounds; Calcium; Cell Proliferation; Chloride Channels; Colorectal Neoplasms; HEK293 Cells; Humans; Lignans

Honokiol (HNK) is a biphenolic compound that has been used in traditional medicine for treating various ailments, including cancers. In this study, we determined the effect of HNK on colon cancer cells in culture and in a colitis-associated cancer model. HNK treatment inhibited proliferation and colony formation while inducing apoptosis. In addition, HNK suppressed colonosphere formation. Molecular docking suggests that HNK interacts with reserve stem cell marker protein DCLK1, with a binding energy of -7.0 Kcal/mol. In vitro kinase assays demonstrated that HNK suppressed the DCLK1 kinase activity. HNK also suppressed the expression of additional cancer stem cell marker proteins LGR5 and CD44. The Hippo signaling pathway is active in intestinal stem cells. In the canonical pathway, YAP1 is phosphorylated at Ser127 by upstream Mst1/2 and Lats1/2. This results in the sequestration of YAP1 in the cytoplasm, thereby not allowing YAP1 to translocate to the nucleus and interact with TEAD1-4 transcription factors to induce gene expression. However, HNK suppressed Ser127 phosphorylation in YAP1, but the protein remains sequestered in the cytoplasm. We further determined that this occurs by YAP1 interacting with PUMA. To determine if this also occurs in vivo, we performed studies in an AOM/DSS induced colitis-associated cancer model. HNK administered by oral gavage at a dose of 5mg/kg bw for 24 weeks demonstrated a significant reduction in the expression of YAP1 and TEAD1 and in the stem marker proteins. Together, these data suggest that HNK prevents colon tumorigenesis in part by inducing PUMA-YAP1 interaction and cytoplasmic sequestration, thereby suppressing the oncogenic YAP1 activity.

Topics: Adaptor Proteins, Signal Transducing; Animals; Apoptosis; Apoptosis Regulatory Proteins; Biomarkers, Tumor; Biphenyl Compounds; Carcinogenesis; Cell Line, Tumor; Cell Proliferation; Cell Survival; Colitis; Colonic Neoplasms; Doublecortin-Like Kinases; Down-Regulation; Hippo Signaling Pathway; Humans; Intracellular Signaling Peptides and Proteins; Lignans; Male; Mice, Inbred ICR; Models, Biological; Neoplastic Stem Cells; Protein Binding; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins; Signal Transduction; Transcription Factors; Tumor Stem Cell Assay; YAP-Signaling Proteins

Honokiol is the dominant biphenolic compound isolated from the Magnolia tree, and has long been considered as the active constituent of the traditional Chinese herb, 'Houpo', which is widely used to treat symptoms due to 'stagnation of qi'. Pharmacological studies have shown that honokiol possesses a wide range of bioactivities without obvious toxicity. Honokiol protects the liver, kidneys, nervous system, and cardiovascular system through reducing oxidative stress and relieving inflammation. Moreover, honokiol shows anti-diabetic property through enhancing insulin sensitivity, and anti-obese property through promoting browning of adipocytes. In vivo and in vitro studies indicated that honokiol functions as an anti-cancer agent through multiple mechanisms: inhibiting angiogenesis, promoting cell apoptosis, and regulating cell cycle. A variety of therapeutic effects of honokiol may be associated with its physiochemical properties, which make honokiol readily cross the blood brain barrier and the blood-cerebrospinal fluid barrier, with high bioavailability. In the future, more clinical researches on honokiol are needed to fully authenticate its therapeutic values.

Topics: Apoptosis; Biphenyl Compounds; Drugs, Chinese Herbal; Humans; Lignans; Magnolia

Topics: Animals; Antineoplastic Agents; Biphenyl Compounds; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Female; Humans; Hydrogen-Ion Concentration; Lignans; Lipids; Lung Neoplasms; Mice; Mice, Inbred BALB C; Nanoparticles

To investigate the mechanism of honokiol (HNK) on bladder cancer cells and its synergistic anticancer effect with hydroxycamptothecin (HCPT).. Control, HNK, HCPT, and HNK plus HCPT groups were established. The morphological characteristics of T24 cells were examined microscopically. The maximal experimental concentration of HNK and HCPT were determined according to IC10 detected by MTT. T24 cell viability and the percentage of apoptotic cells were assessed on the basis of MTT and flow cytometric analysis. The expression of caspase-3, caspase-9, phosphorylated nuclear factor-kappa B (NF-κB)-p65, Akt, and extracellular signal-regulated kinase (ERK) proteins were analyzed by Western blot.. Apoptosis in T24 cells was observed microscopically in both the HNK and HCPT groups and even more obvious in the HNK plus HCPT groups. The percentage of T24 cell viability decreased down to 19.41% , and the percentage of apoptotic cells rose to 54.08% when treated with HNK plus HCPT in an HNK dose-dependent manner. The induction of caspase-3 and caspase-9 proteins and the inhibition of phosphorylation of NF-κB-p65, Akt, and ERK proteins in T24 cells were demonstrated in the HNK groups, and more significantly in the HNK plus HCPT groups, but not in the HCPT group.. The anticancer effect of HNK may be due to the activation of the caspase pathway and inhibition of phosphorylation of NF-κB, Akt, and ERK. HNK in combination with HCPT produces a synergistic cell-killing effect on bladder cancer cells.

Topics: Apoptosis; Biphenyl Compounds; Camptothecin; Cell Line, Tumor; Lignans

Topics: Antineoplastic Agents; Autophagy; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Dose-Response Relationship, Drug; Drug Screening Assays, Antitumor; Humans; Lignans; Molecular Structure; Oxadiazoles; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Structure-Activity Relationship; Thiadiazoles; TOR Serine-Threonine Kinases

Targeting

Topics: Anti-Bacterial Agents; Bacterial Outer Membrane; Biofilms; Biphenyl Compounds; Cetylpyridinium; Lignans; Microbial Sensitivity Tests; Microbiota; Phenols; Streptococcus mutans

Senescence of cardiomyocytes is considered a key factor for the occurrence of doxorubicin (Dox)‑associated cardiomyopathy. The NOD‑like receptor family pyrin domain‑containing 3 (NLRP3) inflammasome is reported to be involved in the process of cellular senescence. Furthermore, thioredoxin‑interactive protein (TXNIP) is required for NLRP3 inflammasome activation and is considered to be a key component in the regulation of the pathogenesis of senescence. Studies have demonstrated that pretreatment with honokiol (Hnk) can alleviate Dox‑induced cardiotoxicity. However, the impact of Hnk on cardiomyocyte senescence elicited by Dox and the underlying mechanisms remain unclear. The present study demonstrated that Hnk was able to prevent Dox‑induced senescence of H9c2 cardiomyocytes, indicated by decreased senescence‑associated β‑galactosidase (SA‑β‑gal) staining, as well as decreased expression of p16INK4A and p21. Hnk also inhibited TXNIP expression and NLRP3 inflammasome activation in Dox‑stimulated H9c2 cardiomyocytes. When TXNIP expression was enforced by adenovirus‑mediated gene overexpression, the NLRP3 inflammasome was activated, which led to inhibition of the anti‑inflammation and anti‑senescence effects of Hnk on H9c2 cardiomyocytes under Dox treatment. Furthermore, adenovirus‑mediated TXNIP‑silencing inhibited the NLRP3 inflammasome. Consistently, TXNIP knockdown enhanced the anti‑inflammation and anti‑senescence effects of Hnk on H9c2 cardiomyocytes under Dox stimulation. In summary, Hnk was found to be effective in protecting cardiomyocytes against Dox‑stimulated senescence. This protective effect was mediated via the inhibition of TXNIP expression and the subsequent suppression of the NLRP3 inflammasome. These results demonstrated that Hnk may be of value as a cardioprotective drug by inhibiting cardiomyocyte senescence.

Topics: Biphenyl Compounds; Carrier Proteins; Cell Line; Cell Survival; Cellular Senescence; Doxorubicin; Drug Antagonism; Humans; Inflammasomes; Lignans; Myocytes, Cardiac; NLR Family, Pyrin Domain-Containing 3 Protein; Oxidative Stress

Aim Liver fibrosis represents a massive global health burden with limited therapeutic options. Thus, the need for curative options is evident. Thus, this study aimed to assess the potential antifibrotic effect of honokiol in Concanavalin A (Con A) induced immunological model of liver fibrosis as well the possible underlying molecular mechanisms.. Male Sprague-Dawley rats were treated with either Con A (20 mg/kg, IV) and/or honokiol (10 mg/kg, orally) for 4 weeks. Hepatotoxicity indices were as well as histopathological evaluation was done. Hepatic fibrosis was assessed by measuring alpha smooth muscle actin (α-SMA) expression and collagen fibers deposition by Masson's trichrome stain and hydroxyproline content. To elucidate the underlying molecular mechanisms, the effect of honokiol on oxidative stress, inflammatory markers as well as transforming growth factor beta (TGF-β)/SMAD and mitogen-activated protein kinase (MAPK) pathways was assessed.. Honokiol effectively reversed the hepatotoxicity indices elevations and abnormal histopathological changes induced by Con A. Besides, honokiol attenuated Con A-induced liver fibrosis by down-regulation of hydroxyproline levels, α-SMA expression together with a marked decrease in collagen fibers deposition. Mechanistically Con A induced oxidative stress, provocation of inflammatory responses and activation of TGF-β/SMAD/MAPK pathways. Contrariwise, honokiol co-treatment significantly restored antioxidant defence mechanisms, down-regulated inflammatory cascades and inhibited TGF-β/SMAD/MAPK signaling pathways.. The results provide an evidence for the promising antifibrotic effect of honokiol that could be partially due to suppressing oxidative stress and inflammatory processes as well as inhibition of TGF-β/SMAD/MAPK signaling pathways.

Topics: Actins; Animals; Biphenyl Compounds; Concanavalin A; Hydroxyproline; Inflammation; Lignans; Liver Cirrhosis; Male; Mitogen-Activated Protein Kinases; Oxidative Stress; Rats; Rats, Sprague-Dawley; Signal Transduction; Smad Proteins; Survival Analysis; Transforming Growth Factor beta

Honokiol is a natural bioactive neolignan and has been widely researched and structural modified as an anticancer agent. In this paper, 18 honokiol derivatives were synthesized and investigated for their antitumor activity. Among these, the promising compound 5a exhibited much higher anti-proliferative activity with IC

Topics: Antineoplastic Agents; Biphenyl Compounds; Humans; Lignans; Neoplasms

The emergence of the plasmid-mediated colistin resistance gene

Topics: Animals; Anti-Bacterial Agents; Bacterial Proteins; Biphenyl Compounds; Colistin; Drug Resistance, Multiple, Bacterial; Enterobacteriaceae; Female; Lignans; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Polymyxins

Topics: A549 Cells; Animals; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Betulinic Acid; Biphenyl Compounds; Cell Proliferation; Cisplatin; Ginsenosides; Humans; Lignans; Liposomes; Lung Neoplasms; Mice; Neoplasm Invasiveness; Pentacyclic Triterpenes; Sesquiterpenes; Triterpenes; Xenograft Model Antitumor Assays

The study mainly aimed to improve the solubility of honokiol (HK) by preparing honokiol nanosuspensions (HNS). In this study, HNS were obtained using Kolliphor®P407 (P407) as a stabilizer through melting method combined with high pressure homogenization (HPH). The crystalline state of HNS was confirmed through differential scanning calorimetry (DSC) and X-ray Diffraction (XRD). In vitro, the dissolution rate of HNS was significantly improved than that of pure HK. In vivo, higher anti-inflammatory activity was achieved after free HK had been made into HNS. There was no significant difference between the degree of edema (DE) of HNS group and that of aspirin group. Consequently, melting method combined with HPH was a potent technique to prepare HNS. Furthermore, nanosuspension was a valid formulation that could be utilized to enhance the anti-inflammatory effect of HK.

Topics: Animals; Anti-Inflammatory Agents; Biphenyl Compounds; Female; Lignans; Male; Mice; Nanoparticles; Solubility; Suspensions

Renal fibrosis is a common feature of all progressive chronic kidney diseases. Sirtuin 3(SIRT3) is one of the mitochondrial sirtuins, and plays a role in the regulation of mitochondrialbiogenesis, oxidative stress, fatty acid metabolism, and aging. Recently, honokiol (HKL), as apharmaceutical SIRT3 activator, has been observed to have a protective effect against pressureoverload-induced cardiac hypertrophy by increasing SIRT3 activity. In this study, we investigatedwhether HKL, as a SIRT3 activator, also has protective effects against unilateral ureteral obstruction(UUO)-induced renal tubulointerstitial fibrosis through SIRT3-dependent regulation ofmitochondrial dynamics and the nuclear factor-κB (NF-κB)/transforming growth factor-β1 (TGF-β1)/Smad signaling pathway. We found that HKL decreased the UUO-induced increase in tubularinjury and extracellular matrix (ECM) deposition in mice. HKL also decreased myofibroblastactivation and proliferation in UUO kidneys and NRK-49F cells. Finally, we showed that HKLtreatment decreased UUO-induced mitochondrial fission and promoted mitochondrial fusionthrough SIRT3-dependent effects. In conclusion, activation of SIRT3 via HKL treatment might havebeneficial effects on UUO-induced renal fibrosis through SIRT3-dependent regulation ofmitochondrial dynamics and the NF-κB/TGF-β1/Smad signaling pathway.

Topics: Animals; Biphenyl Compounds; Cell Line; Disease Models, Animal; Fibrosis; Gene Expression Regulation, Neoplastic; Humans; Inflammation; Kidney Diseases; Lignans; Mice; Mitochondrial Dynamics; NF-kappa B; Signal Transduction; Sirtuin 3; Smad Proteins; Transforming Growth Factor beta1

Fungal pathogens lead to severe quality deterioration and yield loss, making it urgent to explore efficient measures to control fungal diseases at the preharvest and postharvest stages of plants. Therefore, studies on natural substances targeting alternative antimicrobial targets have become hot spots of research. Here, we show that honokiol, a polyphenolic compound obtained from Magnolia officinalis, significantly suppressed mycelial growth and reduced virulence of B. cinerea on harvested fruit by inducing autophagic activities and apoptosis. Moreover, honokiol was capable of abolishing the mitochondrial membrane potential and inducing the accumulation of reactive oxygen species. Some key genes involved in pathogenicity on fruit were also found significantly down-regulated. In summary, honokiol was effective as an alternative agent targeting autophagic and apoptotic machineries to control the incidence of gray mold, which may further enrich the toolkit of crop managers for fighting postharvest diseases caused by this and similar fungi.

Topics: Antifungal Agents; Apoptosis; Autophagy; Biphenyl Compounds; Botrytis; Down-Regulation; Fruit; Genes, Fungal; Lignans; Magnolia; Membrane Potential, Mitochondrial; Mycelium; Plant Diseases; Reactive Oxygen Species; Virulence

The development of acquired resistance to osimertinib (Osim) (AZD9291 or TAGRISSO

Topics: Acrylamides; Aniline Compounds; Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Drug Resistance, Neoplasm; Drug Synergism; ErbB Receptors; Humans; Lignans; Lung Neoplasms; Mice, Nude; Mutation; Protein Kinase Inhibitors

Honokiol has been reported to possess anti-inflammatory and neuroprotective activities. However, the poor aqueous solubility of honokiol limits its clinical application for systemic administration.. This study aims to develop a novel formulation of nanosome-encapsulated honokiol (NHNK) for intravenous therapy against mouse experimental autoimmune encephalomyelitis (EAE) that mimics human multiple sclerosis.. Nanosomes and NHNK were prepared by using an ultra-high pressure homogenization (UHPH) method. Mice were treated with NHNK or empty nanosomes during the peak phase of EAE symptoms. Symptoms of EAE were monitored and samples of the spinal cord were obtained for histopathological examinations.. The stock of NHNK containing honokiol in the nanosome formulation, which showed the structure of single phospholipid bilayer membranes, was well formulated with the particle size of 48.0 ± 0.1 nm and the encapsulation efficiency 58.1 ± 4.2%. Intravenous administration of NHNK ameliorated the severity of EAE accompanied by a significant reduction of demyelination and inflammation in the spinal cord. Furthermore, NHNK decreased the number of IL-6. The UHPH method simplified the preparation of NHNK with uniformly distributed nanosize and high encapsulation efficiency. Intravenous administration of NHNK ameliorated the severity of EAE by suppressing the infiltration of activated microglia and Th1 cells into the spinal cord. Collectively, these results suggest that the formulation of NHNK is a prospective therapeutic approach for inflammatory CNS diseases, such as multiple sclerosis.

Topics: Animals; Anti-Inflammatory Agents; Biphenyl Compounds; Drug Delivery Systems; Encephalomyelitis, Autoimmune, Experimental; Female; Injections, Intravenous; Lignans; Mice, Inbred C57BL; Microglia; Multiple Sclerosis; Myelitis; Nanostructures; Neuroprotective Agents; Spinal Cord; Th1 Cells

Pulmonary fibrosis (PF) is an epithelial/fibroblastic crosstalk disorder of the lungs with highly complex etiopathogenesis. Limited treatment possibilities are responsible for poor prognosis and mean survival rate of 3 to 5 years of PF patients after definite diagnosis. Once thought to be an irreversible disorder, recent evidences have brought into existence the concept of organ fibrosis reversibility due to plastic nature of fibrotic tissues. These findings have kindled interest among the scientific community and given a new direction for research in the arena of fibrosis for developing new anti-fibrotic therapies. The current study is designed to evaluate the anti-fibrotic effects of Honokiol (HNK), a neolignan active constituent from Magnolia officinalis. This study has been conducted in TGF-β1 induced in vitro model and 21 day in vivo murine model of Bleomycin induced PF. The findings of our study suggest that HNK was able to inhibit fundamental pathways of epithelial to mesenchymal transition (EMT) and TGF-β/Smad signaling both in vitro and in vivo. Additionally, HNK also attenuated collagen deposition and inflammation associated with fibrosis. We also hypothesized that HNK interfered with IL-6/CD44/STAT3 axis. As hypothesized, HNK significantly mitigated IL-6/CD44/STAT3 axis both in vitro and in vivo as evident from outcomes of various protein expression studies like western blotting, immunohistochemistry and ELISA. Taken together, it can be concluded that HNK reversed pulmonary fibrotic changes in both in vitro and in vivo experimental models of PF and exerted anti-fibrotic effects majorly by attenuating EMT, TGF-β/Smad signaling and partly by inhibiting IL-6/CD44/STAT3 signaling axis.

Topics: Animals; Biphenyl Compounds; Bleomycin; Bronchoalveolar Lavage Fluid; Cell Line; Cell Movement; Collagen; Cytokines; Epithelial-Mesenchymal Transition; Humans; Hyaluronan Receptors; Interleukin-6; Lignans; Lung; Mice; Pulmonary Fibrosis; Signal Transduction; Smad Proteins; STAT3 Transcription Factor; Transforming Growth Factor beta

Glioblastoma (GBM) responses to bevacizumab are invariably transient with acquired resistance. We profiled paired patient specimens and bevacizumab-resistant xenograft models pre- and post-resistance toward the primary goal of identifying regulators whose targeting could prolong the therapeutic window, and the secondary goal of identifying biomarkers of therapeutic window closure. Bevacizumab-resistant patient specimens and xenografts exhibited decreased vessel density and increased hypoxia versus pre-resistance, suggesting that resistance occurs despite effective therapeutic devascularization. Microarray analysis revealed upregulated mesenchymal genes in resistant tumors correlating with bevacizumab treatment duration and causing three changes enabling resistant tumor growth in hypoxia. First, perivascular invasiveness along remaining blood vessels, which co-opts vessels in a VEGF-independent and neoangiogenesis-independent manner, was upregulated in novel biomimetic 3D bioengineered platforms modeling the bevacizumab-resistant microenvironment. Second, tumor-initiating stem cells housed in the perivascular niche close to remaining blood vessels were enriched. Third, metabolic reprogramming assessed through real-time bioenergetic measurement and metabolomics upregulated glycolysis and suppressed oxidative phosphorylation. Single-cell sequencing of bevacizumab-resistant patient GBMs confirmed upregulated mesenchymal genes, particularly glycoprotein YKL-40 and transcription factor ZEB1, in later clones, implicating these changes as treatment-induced. Serum YKL-40 was elevated in bevacizumab-resistant versus bevacizumab-naïve patients. CRISPR and pharmacologic targeting of ZEB1 with honokiol reversed the mesenchymal gene expression and associated stem cell, invasion, and metabolic changes defining resistance. Honokiol caused greater cell death in bevacizumab-resistant than bevacizumab-responsive tumor cells, with surviving cells losing mesenchymal morphology. Employing YKL-40 as a resistance biomarker and ZEB1 as a target to prevent resistance could fulfill the promise of antiangiogenic therapy. SIGNIFICANCE: Bevacizumab resistance in GBM is associated with mesenchymal/glycolytic shifts involving YKL-40 and ZEB1. Targeting ZEB1 reduces bevacizumab-resistant GBM phenotypes. GRAPHICAL ABSTRACT: http://cancerres.aacrjournals.org/content/canres/80/7/1498/F1.large.jpg.

Topics: Adult; Aged; Angiogenesis Inhibitors; Animals; Antineoplastic Agents, Phytogenic; Antineoplastic Combined Chemotherapy Protocols; Bevacizumab; Biphenyl Compounds; Brain; Brain Neoplasms; Cell Hypoxia; Cell Line, Tumor; Chitinase-3-Like Protein 1; Drug Resistance, Neoplasm; Epithelial-Mesenchymal Transition; Female; Gene Expression Regulation, Neoplastic; Glioblastoma; Human Umbilical Vein Endothelial Cells; Humans; Lignans; Male; Middle Aged; Neoplasm Invasiveness; Neoplastic Stem Cells; Neovascularization, Pathologic; Tumor Microenvironment; Up-Regulation; Xenograft Model Antitumor Assays; Young Adult; Zinc Finger E-box-Binding Homeobox 1

Honokiol (2) is a natural bisphenol neolignan showing a variety of biological properties, including antitumor activity. Some studies pointed out 2 as a potential anticancer agent in view of its antiproliferative and pro-apoptotic activity towards tumor cells. As a further contribution to these studies, we report here the synthesis of a small library of bisphenol neolignans inspired by honokiol and the evaluation of their antiproliferative activity. The natural lead was hence subjected to simple chemical modifications to obtain the derivatives 3-9; further neolignans (12a-c, 13a-c, 14a-c, and 15a) were synthesized employing the Suzuki-Miyaura reaction, thus obtaining bisphenols with a substitution pattern different from honokiol. These compounds and the natural lead were subjected to antiproliferative assay towards HCT-116, HT-29, and PC3 tumor cell lines. Six of the neolignans show GI

Topics: Antineoplastic Agents; Apoptosis; Benzhydryl Compounds; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Drug Screening Assays, Antitumor; HCT116 Cells; HT29 Cells; Humans; Lignans; PC-3 Cells; Phenols

With the aim to discover interesting lead compounds that could be further developed into compounds active against pharmacoresistant epilepsies, we first collected 14 medicinal plants used in traditional Chinese medicine (TCM) against epilepsy. Of the six extracts that tested positive in a pentylenetetrazole (PTZ) behavioral zebrafish model, only the ethanol and acetone extracts from

Topics: Animals; Anticonvulsants; Biphenyl Compounds; Epilepsy; Lignans; Magnolia; Medicine, Chinese Traditional; Mice; Plant Extracts; Zebrafish

This study aimed to develop, evaluate, and optimize the mPEG-PLA/vitamin E-TPGS mixed micelle drug delivery system to encapsulate celecoxib (CXB) and honokiol (HNK) for intravenous treatment of breast cancer. To this end, we formulated CXB-loaded mPEG-PLA/vitamin E-TPGS (PV-CXB) and HNK-loaded mPEG-PLA/vitamin E-TPGS (PV-HNK) mixed micelles and analyzed their characteristics. The 4T1 cell line was used for cytotoxicity determination and cellular uptake experiments, and for establishing a 4T1-bearing mouse model for histopathology, immunofluorescence, terminal deoxynucleotidyl transferase-mediated nick end labeling, and Western blot analysis. The synergistic effects of PV-CXB and PV-HNK combination therapy were investigated in vitro and in vivo using the coefficient of drug interaction values. The mean size of PV-CXB and PV-HNK prepared with optimal formulation was approximately 50 nm, with a spherical shape. PV-CXB and PV-HNK combination therapy exhibited cytotoxicity in 4T1 cells in vitro. However, the toxicity of PV-CXB and PV-HNK combination therapy was not apparent in normal tissues (heart, liver, spleen, lung, and kidney) in vivo and reduced the expression of collagen fibers in tumor tissues. Moreover, the combination therapy reduced the expression of tumor growth biomarkers (Foxp3, CD4, Gr-1, CD11b, CD31, Ki67, FoxM1, and VEGF). In addition, the tumor cell apoptosis rate reached 45.71 ± 0.62%. The combined treatment with PV-CXB and PV-HNK showed synergistic effect both in vitro and in vivo. Thus, the PV-CXB and PV-HNK drug delivery system could be used as a potential combination therapy for breast cancer .

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Breast Neoplasms; Celecoxib; Lignans; Mice; Mice, Inbred BALB C; Polyesters; Polyethylene Glycols; Vitamin E

A facile method for the activation of γ-cyclodextrin metal-organic framework (CD-MOF) without channel blockage and framework collapse was first developed using supercritical carbon dioxide (scCO

Topics: Animals; Biological Availability; Biphenyl Compounds; Caco-2 Cells; Carbon Dioxide; Drug Delivery Systems; Fluorescent Dyes; gamma-Cyclodextrins; Humans; Hydrogen-Ion Concentration; Lignans; Male; Metal-Organic Frameworks; Molecular Structure; Optical Imaging; Particle Size; Rats; Rats, Sprague-Dawley; Solubility; Surface Properties; Tumor Cells, Cultured

Honokiol, a natural bioactive neolignan isolated from the bark and leaf of Magnolia officinalis and Magnolia obovata, exhibits many important biological properties. In continuation of our interest in discovery of the agrochemicals derived from the natural sources, thirty-seven new 8/8'-alkylthiol-benzoxazole and N-alkyl/sulfonyl-benzoxazolone derivatives of honokiol were prepared and their insecticidal activities were evaluated against the larvae of Mythimna separata Walker and Plutella xylostella Linnaeus. The results showed that eleven derivatives exhibited potent insecticidal activity against M. separata when compared with the positive control. Particularly, compound 5h displayed the most promising insecticidal activity against M. separata with the final mortality rate (FMR) of 58.6%. Meanwhile, compounds 7n (FMR = 65.3%), 7p (FMR = 61.5%), and 8c (FMR = 65.3%) demonstrated a greater insecticidal activity against P. xylostella than toosendanin, a well-known botanical insecticide. Additionally, the preliminary structure-activity relationships (SARs) were also discussed. This study indicates that these honokiol derivatives could be used as leads for the further derivation and development of the potential pesticide candidates for crop protection.

Topics: Animals; Benzoxazoles; Biphenyl Compounds; Insecticides; Larva; Lignans; Magnolia; Models, Molecular; Molecular Structure; Moths; Plant Bark; Plant Stems

Temozolomide (TMZ)-induced chemoresistance to human glioblastomas is a critical challenge now. Our previous studies showed that honokiol, a major bioactive constituent of

Topics: Apoptosis; Biphenyl Compounds; Caspase 9; Cell Line, Tumor; Drug Resistance, Neoplasm; Enzyme Activation; Glioblastoma; Humans; Lignans; Mitochondria; Neoplasm Proteins; Temozolomide

Honokiol (HK) is a common herbal medicine extracted from magnolia plants. Low aqueous solubility and limited bioavailability of HK have hindered its clinical application, especially for cancer treatment. Nano-drug delivery system has the potential to enhance HK delivery and therefore, enhance its anti-cancer activity.. The study's aim is to design novel PEGylated-PLGA polymeric nanocapsules (NCs) for HK delivery to breast tumor-bearing mice after systemic administration.. Formulation of different HK-loaded NCs and their physio-chemical characterization were optimized through the use of different formulation variables. The antitumor activity of the HK-loaded NCs was investigated both in vitro using MCF-7 and EAC breast cancer cell lines and in vivo using solid Ehrlich carcinoma (SEC) breast cancer model.. The optimum HK-loaded NCs were prepared from 15% PEG-PLGA diblock copolymer and exhibited the lowest nano size of 125 nm, smooth spherical morphology, highest drug loading of 94% and highest cellular uptake into breast cancer cells. HK-loaded PEGylated NCs can effectively inhibit the in vitro cell growth of breast cancer cells by 80.2% and 58.1% compared to 35% and 31% with free HK in the case of MCF-7 and EAC, respectively. HK-loaded NCs inhibited SEC tumor growth by 2.3 fold significantly higher than free HK, in vivo.. The designed drug delivery system encapsulating HK exhibited a pronounced decrease in tumor growth biomarkers meanwhile proved its safety in animals. Therefore, 15% PEGylated HK-loaded NCs may act as a promising new approach for breast cancer treatment.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Breast Neoplasms; Carcinoma, Ehrlich Tumor; Drug Delivery Systems; Female; Humans; Lignans; MCF-7 Cells; Mice; Micelles; Nanocapsules; Polyethylene Glycols; Polyglactin 910; Solubility; Xenograft Model Antitumor Assays

Magnolia extract (ME) is known to inhibit cancer growth and metastasis in several cell types in vitro and in animal models. However, there is no detailed study on the preventive efficacy of ME for oral cancer, and the key components in ME and their exact mechanisms of action are not clear. The overall goal of this study is to characterize ME preclinically as a potent oral cancer chemopreventive agent and to determine the key components and their molecular mechanism(s) that underlie its chemopreventive efficacy.. The antitumor efficacy of ME in oral cancer was investigated in a 4-nitroquinoline-1-oxide (4NQO)-induced mouse model and in two oral cancer orthotopic models. The effects of ME on mitochondrial electron transport chain activity and ROS production in mouse oral tumors was also investigated.. ME did not cause detectable side effects indicating that it is a promising and safe chemopreventive agent for oral cancer. Three major key active compounds in ME (honokiol, magnolol and 4-O-methylhonokiol) contribute to its chemopreventive effects. ME inhibits mitochondrial respiration at complex I of the electron transport chain, oxidizes peroxiredoxins, activates AMPK, and inhibits STAT3 phosphorylation, resulting in inhibition of the growth and proliferation of oral cancer cells.. Our data using highly relevant preclinical oral cancer models, which share histopathological features seen in human oral carcinogenesis, suggest a novel signaling and regulatory role for mitochondria-generated superoxide and hydrogen peroxide in suppressing oral cancer cell proliferation, progression, and metastasis. Video abstract.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Line, Tumor; Drug Evaluation, Preclinical; Female; Humans; Lignans; Magnolia; Mice; Mice, Nude; Mitochondria; Mouth Neoplasms; Plant Extracts; Reactive Oxygen Species

Fatty acid oxidation (FAO) dysfunction is one of the important mechanisms of renal fibrosis. Sirtuin 3 (Sirt3) has been confirmed to alleviate acute kidney injury (AKI) by improving mitochondrial function and participate in the regulation of FAO in other disease models. However, it is not clear whether Sirt3 is involved in regulating FAO to improve the prognosis of AKI induced by cisplatin. Here, using a murine model of cisplatin-induced AKI, we revealed that there were significantly FAO dysfunction and extensive lipid deposition in the mice with AKI. Metabolomics analysis suggested reprogrammed energy metabolism and decreased ATP production. In addition, fatty acid deposition can increase reactive oxygen species (ROS) production and induce apoptosis. Our data suggested that Sirt3 deletion aggravated FAO dysfunction, resulting in increased apoptosis of kidney tissues and aggravated renal injury. The activation of Sirt3 by honokiol could improve FAO and renal function and reduced fatty acid deposition in wide-type mice, but not Sirt3-defective mice. We concluded that Sirt3 may regulate FAO by deacetylating liver kinase B1 and activating AMP-activated protein kinase. Also, the activation of Sirt3 by honokiol increased ATP production as well as reduced ROS and lipid peroxidation through improving mitochondrial function. Collectively, these results provide new evidence that Sirt3 is protective against AKI. Enhancing Sirt3 to improve FAO may be a potential strategy to prevent kidney injury in the future.

Topics: Acetylation; Acute Kidney Injury; Animals; Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Cisplatin; Fatty Acids; Fatty Acids, Nonesterified; Kidney Function Tests; Lignans; Lipid Metabolism; Lipid Peroxidation; Lipids; Male; Metabolomics; Mice; Mice, Knockout; Mitochondria; Phosphorylation; Prognosis; Reactive Oxygen Species; Sirtuin 3

We characterized the effects of Honokiol (HNK) on Aspergillus fumigatus-caused keratomycosis and the underlying mechanisms. HNK is known to have anti-inflammatory and antifungal properties, but the influence on fungal keratitis (FK) remains unknown.. In ex vivo, minimum inhibitory concentration and Cell Count Kit-8 assay were carried out spectrophotometrically to provide preferred concentration applied in vivo. Time kill assay pointed that HNK was fungicidal and fungistatic chronologically. Adherence assay, crystal violet staining, and membrane permeability assay tested HNK effects on different fungal stages. In vivo, clinical scores reflected the improvement degree of keratitis outcome. Myeloperoxidase (MPO) assay, flow cytometry (FCM), and immunohistofluorescence staining (IFS) were done to evaluate neutrophil infiltration. Plate count detected HNK fungicidal potentiality. RT-PCR, Western blot, and enzyme-linked immunosorbent assay (ELISA) verified the anti-inflammatory activity of HNK collaboratively.. In vitro, MIC90 HNK was 8 µg/mL (no cytotoxicity), and Minimal Fungicidal Concentration (MFC) was 12 µg/mL for A. fumigatus. HNK played the fungistatic and fungicidal roles at 6 and 24 hours, respectively, inhibiting adherence at the beginning, diminishing biofilms formation, and increasing membrane permeability all the time. In vivo, HNK improved C57BL/6 mice outcome by reducing disease severity (clinical scores), neutrophil infiltration (MPO, FCM, and IFS), and fungal loading (plate count). RT-PCR, Western blot, and ELISA revealed that HNK downregulated mRNA and protein expression levels of Toll-like receptor-2 (TLR-2), high mobility group box 1 (HMGB1), IL-1β, and TNF-α.. Our study suggested HNK played antifungal and anti-inflammatory roles on keratomycosis by reducing survival of fungi, infiltration of leucocytes, and expression of HMGB1, TLR-2, and proinflammatory cytokines, providing a potential treatment for FK.

Topics: Animals; Anti-Infective Agents; Aspergillosis; Aspergillus fumigatus; Biphenyl Compounds; Blotting, Western; Cytokines; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Eye Infections, Fungal; Female; Keratitis; Lignans; Mice; Mice, Inbred C57BL; Random Allocation; Real-Time Polymerase Chain Reaction; Toll-Like Receptor 2; Treatment Outcome

Topics: Animals; Atherosclerosis; Biphenyl Compounds; Carotid Arteries; Disease Models, Animal; Down-Regulation; Enzyme Inhibitors; Inflammation; Lignans; Male; Mice; Oxidative Stress; Plaque, Atherosclerotic; Signal Transduction

More effective treatments for diabetic nephropathy remain a major unmet clinical need. Increased oxidative stress is one of the most important pathological mechanisms that lead to kidney damage and functional impairment induced by diabetes. Sirtuin 3 (SIRT3) is the main mitochondrial deacetylase and critically regulates cellular reactive oxygen species (ROS) production and detoxification. Honokiol is a natural biphenolic compound that, by activating mitochondrial SIRT3, can carry out anti-oxidant, anti-inflammatory and anti-fibrotic activities. Here, we sought to investigate the renoprotective effects of honokiol in BTBR ob/ob mice with type 2 diabetes. Diabetic mice were treated with vehicle or honokiol between the ages of 8 and 14 weeks. Wild-type mice served as controls. Renal Sirt3 expression was significantly reduced in BTBR ob/ob mice, and this was associated with a reduction in its activity and increased ROS levels. Selective activation of SIRT3 through honokiol administration translated into the attenuation of albuminuria, amelioration of glomerular damage, and a reduction in podocyte injury. SIRT3 activation preserved mitochondrial wellness through the activation of SOD2 and the restoration of PGC-1α expression in glomerular cells. Additionally, the protective role of SIRT3 in glomerular changes was associated with enhanced tubular Sirt3 expression and upregulated renal Nampt levels, indicating a possible tubule-glomerulus retrograde interplay, which resulted in improved glomerular SIRT3 activity. Our results demonstrate the hitherto unknown renoprotective effect of SIRT3 against diabetic glomerular disease and suggest that the pharmacological modulation of SIRT3 activity is a possible novel approach to treating diabetic nephropathy.

Topics: Albuminuria; Animals; Anti-Inflammatory Agents; Antioxidants; Biphenyl Compounds; Cytokines; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Kidney Glomerulus; Lignans; Male; Mice; Mice, Obese; Mitochondria; Nicotinamide Phosphoribosyltransferase; Oxidative Stress; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; Podocytes; Reactive Oxygen Species; Sirtuin 3; Superoxide Dismutase

Intestinal obstruction (IO) is a kind of acute abdomen with high morbidity and mortality. Patients suffer from poor quality of life and tremendous financial pressure. Da-Cheng-Qi decoction (DCQD), a classical purgation prescription, has clinically been proven to be an effective treatment for IO.. Network pharmacology integrated with bioactive equivalence assessment was used to discover the quality marker (Q-marker) of DCQD against IO.. As there is hardly any targets recorded in database, thus the collection of IO targets was conducted by searching those of alternative diseases which have similar pathological symptoms with IO. In order to improve the reliability of the obtained targets, IO metabolomics data was introduced. Active compounds combination (ACC) was focused as potential Q-markers via component-target network analysis and function query from the identified components corresponding to the common targets. Bioequivalence between ACC and DCQD was assessed from the aspects of intestine motility (somatostatin secretion), inflammation (IL-6 secretion) and injury (wound healing assay) in vitro and was further validated in ileus rat model. PPI network analysis of core targets followed by gene pedigree classification and experimental validation confirmed the potential intervention pathway.. A combination of 11 ingredients, including emodin, physcion, aloe-emodin, rhein, chrysophanol, gallic acid, magnolol, honokiol, naringenin, tangeretin, and nobiletin was finally confirmed bioequivalence with DQCD to some extent and could serve as Q-markers for DCQD to attenuate IO. PI3K/AKT was verified as a possible affected pathway that DCQD exerted the effectiveness against IO.. For the disease with few recorded targets, searching those of alternative diseases which have similar pathological symptoms could be a feasible and effective approach. The proposed network pharmacology integrated bioactive equivalence evaluation paradigm is efficient to discover Q-marker of herbal formulae.

Topics: Algorithms; Animals; Anthraquinones; Biomarkers, Pharmacological; Biphenyl Compounds; Data Mining; Drugs, Chinese Herbal; Flavanones; HT29 Cells; Humans; Intestinal Obstruction; Lignans; Male; Phosphatidylinositol 3-Kinases; Rats, Sprague-Dawley; Reproducibility of Results; Therapeutic Equivalency

Based on the antisolvent and electrostatic deposition methods, we fabricated zein/hyaluronic acid core-shell nanoparticles loaded with honokiol (HA-Zein-HNK), which could target delivery and enhance the therapeutic effect of the HNK. The prepared nanoparticles were found to have a mean size of 210.4 nm and negative surface charge. The HA-Zein-HNK nanoparticles exhibited improved antiproliferative and pro-apoptotic activities against 4T1 cells. Of note, the wound healing and transwell assessments indicated that the migration and invasion of 4T1 cells were markedly weakened by HA-Zein-HNK. Mechanistic insights revealed that HA-Zein-HNK downregulated the expressions of Vimentin and upregulated the expressions of E-cadherin. More importantly, an in vivo tissue distribution study demonstrated the excellent tumor target ability of HA-Zein. And these results correspond with the superior therapeutic efficacy of HA-Zein-HNK in 4T1 tumor bearing mice. In conclusion, we believe that HA-Zein nanoparticles may be served as a promising HNK delivery carrier for metastatic breast cancer therapy.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Breast Neoplasms; Cell Movement; Cell Proliferation; Drug Carriers; Drug Screening Assays, Antitumor; Female; Hyaluronic Acid; Lignans; Mammary Neoplasms, Experimental; Mice; Mice, Inbred BALB C; Nanoparticles; Tumor Cells, Cultured; Zein

Topics: AMP-Activated Protein Kinases; Biphenyl Compounds; Chagas Disease; Humans; Lignans; NF-E2-Related Factor 2; Oxidative Stress; Signal Transduction; Sirtuin 3

Topics: Animals; Biphenyl Compounds; Cell Line; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; Lignans; Magnolia; Membrane Potentials; Patch-Clamp Techniques; Plant Extracts; Rats

Two isomeric biphenyl neolignans, magnolol and honokiol, are considered as constituents responsible for the healing effect of magnolia bark, a traditional Oriental medicine. To survey the increasing number of dietary supplements that contain magnolia bark or its extract, an affordable quantitative thin-layer chromatography (TLC) - densitometry method was developed. The methanol extracts were analyzed on the silica gel plates after manual sample application using n-hexane - ethyl acetate - ethanol (16:3:1, v/v/v) as a mobile phase. For quantitation, the chromatograms were scanned in the absorbance mode at the wavelength λ = 290 nm. The limits of detection and quantitation were 90 and 280 ng/zone for magnolol and 70 and 200 ng/zone for honokiol, respectively. None of the two targeted neolignans were detected in two of the six analyzed supplements. In the other four samples, the measured amounts were between 0.95-114.69 mg g

Topics: Biphenyl Compounds; Chromatography, Thin Layer; Densitometry; Dietary Supplements; Lignans; Limit of Detection; Magnolia; Medicine, East Asian Traditional; Plant Bark

In the present study, a novel strategy based on unidirectional freezing and atom transfer radical polymerization combined with activator regenerated by electron transfer (ARGET-ATRP) was applied to synthesizing orderly macroporous monolithic column with restricted-access (RA) property in a 1000μL pipette tip. The RA column was composed of hydrophobic inner column (poly(styrene-co-ethylene glycol dimethacrylate) and hydrophilic outer layer (poly-hydroxyethyl methacrylate chain) which was grafted on the hydrophobic surface by means of the second ARGET-ATRP reaction. The as-prepared RA monolithic tip was connected to a 2mL syringe for directly extracting magnolol and honokiol from rat plasma just by manually pushing operation. The surface morphology and chemical composition of the column were characterized by scanning electronic microscope, infrared spectroscopy and X-ray photoelectron spectroscopy respectively. The determined results of evaluation experiments based on the optimized solid phase extraction conditions showed that the RA column possessed good protein exclusion power, extraction recovery and reusability. The constructed RA-SPE-HPLC/UV method for simultaneously analyzing magnolol and honokiol in rat plasma was validated with quality control (QC) samples at four concentration levels. Good precision (RSDs, 3.39~11.16%) and acceptable accuracy (relative recoveries, 89.52%~108.42%) were obtained for intra- and inter-day assays. The determined results of real rat plasma as well as the standard-addition samples demonstrated the developed method with good accuracy and precision. It can be extrapolated from the experimental results that this simple and cost-efficient RA-SPE method is also suitable for directly extracting other hydrophobic constituents in biological body fluid for therapeutic drug monitoring or pharmacokinetic study.

Topics: Animals; Biphenyl Compounds; Chromatography, High Pressure Liquid; Freezing; Hydrophobic and Hydrophilic Interactions; Lignans; Methacrylates; Polyethylene Glycols; Quality Control; Rats; Reproducibility of Results; Solid Phase Extraction

Hypertension and endothelial dysfunction are associated with various cardiovascular diseases. Hydrogen sulphide (H

Topics: Acetylation; Angiotensin II; Animals; Aorta; Biphenyl Compounds; Cystathionine gamma-Lyase; Endothelial Cells; HEK293 Cells; Histone Deacetylase 6; Humans; Hydrogen Sulfide; Hypertension; Lignans; Male; Mice; Mice, Inbred C57BL; Proteasome Endopeptidase Complex; Protein Processing, Post-Translational; Proteolysis; Recombinant Proteins

Osteoarthritis is the leading cause of disability worldwide; cartilage degeneration and defects are the central features. Significant progress in tissue engineering holds promise to regenerate damaged cartilage tissue. However, a formidable challenge is to develop a 3-dimensional (3D) tissue construct that can regulate local immune environment to facilitate the intrinsic osteochondral regeneration.. To evaluate efficacy of a 3D-printed decellularized cartilage extracellular matrix (ECM) and polyethylene glycol diacrylate (PEGDA) integrated novel scaffold (PEGDA/ECM) together with the natural compound honokiol (Hon) for regenerating osteochondral defect.. Controlled laboratory study.. We used a stereolithography-based 3D printer for PEGDA/ECM bioprinting. A total of 36 Sprague-Dawley rats with cylindrical osteochondral defect in the trochlear groove of the femur were randomly assigned into 3 different treatments: no scaffold implantation (Defect group), 3D printed PEGDA/ECM scaffold alone (PEGDA/ECM group), or Hon suspended in a 3D-printed PEGDA/ECM scaffold (PEGDA/ECM/Hon group). 12 rats that underwent only medial parapatellar incision surgery were used as normal controls. The femur specimens were postoperatively harvested at 4 and 8 weeks for gross, micro-CT, and histological evaluations. The efficacy of PEGDA/ECM/Hon scaffold on the release of proinflammatory cytokines from the macrophages stimulated by lipopolysaccharide (LPS) was evaluated in-vitro.. In vitro results determined that PEGDA/ECM/Hon scaffold could suppress the release of proinflammatory cytokines from macrophages that were stimulated by LPS. Macroscopic images showed that the PEGDA/ECM/Hon group had significantly higher ICRS scoring than that of defect and PEGDA/ECM groups. Micro-CT evaluation demonstrated that much more bony tissue was formed in the defect sites implanted with the PEGDA/ECM scaffold or PEGDA/ECM/Hon scaffold compared with the untreated defects. Histological analysis showed that the PEGDA/ECM/Hon group had a significant enhancement in osteochondral regeneration at 4 and 8 weeks after surgery in comparison with the ECM/PEGDA or defect group.. This study demonstrated that 3D printing of PEGDA/ECM hydrogel incorporating the anti-inflammatory phytomolecule honokiol could provide a promising scaffold for osteochondral defect repair.

Topics: Animals; Anti-Inflammatory Agents; Biphenyl Compounds; Cartilage, Articular; Extracellular Matrix; Hydrogels; Lignans; Osteoarthritis; Polyethylene Glycols; Printing, Three-Dimensional; Rats; Rats, Sprague-Dawley; Regeneration; Tissue Scaffolds

Honokiol (HK), an active compound derived from Magnolia officinalis Rehd. et Wils, possesses many beneficial biological activities for human beings. However, its poor solubility and low bioavailability severely limits its application. In this way, to improve the pharmaceutical properties, the HK was complexed in hydroxypropyl-β-cyclodextrin (HP-β-CD) and its oral bioavailability and antitumor effects were evaluated. The HK/HP-β-CD inclusion complex (1:1) was prepared by saturated aqueous solution method. The inclusion complex (HK-HP-β-CD) obtained had a higher solubility, about 1497 times that of the free HK. The dissolution rate and the oral bioavailability of HK was also significantly higher from inclusion complex than from free HK. Furthermore, the HK-HP-β-CD exhibited higher antitumor activity against Human Hepatoma Cell Line (HepG2) than free HK. More cells were arrested in the sub-G1 phase of the cell cycle and were induced to undergo late apoptosis when treated with the HK-HP-β-CD than when treated with free HK.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; beta-Cyclodextrins; Biphenyl Compounds; Humans; Lignans; Solubility

The main pathological mechanism of restenosis after percutaneous coronary intervention (PCI) is intimal hyperplasia, which is mainly caused by proliferation and migration of vascular smooth muscle cells (VSMCs). Our previous study found that honokiol (HNK), a small-molecule polyphenol, can inhibit neointimal hyperplasia after balloon injury, but its specific mechanism is still unclear. Moreover, poor water solubility as well as low bioavailability of honokiol has limited its practical use.. We used mesoporous silica nanoparticles (MSNPs) as a standard substance to encapsulate HNK and then assemble into honokiol-mesoporous silica nanoparticles, and we investigated the effect of these nanoparticles on the process of restenosis after common carotid artery injury in rats.. We report a promising delivery system that loads HNK into MSNPs and finally assembles it into a nanocomposite particle. These HNK-MSNPs not merely inhibited proliferation and migration of VSMCs by reducing phosphorylation of Smad3, but also showed a higher suppression of intimal thickening than the free-honokiol-treated group in a rat model of balloon injury.. To sum up, this drug delivery system supplies a potent nano-platform for improving the biological effects of HNK and provides a promising strategy for preventing vascular restenosis.

Topics: Animals; Biphenyl Compounds; Carotid Artery Injuries; Cell Line; Cell Proliferation; Coronary Restenosis; Disease Models, Animal; Drug Delivery Systems; Humans; Lignans; Male; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Nanoparticles; Percutaneous Coronary Intervention; Poloxamer; Rats, Sprague-Dawley; Silicon Dioxide

Using honokiol (HNK), a major anti-inflammatory bioactive compound in Magnolia officinalis, we show a potent therapeutic outcome against an accelerated, severe form of lupus nephritis (ASLN). The latter may follow infectious insults that act as environmental triggers in the patients. In the current study, an ASLN model in NZB/W F1 mice was treated with HNK by daily gavage after onset of the disease. We show that HNK ameliorated the ASLN by improving renal function, albuminuria, and renal pathology, especially reducing cellular crescents, neutrophil influx, fibrinoid necrosis in glomeruli, and glomerulonephritis activity scores. Meanwhile, HNK differentially regulated T cell functions, reduced serum anti-dsDNA autoantibodies, and inhibited NLRP3 inflammasome activation in the mice. The latter involved: (a) suppressed production of reactive oxygen species and NF-κB activation-mediated priming signal of the inflammasome, (b) reduced mitochondrial damage, and (c) enhanced sirtuin 1 (SIRT1)/autophagy axis activation. In conclusion, HNK represents a new drug candidate for acute, severe episodes of LN capable of alleviating renal lesions in ASLN mice by negatively regulating T cell functions and by enhancing SIRT1/autophagy axis-lessened NLRP3 inflammasome activation.

Topics: Animals; Anti-Inflammatory Agents; Autophagy; Biphenyl Compounds; Cells, Cultured; Female; Inflammasomes; Kidney; Lignans; Lupus Nephritis; Mice; NF-kappa B; NLR Family, Pyrin Domain-Containing 3 Protein; Reactive Oxygen Species; Sirtuin 1; T-Lymphocytes

Although radiotherapy, especially carbon-ion radiotherapy, is an effective treatment modality against non-small-cell lung cancer (NSCLC), studies using radiation combined with sensitizer for improving the efficacy of radiotherapy are still needed. In this work, we aimed to investigate in NSCLC A549 and H1299 cell lines the effects of different linear energy transfer (LET) radiations combined with diverse sensitizing compounds. Cells pretreated with the CHK1/CHK2 inhibitor AZD7762, Honokiol or Tunicamycin were irradiated with low-LET X-rays and high-LET carbon ions. Cell survival was assessed using the clonogenic cell survival assay. Cell cycle distribution and apoptosis were measured with flow cytometry, and DNA double strand break (DSB) and repair were detected using γ-H2AX immunofluorescence staining. Our results revealed that AZD7762, Honokiol and Tunicamycin demonstrated low cytotoxicity to NSCLC cells and a pronounced radiosensitizing effect on NSCLC cells exposed to carbon ions than X-rays. Unrepaired DNA DSB damages, the abrogation of G2/M arrest induced by irradiation, and finally apoptotic cell death were the main causes of the radiosensitizing effect. Thus, our data suggest that high-LET carbon ion combined with these compounds may be a potentially effective therapeutic strategy for locally advanced NSCLC.

Topics: Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Carbon; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Proliferation; Checkpoint Kinase 1; Checkpoint Kinase 2; DNA Breaks, Double-Stranded; G2 Phase Cell Cycle Checkpoints; Humans; Ions; Lignans; Linear Energy Transfer; Lung Neoplasms; Protein Kinase Inhibitors; Radiation Tolerance; Radiation-Sensitizing Agents; Thiophenes; Tunicamycin; Urea; X-Rays

BACKGROUND The purpose of our study was to determine the effects and mechanisms of honokiol on human epidermal growth factor receptor 2 (HER2)-positive gastric cancer cells by in vitro study. MATERIAL AND METHODS We measured HER2 expression in different gastric cancer cell lines by real-time quantitative polymerase chain reaction (RT-qPCR) and western blot (WB) assay. Cell proliferation, apoptosis, and cell cycle were evaluated by cell-counting kit 8 and flow cytometry assays. The invading cell numbers and wound-healing rates were measured by transwell and wound-healing assays. Phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), P21, and matrix metalloproteinase (MMP)-9 proteins and messenger ribonucleic acid (mRNA) expression were measured by WB and RT-qPCR assay. HER2 protein expression was evaluated by cellular immunofluorescence. RESULTS Honokiol suppressed cell proliferation via increasing cell apoptosis, invasion, and migration with dose dependence. By WB and RT-qPCR assays, compared with the control group, PI3K, AKT, P21, and MMP-9 proteins and mRNA expression were significantly different (P<0.05). By cellular immunofluorescence, HER2 protein expression was significantly depressed in honokiol-treated groups compared with control groups (P<0.05). CONCLUSIONS Honokiol has suppressive effects on HER2-positive gastric cancer cell biological activities via regulation of HER2/PI3K/AKT pathways in vitro.

Topics: Apoptosis; Biphenyl Compounds; Case-Control Studies; Cell Proliferation; Humans; Lignans; Matrix Metalloproteinase 9; Phosphatidylinositol 3-Kinase; Receptor, ErbB-2; RNA, Messenger; Stomach Neoplasms; Tumor Cells, Cultured

Cardiac energetic dysfunction has been reported in patients with type 2 diabetes (T2D) and is an independent predictor of mortality. Identification of the mechanisms driving mitochondrial dysfunction, and therapeutic strategies to rescue these modifications, will improve myocardial energetics in T2D. We demonstrate using 31P-magnetic resonance spectroscopy (31P-MRS) that decreased cardiac ATP and phosphocreatine (PCr) concentrations occurred before contractile dysfunction or a reduction in PCr/ATP ratio in T2D. Real-time mitochondrial ATP synthesis rates and state 3 respiration rates were similarly depressed in T2D, implicating dysfunctional mitochondrial energy production. Driving this energetic dysfunction in T2D was an increase in mitochondrial protein acetylation, and increased ex vivo acetylation was shown to proportionally decrease mitochondrial respiration rates. Treating T2D rats in vivo with the mitochondrial deacetylase SIRT3 activator honokiol reversed the hyperacetylation of mitochondrial proteins and restored mitochondrial respiration rates to control levels. Using 13C-hyperpolarized MRS, respiration with different substrates, and enzyme assays, we localized this improvement to increased glutamate dehydrogenase activity. Finally, honokiol treatment increased ATP and PCr concentrations and increased total ATP synthesis flux in the T2D heart. In conclusion, hyperacetylation drives energetic dysfunction in T2D, and reversing acetylation with the SIRT3 activator honokiol rescued myocardial and mitochondrial energetics in T2D.

Topics: Acetylation; Adenosine Triphosphate; Animals; Anti-Arrhythmia Agents; Biphenyl Compounds; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diet, High-Fat; Energy Metabolism; Heart Diseases; Lignans; Male; Mitochondria, Heart; Myocardium; Phosphocreatine; Rats; Rats, Wistar

To study the effects of honokiol on proliferation, migration and apoptosis of human tongue carcinoma CAL-27 cells.. Routinely cultured CAL-27 cells were treated with 20, 40, or 60 μmol/L honokiol and the changes in cell proliferation were assessed with MTT assay. The scratch wound healing assay was used to assess the migration ability of the treated cells, and the cell apoptosis was detected with Hoechst33342 fluorescence staining and annexin V-FITC/PI method. The protein expression levels of p-Pi3k, p-Fak, Fak, MMP-2, MMP-9, p-Akt, Akt, Bax, Bcl-2 and cleaved-caspase-3 in the treated cells were detected using Western blotting.. Treatment with honokiol at 20, 40, and 60 μmol/L for 24 h significantly lowered the proliferation and migration ability of CAL-27 cells. The number of apoptotic cells increased with the increase of honokiol concentration, which resulted in a cell apoptosis rate of (15.24±2.06)% at 20 μmol/L, (35.03±2.42)% at 40 μmol/L, and (48.13±4.61)% at 60 μmol/L, as compared with (6.53±1.80)% in the control group. The expressions of p-Pi3k, p-Fak, MMP-2, MMP-9, p-Akt and BCL-2 decreased and those of Bax and cleaved-caspase-3 increased significantly in the cells after the treatment (. Honokiol can inhibit the proliferation and migration and induce apoptosis of CAL-27 cells

Topics: Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Humans; Lignans; Tongue Neoplasms

Honokiol (HNK), a natural pharmaceutically active component extracted from magnolia bark, has been used for clinical treatments and has anti‑inflammatory, antiviral and antioxidative effects. In recent years, anticancer research has become a major hotspot. However, the underlying molecular mechanisms of how HNK inhibits colorectal cancer have remained elusive. The present study focused on elucidating the effects of HNK on the expression of bone morphogenetic protein (BMP)7 and its downstream interaction with transforming growth factor (TGF)‑β1 and p53 in colon cancer. In in vitro assays, cell viability, cell cycle distribution and apoptosis were examined using Cell Counting Kit‑8, flow cytometry and reverse transcription‑quantitative PCR, respectively. In addition, the expression of BMP7, TGF‑β1 and relevant signaling proteins was determined by western blot analysis. In vivo, the anticancer effect of HNK was assessed in xenografts in nude mice. Furthermore, immunohistochemistry was performed to evaluate the association between BMP7 and TGF‑β1 expression in colon cancer. The results indicated that HNK inhibited the proliferation of colon cancer cell lines, with SW620 cells being more sensitive than other colon cancer cell lines. Furthermore, HNK markedly promoted the expression of BMP7 at the mRNA and protein level. Exogenous BMP7 potentiated the effect of HNK on SW620 cells, while knocking down BMP7 inhibited it. As a downstream mechanism, HNK increased the expression of TGF‑β1 and p53, which was enhanced by exogenous BMP7 in SW620 cells. In addition, immunohistochemical analysis indicated a positive association between BMP7 and TGF‑β1 expression. Hence, the present results suggested that HNK is a promising agent for the treatment of colon cancer and enhanced the expression TGF‑β1 and p53 through stimulating BMP7 activity via the non‑canonical TGF‑β signaling pathway.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Bone Morphogenetic Protein 7; Cell Line, Tumor; Cell Proliferation; Cell Survival; Colonic Neoplasms; Female; Humans; Lignans; Mice; Mice, Nude; Signal Transduction; Transforming Growth Factor beta1; Tumor Suppressor Protein p53; Xenograft Model Antitumor Assays

Honokiol (HK) has a variety of biological activities, but its poor solubility limits its application. Rebaudioside A (RA) is able to self-assemble into micelles, which can be used for oral delivery of anticancer drugs. This study aims to create and evaluate a nano-sized anticancer drug delivery system based on RA, as RA micelles are thought to strengthen the therapeutic effects of HK. The results showed that RA and HK can be formulated into self-assembling micelles (RA-HK) with a size of 4.356 ± 0.142 nm and uniform distribution (PDI = 0.1906 ± 0.0184). Moreover, RA-HK could enhance the antitumor activity of HK in vitro. Further, it was shown that RA-HK can induce G0/G1 cycle arrest, apoptosis, and reactive oxygen species (ROS) generation in HuH-7 cells. The results for this mechanism indicate that RA-HK can induce DNA damage as well as changes in cycle and apoptotic-related proteins and activate the ERK signaling pathway. The in vivo antitumor results showed that RA-HK could also enhance the antitumor activity of HK in mice and does not induce any side effects. The pharmacokinetic results illustrate that RA-HK can increase the oral bioavailability of HK that and RA-HK is widely distributed in rats. Taken together, the above results prove that RA is a novel oral nano-drug delivery system with great potential for the delivery of hydrophobic antitumor drugs, such as HK.

Topics: Animals; Biological Availability; Biphenyl Compounds; Diterpenes, Kaurane; Drug Delivery Systems; Lignans; Mice; Micelles; Rats

Topics: Antifungal Agents; Biological Assay; Biphenyl Compounds; Lignans; Magnolia

Mitochondrion is a favorable therapeutic target in cancer, given its regulation of bioenergetics and cell death. Honokiol exhibits antiproliferative effects through mitochondria-mediated death signaling. To enhance its anticancer potential and selectivity, we conjugated honokiol to berberine, a mitochondria-targeting carrier. All designed derivatives displayed 1 order of magnitude increased cytotoxicity compared with the parent compounds, especially with massive cytoplasmic vacuoles. Biological evaluation demonstrated the representative compound

Topics: Antineoplastic Agents; Apoptosis; Berberine; Biphenyl Compounds; Cell Line; Cell Proliferation; Dose-Response Relationship, Drug; Drug Screening Assays, Antitumor; Humans; Lignans; Membrane Potential, Mitochondrial; Mitochondria; Molecular Structure; Reactive Oxygen Species; Structure-Activity Relationship

Depression is closely linked to hypothalamus-pituitary-adrenal axis hyperactivity. Honokiol, a biphenolic lignan compound obtained from the traditional Chinese medicine

Topics: Animals; Antidepressive Agents; Biphenyl Compounds; Corticosterone; Depression; Disease Models, Animal; Hippocampus; Lignans; Male; Mineralocorticoids; Rats, Sprague-Dawley; Receptors, Mineralocorticoid; RNA, Messenger

Adjuvants are added to vaccines to enhance the immune response and provide increased protection against disease. In the last decade, hundreds of synthetic immune adjuvants have been created, but many induce undesirable levels of proinflammatory cytokines including TNF-α and IL-6. Here we present small molecule NF-κB inhibitors that can be used in combination with an immune adjuvant to both decrease markers associated with poor tolerability and improve the protective response of vaccination. Additionally, we synthesize a library of honokiol derivatives identifying several promising candidates for use in vaccine formulations.

Topics: Adjuvants, Immunologic; Animals; Biphenyl Compounds; Lignans; Mice; NF-kappa B; RAW 264.7 Cells; Vaccines

Sirtuin 3 (SIRT3) plays a vital role in regulating oxidative stress in tissue injury. The aim of this study was to evaluate the radioprotective effects of honokiol (HKL) in a zebrafish model of radiation-induced brain injury and in HT22 cells.. The levels of reactive oxygen species (ROS), tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β) were evaluated in the zebrafish brain and HT22 cells. The expression levels of SIRT3 and cyclooxygenase-2 (COX-2) were measured using western blot assays and real-time polymerase chain reaction (RT-PCR).. HKL treatment attenuated the levels of ROS, TNF-α, and IL-1β in both the. HKL attenuated oxidative stress and pro-inflammatory responses in a SIRT3-dependent manner in radiation-induced brain injury.

Topics: Animals; Biphenyl Compounds; Brain; Brain Injuries; Lignans; Oxidative Stress; Reactive Oxygen Species; Sirtuin 3; Zebrafish

BACKGROUND Acute pancreatitis (AP) is generally a self-limiting inflammatory disease, but is associated with a high mortality rate when severe. The present study aimed to investigate the effects of rhein and honokiol on AP. MATERIAL AND METHODS Thirty mice were randomly divided into 5 groups (n=6 per group): blank control, AP model, AP+rhein, AP+honokiol, and AP+rhein+honokiol. The AP model was prepared by intraperitoneal injection of cerulein and lipopolysaccharide (LPS). We observed the pathological changes of the pancreas by hematoxylin and eosin (H&E) staining. A mouse amylase kit was utilized to detect the level of amylase content in serum. Gas chromatography mass spectrometer analysis was performed to detect the differences in metabolites among the blank control, AP model, and AP+rhein+honokiol groups. RESULTS The serum amylase level was significantly higher in the AP model, which suggested that the AP model was constructed successfully. The AP+rhein+honokiol group had significantly reduced interstitial edema, inflammatory cell infiltration, hemorrhage, and necrosis. In addition, the rhein and honokiol treatment influenced some of the metabolic pathways in AP, including riboflavin metabolism, glycerophospholipid metabolism, linoleic acid metabolism, and the pentose and glucuronate interconversions pathway. CONCLUSIONS This study showed that the combination of rhein and honokiol ameliorated pathological changes in the pancreas of mice with AP.

Topics: Amylases; Animals; Anthraquinones; Biphenyl Compounds; Disease Models, Animal; Enzyme Inhibitors; Lignans; Male; Metabolome; Mice; Mice, Inbred C57BL; Pancreas; Pancreatitis

Epidural fibrosis (EF)‑induced failed back surgery syndrome (FBSS) in patients post‑laminectomy remains a medical challenge. Although the scarring mechanisms remain unclear, the majority of aetiological studies have reported fibroblast dysfunction. Honokiol, the major bioactive constituent of the magnolia tree, exerts a variety of pharmacological effects, including anti‑proliferative and anti‑fibrotic effects, on various cell types. The present study investigated whether honokiol attenuates EF progression. In vitro, it was found that honokiol inhibited excessive fibroblast proliferation induced by transforming growth factor‑β1 (TGF‑β1) and the synthesis of extracellular matrix (ECM) components, including fibronectin and type I collagen, in a dose‑dependent manner. These effects were attributed to the ability of honokiol to suppress the activity of connective tissue growth factor (CTGF), which is indispensable for the progression of fibrosis. Mechanistically, honokiol attenuated the TGF‑β1‑induced activation of the Smad2/3 and mitogen‑activated protein kinase (MAPK) signalling pathways in fibroblasts. In vivo, honokiol reduced the proliferation of fibroblasts and the synthesis of ECM components, thus ameliorating EF in a rat model post‑laminectomy. Taken together, these preclinical findings suggest that honokiol deserves further consideration as a candidate therapeutic agent for EF.

Topics: Animals; Biphenyl Compounds; Cell Proliferation; Cicatrix; Connective Tissue Growth Factor; Epidural Space; Extracellular Matrix; Fibroblasts; Fibrosis; Laminectomy; Lignans; Male; MAP Kinase Signaling System; Neuroprotective Agents; Rats, Sprague-Dawley; Smad2 Protein; Smad3 Protein; Transforming Growth Factor beta1

The association between protein aggregation and neurodegenerative diseases such as Parkinson's disease continues to be well interrogated but poorly elucidated at a mechanistic level. Nevertheless, the formation of amyloid fibrils from the destabilization and misfolding of native proteins is a molecular hallmark of disease. Consequently, there is ongoing demand for the identification and development of small molecules which prevent fibril formation. This study comprehensively assesses the inhibitory properties of two small molecules, the lignan polyphenol honokiol and the flavonoid 2',3',4'-trihydroxyflavone, in preventing α-synuclein fibrilization. The data shows that honokiol does not prevent α-synuclein fibril elongation, while 2',3',4'-trihydroxyflavone is effective at inhibiting fibril elongation and induces oligomer formation (for both wild-type α-synuclein and the disease-associated A53T mutation). Moreover, the exposed hydrophobicity of α-synuclein fibrils is reduced in the presence of 2',3',4'-trihydroxyflavone, whereas the addition of honokiol did not reduce the hydrophobicity of fibrils. In addition, ion mobility-mass spectrometry revealed that the conformation of α-synuclein wild-type and A53T monomers after disassembly is restored to a nonaggregation-prone state upon 2',3',4'-trihydroxyflavone treatment. Collectively, this study shows that the mechanisms by which these polyphenols and flavonoids prevent fibril formation are distinct by their interactions at various phases of the fibril-forming pathway. Furthermore, this study highlights how thorough biophysical interrogation of the interaction is required for understanding the ability of inhibitors to prevent protein aggregation associated with disease.

Topics: alpha-Synuclein; Amyloid; Biphenyl Compounds; Flavones; Lignans; Polyphenols

Honokiol and magnolol were considered as markers for the analysis of Cortex Magnoliae Officinalis, its related Chinese Patent Medicines and their metabolites. However, the determination of these two analytes in a water-soluble sample is difficult and therefore requires a more efficient method.. To develop a sensitive method for the determination of honokiol and magnolol in a water-soluble sample for better quality control of Cortex Magnoliae Officinalis and its related Chinese Patent Medicines.. In this work, a combination of dispersive micro-solid-phase extraction (DMSPE) and high-performance liquid chromatography (HPLC) has been developed for simultaneous preconcentration and determination of honokiol and magnolol in complex bio-samples. Several experimental factors affecting the extraction efficiency were optimized by single factor test.. Under the optimized extraction conditions, the proposed method exhibited good linearity of not less than 0.9998, satisfactory precision with relative standard deviation of less than 1.3%, and acceptable mean recoveries of 97.3% and 101.5% for honokiol and magnolol, respectively. Furthermore, the method exhibits extremely high sensitivity with detection limits of 0.0097 and 0.0231 ng/mL, which is even more sensitive than those methods developed by MS.. The method established in this study is fast, economic, accurate, easy to operate, and importantly well suited to the extraction and analysis of honokiol and magnolol in a real complex sample matrix.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Lignans; Magnolia; Solid Phase Extraction

Ergosterol, an essential constituent of membrane lipids of yeast, is distributed in both the cell membrane and intracellular endomembrane components such as vacuoles. Honokiol, a major polyphenol isolated from

Topics: Antifungal Agents; Biphenyl Compounds; Calcineurin; Candida albicans; Drug Resistance, Fungal; Ergosterol; Lignans; Magnolia; Microbial Sensitivity Tests; Plant Extracts; Vacuoles

Herein, we sought to evaluate the contribution of the 1,3,5-triazine ring through the metformin cyclization unit to the biological activity of magnolol and honokiol-conjugates. One of the phenolic OH groups of magnolol or honokiol was replaced by a 1,3,5-triazine ring to further explore their synthesis and medicinal versatility. In this study, a robust procedure of three steps was adopted for the synthesis of magnolol and honokiol derivatives by alkylation of potassium carbonate with a 1,3,5-triazine ring. To our knowledge, this is the first report to connect one of the phenolic OH positions of magnolol or honokiol to a 1,3,5-triazine ring cyclized by metformin. The structural characterization of three new compounds was carried out via spectroscopic techniques, i.e.,

Topics: Animals; Antineoplastic Agents; Biphenyl Compounds; Cell Line, Tumor; Cell Survival; Chemistry Techniques, Synthetic; Cyclization; Cytokines; Gene Expression Regulation; Humans; Lignans; Macrophages; Metformin; Mice; Molecular Structure; RAW 264.7 Cells; Structure-Activity Relationship; Triazines

Renal Cell Carcinoma (RCC) often becomes resistant to targeted therapies, and in addition, dose-dependent toxicities limit the effectiveness of therapeutic agents. Therefore, identifying novel drug delivery approaches to achieve optimal dosing of therapeutic agents can be beneficial in managing toxicities and to attain optimal therapeutic effects. Previously, we have demonstrated that Honokiol, a natural compound with potent anti-tumorigenic and anti-inflammatory effects, can induce cancer cell apoptosis and inhibit the growth of renal tumors in vivo. In cancer treatment, implant-based drug delivery systems can be used for gradual and sustained delivery of therapeutic agents like Honokiol to minimize systemic toxicity. Electrospun polymeric fibrous scaffolds are ideal candidates to be used as drug implants due to their favorable morphological properties such as high surface to volume ratio, flexibility and ease of fabrication. In this study, we fabricated Honokiol-loaded Poly(lactide-co-glycolide) (PLGA) electrospun scaffolds; and evaluated their structural characterization and biological activity. Proton nuclear magnetic resonance data proved the existence of Honokiol in the drug loaded polymeric scaffolds. The release kinetics showed that only 24% of the loaded Honokiol were released in 24hr, suggesting that sustained delivery of Honokiol is feasible. We calculated the cumulative concentration of the Honokiol released from the scaffold in 24hr; and the extent of renal cancer cell apoptosis induced with the released Honokiol is similar to an equivalent concentration of direct application of Honokiol. Also, Honokiol-loaded scaffolds placed directly in renal cell culture inhibited renal cancer cell proliferation and migration. Together, we demonstrate that Honokiol delivered through electrospun PLGA-based scaffolds is effective in inhibiting the growth of renal cancer cells; and our data necessitates further in vivo studies to explore the potential of sustained release of therapeutic agents-loaded electrospun scaffolds in the treatment of RCC and other cancer types.

Topics: Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Movement; Cell Proliferation; Drug Liberation; Humans; Kidney Neoplasms; Lignans; Polylactic Acid-Polyglycolic Acid Copolymer; Tissue Scaffolds

Honokiol (HNK) has been reported to possess various beneficial effects in the context of metabolic disorders, including fatty liver, insulin resistance, and oxidative stress which are closely related to nonalcoholic steatohepatitis (NASH), however with no particular reference to CFLAR or JNK.. The. HNK ameliorated NASH mainly by activating the CFLAR-JNK pathway, which not only alleviated fat deposition by promoting the efflux and

Topics: Animals; Biphenyl Compounds; CASP8 and FADD-Like Apoptosis Regulating Protein; Fatty Liver; Food, Formulated; Lignans; Liver; Male; MAP Kinase Kinase 4; MAP Kinase Signaling System; Methionine; Mice; Oxidative Stress

Honokiol (HK) is a natural product isolated from the bark, cones, seeds and leaves of plants belonging to the genus Magnolia. It possesses anti-cancer activity which can efficiently impede the growth and bring about apoptosis of a diversity of cancer cells. The major concerns of using HK are its poor solubility and lack of targeted drug delivery. In this study, a combinatorial drug is prepared by combining HK and camptothecin (CPT). Both CPT and HK belong to the

Topics: Antineoplastic Agents; Biphenyl Compounds; Camptothecin; Drug Carriers; Graphite; Humans; Lignans; Nanoparticles

Losartan and honokiol are small molecules which have been described to inhibit aggregation of platelets by collagen. Losartan has been proposed to block clustering of GPVI but not to affect binding of collagen. Honokiol has been reported to bind directly to GPVI but only at a concentration that is three orders of magnitude higher than that needed for inhibition of aggregation. The mechanism of action of both inhibitors is so far unclear. In the present study, we confirm the inhibitory effects of both agents on platelet aggregation by collagen and show that both also block the aggregation induced by the activation of CLEC-2 or the low affinity immune receptor FcγRIIa at similar concentrations. For GPVI and CLEC-2, this inhibition is associated with a reduction in protein tyrosine phosphorylation of multiple proteins including Syk. In contrast, on a collagen surface, spreading of platelets and clustering of GPVI (measured by single molecule localisation microscopy) was not altered by losartan or honokiol. Furthermore, in flow whole-blood, both inhibitors suppressed the formation of multi-layered platelet thrombi at arteriolar shear rates at concentrations that hardly affect collagen-induced platelet aggregation in platelet rich plasma. Together, these results demonstrate that losartan and honokiol have multiple effects on platelets which should be considered in the use of these compounds as anti-platelet agents.

Topics: Biphenyl Compounds; Blood Platelets; Collagen; Humans; Lectins, C-Type; Lignans; Losartan; Membrane Glycoproteins; Phosphorylation; Platelet Aggregation; Platelet Aggregation Inhibitors; Platelet Membrane Glycoproteins; Platelet-Rich Plasma; Receptors, IgG; Syk Kinase; Thrombosis

Cholangiocarcinoma or biliary tract cancer has a high mortality rate resulting from late presentation and ineffective treatment strategy. Since immunotherapy by dendritic cells (DC) may be beneficial for cholangiocarcinoma treatment but their efficacy against cholangiocarcinoma was low. We suggest how such anti-tumor activity can be increased using cell lysates derived from an honokiol-treated cholangiocarcinoma cell line (KKU-213L5).. To increase antitumour activity of DCs pulsed with cell lysates derived from honokiol-treated cholangiocarcinoma cell line (KKU-213L5).. The effect of honokiol, a phenolic compound isolated from. Honokiol can effectively activate cholangiocarcinoma apoptosis and increase the release of damage-associated molecular patterns. DCs loaded with cell lysates derived from honokiol-treated tumour cells enhanced priming and stimulated T lymphocyte proliferation and type I cytokine production. T lymphocytes stimulated with DCs pulsed with cell lysates of honokiol-treated tumour cells significantly increased specific killing of human cholangiocarcinoma cells compared to those associated with DCs pulsed with cell lysates of untreated cholangiocarcinoma cells.. The present findings suggested that honokiol was able to enhance the immunogenicity of cholangiocarcinoma cells associated with increased effectiveness of DC-based vaccine formulation. Treatment of tumour cells with honokiol offers a promising approach as an

Topics: Bile Duct Neoplasms; Biphenyl Compounds; Cancer Vaccines; Cell Line, Tumor; Cell Proliferation; Cholangiocarcinoma; Dendritic Cells; Healthy Volunteers; Humans; Immunogenicity, Vaccine; Immunotherapy; Lignans; Lymphocyte Activation; T-Lymphocytes, Cytotoxic; Transplantation, Autologous

The antifungal activity of magnolol and honokiol, two naturally occurring hydroxylated biphenyls, and of their synthetic derivatives was evaluated on a collection of representative isolates of Fusarium oxysporum, F. solani and F. verticillioides of clinical and ecological concern. The tested compounds were proposed as a 'natural' alternative to conventional fungicides, even though a larger range of concentrations (5-400 μg/ml) was applied. The activity of magnolol and honokiol was compared with that of terbinafine (0.1-10 μg/ml), and fluconazole (1-50 μg/ml), two fungicides widely used in treating fungal infections on humans. Magnolol showed similar fungicidal activity compared to fluconazole, whereas honokiol was more effective in inhibiting mycelium growth compared to this fungicide on all tested clinical Fusarium spp. isolates. Compared to terbinafine, honokiol showed similar antifungal activity when tested on clinical F. solani isolates, whereas magnolol was less effective at all selected concentrations (5-400 μg/ml). The different position of the phenol-OH group, as well as its protection, explain different in vitro activities between magnolol, honokiol, and their derivatives. Furthermore, magnolol showed mycelium dry weight reduction at a concentration of 0.5 mM when tested on a set of agricultural isolates of Fusaria, leading to complete inhibition of some of them. Magnolol and honokiol are proposed as efficient and safe candidates for treating clinically relevant Fusaria.

Topics: Antifungal Agents; Biphenyl Compounds; Fusariosis; Fusarium; Humans; Lignans; Microbial Sensitivity Tests; Plant Diseases

The link between Neuraminidase 1 (Neu1) and cancer development has been highlighted in numerous studies. In an effort to understand the role of Neu1 in mammary carcinoma cells, we evaluated the effect of Neu1 on controlling cell proliferation and apoptosis, as well as regulating the expression of cadherins. By blocking the activity of Neu1 with oseltamivir phosphate or using siRNA to silence the Neu1 protein, we observed suppression of cell growth in MCF-7 and MDA-MB-231 cells. Enhanced cleaved caspase 3 expression was demonstrated in breast cancer cells treated with oseltamivir phosphate or in Neu1 knockdown mammary carcinoma cells. We also provided evidence of Neu1 reversing the epithelial-mesenchymal properties with associated changes to the respective cadherin family. Additional observations indicated that the phytochemical, honokiol downregulates the expression of Neu1. As a consequence of blocking Neu1, honokiol reduced the levels of sialic acid in the two subtypes of breast cancer. These findings provide evidence that Neu1 regulates cell growth and death, and facilitates cancer progression by modulating the expression levels of cadherins.

Topics: Apoptosis; Biphenyl Compounds; Breast Neoplasms; Cadherins; Cell Line, Tumor; Cell Proliferation; Female; Humans; Lignans; N-Acetylneuraminic Acid; Neuraminidase

Magnoliae Cortex contains a range of bioactive components including terpenes (e.g. α-, β- and γ-eudesmol), phenylpropanoids (e.g. honokiol and magnolol) and alkaloids (e.g. magnocurarine). We recently reported that pretreatment of PC12 cells with Magnoliae Cortex extract significantly suppresses cytotoxicity induced by H

Topics: Animals; Biphenyl Compounds; Catalase; Cell Death; Cell Survival; Hydrogen Peroxide; Lignans; NAD(P)H Dehydrogenase (Quinone); Oxidopamine; PC12 Cells; Protective Agents; Rats

Honokiol is a natural biphenolic compound extracted from traditional Chinese medicine

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Apoptosis Regulatory Proteins; Biphenyl Compounds; CASP8 and FADD-Like Apoptosis Regulating Protein; Cell Line, Tumor; Gene Knockdown Techniques; Humans; Lignans; Peptide Hydrolases; Plant Extracts; Survivin; TNF-Related Apoptosis-Inducing Ligand; Ubiquitination

Increasing evidence indicates that neuroinflammatory and oxidative stress play two pivotal roles in cognitive impairment after surgery. Honokiol (HNK), as an activator of Sirtuin3 (SIRT3), has potential multiple biological functions. The aim of these experiments is to evaluate the effects of HNK on surgery/anesthesia-induced cognitive decline in mice.. Adult C57BL/6 mice received a laparotomy under sevoflurane anesthesia and HNK or SIRT3 inhibitor (3-TYP) treatment. Cognitive function and locomotor activity of mice were evaluated using fear conditioning test and open field test on postoperative 1 and 3 days. Neuronal apoptosis in CA1 and CA3 area of hippocampus was examined using TUNEL assay. And Western blot was applied to measure the expression of pro-inflammatory cytokines and SIRT3/SOD2 signaling-associated proteins in hippocampus. Meanwhile, SIRT3 positive cells were calculated by immunohistochemistry. The mitochondrial membrane potential, malondialdehyde (MDA), and mitochondrial radical oxygen species (mtROS) were detected using standard methods.. Honokiol attenuated surgery-induced memory loss and neuronal apoptosis, decreased neuroinflammatory response, and ameliorated oxidative damage in hippocampus. Notably, surgery/anesthesia induced an obviously decrease in hippocampal SIRT3 expression, whereas the HNK increased SIRT3 expression and thus decreased the acetylation of superoxide dismutase 2 (SOD2). However, 3-TYP treatment inhibited the HNK's rescuing effects.. These results suggested that activation of SIRT3 by honokiol may attenuate surgery/anesthesia-induced cognitive impairment in mice through regulation of oxidative stress and neuroinflammatory in hippocampus.

Topics: Anesthesia; Anesthetics, Inhalation; Animals; Anti-Inflammatory Agents; Biphenyl Compounds; Cognitive Dysfunction; Female; Hippocampus; Inflammation; Laparotomy; Lignans; Mice, Inbred C57BL; Nootropic Agents; Oxidative Stress; Postoperative Complications; Random Allocation; Sevoflurane; Sirtuin 3

Magnolol and honokiol are the main active components of Magnolia officinalis Rehd. et Wils. The study of their interactions with liver microsomes is very important for the clinical safety of M. officinalis Rehd. et Wils.. The main metabolites of magnolol and honokiol in rat liver microsomes were investigated using ultrahigh-performance liquid chromatography/mass spectrometry and their possible structures were identified. In addition, cytochrome P450 (CYP450) isoenzymes of the major rat metabolites of magnolol and honokiol were identified using a specific inhibitor.. This study suggests that the CYP2E1 subtype is responsible for the oxidation of magnolol and honokiol terminal double bonds to epoxy metabolites. CYP3A4 appears to be the major subtype responsible for further hydrolytic metabolism, while CYP1A2 may promote decarboxylation of the metabolites. CYP2A6 may be the main subtype responsible for the hydrogenation of magnolol (p < 0.05).. This study demonstrated that different CYP450 enzyme isoforms showed different activities in the in vitro metabolism of magnolol and honokiol in rat liver microsomes. It has certain practical applications in that we should pay attention to drug-drug interactions in clinical medications and also to drug-enzyme interactions.

Topics: Animals; Biphenyl Compounds; Chromatography, High Pressure Liquid; Cytochrome P-450 Enzyme Inhibitors; Cytochrome P-450 Enzyme System; Inactivation, Metabolic; Inhibitory Concentration 50; Lignans; Male; Mass Spectrometry; Microsomes, Liver; Rats, Sprague-Dawley; Substrate Specificity

There is increasing global incidence of highly metastatic melanoma and therapeutic strategies like those focusing on the downstream beta-catenin/MITF axis of invading melanoma cells are urgently needed. Targeting endoplasmic reticulum (ER) stress can promote cancer cell death and inhibit epithelial mesenchymal transition (EMT) in metastatic tumors. This study aimed to determine if Honokiol could promote ER stress-dependent apoptosis and regulate metastatic melanoma. The therapeutic efficacy of Honokiol was assessed using the highly metastatic melanoma xenograft mouse model for peritoneal metastasis and evaluated by computed tomography imaging. The ER stress marker, Calpain-10, delineated a novel proteolytic cleavage enzyme, while CHOP/GADD153-regulated apoptosis was used for gene silencing to determine the role of the β-catenin/MITF axis in melanoma cells. The results showed that Honokiol effectively decreased peritoneal dissemination and organ metastasis via ER stress activation and EMT marker inhibition. Knockdown Calpain-10 or CHOP/GADD153 blocked all of the biological effects in Honokiol-induced β-catenin/MITF cleavage, ERSE or TCF/LEF luciferase activity, and β-catenin kinase activity. These findings suggest that Honokiol can significantly thwart the progression of highly metastatic melanoma using the β-catenin/MITF axis via prompt Calpain-10 and CHOP/GADD153 regulated cascades.

Topics: Animals; Antineoplastic Agents, Phytogenic; beta Catenin; Biphenyl Compounds; Calpain; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cyclin-Dependent Kinase 2; Endoplasmic Reticulum Stress; Gene Expression Regulation, Neoplastic; Humans; Lignans; Male; Melanoma; Mice, Inbred BALB C; Mice, Nude; Microphthalmia-Associated Transcription Factor; Peritoneal Neoplasms; Skin Neoplasms; Transcription Factor CHOP; Wnt Signaling Pathway; Xenograft Model Antitumor Assays

Novel isoxazole and pyrazole analogs based on natural biphenyl-neolignan honokiol were synthesized and evaluated for their inhibitory activities against nitric oxide production in lipopolysaccharide-activated BV-2 microglial cells. The isoxazole skeleton was constructed via nitrile oxide cycloaddition from oxime 3 and pyrazole was generated by condensation of 4-chromone and alkylhydrazine. Among the analogs, 13b and 14a showed stronger inhibitory activities with IC

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Biological Products; Biphenyl Compounds; Cell Line; Dose-Response Relationship, Drug; Heterocyclic Compounds; Lignans; Lipopolysaccharides; Mice; Molecular Structure; Nitric Oxide; Structure-Activity Relationship

Honokiol, a natural polyphenol, which was reported to have satisfactory influenza neuraminidase (NA) inhibitory activity, was structurally modified. Twenty-three compounds were synthesized and the

Topics: Antiviral Agents; Biphenyl Compounds; Drug Design; Humans; Influenza, Human; Lignans; Molecular Structure; Neuraminidase

Topics: Administration, Oral; Animals; Benzodiazepines; Biphenyl Compounds; Drug Carriers; Electroencephalography; Eye Movements; Female; Lignans; Rats; Rats, Sprague-Dawley; Receptors, GABA-A; Sleep, Slow-Wave

Acute kidney injury (AKI) is one of the most common complications of sepsis, which largely contributes to the high mortality rate of sepsis. Honokiol, a natural polyphenol from the traditional Chinese herb Magnolia officinalis, is known to possess anti-inflammatory and antioxidant activity. Here, the underlying mechanism of honokiol-induced amelioration of sepsis-associated AKI was analyzed. The expression patterns of oxidative stress moleculars and TLRs-mediated inflammation pathway were examined to identify the response of NRK-52E cells incubated with septic rats' serum to honokiol. The levels of iNOS, NO, and myeloperoxidase in NRK-52E cells were increased during sepsis, which could be reversed by honokiol. The production of GSH and SOD as in vivo antioxidant was increased after honokiol treatment. The administration of honokiol significantly inhibited TLR2/4/MyD88 signaling pathway in AKI-induced NRK-52E cells. Furthermore, ZnPPIX, the HO-1 inhibitor, weakened honokiol-mediated morphological amelioration, and the reduced level of TNF-α, IL-1β, and IL-6 in kidneys of rats subjected to CLP. Finally, Honokiol was shown to connect with the Nrf2-Keap1 dimensionally. These findings suggest that honokiol plays its protective role on sepsis-associated AKI against oxidative stress and inflammatory signals.

Topics: Acute Kidney Injury; Animals; Anti-Infective Agents; Biphenyl Compounds; Cell Line; Inflammation; Lignans; Oxidative Stress; Rats; Sepsis

At present, there is a higher demand for the efficacy of nanoparticle drugs. It is hoped that more drugs will reach the tumor site and that the drug will be less harmful to other normal cells of the body before reaching the tumor site. Most target research for nanomedicine can achieve better positioning through complex processes, such as synthesis. To overcome these difficulties, such as the complexity of the preparation method and lack of good targeting, we used simple polydopamine (PDA) as a pH-sensitive targeting anchor for nanoparticles (NPs). We successfully conjugated folic acid (FA) to the surface of honokiol (HK) nanoparticles coated with PDA using a typical surface modifier. After preparation into HK-PDA-FA-NPs, we characterized the particle size, potential and transmission electron microscope (TEM). The targeted nanoparticles (HK-PDA-FA-NPs) can be stably present in various physiological media and exhibit pH sensitivity during drug release in vitro. HK-PDA-FA-NPs have better targeting ability to 4T1 cells than HK-NPs. Targeted nanoparticles have a tumor inhibition rate of greater than 80% in vivo, which is significantly higher than ordinary HK-NPs. This experiment shows that surface modification of HK-NPs coated with PDA is a promising preparation method for targeted therapy.

Topics: Animals; Antineoplastic Agents; Biphenyl Compounds; Breast Neoplasms; Cell Proliferation; Drug Carriers; Drug Screening Assays, Antitumor; Female; Folic Acid; Hydrogen-Ion Concentration; Indoles; Lignans; Mammary Neoplasms, Experimental; Mice; Mice, Inbred BALB C; Nanoparticles; Optical Imaging; Particle Size; Polymers; Surface Properties

Honokiol is a hydroxylated biphenyl natural product and displays potent antitumor activity against several cancers including prostate cancer, melanoma, leukemia, and colorectal cancer. The present study was to investigate the in vitro activity of honokiol against A549 and 95-D human lung cancer cells.. A549 and 95-D cells were used with honokiol treatment. Cell viability was determined by CCK-8 assay. The cell migration and apoptosis were evaluated by wound healing assay and TUNEL staining method respectively. The expressions of ER-related proteins were analyzed by western blot and the CHOP siRNA was used to downregulate the CHOP expression.. The results demonstrated that treatment of A549 and 95-D cells with honokiol significantly reduced cell viability in a dose- and time-dependent manner. Furthermore, honokiol treatment decreased cell migration and enhanced cell apoptosis, which is accompanied by the upregulation of the expressions of ER stress-induced apoptotic signaling molecules such as GRP78, phosphorylated PERK, phosphorylated eIF2α, CHOP, Bcl-2, Bax, and cleaved Caspase 9. Honokiol treatment-induced increase of ER stress-related signaling molecules and apoptotic proteins in A549 and 95-D cells were reversed by CHOP siRNA.. Collectively, we conclude that ER stress may participate in the action of the anticancer activity of honokiol in A549 and 95-D cells and induction of ER stress-related apoptosis may represent a novel therapeutic intervention for human lung cancer.

Topics: A549 Cells; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Movement; Cell Survival; Endoplasmic Reticulum Chaperone BiP; Endoplasmic Reticulum Stress; Humans; Lignans; Lung Neoplasms; RNA, Small Interfering; Signal Transduction; Transcription Factor CHOP

Abnormal autophagy plays critical roles in the structure and function of the pulmonary vasculature. Cyclophilin A (CyPA) can be secreted from cells in response to hypoxia and oxidative stress, which are involved in inducing autophagy and regulating the function of endothelial cells in pulmonary arterial hypertension. Honokiol is a small molecule natural compound; it has many bioactivities, such as antitumor, anti-inflammatory, antioxidant and antiangiogenic properties, but how honokiol mediates autophagy in pulmonary arterial hypertension is unclear. Rat' lungs gavaged with honokiol were examined for autophagy via western blot and fluorescence microscopy. In addition, western blot, quantitative RT-PCR and immunofluorescence were employed to test the expression of CyPA and autophagy markers in pulmonary artery endothelial cells (PAECs). Small interfering RNA targeting CyPA (si-CyPA) was used to knockdown the expression of CyPA, and then autophagy was tested with mRFP-GFP-LC3 fluorescence microscopy and western blot. We found that honokiol could reduce the expression of CyPA and autophagy markers in vivo and in vitro. Furthermore, autophagy was also down-regulated by si-CyPA. Taken together, we revealed a novel mechanism by which honokiol regulates autophagy. The results revealed that honokiol can alleviate autophagy and pulmonary arterial hypertension regulated by CyPA in PAECs.

Topics: Animals; Autophagy; Biphenyl Compounds; Blotting, Western; Cattle; Cells, Cultured; Cyclophilin A; Endothelial Cells; Endothelium, Vascular; Hypertension, Pulmonary; Lignans; Male; Microscopy, Fluorescence; Pulmonary Artery; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction

Honokiol, a biphenolic neolignan with inappreciable toxicity isolated from Magnolia officinalis, has been reported to have antiangiogenic and antitumor properties in several tumor cell lines and tumor xenograft models. In our previous study, structural modification by chemical synthesis has been carried out to develop novel honokiol derivatives to improve antitumor activity and clarify the structure-activity relationship. Honokiol analogs, especially 3,5'-diformylated honokiol HK-(CHO)2, have been found to moderately block the newly grown segmental vessels from the dorsal aorta in the transgenic zebrafish-based assay, show antiangiogenic property, and exert medium cytotoxicity against two lung cell lines (Lewis lung carcinoma LL/2 cells and human non-small-cell lung cancer A549 cells). However, the in-vitro and in-vivo antitumor effects of formylated honokiol derivatives against lung carcinoma remained poorly understood. In the study, two formylated honokiol derivatives also showed potent antitumor effects in the Lewis lung carcinoma cells, K-ras-dirived lung adenocarcinoma mice, and a mouse lung tumor xenograft model, with HK-(CHO)2 being most efficacious. The potential mechanism was inhibiting cell proliferation and inducing apoptosis in lung cancer by the regulation of vascular endothelial growth factor A expression. These results further suggested that HK-(CHO)2 might be potential candidates for the treatment of lung carcinoma.

Topics: Adenocarcinoma of Lung; Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Carcinoma, Lewis Lung; Cell Proliferation; Female; Formates; Humans; Lignans; Lung Neoplasms; Mice; Mice, Inbred C57BL; Mice, Nude; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

Honokiol is a low-molecular-weight natural product and has been reported to exhibit anti-inflammatory activity.. Our study aimed to investigate the influence of honokiol on sepsis-induced acute kidney injury (AKI) in a mouse model.. A cecal ligation and puncture (CLP) surgical operation was performed to establish a sepsis-induced acute kidney injury model in mice. Renal histomorphological analysis was performed with periodic acid-Schiff (PAS) staining. The levels of inflammatory markers in serum were measured by ELISA assay. The mRNA and protein levels were assayed by RT-qPCR and western blotting, respectively. Annexin V-FITC/PI staining was used to evaluate glomerular mesangial cell (GMC) apoptosis.. The results revealed that honokiol significantly increased the survival rate in mice undergoing a CLP operation. Inflammatory cytokines, such as TNF-α, IL-6 and IL-1β, were significantly inhibited in honokiol-treated septic mice compared with the CLP group. In addition, honokiol showed the ability to reverse CLP-induced AKI in septic mice. Furthermore, heme oxygenase-1 (HO-1) expression levels were significantly up-regulated and miR-218-5p was markedly down-regulated in honokiol-treated septic mice as compared to CLP-operated mice. Bioinformatics and experimental measurements showed that HO-1 was a direct target of miR-218-5p. In vitro experiments showed that both honokiol and miR-218-5p inhibitors blocked lipopolysaccharide (LPS)-induced cell growth inhibition and GMC apoptosis by increasing the expression of HO-1.. Honokiol ameliorated AKI in septic mice and LPS-induced GMC dysfunction, and the underlying mechanism was mediated, at least partially, through the regulation of miR-218-5p/HO-1 signaling.

Topics: Acute Kidney Injury; Animals; Biphenyl Compounds; Disease Models, Animal; Heme Oxygenase-1; Kidney; Lignans; Male; Mice; MicroRNAs; Protective Agents; Sepsis; Signal Transduction

Traumatic brain injury (TBI) leads to neuronal damage and neurological dysfunction. The aim of our study was to investigate the antioxidative effect of honokiol on TBI in rats with biochemical, histopathological and immunohistochemical methods.. Sprague-Dawley rats were subjected to TBI with a weight-drop device using 300 g/1 m weight/height impact. Forty-five rats were divided into three groups as control group, TBI group and TBI + honokiol group (5 mg/kg/day, i.p.). Honokiol (5 mg/kg) dissolved in dimethyl sulfoxide (DMSO) was intraperitoneally administered to rats for 7 days after the trauma. At the end of experiment, blood samples were taken from the animals and analysed with various biochemical markers.. Histopathological examination of the trauma group revealed some degenerated pyramidal cells, dilatation and congestion in blood vessels, hyperplasia in endothelial cells, inflammatory cell infiltration around the vein and disruptions in glial extensions. In TBI + honokiol group, pyramidal neurons showed a decrease in degeneration, slight dilatation in blood vessels, improvement of endothelial cells towards the lumen, and reduction of inflammatory cells in the vessel. In TBI + honokiol group, vascular endothelial growth factor expression was positive in the endothelial and few inflammatory cells of the mildly dilated blood vessels. In the blood brain barrier deteriorated after trauma, it was observed that the glial foot processes were positive expression and extended to the endothelial cells in the TBI + honokiol group.. Glial fibrillary acidic protein expression showed a positive reaction in these processes. Considering the important role of antioxidants and inflammatory responses in cerebral damage induced by traumatic head injury, honokiol is thought to be important in decreasing lipid peroxidation, protecting the membrane structure of blood brain barrier, degeneration of neurons and glial cells.

Topics: Animals; Biphenyl Compounds; Blood-Brain Barrier; Brain Injuries, Traumatic; Disease Models, Animal; Glial Fibrillary Acidic Protein; Glutathione Peroxidase; Immunohistochemistry; Lignans; Male; Malondialdehyde; Permeability; Peroxidase; Rats, Sprague-Dawley; Vascular Endothelial Growth Factor A

Honokiol, a natural biphenolic compound, exerts anticancer effects through a variety of mechanisms on multiple types of cancer with relatively low toxicity. Adenosine 5'‑phosphate‑activated protein kinase (AMPK), an essential regulator of cellular homeostasis, may control cancer progression. The present study aimed to investigate whether the anticancer activities of honokiol in ovarian cancer cells were mediated through the activation of AMPK. Honokiol decreased cell viability of 2 ovarian cancer cell lines, with an half‑maximal inhibitory concentration value of 48.71±11.31 µM for SKOV3 cells and 46.42±5.37 µM for Caov‑3 cells. Honokiol induced apoptosis via activation of caspase‑3, caspase‑7 and caspase‑9, and cleavage of poly‑(adenosine 5'‑diphosphate‑ribose) polymerase. Apoptosis induced by honokiol was weakened by compound C, an AMPK inhibitor, suggesting that honokiol‑induced apoptosis was dependent on the AMPK/mechanistic target of rapamycin signaling pathway. Additionally, honokiol inhibited the migration and invasion of ovarian cancer cells. The combined treatment of honokiol with compound C reversed the activities of honokiol in wound healing and Matrigel invasion assays. These results indicated that honokiol may have therapeutic potential in ovarian cancer by targeting AMPK activation.

Topics: AMP-Activated Protein Kinases; Animals; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Enzyme Activation; Female; Humans; Lignans; Mice; Models, Biological; Neoplasm Invasiveness; NIH 3T3 Cells; Ovarian Neoplasms; Signal Transduction; TOR Serine-Threonine Kinases; Tumor Stem Cell Assay

Honokiol is a pleiotropic natural compound isolated from Magnolia and has multiple biological and clinically relevant effects, including anticancer and antimicrobial function. However, the antiviral activity of honokiol has not yet been well studied. Here we showed that honokiol had no effect on herpes simplex virus-1 (HSV-1) entry, but inhibited HSV-1 viral DNA replication, gene expression and the production of new progeny viruses. The combination of honokiol and clinical drug acyclovir augmented inhibition of HSV-1 infection. Our results illustrate that honokiol could be a potential new candidate for clinical consideration in the treatment of HSV-1 infection alone or combination with other therapeutics.

Topics: Acyclovir; Animals; Antiviral Agents; Biphenyl Compounds; Chlorocebus aethiops; DNA Replication; Genome, Viral; Herpesvirus 1, Human; Lignans; Magnolia; Mice; Mice, Inbred C57BL; Phytochemicals; Vero Cells; Virus Replication

The potential mechanism of postoperative cognitive impairment is still largely unclear. The activation of NLRP3 inflammasome had been reported to be involved in neurodegenerative diseases, including postoperative cognitive change, and is closely related to mitochondrial ROS and mitophagy. Honokiol (HNK) owns multiple organic protective effects. This study is aimed at observing the neuroprotective effect of HNK in postoperative cognitive change and examining the role of HNK in the regulation of mitophagy and the relationship between these effects and NLRP3 inflammasome activation in mice induced by surgery/anesthesia.. In this study, mice were divided into several groups: control group, surgery group, surgery+HNK group, and surgery+HNK+3-methyladenine (3-MA) group. Hippocampal tissue samples were harvested and used for proinflammatory cytokines, mitochondrial ROS, and malondialdehyde (MDA) assay. The process of mitophagy and the activation of NLRP3 inflammasome were observed by Western blot, immunohistochemistry, and transmission electron microscopy.. The results showed that HNK treatment obviously recovered the postoperative decline and enhanced the expressions of LC3-II, Beclin-1, Parkin, and PINK1 at protein levels after surgery/sevoflurane treatment, which are both an autophagy marker and a mitophagy marker. In addition, HNK attenuated mitochondrial structure damage and reduced mtROS and MDA generation, which are closely associated with NLRP3 inflammasome activation. Honokiol-mediated mitophagy inhibited the activation of NLRP3 inflammasome and neuroinflammation in the hippocampus. Using 3-MA, an autophagy inhibitor, the neuroprotective effects of HNK on mitophagy and NLRP3 inflammasome activation were eliminated.. These results indicated that HNK-mediated mitophagy ameliorates postoperative cognitive impairment induced by surgery/sevoflurane. This neuroprotective effect may be involved in inhibiting the activation of NLRP3 inflammasome and suppressing inflammatory responses in the hippocampus.

Topics: Animals; Apoptosis; Autophagy; Biphenyl Compounds; Cognitive Dysfunction; Dentate Gyrus; Female; Hippocampus; Inflammasomes; Lignans; Mice, Inbred C57BL; Microglia; Mitochondria; Mitophagy; Neurons; NLR Family, Pyrin Domain-Containing 3 Protein; Oxidative Stress; Postoperative Complications; Reactive Oxygen Species; Sevoflurane

An effective strategy based on high-speed counter-current chromatography (HSCCC) knockout combination with HPLC-DAD-QTOF-MS/MS analysis were developed to identify minor lignans, alkaloids, and phenylpropanoid glycosides in M. officinalis. Petroleum ether/ethyl acetate/methanol/water (8:4:7:5, v/v/v/v) as solvent system was firstly selected to separate the crude extract of M. officinalis. Two major lignans, honokiol and magnolol were knocked out, and minor components were enriched. Then, five standards (honokiol, magnolol, magnocurarine, magnoflorine and acteoside) were used as examples to discuss their fragmentation patterns for structural identification. By comprehensive screening, sixteen lignans, nine alkaloids, six phenylpropanoid glycosides were unambiguously or tentatively identified by comparing their retention time, UV spectra, accurate mass and fragmentation patterns with standards or reported components. Eight of them, as far as was known, were discovered from M. officinalis for the first time. The proposed method might provide a model for the effective identification of minor components from complex herbs. Additionally, this study laid a foundation for the study of quality control, and clinical applications of M. officinalis.

Topics: Alkaloids; Aporphines; Biphenyl Compounds; Chromatography, High Pressure Liquid; Glucosides; Glycosides; Isoquinolines; Lignans; Magnolia; Methanol; Phenols; Propanols; Tandem Mass Spectrometry

Burn pain is one of the worst imaginable pain, associated with considerable morbidity and mortality worldwide. The management of pain made significant progress; however, more research is needed for burn pain. In the present study, the antinociceptive effect of honokiol extracted from Magnolia officinalis was assessed for 3 consecutive days. The third-degree burns were induced by the hot water method. The honokiol both by intraperitoneal (i.p) and intra plantar (i.pl) route and in combination with tramadol (i.p) was found to be effective in significantly reducing the mechanical allodynia, hyperalgesia, thermal hyperalgesia and paw edema. Honokiol also succeeded in reducing weight loss and spontaneous pain behavior in mice. Honokiol treatment both i.p and ipl decrease significantly the loss of total protein (3.3 and 3.4 g/dl of total protein) and albumin (2.2 and 2.6 g/dl of total albumin) respectively. It also significantly recovers the normal balance of blood electrolytes and normalizes blood profile. Effect of honokiol on cytokines and mRNA expression levels of TRPV1 and P2Y were also assessed. Honokiol significantly decreases the expression of TNF-α, IL-1β and IL-6 and decreases expression level of TRPV1 and P2Y. Additionally, TRPV1 and P2Y proteins expression levels were also assessed by Western blot in paw skin tissue, sciatic nerve and spinal cord which were remarkably down-regulated by honokiol. Histological analysis of vehicle control and drug-treated paws were also performed through hematoxylin and eosin (H&E) staining which exhibited that honokiol significantly reduced the dermal layers distortion and inflammation associated with the burn. The antioxidant enzymes and nitric oxide (NO) were also determined through ELISA. Honokiol treatment also potentiates the expression of reduced glutathione and glutathione S-transferase, and catalase levels and reduced significantly the nitric oxide (NO) as compared to the burn-induced group. It can be concluded on the base of the results that honokiol has a significant analgesic activity through its action on cytokines and by downregulating TRPV1 and P2Y receptors. It also has a protective role against burn damage by upregulation of antioxidants.

Topics: Analgesics; Animals; Biphenyl Compounds; Burns; Cytokines; Edema; Hyperalgesia; Inflammation; Inflammation Mediators; Interleukin-1beta; Lignans; Magnolia; Male; Mice; Mice, Inbred BALB C; Pain; Receptors, Purinergic P2Y; Signal Transduction; Spinal Cord; TRPV Cation Channels; Tumor Necrosis Factor-alpha; Up-Regulation

The aim of the present study was to evaluate the effects of two well-known natural antioxidants vitamin C (VC) and vitamin E (VE) on the antifungal activity of honokiol against

Topics: Antifungal Agents; Antioxidants; Ascorbic Acid; Biphenyl Compounds; Candida albicans; DNA, Mitochondrial; Drug Antagonism; Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); Glycolysis; Lignans; Lipid Peroxidation; Membrane Potential, Mitochondrial; Microbial Sensitivity Tests; Mitochondria; Reactive Oxygen Species; Vitamin E

Honokiol, the main bioactive component of Magnolia officinalis, has a variety of pharmacological actions. However, its toxicity has rarely been reported. According to previous studies performed in our laboratory, honokiol microemulsion has embryo developmental toxicity. For further exploration, Zebrafish embryos were exposed to different doses of honokiol microemulsion to record the rates of mortality, malformation, and hatching. We found that high doses of honokiol microemulsion (0.6 and 0.9 μg/ml) increased mortality, inhibited hatching, caused malformation and reduced swimming activities. The low-dose group (0.15 and 0.30 μg/ml) had decreased production of reactive oxygen species (ROS), but the high-dose group had inhibited superoxide dismutase (SOD) enzyme activity and increased ROS content. The mRNA expression of sod1, sod2, catalase(cat), and heme oxygenase 1 (ho1) was up-regulated at low doses but down-regulated at high doses. The nuclear factor E2-related factor 2 (Nrf2) mRNA expression increased at low doses but decreased at high doses. After knocking down Nrf2 in zebrafish embryos, the rates of mortality and malformation were markedly increased and the hatching rate was significantly decreased. These results suggest that honokiol has antioxidative effects at low doses but causes embryo-developmental toxicity at high doses, and the Nrf2 gene may play a pivotal role in regulating these processes.

Topics: Animals; Antioxidants; Biphenyl Compounds; Catalase; Dose-Response Relationship, Drug; Embryo, Nonmammalian; Gene Expression Regulation, Developmental; Heme Oxygenase-1; Lethal Dose 50; Lignans; Locomotion; NF-E2-Related Factor 2; Oxidative Stress; Signal Transduction; Superoxide Dismutase; Superoxide Dismutase-1; Swimming; Zebrafish; Zebrafish Proteins

Honokiol (HKL) is a natural low-molecular-weight biphenolic compound derived from the bark of magnolia trees. Previous studies indicate that HKL exerts potent cardioprotective effects on ischemia/reperfusion (I/R) injury; however, evidence of the further relationship between HKL posttreatment and myocardial I/R injury has not been clearly found. In our study, we explored the protective effect of HKL post treatment on myocardial I/R injury in C57BL/6 mice. We also demonstrated that HKL significantly reduced cellular reactive oxygen species production and attenuated mitochondrial damage in neonatal rat cardiomyocytes exposed to hypoxia/reoxygenation (H/R). In addition, HKL was found to enhance autophagy during I/R or H/R; these effects could be partially blocked by the autophagic flux inhibitor chloroquine. Moreover, our results suggested that enhanced autophagic flux is associated with the Akt signaling pathway. Collectively, our results indicate that HKL posttreatment alleviates myocardial I/R injury and suggest a critical cardioprotective role of HKL in promoting autophagic flux.

Topics: Animals; Apoptosis; Autophagy; Biphenyl Compounds; Chloroquine; Disease Models, Animal; Lignans; Male; Mice; Mice, Inbred C57BL; Mitochondria; Myocardial Reperfusion Injury; Myocytes, Cardiac; Proto-Oncogene Proteins c-akt; Rats; Reactive Oxygen Species; Signal Transduction

Topics: Apoptosis; Autophagy; Biphenyl Compounds; Caspase 3; Cell Survival; Humans; Lignans; Neuroblastoma; Signal Transduction; Time Factors; Tumor Cells, Cultured; Tumor Suppressor Protein p53

Topics: A549 Cells; Animals; Anti-Bacterial Agents; Anti-Inflammatory Agents; Bacterial Toxins; Biphenyl Compounds; Cell Survival; Disease Models, Animal; Hemolysin Proteins; Histocytochemistry; Humans; Inflammasomes; Lignans; Liver; Mice, Inbred C57BL; Molecular Docking Simulation; Protein Binding; Receptors, Cell Surface; Staphylococcal Infections; Staphylococcus aureus; Treatment Outcome

Acetaminophen (APAP) overdose is the most frequent cause of drug-induced liver damage. Magnolia officinalis is a traditional hepatoprotective Chinese medicine and Honokiol (HO) is the major active constituent. The present study was to investigate the effect of HO on APAP-induced hepatotoxicity and related mechanisms.. Four groups of mice were subjected to treatment as vehicle, APAP, APAP + HO and APAP + HO + NRF2 inhibitor. The morphological and biochemical assessments were used to evaluate the hepatoprotective effects. The extent of APAP-protein adducts was determined through evaluate the hepatic content 3‑(cystein‑S‑yl)acetaminophen (APAP-Cys), the hydrolysis products of APAP-protein adducts. The activities of CYP2E1, CYP1A2 and CYP3A4 were evaluated by cocktail incubation, and the protein expression levels of NRF2, GCLC, GCLM, GS and GST were evaluated by western blot analysis.. Morphological and biochemical assessments clearly demonstrated that HO could alleviate APAP-induced liver damage. The hepatoprotective effect of HO was positively associated with the reduction of APAP-protein adducts. Further investigation suggested that HO induced inhibition of CYP 2E1 and CYP2A1 as well as upregulation of GSH co-contributed to the reduction of APAP-protein adducts. Furthermore, HO induced activations of NRF2 and its target enzymes, such as GCLC, GCLM and GST, gave rise to the upregulation of GSH.. Our results suggested that HO could alleviate APAP-induced liver damage through reducing the generation of APAP-protein adducts, which might be mediated by inhibiting the activity of CYP 2E1 and CYP2A1 as well as enhancing the generation of GSH via NRF2 pathway.

Topics: Acetaminophen; Animals; Biphenyl Compounds; Chemical and Drug Induced Liver Injury; Glutathione; Lignans; Liver; Male; Mice; Mice, Inbred BALB C; NF-E2-Related Factor 2; Oxidative Stress; Protective Agents; Signal Transduction

Topics: Animals; Antidepressive Agents; Autonomic Nervous System; Biphenyl Compounds; Brain; Calcium; Cytokines; Depression; Disease Models, Animal; Hindlimb Suspension; Immobilization; Indoleamine-Pyrrole 2,3,-Dioxygenase; Inflammation Mediators; Kynurenine; Lignans; Lipopolysaccharides; Mice, Inbred ICR; NF-kappa B; RNA, Messenger; Swimming; Tryptophan

In this study, two phenol compounds, magnolol and honokiol, were extracted from

Topics: Alternaria; Antifungal Agents; Biphenyl Compounds; Lignans; Magnolia; Nicotiana; Plant Diseases

In this study, we aimed to explore key micro(mi)RNAs and their potential regulatory mechanisms induced by honokiol treatment in non-small cell lung cancer (NSCLC) cells.. NSCLC A549 cells were treated with 0 (control) or 45 μM honokiol. Cell proliferation and migration were determined using CCK-8 and transwell assay, respectively, and apoptosis was determined using flow cytometry. RNA-sequencing was performed to detect the transcript expression levels. The differentially expressed miRNAs (DE-miRNAs) between the honokiol group and the control group were screened and analyzed for their functions and pathways. Then, protein-protein interaction (PPI) networks and miRNA-mRNA regulatory networks were constructed. In addition, survival analysis based on the key miRNAs was performed. Finally, the expression of the key miRNAs and their target genes were determined, and their effects on drug sensitivity were validated using their inhibitors.. Cell proliferation and migration were inhibited (P < 0.01), and the apoptosis rate was increased (P < 0.01) after honokiol treatment compared to that in the control group. A total of 26 upregulated and 20 downregulated DE-miRNAs were screened. DE-miRNAs were enriched in 10 pathways and 48 biological processes, such as the PI3K/AKT signaling pathway (involving miR-148a-3p). The miRNA-mRNA regulatory networks involved eight upregulated (including miR-148a-3p and let-7c-5p) and seven downregulated miRNAs (including miR-7-5p) and 190 target mRNAs. Survival analysis revealed that let-7c-5p, miR-148a-3p, and miR-148a-5p levels correlated with NSCLC prognosis. The expression of let-7c-5p, miR-148a-3p, and miR-148a-5p was significantly increased and negatively correlated with the expression of their target genes. The cytological effects of honokiol on A549 cells was partly reversed by treatment with the inhibitors of Let-7c-5p and miR-148a-3p.. Let-7c-5p, miR-148a-3p, miR-148a-5p, and miR-7-5p are favorable indicators of NSCLC patients treated with honokiol.

Topics: A549 Cells; Apoptosis; Biphenyl Compounds; Carcinoma, Non-Small-Cell Lung; Cell Movement; Cell Proliferation; Gene Expression Regulation, Neoplastic; Gene Regulatory Networks; Humans; Lignans; Lung Neoplasms; MicroRNAs; Principal Component Analysis; RNA, Messenger

Honokiol (HNK), an active compound isolated from traditional Chinese medicine Magnolia officinalis, has shown potent anticancer activities. In the present study, we investigated the effects of HNK on breast cancer metastasis in vitro and in vivo, as well as the underlying molecular mechanisms. We showed that HNK (10-70 μmol/L) dose-dependently inhibited the viability of human mammary epithelial tumor cell lines MCF7, MDA-MB-231, and mouse mammary tumor cell line 4T1. In the transwell and scratch migration assays, HNK (10, 20, 30 μmol/L) dose-dependently suppressed the invasion and migration of the breast cancer cells. We demonstrated that HNK (10-50 μmol/L) dose-dependently upregulated the epithelial marker E-cadherin and downregulated the mesenchymal markers such as Snail, Slug, and vimentin at the protein level in breast cancer cells. Using a puromycin incorporation assay, we showed that HNK decreased the Snail translation efficiency in the breast cancer cells. In a mouse model of tumor metastasis, administration of HNK (50 mg/kg every day, intraperitoneal (i.p.), 6 times per week for 30 days) significantly decreased the number of metastatic 4T1 cell-derived nodules and ameliorated the histological alterations in the lungs. In addition, HNK-treated mice showed decreased Snail expression and increased E-cadherin expression in metastatic nodules. In conclusion, HNK inhibits EMT in the breast cancer cells by downregulating Snail and Slug protein expression at the mRNA translation level. HNK has potential as an integrative medicine for combating breast cancer by targeting EMT.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Breast Neoplasms; Cell Line, Tumor; Cell Movement; Down-Regulation; Epithelial-Mesenchymal Transition; Female; Humans; Lignans; Lung Neoplasms; Mice, Inbred BALB C; Snail Family Transcription Factors

Honokiol, a biphenolic neolignan isolated from Magnolia officinalis, was reported to have a promising anti-inflammatory activity for the treatment of various diseases. There are many efforts on the synthesis and structure-activity relationship of honokiol derivatives. However, regioselective O-alkylation of honokiol remains a challenge and serves as a tool to provide not only some derivatives but also chemical probes for target identification and mode of action. In this study, we examined the reaction condition for regioselective O-alkylation, in which C2 and C4'-alkylated analogs of honokiol were synthesized and evaluated for inhibitory activity on nitric oxide production and cyclooxygenase-2 expression. Furthermore, we successfully synthesized a potential photoaffinity probe consisting of biotin and benzophenone based on a C4'-alkylated derivative.

Topics: Alkylation; Animals; Anti-Inflammatory Agents, Non-Steroidal; Biphenyl Compounds; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Humans; Inflammation; Lignans; Lipopolysaccharides; Mice; Molecular Structure; Nitric Oxide; RAW 264.7 Cells; Stereoisomerism

As a known natural product with anti-tumor activity, honokiol has been widely researched and structural modified. Lots of honokiol derivatives have been found to possess good anti-proliferative activity and showed great potential in cancer therapy, but the SAR (structure-activity relationship) was still confused. Here in, the SAR were comprehensively researched by summary of reported derivatives and synthesis of novel derivatives. Amongst novel derivatives, the promising compounds A6 and A10 exhibited potent and selective anti-proliferative activities against K562 cell line with the IC

Topics: Biphenyl Compounds; Humans; Lignans; Structure-Activity Relationship

Vascular dementia (VaD) is caused by chronic decreases in brain blood flow and accounts for 15-20% of dementia cases worldwide. In contrast to Alzheimer's disease (AD), no effective drug treatments are currently available for VaD. Previous studies have suggested that oxidative stress and neuroinflammation in the brain play important roles in the pathogenesis of VaD. Honokiol (HKL) is a well-known bioactive and nutraceutical compound that can act as an antioxidant and anti-inflammatory molecule. HKL can protect against memory impairments in AD mouse models. In this study, we explored whether the application of HKL was also protective against the insult of chronic cerebral hypoperfusion (CCH) in rats. We found that HKL supplementation prevented the memory impairments in the inhibitory avoidance step-down and Morris water maze tasks in CCH rats. HKL also suppressed the levels of oxidative stress and inflammation in CCH rats. Moreover, HKL prevented dendritic spines abnormalities in CCH rats. We also found that HKL inhibited the activity of GSK-3β, which may be critical for the neuroprotective activity of HKL. Thus, our study demonstrated the protective role of HKL in VaD.

Topics: Animals; Biphenyl Compounds; Brain; Dementia, Vascular; Disease Models, Animal; Enzyme Activation; Glycogen Synthase Kinase 3 beta; Inflammation; Lignans; Maze Learning; Memory Disorders; Neuroprotective Agents; Oxidative Stress; Rats; Rats, Wistar

Retinal neovascularization (NV) is the leading cause of blindness in the majority of ocular diseases. Several treatment approaches have been developed for retinal NV; of these methods, instillation of nanoparticles into the conjunctival sac has shown potential for retinal NV treatment because it does not cause physical damage and is easy to operate.. In this study, honokiol-loaded chitosan/sulfobutylether-β-cyclodextrin nanoparticles (HKCS- NPs) were prepared for ophthalmic drug delivery systems. An inclusion complex of honokiol and sulfobutylether-β-cyclodextrin was used to incorporated insoluble honokiol into chitosan nanoparticles, which were prepared through ionotropic gelation.. HK-CS-NPs featured a spherical surface with a narrow size distribution of polydispersity index less than 0.250, a mean size range of 373-523 nm, a positive surface charge of +19.9 to +24.2 mV, and an entrapment efficiency of 84.92%. In vitro release studies showed an initial burst release phase and a sustained release phase of nanoparticles. Moreover, in vivo study showed that HK-CS-NPs exhibited good ocular tolerability and could improve ophthalmic bioavailability of honokiol. In particular, the maximum concentration of honokiol after administration of HK-CS-NPs was enhanced by 1.65 times compared with that after instillation of the honokiol suspension alone.. This study proposes HK-CS-NPs as a potential ophthalmic delivery system.

Topics: Administration, Ophthalmic; Animals; beta-Cyclodextrins; Biological Availability; Biphenyl Compounds; Chitosan; Drug Delivery Systems; Drug Liberation; Eye; Lignans; Nanoparticles; Particle Size; Rabbits; Surface Properties

Non-alcoholic steatohepatitis (NASH) may develop into hepatic cirrhosis. This study aimed to investigate whether honokiol could prevent NASH induced by high-cholesterol and high-fat (CL) diet in mice and the possible mechanism involved.. Mice were fed with CL diet for 12 weeks to establish a NASH model; honokiol (0.02% w/w in diet) was added to evaluate its effect on NASH. Murine peritoneal macrophages, RAW264.7 and ANA-1 cells, were used to explore the possible mechanisms of honokiol on macrophage polarization.. Mice developed NASH after fed with CL diet for 12 weeks. Honokiol supplementation alleviated insulin resistance, hepatic steatosis, inflammation, and fibrosis induced by CL diet. Immunohistochemistry showed that honokiol induced more M2 macrophages in livers compared with CL diet alone. Honokiol decreased M1 marker genes (TNFα and MCP-1) and increased M2 marker gene (YM-1, IL-10, IL-4R and IL-13) expression in mice liver compared with CL diet. Moreover, treatment with honokiol lowered alanine aminotransferase and aspartate aminotransferase in serum and preserved liver from lipid peroxidation, evidenced by lowered hepatic malondialdehyde level. Honokiol has antioxidant function, as honokiol upregulated hepatic glutathione and superoxide dismutase level and downregulated hepatic CYP2E1 protein level. Hepatic peroxisome proliferator-activated receptor γ (PPARγ) and its target genes were upregulated by honokiol. Furthermore, honokiol (10 μM) treatment in mouse peritoneal cells, RAW264.7 cells and ANA-1 cells, led to M2 macrophage polarization, whereas a PPARγ antagonist, GW9662, abolished this effect of honokiol.. Honokiol can attenuate CL diet-induced NASH and the mechanism in which possibly is polarizing macrophages to M2 phenotype via PPARγ activation.

Topics: Animals; Biphenyl Compounds; Cell Polarity; Diet, High-Fat; Lignans; Liver; Macrophages; Male; Mice; Mice, Inbred C57BL; Non-alcoholic Fatty Liver Disease; PPAR gamma; RAW 264.7 Cells

Honokiol is a biphenolic isolate extracted from the bark of the magnolia tree that has been used in traditional Chinese and Japanese medicine, and has more recently been investigated for its anti-inflammatory and antibacterial properties. Honokiol has previously been demonstrated to improve survival in sepsis models that have rapid 100% lethality. The purpose of this study was to determine the impact of Honokiol on the host response in a model of sepsis that more closely approximates human disease. Male and female C57BL/6 mice underwent cecal ligation and puncture to induce polymicrobial intra-abdominal sepsis. Mice were then randomized to receive an injection of either Honokiol (120 mg/kg/day) or vehicle and were sacrificed after 24 h for functional studies or followed 7 days for survival. Honokiol treatment after sepsis increased the frequency of CD4 T cells and increased activation of CD4 T cells as measured by the activation marker CD69. Honokiol also increased splenic dendritic cells. Honokiol simultaneously decreased frequency and number of CD8 T cells. Honokiol decreased systemic tumor necrosis factor without impacting other systemic cytokines. Honokiol did not have a detectable effect on kidney function, lung physiology, liver function, or intestinal integrity. In contrast to prior studies of Honokiol in a lethal model of sepsis, Honokiol did not alter survival at 7 days (70% mortality for Honokiol vs. 60% mortality for vehicle). Honokiol is thus effective in modulating the host immune response and inflammation following a clinically relevant model of sepsis but is not sufficient to alter survival.

Topics: Animals; Antigens, CD; Antigens, Differentiation, T-Lymphocyte; Biphenyl Compounds; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Disease Models, Animal; Female; Inflammation; Lectins, C-Type; Lignans; Lymphocyte Activation; Lymphocyte Count; Male; Mice; Random Allocation; Sepsis; Tumor Necrosis Factor-alpha

Overexpression and activation of the EGFR have been linked to poor prognosis in several human cancers. Cetuximab is a mAb against EGFR that is used for the treatment in head and neck squamous cell carcinoma (HNSCC) and metastatic colorectal cancer. Unfortunately, most tumors have intrinsic or will acquire resistance to cetuximab during the course of therapy. Honokiol is a natural compound found in the bark and leaves of the Chinese Magnolia tree and is established to have several anticancer properties without appreciable toxicity. In this study, we hypothesized that combining cetuximab and honokiol treatments could overcome acquired resistance to cetuximab. We previously developed a model of acquired resistance to cetuximab in non-small cell lung cancer H226 cell line. Treatment of cetuximab-resistant clones with honokiol and cetuximab resulted in a robust antiproliferative response. Immunoblot analysis revealed the HER family and their signaling pathways were downregulated after combination treatment, most notably the proliferation (MAPK) and survival (AKT) pathways. In addition, we found a decrease in phosphorylation of DRP1 and reactive oxygen species after combination treatment in cetuximab-resistant clones, which may signify a change in mitochondrial function. Furthermore, we utilized cetuximab-resistant HNSCC patient-derived xenografts (PDX) to test the benefit of combinatorial treatment

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Proliferation; Cetuximab; Drug Resistance, Neoplasm; Female; Humans; Lignans

H2-P, a derivative of honokiol, was first synthesized in our laboratory. Compared with honokiol, H2-P has even high anti-tumor activity. In the present study, we evaluated the ability of H2-P to inhibit the survival rate in four gliomas cell lines. The result showed that H2-P could significantly inhibit proliferation of gliomas cells in a dose-dependent manner (IC50

Topics: Angiogenesis Inhibitors; Animals; Biphenyl Compounds; Brain Neoplasms; Cell Line, Tumor; Cell Proliferation; Chickens; Chorioallantoic Membrane; Female; Glioblastoma; Humans; Lignans; Mice; Proto-Oncogene Proteins c-myc; Rats; Signal Transduction; Xenograft Model Antitumor Assays

Obstructive sleep apnea hypopnea syndrome (OSAHS) is associated with glucose intolerance, insulin resistance and type 2 diabetes mellitus (T2DM). Although several studies have revealed that intermittent hypoxia (IH) in OSAHS may further aggravate pancreatic β cell damage and promote the evolution of type 2 diabetes (T2DM) by increasing oxidative stress, the underlying mechanisms are unclear. Honokiol, a potent radical scavenger, has been demonstrated to ameliorate oxidative stress in many cases. The present study aimed to explore the potential mechanism of IH and diabetes synergistically damage and destruct the pancreatic β cell, examine the effects of honokiol on ameliorating pancreatic β cell injury in this context and explore the mechanism of such effects. High glucose (HG) cultured INS-1 cells were exposed to 50 μM of honokiol for 24, 48 and 72 h with or without IH intervention. T2DM rats were treated with honokiol and exposed to 80 s of IH followed by 160 s of normoxia for 8 weeks. The cell proliferation, apoptosis and oxidative stress were measured. Blood glucose, insulin, glucagon and HOMA-IR (Homeostasis model assessment -insulin resistence) were also detected, and the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were detected by immunofluorescence staining and western blotting. Honokiol can reduce oxidative stress, cytotoxicity and apoptosis in the INS-1 cells of rats receiving HG treatment or both HG and IH treatment. IH can further aggravate pancreas dysfunction, cause a marked elevation in fasting blood glucose, glucagon, HOMA-IR and oxidative stress levels in DM rats. In addition, honokiol can effectively activate the Nrf2/ARE pathway and reverse this pancreatic dysfunction in vivo and in vitro. These findings indicate that honokiol acts as a potent ROS scavenger via Nrf2/ARE pathway and effectively attenuates oxidative stress and improves pancreatic β cell function of DM rats under IH treatment.

Topics: Animals; Apoptosis; Biphenyl Compounds; Blood Glucose; Cell Proliferation; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Free Radical Scavengers; Glucagon; Hypoxia; Insulin; Insulin Resistance; Insulin-Secreting Cells; Lignans; NF-E2-Related Factor 2; Oxidative Stress; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Sleep Apnea, Obstructive

The goal of the study was to investigate the cellular tolerance mechanism in response to honokiol exposure.. The broth microdilution method was employed to test the sensitivity of different Saccharomyces cerevisiae strains to honokiol. Intracellular levels of reactive oxygen species (ROSs) were determined by DCFH-DA staining. The phosphorylation of Hog1 was evaluated by Western blot analysis. The mRNA expressions of genes involved in the Ras-cyclic AMP (cAMP) pathway were analysed by real-time reverse transcription polymerase chain reaction. We found that the sod1▵ mutant was hypersensitive to honokiol and produced more ROS compared with wild-type and sod2▵ cells. Hog1 was phosphorylated in response to honokiol exposure and deletion of HOG1 increased the sensitivity to honokiol. The expressions of genes involved in the Ras-cAMP pathway were down-regulated after honokiol exposure; exogenous cAMP significantly reduced the phosphorylation of Hog1, although the level was higher than the control level.. In addition to SOD1, the Ras-cAMP cascade and Hog1 MAP kinase pathway is essential for protecting against honokiol-induced oxidative stress.. Our results provide insight into the understanding of the action mechanism of honokiol.

Topics: Biphenyl Compounds; Lignans; Mitogen-Activated Protein Kinases; Mutation; Oxidative Stress; Phosphorylation; Reactive Oxygen Species; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins

Voltage-gated potassium (K

Topics: Animals; Biphenyl Compounds; Cells, Cultured; Ganglia, Spinal; Ion Channel Gating; Lignans; Membrane Potentials; Mice; Neurons; Patch-Clamp Techniques; Potassium; Potassium Channels; Potassium Channels, Voltage-Gated

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Carcinoma, Squamous Cell; Cell Line, Tumor; Chemoradiotherapy; Down-Regulation; Female; Head and Neck Neoplasms; Humans; Kaplan-Meier Estimate; Lignans; Mice, Nude; Radiation-Sensitizing Agents; Survivin; Tumor Burden; Xenograft Model Antitumor Assays

Honokiol is the main active constituent of Magnolia officinalis. With effective and long‑term pharmacological functions of being antibacterial, anti‑oxidative, anti‑inflammatory, antitumor, anti‑spasmic, anti‑anxiety and anti‑viral, Honokiol is clinically used in the treatment of acute enteritis and chronic gastritis. The aim of the present study was to observe the possible anti‑effects of honokiol on autophagy and apoptosis of osteosarcoma, and to investigate the role of the PI3K/Akt/mTOR signaling pathway in its anticancer effects. MTT assay was used to evaluate cell proliferation and Annexin V‑fluorescein isothiocyanate/propidium iodide staining flow cytometry was used to analyze the apoptotic rate. The authors identified that honokiol could inhibit cell proliferation and induce the apoptotic rate of osteosarcoma cells. The expression level of Bcl‑2‑like protein 4, caspase‑3 and p53 protein expression were induced and cyclin D1 protein expression was suppressed in osteosarcoma cells by honokiol. Autophagy‑associated LC3II protein expression level was promoted, and PI3K, p‑Akt and p‑mTOR protein expression level was suppressed in osteosarcoma cells by honokiol. The present study demonstrated, to the best of the authors' knowledge, for the first time that honokiol induces autophagy and apoptosis of osteosarcoma cells through the PI3K/Akt/mTOR signaling pathway.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; bcl-2-Associated X Protein; Biphenyl Compounds; Bone Neoplasms; Caspase 3; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Humans; Lignans; Osteosarcoma; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction; TOR Serine-Threonine Kinases; Tumor Suppressor Protein p53

The oral microbiome is a complex ecological niche where both commensal and pathogenic bacteria coexist. Previous reports have cited that the plant isolate honokiol is a potent inhibitor of S. mutans biofilms. Herein we report a cross-coupling method that provides access to a concise library of honokiol-inspired analogs. Through this work we determined that the inhibitory activity of honokiol is highly dependent on the growth conditions. Further, we identify a series of analogs that display significant potency against oral bacteria leading to the discovery of a potent antimicrobial.

Topics: Anti-Bacterial Agents; Bacteria; Bacterial Infections; Biofilms; Biphenyl Compounds; Humans; Lignans; Microbial Sensitivity Tests; Stomatitis

Honokiol is a natural bioactive product with anti-tumor, anti-inflammatory, anti-oxidative, anti-angiogenic and neuroprotective properties. The present study aimed to investigate the effects of honokiol treatment on intimal thickening following vascular balloon injury. The current study determined that perivascular honokiol application reduced intimal thickening in rabbits 14 days after carotid artery injury, it may inhibit vascular smooth muscle cell (VSMCs) proliferation and reduce collagen deposition in local arteries. The findings of the presents study also suggested that honikiol may increase the mRNA expression levels of matrix metalloproteinase‑1 (MMP‑1), MMP‑2 and MMP‑9 and decrease tissue inhibitor of metalloproteinase‑1 (TIMP‑1) mRNA expression in the rabbit arteries. Additionally, perivascular honokiol application inhibited intimal thickening, possibly via inhibition of the phosphorylation of SMAD family member 2/3.

Topics: Animals; Biphenyl Compounds; Carotid Arteries; Carotid Artery Injuries; Cell Proliferation; Collagen; Gene Expression Regulation; Lignans; Male; Matrix Metalloproteinase 1; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Muscle, Smooth, Vascular; Protective Agents; Rabbits; Tissue Inhibitor of Metalloproteinase-1; Tunica Intima

Honokiol (poly-phenolic lignan from Magnolia grandiflora) is a Sirtuin-3 (SIRT3) activator which exhibit antioxidant activity and augment mitochondrial functions in several experimental models. Modern evidence suggests the critical role of SIRT3 in the progression of several metabolic and neurodegenerative diseases. Amyloid beta (Aβ), the precursor to extracellular senile plaques, accumulates in the brains of patients with Alzheimer's disease (AD) and is related to the development of cognitive impairment and neuronal cell death. Aβ is generated from amyloid-β precursor protein (APP) through sequential cleavages, first by β-secretase and then by γ-secretase. Drugs modulating this pathway are believed to be one of the most promising strategies for AD treatment. In the present study, we found that Honokiol significantly enhanced SIRT3 expression, reduced reactive oxygen species generation and lipid peroxidation, enhanced antioxidant activities, and mitochondrial function thereby reducing Aβ and sAPPβ levels in Chinese Hamster Ovarian (CHO) cells (carrying the amyloid precursor protein-APP and Presenilin PS1 mutation). Mechanistic studies revealed that Honokiol affects neither protein levels of APP nor α-secretase activity. In contrast, Honokiol increased the expression of AMPK, CREB, and PGC-1α, thereby inhibiting β-secretase activity leading to reduced Aβ levels. These results suggest that Honokiol is an activator of SIRT3 capable of improving antioxidant activity, mitochondrial energy regulation, while decreasing Aβ, thereby indicating it to be a lead compound for AD drug development.

Topics: Adenylate Kinase; Amyloid beta-Peptides; Animals; Biphenyl Compounds; CHO Cells; Cricetulus; Cyclic AMP Response Element-Binding Protein; Humans; Lignans; Lipid Peroxidation; Mitochondria; Reactive Oxygen Species; Sirtuin 3

Breast cancer is an alarming global public health problem and a main cause of cancer-related death in women. Systemic chemotherapy is the most widely used treatment for breast cancer. However, current chemotherapy treatments are far from desirable due to poor targeting specificity, severe side effects and vasculogenic mimicry (VM).. Hyaluronic acid (HA)-modified daunorubicin plus honokiol (HNK) cationic liposomes were prepared and characterised for treatment of breast cancer by eliminating VM.. HA-modified daunorubicin plus HNK cationic liposomes were prepared by a thin-film hydration method. Evaluations were performed on MCF-7 cells and MDA-MB-435S cells, which are human breast cancer cells, and xenografts of MDA-MB-435S cells.. In vitro results revealed that the HA-modified daunorubicin plus HNK cationic liposomes enhanced the cellular uptake and destroyed VM channels. In vivo results demonstrated that the liposomes prolonged the circulation time in the blood, obviously accumulated in the tumour region, and enhanced the overall anticancer effects. Action mechanisms were related to down-regulation of VM protein indicators including FAK, EphA2, MMP-2 and MMP-9.. The prepared HA-modified daunorubicin plus HNK cationic liposomes may serve as a promising therapeutic strategy for the treatment of breast cancer.

Topics: Animals; Biphenyl Compounds; Breast Neoplasms; Cations; Daunorubicin; Female; Humans; Hyaluronic Acid; Lignans; Liposomes; MCF-7 Cells; Mice; Neovascularization, Pathologic

Nonalcoholic fatty liver disease (NAFLD) is the most common liver disease, and is associated with the development of metabolic syndrome. Postmenopausal women with estrogen deficiency are at a higher risk of progression to NAFLD. Estrogen has a protective effect against the progression of the disease. Currently, there are no safe and effective treatments for these liver diseases in postmenopausal women. Honokiol (Ho), a bioactive natural product derived from Magnolia spp, has anti-inflammatory, anti-angiogenic, and anti-oxidative properties. In our study, we investigated the beneficial effects of Ho on NAFLD in ovariectomized (OVX) mice. We divided the mice into four groups, as follows: SHAM, OVX, OVX+β-estradiol (0.4 mg/kg of bodyweight), and OVX+Ho (50 mg/kg of diet). Mice were fed diets with/without Ho for 12 weeks. The bodyweight, epidermal fat, and weights of liver tissue were lower in the OVX group than in the other groups. Ho improved hepatic steatosis and reduced proinflammatory cytokine levels. Moreover, Ho markedly downregulated plasma lipid levels. Our results indicate that Ho ameliorated OVX-induced fatty liver and inflammation, as well as associated lipid metabolism. These findings suggest that Ho may be hepatoprotective against NAFLD in postmenopausal women.

Topics: Adiposity; Animals; Biomarkers; Biphenyl Compounds; Body Weight; Cytokines; Disease Models, Animal; Fatty Liver; Gene Expression Profiling; Inflammation Mediators; Lignans; Lipid Metabolism; Liver; Mice; Non-alcoholic Fatty Liver Disease; Organ Size; Ovariectomy

Honokiol, an active natural product derived from Magnolia officinalis, exerted anticancer effects through a variety of mechanisms on multiple types of cancers. In this study, the molecular mechanisms of honokiol in suppressing the human oral squamous cell carcinoma (OSCC) cells were evaluated. Treatment of two OSCC cell lines with honokiol resulted in reducing the cell proliferation and arresting the cell cycle at G1 stage which was correlated with the down-regulation of Cdk2 and Cdk4 and the up-regulation of cell cycle suppressors, p21 and p27. In addition, the caspase-dependent programmed cell death was substantially detected, and the autophagy was induced as the autophagosome formation and autophagic flux proceeded. Modulation of autophagy by autophagic inducer, rapamycin or inhibitors, 3-MA or bafilomycin, potentiated the honokiol-mediated anti-OSCC effects where honokiol exerted multiple actions in suppression of MAPK pathway and regulation of Akt/mTOR or AMPK pathways. As compared to clinical therapeutic agent, 5-FU, honokiol exhibited more potent activity against OSCC cells and synergistically enhanced the cytotoxic effect of 5-FU. Furthermore, orally administrated honokiol exerted effective antitumour activity in vivo in OSCC-xenografted mice. Thus, this study revealed that honokiol could be a promising candidate in preventing human OSCCs.

Topics: Adenine; Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Autophagy; Biphenyl Compounds; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Cyclin-Dependent Kinase 2; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinase Inhibitor p21; Cyclin-Dependent Kinase Inhibitor p27; Fluorouracil; G1 Phase Cell Cycle Checkpoints; Gene Expression Regulation, Neoplastic; Humans; Lignans; Macrolides; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Mouth Neoplasms; Proto-Oncogene Proteins c-akt; Signal Transduction; Sirolimus; TOR Serine-Threonine Kinases; Tumor Burden; Xenograft Model Antitumor Assays

Honokiol is a natural product and an emerging drug for a wide variety of malignancies, including hematopoietic malignancies, sarcomas, and common epithelial tumors. The broad range of activity of honokiol against numerous malignancies with diverse genetic backgrounds suggests that honokiol is inhibiting an activity that is common to multiple malignancies. Oncogenic transcription factor FOXM1 is one of the most overexpressed oncoproteins in human cancer. Here we found that honokiol inhibits FOXM1-mediated transcription and FOXM1 protein expression. More importantly, we found that honokiol's inhibitory effect on FOXM1 is a result of binding of honokiol to FOXM1. This binding is specific to honokiol, a dimerized allylphenol, and was not observed in compounds that either were monomeric allylphenols or un-substituted dihydroxy phenols. This indicates that both substitution and dimerization of allylphenols are required for physical interaction with FOXM1. We thus demonstrate a novel and specific mechanism for FOXM1 inhibition by honokiol, which partially may explain its anticancer activity in cancer cells.

Topics: Animals; Biphenyl Compounds; Cell Line, Tumor; Down-Regulation; Forkhead Box Protein M1; Humans; Lignans; Mice; Proteasome Inhibitors; Transcriptional Activation

Honokiol (HNK) is a small biphenolic compound, which exerts antineoplastic effects in various types of cancer. However, the mechanism underlying the antitumor effects of HNK in osteosarcoma (OS) cells is not yet fully understood. Emerging evidence has indicated that microRNAs (miRNAs/miRs) serve key roles in numerous pathological processes, including cancer. It has previously been reported that Chinese medicinal herbs harbor anticancer properties via modulating miRNA expression. Therefore, the present study aimed to determine whether HNK could suppress OS cell growth by regulating miRNA expression. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometric analysis were used to evaluate the cell proliferation and apoptosis in human OS cells after treatment with HNK, respectively. The results demonstrated that HNK inhibits proliferation and induces apoptosis of human OS cells in a dose‑dependent manner. Furthermore, HNK‑induced apoptosis was characterized by upregulation of proapoptotic proteins, including cleaved‑caspase‑3, cleaved‑poly (ADP‑ribose) polymerase and B‑cell lymphoma 2 (Bcl‑2)‑associated X protein, and downregulation of the anti‑apoptotic protein Bcl‑2. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) verified that HNK was able to induce aberrant expression of miRNAs in human OS cells, and miR‑21 was one of the miRNAs that was most significantly downregulated. To further investigate miR‑21 function, the present study validated that HNK reduces miR‑21 levels in a dose‑dependent manner. In addition, restoration of miR‑21 expression abrogated the suppressive effects of HNK on OS cells. Luciferase assay and western blot analysis identified that miR‑21 inhibits the expression of phosphatase and tensin homolog (PTEN) by directly targeting its 3'-UTR. Notably, HNK was able to suppress the phosphoinositide 3‑kinase (PI3K)/protein kinase B (AKT) signaling pathway; however, it was reactivated by miR‑21 overexpression. Taken together, these data indicated that HNK may inhibit proliferation and induce apoptosis of human OS cells by modulating the miR‑21/PTEN/PI3K/AKT signaling pathway. Therefore, miR‑21 may be considered a potential therapeutic target for the treatment of osteosarcoma with HNK.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Bone Neoplasms; Cell Line, Tumor; Cell Proliferation; Gene Expression Regulation, Neoplastic; Humans; Lignans; MicroRNAs; Osteosarcoma; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; PTEN Phosphohydrolase; Signal Transduction

Glioblastoma multiforme (GBM), a prevalent brain cancer with high mortality, is resistant to the conventional single-agent chemotherapy. In this study, we employed a combination chemotherapy strategy to inhibit GBM growth and addressed its possible beneficial effects. The synergistic effect of lauroyl-gemcitabine (Gem-C

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Biphenyl Compounds; Brain Neoplasms; Cell Line, Tumor; Deoxycytidine; Drug Carriers; Drug Compounding; Gemcitabine; Glioblastoma; Humans; Hyaluronan Receptors; Hyaluronic Acid; Lignans; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Micelles; Nanoparticles; Rats; Rats, Sprague-Dawley; Spheroids, Cellular; Survival Rate; Treatment Outcome; Xenograft Model Antitumor Assays

Honokiol-loaded micelles were prepared by emulsion-solvent evaporation procedure when oligochitosan-pluronic conjugate (CS-F127) as carrier. Differential scanning calorimetry (DSC) indicated that honokiol existed in amorphous form when it was encapsulated into the micelles with 87.54 ± 1.52% of encapsulation efficiency (EE) and 12.51 ± 0.22% of drug loading (DL) capacity. The water-solubility was increased to 1.46 mg/mL, being >27-folds higher than pure honokiol. The in vitro release study demonstrated a slow and sustained ± release of honokiol from the drug-loaded micelles with pure honokiol as control. The in vitro antifungal and cellular uptake tests indicated that the drug-loaded micelles showed the same activity as pure honokiol against Candida albicans due to its good cellular uptake although it slowly released honokiol. The pharmacokinetic test results showed that the honokiol-loaded micelles increased area under curves and mean retention time of honokiol with low clearance rate and apparent distribution volume when compared with pure honokiol, showing its ability to improve honokiol's pharmacokinetic properties. The honokiol-loaded micelles also showed good bio-security to normal cells and main organs of mice. In conclusion, the CS-F127 conjugate should be a potential carrier for honokiol or other antifungal agents in the treatment of fungal infections.

Topics: Animals; Antifungal Agents; Biological Transport; Biphenyl Compounds; Candida albicans; Cell Line; Chitin; Chitosan; Drug Carriers; Drug Liberation; Hemolysis; Lignans; Materials Testing; Micelles; Oligosaccharides; Poloxamer; Rats; Solubility; Solvents; Volatilization

Staphylococcus aureus is a major and dangerous human pathogen that causes a range of clinical manifestations of varying severity, and is the most commonly isolated pathogen in the setting of skin and soft tissue infections, pneumonia, suppurative arthritis, endovascular infections, foreign-body associated infections, septicemia, osteomyelitis, and toxic shocksyndrome. Honokiol, a pharmacologically active natural compound derived from the bark of Magnolia officinalis, has antibacterial activity against Staphylococcus aureus which provides a great inspiration for the discovery of potential antibacterial agents. Herein, honokiol derivatives were designed, synthesized and evaluated for their antibacterial activity by determining the minimum inhibitory concentration (MIC) against S. aureus ATCC25923 and Escherichia coli ATCC25922 in vitro. 7c exhibited better antibacterial activity than other derivatives and honokiol. The structure-activity relationships indicated piperidine ring with amino group is helpful to improve antibacterial activity. Further more, 7c showed broad spectrum antibacterial efficiency against various bacterial strains including eleven gram-positive and seven gram-negative species. Time-kill kinetics against S. aureus ATCC25923 in vitro revealed that 7c displayed a concentration-dependent effect and more rapid bactericidal kinetics better than linezolid and vancomycin with the same concentration. Gram staining assays of S. aureus ATCC25923 suggested that 7c could destroy the cell walls of bacteria at 1×MIC and 4×MIC.

Topics: Anti-Bacterial Agents; Biphenyl Compounds; Cell Wall; Cyclization; Drug Design; Escherichia coli; Kinetics; Lignans; Linezolid; Microbial Sensitivity Tests; Staphylococcus aureus; Structure-Activity Relationship; Vancomycin

Osteosarcoma is the most common primary malignant tumor of bone, the long-term survival of which has stagnated in the past several decades. In the present study, we investigated the anticancer effect of honokiol (HNK), an active component isolated and purified from the magnolia officinalis on human osteosarcoma cells. Our results showed that honokiol caused dose-dependent and time-dependent cell death in human osteosarcoma cells. The types of cell death induced by honokiol were primarily autophagy and apoptosis. Furthermore, honokiol induced G0/G1 phase arrest, elevated the levels of glucose-regulated protein (GRP)-78, an endoplasmic reticular stress (ERS)-associated protein, and increased the production of intracellular reactive oxygen species (ROS). In contrast, reducing production of intracellular ROS using N-acetylcysteine, a scavenger of ROS, concurrently suppressed honokiol-induced cellular apoptosis, autophagy, and cell cycle arrest. Consequently, honokiol stimulated phosphorylation of extracellular signal-regulated kinase (ERK)1/2. Furthermore, pretreatment of osteosarcoma cells with PD98059, an inhibitor of ERK1/2, inhibited honokiol-induced apoptosis and autophagy. Finally, honokiol suppressed tumor growth in the mouse xenograft model. Taken together, our results revealed that honokiol caused G0/G1 phase arrest, induced apoptosis, and autophagy via the ROS/ERK1/2 signaling pathway in human osteosarcoma cells. Honokiol is therefore a promising candidate for development of antitumor drugs targeting osteosarcoma.

Topics: Animals; Apoptosis; Autophagy; Autophagy-Related Protein 7; Biphenyl Compounds; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Proliferation; Endoplasmic Reticulum Stress; Enzyme Activation; Fibroblasts; Humans; Lignans; MAP Kinase Signaling System; Mice, Inbred BALB C; Mice, Nude; Osteosarcoma; Reactive Oxygen Species; Xenograft Model Antitumor Assays

The hepatocellular carcinoma is one of the most common malignant tumour with high level of mortality rate due to its rapid progression and high resistance to conventional chemotherapies. Thus, the search for novel therapeutic leads is of global interest. Herein, a small set of derivatives of magnolol 1 and honokiol 2, the main components of Magnolia grandiflora and Magnolia obovata, were evaluated in in vitro assay using tumoral hepatocytes. The pro-drug approach was applied as versatile strategy to the improve bioactivity of the compounds by careful transformation of the hydroxyl groups of magnolol 1 and honokiol 2 in suitable ester derivatives. Compounds 10 and 11 resulted to be more potent than the parental honokiol 2 at concentration down to 1 μM with complete viability of treated fibroblast cells up to concentrations of 80 μM. The combination of a butyrate ester and a bare phenol-OH group in the honokiol structure seemed to play a significant role in the antiproliferative activity identifying an interesting pharmacological clue against hepatocellular carcinoma.

Topics: Biphenyl Compounds; Carbon-13 Magnetic Resonance Spectroscopy; Carcinoma, Hepatocellular; Cell Line; Cell Line, Tumor; Drug Screening Assays, Antitumor; Humans; Lignans; Liver Neoplasms; Proton Magnetic Resonance Spectroscopy

A novel method based on amino acid ionic liquids (AAILs) as an additive synchronous fluorescence spectrometry is proposed for simultaneous determination of magnolol (MN) and honokiol (HN) in traditional Chinese medicine Houpu. The overlapping fluorescence spectrum of MN and HN could be completely separated in the AAILs medium. Experiment parameters (the type and concentration of AAILs, pH values and temperature) were discussed. The detection limits of MN and HN reached 1.46ng/mL, 0.92ng/mL and the recovery rates ranged from 98.6%-100.7%, 99.7%-100.6%, respectively. This methods was successfully employed for simultaneously determination of MN and HN in real samples. No significant differences could be found in the results of this method and the pharmacopoeia of People's Republic of China 2015 (Ch.P.2015). The experiment mechanisms were discussed by the Gaussian simulation and fluorescence quantum yield.

Topics: Amino Acids; Biphenyl Compounds; Hydrogen-Ion Concentration; Imidazoles; Ionic Liquids; Lignans; Limit of Detection; Linear Models; Models, Molecular; Reproducibility of Results; Spectrometry, Fluorescence; Temperature

Topics: Antineoplastic Agents; Biological Products; Biphenyl Compounds; China; Chromatography, High Pressure Liquid; Clinical Trials as Topic; Drug Contamination; Drugs, Chinese Herbal; Lignans; Magnetic Resonance Spectroscopy; Reproducibility of Results; Sensitivity and Specificity; Tandem Mass Spectrometry

This study was aimed to clarify the effect of honokiol (Hon) on the activity of Cytochrome P450 (CYP450) enzymes, and the level of mRNA expression of liver and kidney transporters in type 2 diabetic rats induced by high-fat diet and strepotozotocin. Rats were randomly divided into normal control (NC) group, diabetic control (DC) group and Hon groups (

Topics: Animals; Anion Transport Proteins; Biphenyl Compounds; Cytochrome P-450 Enzyme System; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Gastrointestinal Agents; Kidney; Lignans; Liver; Male; Rats; Rats, Sprague-Dawley; RNA, Messenger

Temozolomide (TMZ)-induced side effects and drug tolerance to human gliomas are still challenging issues now. Our previous studies showed that honokiol, a major bioactive constituent of Magnolia officinalis (Houpo), is safe for normal brain cells and can kill human glioma cells. This study was further aimed to evaluate the improved effects of honokiol and TMZ on drug-sensitive and -resistant glioma cells and the possible mechanisms.. TMZ-sensitive human U87-MG and murine GL261 glioma cells and TMZ-resistant human U87-MR-R9 glioma cells were exposed to honokiol and TMZ, and cell viability and LC50 of honokiol were assayed. To determine the death mechanisms, caspase-3 activity, DNA fragmentation, apoptotic cells, necrotic cells, cell cycle, and autophagic cells. The glioma cells were pretreated with 3-methyladenine (3-MA) and chloroquine (CLQ), two inhibitors of autophagy, and then exposed to honokiol or TMZ.. Exposure of human U87-MG glioma cells to honokiol caused cell death and significantly enhanced TMZ-induced insults. As to the mechanism, combined treatment of human U87-MG cells with honokiol and TMZ induced greater caspase-3 activation, DNA fragmentation, cell apoptosis, and cell-cycle arrest at the G. Taken together, this study demonstrated the improved effects of honokiol with TMZ on autophagy and subsequent apoptosis of drug-sensitive and -tolerant glioma cells. Thus, honokiol has the potential to be a drug candidate for treating human gliomas.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Autophagy; Biphenyl Compounds; Caspase 3; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Survival; Drug Resistance, Neoplasm; Drug Synergism; Glioma; Humans; Lignans; Temozolomide

Topics: Animals; Antioxidants; Biphenyl Compounds; Carrier Proteins; Cell Cycle Proteins; Inflammasomes; Intervertebral Disc Degeneration; Lignans; Male; NLR Family, Pyrin Domain-Containing 3 Protein; Nucleus Pulposus; Rats; Rats, Sprague-Dawley; Signal Transduction

This article aims to conduct toxicity test research on honokiol microemulsion(HM) to provide reference frame for the safe dose design as well as the toxic and adverse reaction monitoring in clinic. High performance liquid chromatography (HPLC) method was adopted to determine the concentration, stability and uniformity of HM and the results indicated that the test sample was conformed to the toxicity test requirements. In the acute toxicity test, six intravenous drip dosages, namely, 100.0, 66.7, 44.4, 19.8, 8.8, and 3.9 mg/kg were set, with one beagle dog in each dosage, respectively. In addition, the results also demonstrated that the approximate lethal dose range of HM was 66.7-100.0 mg/kg. In the subchronic toxicity test, beagle dogs were intravenously dripped with HM at doses of 1.25, 0.25 and 0.05 mg/kg for 30 days. During the test period, signs of gross toxicity, behavioral changes, body weight, rectal temperature, food consumption, ophthalmoscopy, electrocardiography, urinalysis, blood biochemistry, coagulation, hematology, organ weights and histopathology were examined. Under the present study conditions, the no-observed-adverse-effect level for HM was estimated to be 0.25 mg/kg. According to the results of bacterial reverse mutation, chromosomal aberration and micronucleus assays, HM exhibited no notable genotoxicity both in vivo and in vitro.

Topics: Animals; Biphenyl Compounds; Dogs; Emulsions; Female; Lignans; Male; Mice, Inbred ICR; Mutagenicity Tests; No-Observed-Adverse-Effect Level; Toxicity Tests, Acute; Toxicity Tests, Subchronic

Reducing oxidative stress is a crucial therapeutic strategy for ameliorating diabetic myocardial ischemia/reperfusion (MI/R) injury. Honokiol (HKL) acts as an effective cardioprotective agent for its strong antioxidative activity. However, its roles and underlying mechanisms against MI/R injury in type 1 diabetes (T1D) remain unknown. Since SIRT1 and Nrf2 are pivotal regulators in diabetes mellitus patients suffering from MI/R injury, we hypothesized that HKL ameliorates diabetic MI/R injury via the SIRT1-Nrf2 signaling pathway. Streptozotocin-induced T1D rats and high-glucose-treated H9c2 cells were exposed to HKL, with or without administration of the SIRT1 inhibitor EX527, SIRT1 siRNA, or Nrf2 siRNA, and then subjected to I/R operation. We found that HKL markedly improved the postischemic cardiac function, decreased the infarct size, reduced the myocardial apoptosis, and diminished the reactive oxygen species generation. Intriguingly, HKL remarkably activated SIRT1 signaling, enhanced Nrf2 nuclear translocation, increased antioxidative signaling, and decreased apoptotic signaling. However, these effects were largely abolished by EX527 or SIRT1 siRNA. Additionally, our cellular experiments showed that Nrf2 siRNA blunted the cytoprotective effects of HKL, without affecting SIRT1 expression and activity. Collectively, these novel findings indicate that HKL abates MI/R injury in T1D by ameliorating myocardial oxidative damage and apoptosis via the SIRT1-Nrf2 signaling pathway.

Topics: Animals; Apoptosis; Biphenyl Compounds; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 1; Echocardiography; In Situ Nick-End Labeling; Lignans; Myocardial Reperfusion Injury; Myocardium; NF-E2-Related Factor 2; Oxidative Stress; Rats; Signal Transduction; Sirtuin 1; Superoxide Dismutase

This study aims to design a novel nano-sized anticancer drug delivery system that can enhance the therapeutic effects of the loaded drug. With this idea in mind, this work reported the design and characterization of epigallocatechin-3-gallate (EGCG) functionalized chitin (CH) derivative, and its application in nano-drug delivery system. The EGCG-functionalized CH (CE) polymer was firstly prepared and characterized. The nanoparticles (NPs) of CE-loaded honokiol (HK), which was prepared by ionic crosslinking, exhibited a size of 80 nm, zeta potential of +33.8 mV, and spherical morphology. The antitumor activity of the CE-HK NPs in vitro and in vivo was investigated and compared to free HK. As a result, the CE-HK NPs can effectively inhibited cell proliferation of HepG2 cell by inhibiting more cells in the G2/M phase and decreasing mitochondrial membrane potential. The CE-HK NPs (40 mg/kg) inhibited tumor growth by 83.55% (p < 0.05), which was far higher than the 30.15% inhibition of free HK (40 mg/kg). The proposed delivery system exhibits better tumor selectivity and growth reduction both in vitro and in vivo, and does not induce any side effects. Therefore, the CE-HK NPs may act as an effective delivery system of liver cancer agent HK.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Carcinoma, Hepatocellular; Catechin; Cell Proliferation; Chitin; Cross-Linking Reagents; Dose-Response Relationship, Drug; Drug Carriers; Drug Compounding; Drug Liberation; G2 Phase Cell Cycle Checkpoints; Hep G2 Cells; Humans; Kinetics; Lignans; Liver Neoplasms; Male; Membrane Potential, Mitochondrial; Mice, Inbred BALB C; Nanoparticles; Nanotechnology; Particle Size; Solubility; Surface Properties; Technology, Pharmaceutical; Tumor Burden; Xenograft Model Antitumor Assays

In an effort to prevent metastasis of breast tumor cells- at the same time of inhibiting tumor growth with less toxic side effects, honokiol (HNK) was encapsulated into pH-sensitive polymeric micelles based on the conjugate of poly(2-ethyl-2-oxazoline)-poly(d,l-lactide) (PEOz-PLA) with doxorubicin (DOX), denoted as PEOz-PLA-imi-DOX. PEOz-PLA-imi-DOX was successfully synthesized by connecting DOX to the hydrophobic end of PEOz-PLA via acid-cleavable benzoic imine linker. HNK-loaded conjugate micelles (HNK/PP-DOX-PM) with a size of 21 nm and homogeneous spherical shape exhibited high drug-loading capacity. PEOz-PLA-imi-DOX and HNK/PP-DOX-PM displayed faster release of DOX at pH 5.0 than at pH 7.4. As anticipated, PEOz-PLA-imi-DOX maintained cytotoxicity of DOX against MDA-MB-231 cells. The synergistically enhanced in vitro antitumor effect of HNK/PP-DOX-PM was confirmed by their synergetic inhibition of MDA-MB-231 cell growth. Furthermore, the efficient prevention of tumor metastasis by HNK/PP-DOX-PM was testified by in vitro anti-invasion, wound healing and antimigration assessment in MDA-MB-231 cells, and in vivo bioluminescence imaging in nude mice. The suppression of growth and metastasis of tumor cells by HNK/PP-DOX-PM was attributed to the synergistic effect of pH-triggered drug release and HNK-aroused inhibition of matrix metalloproteinases and epithelial-mesenchymal transition, respectively. In addition, HNK/PP-DOX-PM exhibited superior biosafety than physically encapsulated dual-drug micelles. Consequently, the fabricated HNK/PP-DOX-PM may have great potential for safe and effective suppression of tumor growth and metastasis.

Topics: Animals; Biphenyl Compounds; Cell Line, Tumor; Doxorubicin; Drug Carriers; Humans; Hydrogen-Ion Concentration; Lignans; Mice; Mice, Nude; Micelles; Polymers

BACKGROUND The cortex of Magnolia officinalis has long been used as an element of traditional Chinese medicine for the treatment of anxiety, chronic bronchitis, and gastrointestinal dysfunction. This study aimed to elucidate the underlying mechanism of its functional ingredients (magnolol and honokiol) in modifying the secretion and absorption homeostasis and protecting mucosal integrity in an Enterotoxigenic Escherichia coli (ETEC)-induced diarrhea mouse model. MATERIAL AND METHODS This study established a diarrhea mouse model infected by ETEC at a dosage of 0.02 ml/g live body weight (BW) in vivo. Magnolol or honokiol was followed by an intraperitoneal administration at dosages of 100, 300, and 500 mg/kg BW according to a 3×3 factorial arrangement. The useful biomarkers for evaluating the integrity of intestinal tract and histologic injury were analyzed and morphological development (including villus height, crypt depth, and ratio of villus height to crypt depth) and the expressions of inflammatory cytokines were determined by real-time PCR. RESULTS The results showed that magnolol and honokiol (500 mg/kg BW) reduced the concentrations of NO, DAO, and DLA, and iNOS activity, and the mRNA expressions of the interferon gamma (IFN-γ) and interleukin 10 (IL-10), and inhibited intestinal epithelial cell apoptosis. Magnolol and honokiol (300 mg/kg BW) elongated the villus height and crypt depth and decreased the number of goblet cells and the ratio of villus height to crypt depth. CONCLUSIONS The current results indicate that magnolol and honokiol enhance the intestinal anti-inflammatory capacities, elongate the villus height and crypt depth, and reduce goblet cell numbers to inhibit the intestinal epithelium apoptosis and effectively protect the intestinal mucosa. These results show that magnolol and honokiol protect the intestinal mucosal integrity and regulate gastrointestinal dysfunction.

Topics: Administration, Oral; Animals; Apoptosis; Biphenyl Compounds; Cytokines; Enterotoxigenic Escherichia coli; Homeostasis; Inflammation Mediators; Intestinal Absorption; Intestinal Mucosa; Lignans; Mice; Nitric Oxide Synthase

Honokiol (HNK), the main active component of Magnolia officinalis, has shown a variety of pharmacological activities. In the present study, we measured the antidepressant-like effects of HNK in a rat model of chronic unpredictable mild stress (CUMS) and explored its possible mechanisms. The antidepressant-like effects of HNK were assessed in rats by an open field test (OFT), sucrose preference test (SPT) and forced swimming test (FST). Then, serum levels of corticotrophin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and corticosterone (CORT) and hippocampal brain-derived neurotrophic factor (BDNF) and glucocorticoid receptor α (GRα) levels were assessed to explore the possible mechanisms. We identified that HNK treatment (2, 4, and 8 mg/kg) alleviated the CUMS-induced behavioural deficits. Treatment with HNK also normalized the CUMS-induced hyperactivity of the limbic hypothalamic-pituitary-adrenal (HPA) axis, as indicated by reduced CRH, ACTH and CORT serum levels. In addition, HNK increased the expression of GRα (mRNA and protein) and BDNF (mRNA and protein) in the hippocampus. These data confirmed the antidepressant-like effects of HNK, which may be related to its normalizing the function of the HPA axis and increasing the BDNF level in the hippocampus.

Topics: Animals; Antidepressive Agents; Biphenyl Compounds; Brain-Derived Neurotrophic Factor; Chronic Disease; Dose-Response Relationship, Drug; Hippocampus; Hypothalamo-Hypophyseal System; Lignans; Male; Pituitary-Adrenal System; Rats; Rats, Wistar; Stress, Psychological; Treatment Outcome

Accumulating evidence has demonstrated that mitochondrial dysfunction is a prominent early event in the progression of Alzheimer's disease (AD). Whether protecting mitochondrial function can reduce amyloid-β oligomer (AβO)-induced neurotoxicity in PS1V97L transgenic mice remains unknown. In this study, we examined the possible protective effects of honokiol (HKL) on mitochondrial dysfunction induced by AβOs in neurons, and cognitive function in AD PS1V97Ltransgenic mice. We determined that HKL increased mitochondrial sirtuin 3 (SIRT3) expression levels and activity, which in turn markedly improved ATP production and weakened mitochondrial reactive oxygen species production. We demonstrated that the enhanced energy metabolism and attenuated oxidative stress of HKL restores AβO-mediated mitochondrial dysfunction in vitro and in vivo. Consequently, memory deficits in the PS1V97L transgenic mice were rescued by HKL in the early stages. These results suggest that HKL has therapeutic potential for delaying the onset of AD symptoms by alleviating mitochondrial impairment and increasing hyperactivation of SIRT3 in the pathogenesis of preclinical AD.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Apoptosis; Biphenyl Compounds; Cells, Cultured; Cognition Disorders; Disease Models, Animal; Embryo, Mammalian; Enzyme Inhibitors; Female; Hippocampus; Lignans; Male; Membrane Potential, Mitochondrial; Mice; Mice, Inbred C57BL; Mice, Transgenic; Mitochondria; Neurons; Oligoribonucleotides; Oxidative Stress; Presenilin-1; Rats; Rats, Sprague-Dawley; Sirtuin 3

Honokiol is a natural product extracted from herbal plants such as the Magnolia species which have been shown to exhibit anti-tumor and anti-metastatic properties. However, the effects of honokiol on thyroid cancers are largely unknown.. To determine whether honokiol might be useful for the treatment of thyroid cancer and to elucidate the mechanism of toxicity of honokiol, we analyzed the impact of honokiol treatment on differential protein expression in human thyroid cancer cell line ARO using lysine-labeling two-dimensional difference gel electrophoresis (2D-DIGE) combined with mass spectrometry (MS).. This study revealed 178 proteins that showed a significant change in expression levels and also revealed that honokiol-induced cytotoxicity in thyroid cancer cells involves dysregulation of cytoskeleton, protein folding, transcription control and glycolysis.. Our work shows that combined proteomic strategy provides a rapid method to study the molecular mechanisms of honokiol-induced cytotoxicity in thyroid cancer cells. The identified targets may be useful for further evaluation as potential targets in thyroid cancer therapy.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Survival; Cytoskeleton; Electrophoresis, Gel, Two-Dimensional; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Glycolysis; Humans; Lignans; Magnolia; Mass Spectrometry; Neoplasm Metastasis; Plant Extracts; Protein Processing, Post-Translational; Proteome; Proteomics; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Thyroid Neoplasms

Parkinson's disease (PD) is a progressive and profound movement disorder resulting from neurodegeneration in the nigrostriatal dopaminergic system, but current treatment neither cures nor stops PD from advancing. Based on the ability to suppress oxidative stress, excitotoxicity, and neuroinflammation, the potential of honokiol as a novel neuroprotective agent for PD treatment was determined.. The hemi-parkinsonian model was used to investigate the protective and therapeutic effects of honokiol on motor dysfunctions and dopaminergic neurodegeneration in mice, with a single unilateral striatal injection of 6-hydroxydopamine (6-OHDA).. One day after 6-OHDA-induced lesion, the mice exhibited spontaneous ipsilateral turning, motor imbalance, and incoordination which were mild with a single administration of honokiol prior to 6-OHDA injection. Thereafter, honokiol was continually applied daily for 14 days, which ameliorated apomorphine-induced contralateral rotation and reduced the loss of tyrosine hydroxylase-immunoreactive (TH-ir) fibers in the lesioned striatum. In addition, honokiol posttreatment, beginning on day 8 after 6-OHDA lesion, for 14 days efficiently rescued motor deficits and recovered the TH-ir neuronal loss in both the lesioned striatum and the ipsilateral substantia nigra. The 6-OHDA-induced increases in nigrostriatal expression of inducible nitric oxide synthase (iNOS) and decreases in that of nNOS were also reversed by honokiol posttreatment.. These findings revealed that honokiol has both protective and therapeutic effects on motor impairments and dopaminergic progressive damage, at least in part through modulation of NOS signaling, in 6-OHDA-lesioned mice. Honokiol may represent a potential therapeutic candidate for the management of motor symptoms and neurodegeneration in PD.

Topics: Animals; Apomorphine; Biphenyl Compounds; Corpus Striatum; Disease Models, Animal; Lignans; Male; Mice; Microinjections; Motor Activity; Neuroprotective Agents; Nitric Oxide Synthase Type I; Nitric Oxide Synthase Type II; Oxidopamine; Parkinson Disease, Secondary; Postural Balance; Substantia Nigra; Tyrosine 3-Monooxygenase

Honokiol, a bioactive compound isolated from the cone and bark of

Topics: Antifungal Agents; Apoptosis; Biphenyl Compounds; Calcineurin; Calcineurin Inhibitors; Candida albicans; Cyclosporine; HSP90 Heat-Shock Proteins; Lignans; Magnolia; Microbial Sensitivity Tests; Mitochondria

Honokiol (2-(4-hydroxy-3-prop-2-enyl-phenyl)-4-prop-2-enyl-phenol) and magnolol (4-Allyl-2-(5-allyl-2-hydroxy-phenyl)phenol) are the major active polyphenol constituents of

Topics: Administration, Intravenous; Animals; Biphenyl Compounds; Chromatography, High Pressure Liquid; Cytochrome P-450 CYP1A1; Cytochrome P-450 Enzyme System; Diclofenac; Drug Interactions; Gene Expression Regulation; Lignans; Liver; Microsomes, Liver; Molecular Structure; Phenacetin; Rats; Rats, Sprague-Dawley

Honokiol (Hon), a polyphenol and main active ingredient from the bark of Magnolia officinalis, has been documented as having multiple pharmacological functions, including neuroprotection. However, the mechanisms underlying its neuroprotective effects are not well-defined. In this study, we reported that Hon attenuates the H

Topics: Adrenergic Agents; Animals; Apoptosis; Biphenyl Compounds; Glutathione; Heme Oxygenase (Decyclizing); Hydrogen Peroxide; Lignans; NAD(P)H Dehydrogenase (Quinone); Neurons; Neuroprotective Agents; NF-E2-Related Factor 2; Oxidants; Oxidative Stress; Oxidopamine; PC12 Cells; Rats; Thioredoxin Reductase 1; Thioredoxins

Allergic asthma is a disease that pathologically characterized by eosinophilia infiltration, airway inflammation and hyper responsiveness. In this study, we evaluated the anti-inflammatory and anti-allergy possibilities of honokiol, a bi-phenolic compound obtained from species of the genus Magnolia, which has long been involved in traditional Chinese prescriptions for asthma-related lung diseases, in an ovalbumin-induced mouse model of allergic asthma. We found honokiol significantly inhibited the eosinophilia infiltration, reduced the airway inflammation and suppressed the production of inflammatory cytokines) as well as the IgE in serum. Moreover, MMP-9 and? (IL-4 and IFN- NF-κB were found to be involved in the honokiol induced biological process. These results suggested that honokiol may be a possible candidate in the treatment of lung asthma related diseases.

Topics: Administration, Oral; Animals; Anti-Allergic Agents; Anti-Inflammatory Agents; Asthma; Biphenyl Compounds; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Immunoglobulin E; Inflammation; Lignans; Mice, Inbred BALB C; Respiratory System

Oxidative stress contributes to the initiation and progression of liver damage. SIRT3 is a member of nicotinamide adenine dinucleotide-dependent deacetylases that plays a key role in anti-oxidative defense and mitochondrial function in the liver. Honokiol is a natural lignan from the plants of Magnolia genus that exhibits potent anti-oxidative property. This study aims to evaluate the hepatoprotective potential of honokiol against oxidative injury in tert-butyl hydroperoxide (t-BHP)-injured AML12 hepatocytes in vitro and carbon tetrachloride (CCl

Topics: AMP-Activated Protein Kinases; Animals; Biphenyl Compounds; Cell Line; Cytoprotection; Enzyme Activation; Hepatocytes; Lignans; Mice; Mitochondria; Oxidative Stress; Protein Serine-Threonine Kinases; Signal Transduction; Sirtuin 3

Honokiol and magnolol, which are lignans isolated from Magnolia quinquepeta, have some pharmacological effects. However, the anti-inflammatory effects of honokiol and magnolol on periodontal disease are still uncertain. The aim of this study was to examine the effect of honokiol and magnolol on CXC chemokine receptor 3 (CXCR3) ligands, which are related with Th1 cell migration, production in interleukin (IL)-27-stimulated human oral epithelial cells (TR146 cells). Honokiol and magnolol inhibited CXC chemokine ligand (CXCL)10 and CXCL11 production in IL-27-stimulated TR146 cells in a dose-dependent manner. Moreover, we revealed that honokiol and magnolol could suppress signal transducer and activator of transcription (STAT)3 and protein kinase B (Akt) phosphorylation in IL-27-stimulated TR146 cells though STAT1 phosphorylation was not suppressed by honokiol and magnolol treatment. Furthermore, STAT3 and Akt inhibitors could suppress CXCR3 ligand production in TR146 cells. In summary, honokiol and magnolol could reduce CXCR3 ligand production in oral epithelial cell by inhibiting STAT3 and Akt activation.

Topics: Anti-Inflammatory Agents; Biphenyl Compounds; Chemokine CXCL10; Chemokine CXCL11; Epithelial Cells; Humans; Interleukin-27; Ligands; Lignans; Mouth; Periodontal Diseases; Proto-Oncogene Proteins c-akt; Receptors, CXCR3; STAT3 Transcription Factor

To observe the growth inhibition of subcutaneous ovarian cancer xenografts in nude mice and mechanisms by ultrasound irradiation with honokiol-loaded poly(ε-caprolactone)-poly(ethylene glycol)-poly(ε-caprolactone) (PCL-PEG-PCL, PCEC) microbubbles (HK-PCEC-MB) as a promising therapeutic approach to ovarian tumor. HK-PCEC-MB were prepared using the double-emulsion solvent evaporation technique. Particles were characterized with regard to shape, size, distribution and surface potential using dynamic light scattering and the Malvern Zetasizer ZS90 system. Entrapment efficiency and loading amounts of honokiol were determined via high-performance liquid chromatography (HPLC). Pharmacological activity and optimal mode of action were examined using MTT assay. Nude mice were sorted into five groups of cisplatin-sensitive (A2780s) and cisplatin-resistant (A2780cp) ovarian cancer cell subcutaneous xenograft models: (1) HK-PCEC-MB+ultrasound; (2) HK; (3) PCEC-MB+ultrasound; (4) HK-PCEC-MB; (5) controls. We observed rates of growth inhibition, necrosis, and apoptosis, as well as microvessel density (MVD) changes. Physical properties of HK-PCEC-MB followed the normal pattern. The drug entrapment efficiency of HK-PCEC-MB was 65.08±2.31% and drug loading amount was 6.51±0.23%. Cancer cell survival rate was lowest in vitro at a drug concentration of 10 μg/mL with exposure to 2.58 W ultrasonic wave. In ultrasound irradiation combined with HK-PCEC-MB group, rates of necrosis and apoptosis were the highest, while the number of microvessel was the least. We have successfully prepared self-assembled HK-PCEC-MB that inhibits resistant human ovarian tumor growth by ultrasound irradiation.

Topics: Animals; Biphenyl Compounds; Female; Humans; Lignans; Mice; Mice, Nude; Microbubbles; Ovarian Neoplasms; Particle Size; Polyesters; Polyethylene Glycols; Ultrasonic Waves

Treatment effect of chemotherapy for aggressive non-small-cell lung cancer (NSCLC) is usually unsatisfactory for non-selective distributions of anticancer drugs, generation of vasculogenic mimicry (VM) channels, high metastasis and recurrence rate. Therefore, we developed a kind of dequalinium (DQA) modified paclitaxel plus honokiol micelles in this study to destroy VM channels and inhibit tumour metastasis. In vitro assays indicated that the targeting paclitaxel micelles with ideal physicochemical characteristics could exhibit not only the powerful cytotoxicity on Lewis lung tumour (LLT) cells but also the effective suppression on VM channels and tumour metastasis. Action mechanism studies manifested that DQA modified paclitaxel plus honokiol micelles could activate apoptotic enzymes caspase-3 and caspase-9 as well as down-regulate FAK, PI3K, MMP-2 and MMP-9. In vivo assays indicated that polymeric micelles could increase selective accumulation of chemotherapeutic drugs at tumour sites and showed a conspicuous antitumour efficacy. Hence, the DQA modified paclitaxel plus honokiol micelles prepared in this study provided a potential treatment strategy for NSCLC.

Topics: Animals; Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Carcinoma, Non-Small-Cell Lung; Cell Adhesion; Cell Line, Tumor; Cell Survival; Dequalinium; Drug Interactions; Drug Liberation; Humans; Lignans; Lung Neoplasms; Mice; Micelles; Neoplasm Metastasis; Paclitaxel; Temperature; Xenograft Model Antitumor Assays

Oral squamous cell carcinoma (OSCC) is one of the most common cancers worldwide with poor prognosis. Numerous studies have attempted to explore alternative regimens aimed at reducing cancer stem cells (CSCs) without compromising the efficacy of conventional chemoradiotherapy. The present study sought to assess the effect of a natural compound honokiol on the reduction of elevated cancer stemness, metastatic capacity, and chemoresistance of oral carcinoma stem cells (OCSCs). Our results demonstrated that honokiol attenuated the cell survival and self-renewal of OCSCs in a dose-dependent manner. Moreover, honokiol downregulated the expression of 2 selective markers of OCSCs, ALDH1, and CD44, as well as the migration and invasion abilities, indicating its potential to suppress cancer stemness. We showed that honokiol reduced the secretion of IL-6 and phosphorylation of STAT3, and the honokiol-inhibited self-renewal, invasion and colony formation were reversed by administration of IL-6. Most importantly, our data demonstrated that honokiol was able to potentiate the effect of Cisplatin, leading to a lower proportion of OCSCs and the decreased cancer stemness features. Taken together, this study demonstrated the benefits of utilizing honokiol as an adjunct therapy for OSCC treatment.

Topics: Antineoplastic Combined Chemotherapy Protocols; Biphenyl Compounds; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Survival; Cisplatin; Down-Regulation; Drug Resistance, Neoplasm; Drug Synergism; Humans; Interleukin-6; Lignans; Mouth Neoplasms; Neoplastic Stem Cells; Signal Transduction; STAT3 Transcription Factor

Honokiol (HNK), an important medicinal component of Magnolia officinalis, is reported to possess pharmacological activities against a variety of diseases. However, the molecular mechanisms of HNK medicinal functions are not fully clear. To systematically study the mechanisms of HNK action, we screened a yeast mutant library based on the conserved nature of its genes among eukaryotes. We identified genes associated with increased resistance or sensitivity to HNK after mutation. After functional classification of these genes, we found that most HNK-resistant strains in the largest functional category were petites with mutations in mitochondrial genes, indicating that mitochondria were related to HNK resistance. Additional analysis showed that resistance of petite mutants to HNK was associated with upregulation of the ATP-binding cassette transporter Pdr5, which pumps out HNK. We also found that several HNK-sensitive mitochondria mutants were not petites, and had larger lipid droplets (LDs). Furthermore, HNK treatment on wild-type yeast cells seemed to disrupt mitochondrial morphology, induced triacylglycerol synthesis, and generated supersized LDs surrounded by mitochondria and endoplasmic reticulum (ER). These changes are also applied to atp7Δ mutant if no carbon resource was available. These results suggested that HNK treatment partly impaired normal mitochondrial function to form larger LDs by altering lipid metabolism.

Topics: ATP-Binding Cassette Transporters; Biphenyl Compounds; Drug Resistance; Enzyme Inhibitors; Genes, Fungal; Lignans; Lipid Metabolism; Mitochondria; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins

To explore the therapeutic effect of honokiol on particulate matter 2.5 (PM2.5)-induced lung injury in asthmatic mice and the possible mechanisms.
 Methods: A total of 32 BALB/C mice were randomly divided into four groups: a normal saline group, a model group, a PM2.5 group and a honokiol group (n=8 in each group). The asthma mouse model was established by ovalbumin treatment. The mice were treated with physiological saline, ovalbumin, PM2.5 and honokiol, respectively. Lung tissues and serum were collected. The pathological changes of lung tissues were evaluated. The levels of inflammatory cytokines in bronchoalveolar lavage fluid (BALF) and serum were measured and the expressions of Toll like receptor 4 (TLR4), nuclear factor kappa B (NF-κB), retinoid-related orphan receptor gamma-t (RORγt) and forkhead box protein 3 (Foxp3) in lung tissues were detected.
 Results: 1) The lung tissues of mice in the asthma group showed obvious pathological changes and inflammatory state, suggesting that the asthma model was established successfully. PM2.5 could aggravate the pathological condition of inflammatory injury in lung tissues in asthmatic mice. 2) Compared to the PM2.5 group, the pathological symptoms in the lung tissues were alleviated in the honokiol group and the percentage of inflammatory cells in BALF and the levels of inflammatory cytokines in BALF and serum were significantly reduced (all P<0.05). 3) Compared to the PM2.5 group, the expressions of TLR4, NF-κB (p-p65) and RORγt in lung tissues were significantly decreased, while the expression of Foxp3 was increased; the ratio of RORγt/Foxp3 was also decreased in the honokiol group (all P<0.05).
 Conclusion: Honokiol can resist lung injury induced by PM2.5 in asthmatic mice. These effects are through inhibiting TLR4-NF-κB pathway-mediated inflammatory response or regulating the balance of Th17/Treg cells.. 目的：探究和厚朴酚对颗粒物2.5(particulate matter 2.5，PM2.5)诱导的哮喘小鼠肺损伤的治疗作用及其可能的作用机制。方法：32只BALB/C小鼠随机分为生理盐水组、模型组、PM2.5组和厚朴酚组，每组8只。使用卵清蛋白诱导哮喘小鼠模型，分别采用生理盐水、卵清蛋白、PM2.5和和厚朴酚处理，收集各组小鼠肺组织和血清，检测小鼠肺组织的病理损伤状态、支气管肺泡灌洗液(bronchoalveolar lavage fluid，BALF)和血清中炎性因子的表达水平，以及肺组织中Toll样受体4(Toll like receptor 4，TLR4)、核因子κB(nuclear factor kappa B，NF-κB)、维甲酸相关核孤儿受体γt(retinoid-related orphan receptor gamma-t，RORγt)和叉头状转录蛋白3(forkhead box protein 3，Foxp3)的蛋白表达水平。结果：1)模型组小鼠肺组织出现明显病理损伤和炎性状态，提示哮喘模型构建成功。PM2.5能够加重哮喘小鼠的肺组织病理损伤和炎性状态；2)与PM2.5组比较，和厚朴酚组小鼠肺组织的病理损伤状态得到缓解，BALF中炎性细胞减少，炎性因子水平降低(均P<0.05)。3)与PM2.5组比较，和厚朴酚组小鼠肺组织中TLR4，NF-κB(p-p65)和RORγt的表达减少，Foxp3表达水平增加，且RORγt/Foxp3比值减少(均P<0.05)。结论：和厚朴酚能够抵抗PM2.5诱导哮喘小鼠的肺损伤，这一方面可能是通过抑制TLR4-NF-κB信号通路介导的炎症反应，另一方面可能是通过影响T辅助细胞17/调节性T细胞平衡的方式来实现的。.

Topics: Animals; Asthma; Biphenyl Compounds; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Drugs, Chinese Herbal; Inflammation Mediators; Lignans; Lung; Lung Injury; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Particulate Matter; Random Allocation; Toll-Like Receptor 4

Acute respiratory distress syndrome (ARDS) is characterized by acute hypoxemia with diffuse alveolar damage and increased pulmonary microvascular permeability. Honokiol (HKL), the principal active ingredient of Chinese herb magnolia officinalis, protected the lung of experimental ARDS models via attenuation of inflammation and oxidative stress. However, whether HKL has protective effects against the dysfunction of pulmonary microvascular endothelial barrier and the potential mechanisms remain unclear.. In the present study, we examined the levels of plasma Angiopoietin-2 (Ang-2) in ARDS patients, explored the effects of HKL on the vascular endothelial barrier at the ARDS animal and cell levels.. Our data showed that compared with the healthy controls, circulating Ang-2 level was higher in the patients with ARDS, and were usually supposed to be positively related to the severity of ARDS. Moreover, HKL effectively inhibited lung inflammatory injury and microvascular leakage, and improved ARDS mice survival. HKL also inhibited the expression of Ang-2, ICAM-1 and VCAM-1, and restored the expression of Sirt3, β-Catenin and VE-Cadherin. Furthermore, HKL improved ECs survival and inhibited the apoptosis of ECs. The inhibition of Ang-2 expression in vitro by HKL is accompanied by the upregulation of Sirt3 and AMPK phosphorylation.. Our data demonstrated that HKL protected pulmonary microvascular endothelial barrier against LPS-induced ARDS at least in part through activating the Sirt3/AMPK signaling and inhibiting the Ang-2 expression. Thus, our findings show that the activation of Sirt3 signaling is a potential mechanism for the protective effects of HKL on vascular barrier.

Topics: AMP-Activated Protein Kinase Kinases; Angiopoietin-2; Animals; Apoptosis; Biphenyl Compounds; Capillary Permeability; Case-Control Studies; Cell Proliferation; Cells, Cultured; Endothelium, Vascular; Enzyme Inhibitors; Humans; Lignans; Lipopolysaccharides; Lung; Male; Mice; Mice, Inbred C57BL; Protein Kinases; Respiratory Distress Syndrome; Signal Transduction; Sirtuin 3

Honokiol has been used in traditional medicine for the treatment of inflammatory diseases. Activation of glial cells plays an essential role in neurodegenerative disorders. In this study, we show that Honokiol reduces the inflammatory response to LPS of primary cultures of microglia and astrocytes through the inhibition of pro-inflammatory mediators (iNOS, IL-6, IL-1β and TNF-α) and the simultaneous stimulation of anti-inflammatory cytokines (IL-10). Expression of KLF4 was induced in microglia and astrocytes after treatment with LPS and this response was mitigated by Honokiol. These findings extend our understanding of the anti-inflammatory properties of Honokiol on central glial cells and support its use as a therapeutic compound in neuroinflammatory disorders.

Topics: Animals; Anti-Inflammatory Agents; Astrocytes; Biphenyl Compounds; Cells, Cultured; Inflammation Mediators; Kruppel-Like Factor 4; Lignans; Lipopolysaccharides; Microglia; Rats; Rats, Sprague-Dawley; Rats, Wistar

Preterm birth is the most prominent complication attributing to poor pregnancy and neonatal outcome. Infection is most commonly implicated in preterm birth; it initiates a cascade of inflammatory events that leads to the rupture of fetal membranes and spontaneous uterine contractions. Anti-inflammatory agents may thus be a therapeutic approach to prevent the premature rupture of fetal membranes and block contractions. In non-gestational tissues, the polyphenol honokiol has been shown to possess potent anti-inflammatory properties.. The aim of this study was to investigate the effect of honokiol on pro-inflammatory mediators in human gestational tissues.. Fetal membranes, myometrium and freshly isolated amnion cells and primary myometrial cells were treated with honokiol in the absence or presence of the products lipopolysaccharide (LPS) and fibroblast-stimulating lipopeptide-1 (fsl-1), the viral dsRNA analogue polyinosinic:polycytidylic acid (poly(I:C)) or the pro-inflammatory cytokines TNF or IL1B. A luciferase assay was used to determine the effect of honokiol on nuclear factor kappa B (NF-κB) RelA transcriptional activity.. Honokiol significantly decreased pro-inflammatory cytokine (IL1A, IL6) and chemokine (CXCL8, CXCL1, CCL2) mRNA expression and secretion from fetal membranes (amnion and choriodecidua) and myometrium stimulated with LPS, fsl-1 or poly(I:C). In amnion cells, honokiol also significantly decreased the expression and secretion of the extracellular matrix degrading enzyme MMP9. Moreover, in myometrium, honokiol significantly suppressed the expression of the contraction associated protein PTGFR, the secretion of the uterotonic prostaglandins PGE. Honokiol reduced the expression of pro-inflammatory and pro-labour mediators in human amnion, choriodecidua and myometrium and that this may be facilitated through the suppression of NF-κB activation. These results indicate that the polyphenol honokiol may be a potent therapeutic for the prevention of preterm birth.

Topics: Anti-Inflammatory Agents; Biphenyl Compounds; Chemokine CCL2; Chemokine CXCL1; Dinoprost; Dinoprostone; Extraembryonic Membranes; Female; Humans; Interleukin-1beta; Interleukin-8; Lignans; Myometrium; Pregnancy; Premature Birth; Primary Cell Culture; Transcription Factor RelA; Tumor Necrosis Factor-alpha

Temozolomide (TMZ) is a first-line chemotherapeutic drug for malignant gliomas. Nonetheless, TMZ-induced side effects and drug resistance remain challenges. Our previous study showed the suppressive effects of honokiol on growth of gliomas.. This study was further aimed to evaluate if honokiol could enhance TMZ-induced insults toward malignant glioma cells and its possible mechanisms.. Human U87 MG glioma cells were exposed to TMZ, honokiol, and a combination of TMZ and honokiol. Cell survival, apoptosis, necrosis, and proliferation were successively assayed. Fluorometric substrate assays were conducted to determine activities of caspase-3, -6, -8, and -9. Levels of Fas ligand, Bax, and cytochrome c were immunodetected. Translocation of Bax to mitochondria were examined using confocal microscopy. Mitochondrial function was evaluated by assaying the mitochondrial membrane potential (MMP), reactive oxygen species (ROS), and complex I enzyme activity. Caspase-6 activity was suppressed using specific peptide inhibitors. The honokiol-induced effects were further confirmed using human U373 MG and murine GL261 cells.. Exposure of human U87 MG glioma cells to honokiol significantly increased TMZ-induced DNA fragmentation and cell apoptosis. Interestingly, honokiol enhanced intrinsic caspase-9 activity without affecting extrinsic Fas ligand levels and caspase-8 activity. Sequentially, TMZ-induced changes in Bax translocation, the MMP, mitochondrial complex I enzyme activity, intracellular ROS levels, and cytochrome c release were enhanced by honokiol. Consequently, honokiol amplified TMZ-induced activation of caspases-3 and -6 in human U87 MG cells. Fascinatingly, suppressing caspase-6 activity concurrently decreased honokiol-induced DNA fragmentation and cell apoptosis. The honokiol-involved improvement in TMZ-induced intrinsic apoptosis was also confirmed in human U373 MG and murine GL261 glioma cells.. This study showed that honokiol can enhance TMZ-induced apoptotic insults to glioma cells via an intrinsic mitochondrion-dependent mechanism. Our results suggest the therapeutic potential of honokiol to attenuate TMZ-induced side effects.

Topics: Animals; Apoptosis; Biphenyl Compounds; Caspases; Cell Line, Tumor; Cell Survival; Cytochromes c; Dacarbazine; DNA Fragmentation; Drugs, Chinese Herbal; Fas Ligand Protein; Glioma; Humans; Lignans; Membrane Potential, Mitochondrial; Mice; Mitochondria; Reactive Oxygen Species; Signal Transduction; Temozolomide

Parkinson's disease (PD) is a profound neurodegenerative disorder with gradual loss of dopamine nigrostriatal neurons linked to serious behavioral symptoms. While the current treatment strategies present limitations on halting the progression of PD, this study aimed to investigate the therapeutic potential of honokiol, as a partial peroxisome proliferator-activated receptor-gamma (PPARγ) mimic, on the proceeding behavioral and biochemical alterations in hemiparkinsonian mice. Results showed that unilateral striatal 6-hydroxydopamine (6-OHDA)-lesioned mice exhibited motor impairment, reflecting the contralateral rotation induced by apomorphine at 1-3 weeks post-lesion. Subchronic honokiol administration for 1-2 weeks, beginning 7 days after 6-OHDA-lesion, dose-dependently ameliorated motor dysfunction in hemiparkinsonian mice. Recovery of motor function was correlated with reversal of nigrostriatal dopaminergic neuronal loss, accompanied by higher tyrosine hydroxylase (TH) density, dopamine transporter (DAT) expression and vesicular monoamine transporter-2 (VMAT2) levels. Furthermore, honokiol attenuated oxidative stress and reactive astrocyte induction via decreasing NADPH-oxidase and glial fibrillary acidic protein (GFAP) expressions in 6-OHDA-lesioned striatum. The reversal effects of honokiol on behavioral impairment and striatal PPARγ expression were impeded by PPARγ antagonist GW9662. Notably, subchronic honokiol treatment extended the lifespan of these hemiparkinsonian mice. The present findings demonstrate the therapeutic activities of honokiol in ameliorating motor impairment and progressive dopaminergic damage that could be associated with regulating PPARγ signaling. Therefore, honokiol may potentially exert as a novel therapeutic candidate through PPARγ activation for management of motor symptoms and progressive neurodegeneration in PD.

Topics: Animals; Biphenyl Compounds; Gliosis; Lignans; Longevity; Male; Mice; Motor Activity; NADPH Oxidases; Neostriatum; Nerve Degeneration; Oxidation-Reduction; Oxidopamine; Parkinson Disease; PPAR gamma

Honokiol is a bioactive lignanoid and has been utilized in traditional Chinese medicine for a long time. It exhibits several pharmacological properties, such as anticancer effects, anti-inflammatory effects, and antianxiety effects. However, the poor aqueous solubility of honokiol has impeded clinical applications.. In the present study, we adopted the liquid antisolvent precipitation (LAP) technique to prepare nanoparticles of honokiol for enhancement of solubility and bioavailability. Moreover, the honokiol nanoparticles obtained were investigated and evaluated in terms of morphology, physicochemical properties, saturation solubility, dissolution in vitro, bioavailability in vivo, toxicity, and the inhibitory effect on growth of HepG2 cells.. The obtained honokiol nanoparticles existed nearly in spherical shape and could be turned into amorphous structure by the LAP method. Moreover, the solubility of the honokiol nanoparticles was extremely higher than that of free honokiol, and the nanoparticle dissolution rate was also higher than that of free honokiol, which was about 20.41 times and 26.2 times than that of free honokiol in artificial gastric juice and in artificial intestinal juice. The area under the curve [AUC(0-t)] value of honokiol nanoparticles was about 6.52 times greater than that of free honokiol; therefore, the honokiol nanoparticles had a higher bioavailability than free honokiol but were innoxious to the organs of rats. Additionally, the honokiol nanoparticles exhibited a higher inhibition of HepG2 cells due to their lower IC. Honokiol nanoparticles have high solubility and bioavailability, and can become a new oral drug formulation and produce a better response for its clinical applications.

Topics: Administration, Oral; Animals; Biological Availability; Biphenyl Compounds; Calorimetry, Differential Scanning; Chemical Precipitation; Female; Hep G2 Cells; Humans; Lignans; Nanoparticles; Organ Specificity; Particle Size; Rats, Sprague-Dawley; Solubility; Solvents; Thermogravimetry; Toxicity Tests; X-Ray Diffraction

In traditional Asian medicinal systems, preparations of the root and stem bark of

Topics: Adrenergic Neurons; Anti-Anxiety Agents; Benzhydryl Compounds; Biphenyl Compounds; Central Nervous System; HEK293 Cells; Humans; Lignans; Magnolia; Norepinephrine; Norepinephrine Plasma Membrane Transport Proteins; Plant Bark; Plant Extracts; Receptors, GABA-A

Topics: Animals; Apoptosis; Biphenyl Compounds; Carcinoma, Hepatocellular; Cell Line, Tumor; Hep G2 Cells; Humans; Lignans; Liver Neoplasms; MAP Kinase Signaling System; Mice; Mice, Nude; TNF-Related Apoptosis-Inducing Ligand

Oxidative stress-induced mitochondrial dysfunction is implicated in the pathogenesis of intervertebral disc degeneration (IVDD). Sirtuin 3 (SIRT3), a sirtuin family protein located in mitochondria, is essential for mitochondrial homeostasis; however, the role of SIRT3 in the process of IVDD has remained elusive. Here, we explored the expression of SIRT3 in IVDD in vivo and in vitro; we also explored the role of SIRT3 in senescence, apoptosis, and mitochondrial homeostasis under oxidative stress. We subsequently activated SIRT3 using honokiol to evaluate its therapeutic potential for IVDD. We assessed SIRT3 expression in degenerative nucleus pulposus (NP) tissues and oxidative stress-induced nucleus pulposus cells (NPCs). SIRT3 was knocked down by lentivirus and activated by honokiol to determine its role in oxidative stress-induced NPCs. The mechanism by which honokiol affected SIRT3 regulation was investigated in vitro, and the therapeutic potential of honokiol was assessed in vitro and in vivo. We found that the expression of SIRT3 decreased with IVDD, and SIRT3 knockdown reduced the tolerance of NPCs to oxidative stress. Honokiol (10 μM) improved the viability of NPCs under oxidative stress and promoted their properties of anti-oxidation, mitochondrial dynamics and mitophagy in a SIRT3-dependent manner. Furthermore, honokiol activated SIRT3 through the AMPK-PGC-1α signaling pathway. Moreover, honokiol treatment ameliorated IVDD in rats. Our study indicated that SIRT3 is involved in IVDD and showed the potential of the SIRT3 agonist honokiol for the treatment of IVDD.

Topics: Animals; Antioxidants; Biphenyl Compounds; Cells, Cultured; Female; Humans; Intervertebral Disc Degeneration; Lignans; Male; Middle Aged; Mitochondrial Dynamics; Nucleus Pulposus; Oxidative Stress; Rats; Rats, Sprague-Dawley; Sirtuin 3

This paper builds on the fact that high aspect ratio rectilinear tubing columns of the same length and outside dimensions can double column efficiency. It demonstrates that further improvements in efficiency can be made by using rectilinear tubing columns with half the wall thickness thus replacing heavy PTFE with light solvent systems and producing lighter higher capacity columns. Increases in sample loading/throughput of up to 55x are demonstrated by comparing the separation of Honokiol and Magnolol using a Hexane: Ethyl Acetate: Methanol: Water (5:2:5:2) phase system with the new thin wall rectilinear column (56 mL, 30 mL/min, 2.1 g/h in 6.5 min.) with the original optimization performed using a conventional DE-Mini column (18 mL, 0.8 mm bore circular PTFE tubing, 2.5 mL/min, 0.038 g/h in 45 min.). Honokiol is currently going through first phase clinical trials as an anti-lung cancer therapy where preparative countercurrent chromatography was used for its manufacture. To be competitive in the future it is important for the technology to become more efficient. This is the first big step in that direction.

Topics: Biphenyl Compounds; Chemistry, Pharmaceutical; Countercurrent Distribution; Hexanes; Lignans; Methanol; Solvents; Water

Lung squamous cell carcinoma (SCC) accounts for a considerable proportion of lung cancer cases, but there is still a lack of effective therapies. FGFR1 amplification is generally considered a promising therapeutic target. Honokiol is a chemical compound that has been proven to be effective against various malignancies and whose analog has been reported to target the mitogen-activated protein kinase family, members of a downstream signaling pathway of FGFR1. This was an explorative study to determine the mechanism of honokiol in lung SCC. We found that honokiol induced apoptosis and cell cycle arrest in lung SCC cell lines in a time- and dose-dependent manner. Honokiol also restricted cell migration in lung SCC cell lines. Moreover, the expression of FGF2 and the activation of FGFR1 were both downregulated by honokiol. Pharmacological inhibition and siRNA knockdown of FGFR1 induced apoptosis in lung SCC cells. Our in vivo study indicated that honokiol could suppress the growth of xenograft tumors, and this effect was associated with the inhibition of the FGF2-FGFR1 signaling pathway. In conclusion, honokiol induced cell apoptosis in lung SCC by targeting the FGF2-FGFR1 autocrine loop.

Topics: Animals; Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Movement; Cell Survival; Female; Fibroblast Growth Factor 2; Humans; Lignans; Lung Neoplasms; Mice, Inbred BALB C; Mice, Nude; Receptor, Fibroblast Growth Factor, Type 1

To explore the effects of honokiol (HKL) on pulmonary microvascular endothelial cells in lipopolysaccharide (LPS)-induced acute respiratory distress syndrome (ARDS) and the underlying mechanisms.
 Methods: In animal experiment, a total of 40 C57BL/6J mice were randomly divided into a control group (Con group), a LPS intervention group (LPS group), a LPS+honokiol (HKL) intervention group (HKL group) and a LPS+HKL+nicotinamide (NAM) intervention group (NAM group) (n=10 in each group). In the cell experiment, the experiment cells were divided into a control group (Con group), a LPS intervention group (LPS group), a LPS+HKL intervention group (HKL group), a LPS+HKL+NAM intervention group (NAM group), and a LPS+HKL+compound C (CMC) intervention group (CMC group). The pathological changes of the lung tissues were evaluated by hematoxylin and eosin (HE) staining; the protein concentration, total cells and neutrophils in the bronchoalveolar lavage fluid (BALF) and myeloperoxidase (MPO) activity in the lung tissues were detected; the changes of pulmonary microvascular permeability were determined by Evans blue assay; the effect of HKL on the vitality of human pulmonary microvascular endothelial cells were examined by cell counting kit-8 (CCK-8); the inhibitors including NAM and CMC were applied to explore the molecular mechanism of the protective effects of HKL. The expression levels of Sirt3, caspase-3, cleaved caspase-3, Bcl-2, Bax, p-adenosine monophosphate activated protein kinase (p-AMPK) and AMPK in lung tissues or cells were detected by Western blot.
 Results: In animal models, compared with the Con group, the mice in the LPS group displayed typical ARDS pathological changes, and the ratio of lung wet/dry weight (W/D) and MPO activity in the lung tissues, protein concentration, total cells and neutrophils in BALF, Evans blue leaking index (ELI), expression levels of cleaved caspase-3 were significantly increased (all P<0.05), while the expression levels of Sirt3 was obviously decreased (P<0.05). Compared with the LPS group, the above changes in the LPS group were significantly improved in the HKL group (all P<0.05); Compared with the HKL group, the curative effect of HKL intervention could be partly inhibited in the NAM group (P<0.05). In cell experiments, compared with the LPS group, the HPMECs viability in the HKL group was markedly improved (P<0.05), while the expression levels of Bcl-2 and Sirt3 were significantly upregulated (P<0.05), and the expressio. 目的：探讨和厚朴酚(honokiol，HKL)对脂多糖(lipopolysaccharide，LPS)所致的急性呼吸窘迫综合征(acute respiratory distress syndrome，ARDS)肺微血管内皮细胞的作用及潜在机制。方法：动物实验中，40只C57BL/6J小鼠随机分为对照组(Con组)、LPS干预组(LPS组)、LPS+HKL干预组(HKL组)、LPS+HKL+尼克酰胺(nicotinamide，NAM)干预组(NAM组)，每组10只。细胞实验中，实验分为对照组(Con组)、LPS干预组(LPS组)、LPS+HKL干预组(HKL组)、LPS+HKL+NAM干预组(NAM组)和LPS+HKL+混合抑制剂(compound C，CMC)干预组(CMC组)。采用HE染色观察肺组织病理改变；检测支气管肺泡灌洗液(bronchoalveolar lavage fluid，BALF)中蛋白浓度、细胞总数、中性粒细胞数及肺组织髓过氧化物酶(myeloperoxidase，MPO)活性；伊文思蓝实验检测肺微血管渗透性的改变；采用细胞计数试剂盒(cell counting kit-8，CCK-8)检测HKL对人肺微血管内皮细胞(human pulmonary microvascular endothelial cells，HPMECs) 活力的影响；采用抑制剂NAM和CMC干预探讨HKL保护性作用的分子机制；采用Western印迹检测组织或细胞Sirt3，caspase-3，cleaved caspase-3，Bcl-2，Bax，p-腺苷酸活化蛋白激酶(p-adenosine monophosphate activated protein kinase，p-AMPK)和AMPK蛋白表达。结果：动物实验中，与Con组比较，LPS组小鼠肺组织呈典型的ARDS病理改变，肺组织湿干重比(W/D)值及MPO活性、BALF蛋白浓度、细胞总数和中性粒细胞数、伊文思蓝渗漏指数(evans blue leaking index，ELI)和肺组织cleaved caspase-3表达均显著升高(均P<0.05)，而Sirt3表达明显下调(P<0.05)；与LPS组比较，HKL组的上述改变显著改善(均P<0.05)；与HKL组比较，NAM组中HKL干预的疗效可部分抑制(均P<0.05)。细胞实验中，与LPS组比较，HKL组HPMECs的存活率升高(P<0.05)，Bcl-2和Sirt3蛋白表达显著上调(均P<0.05)， Bax和cleaved caspase-3蛋白表达下调(均P<0.05)，AMPK通路活化加强(P<0.05)；与HKL组比较，CMC组HKL干预的疗效被部分抑制(均P<0.05)。结论：HKL能够显著减轻LPS所致的ARDS，抑制肺微血管内皮凋亡，其作用可能是通过线粒体依赖的Sirt3/AMPK途径实现的。.

Topics: Acute Lung Injury; AMP-Activated Protein Kinases; Animals; Biphenyl Compounds; Humans; Lignans; Lipopolysaccharides; Lung; Mice; Mice, Inbred C57BL; Mitochondria; Signal Transduction; Sirtuin 3

Doxorubicin is the chemotherapeutic drug of choice for a wide variety of cancers, and cardiotoxicity is one of the major side effects of doxorubicin treatment. One of the main cellular targets of doxorubicin in the heart is mitochondria. Mitochondrial sirtuin, SIRT3 has been shown to protect against doxorubicin-induced cardiotoxicity. We have recently identified honokiol (HKL) as an activator of SIRT3, which protects the heart from developing pressure overload hypertrophy. Here, we show that HKL-mediated activation of SIRT3 also protects the heart from doxorubicin-induced cardiac damage without compromising the tumor killing potential of doxorubicin. Doxorubicin-induced cardiotoxicity is associated with increased ROS production and consequent fragmentation of mitochondria and cell death. HKL-mediated activation of SIRT3 prevented Doxorubicin induced ROS production, mitochondrial damage and cell death in rat neonatal cardiomyocytes. HKL also promoted mitochondrial fusion. We also show that treatment with HKL blocked doxorubicin-induced cardiac toxicity in mice. This was associated with reduced mitochondrial DNA damage and improved mitochondrial function. Furthermore, treatments of mice, bearing prostrate tumor-xenografts, with HKL and doxorubicin showed inhibition of tumor growth with significantly reduced cardiac toxicity. Our results suggest that HKL-mediated activation of SIRT3 protects the heart from doxorubicin-induced cardiotoxicity and represents a potentially novel adjunct for chemotherapy treatments.

Topics: Animals; Biphenyl Compounds; Cardiomyopathies; Cell Line, Tumor; Cells, Cultured; Disease Models, Animal; Doxorubicin; Lignans; Mice; Mitochondria, Heart; Myocytes, Cardiac; Rats; Reactive Oxygen Species; Sirtuin 3; Up-Regulation

Renal fibrosis participates in the progression of hypertension-induced kidney injury. The effect of SIRT3, a member of the NAD+-dependent deacetylase family, in hypertensive nephropathy remains unclear. In this study, we found that SIRT3 was reduced after angiotensin II (AngII) treatment both in vivo and in vitro. Furthermore, SIRT3-knockout mice aggravated hypertension-induced renal dysfunction and renal fibrosis via chronic AngII infusion (2000 ng/kg per minute for 42 days). On the contrary, SIRT3-overexpression mice attenuated AngII-induced kidney injury compared with wild-type mice. Remarkably, a co-localization of SIRT3 and KLF15, a kidney-enriched nuclear transcription factor, led to SIRT3 directly deacetylating KLF15, followed by decreased expression of fibronectin and collagen type IV in cultured MPC-5 podocytes. In addition, honokiol (HKL), a major bioactive compound isolated from Magnolia officinalis (Houpo), suppressed AngII-induced renal fibrosis through activating SIRT3-KLF15 signaling. Taken together, our findings implicate that a novel SIRT3-KLF15 signaling may prevent kidney injury from hypertension and HKL can act as a SIRT3-KLF15 signaling activator to protect against hypertensive nephropathy.

Topics: Acetylation; Angiotensin II; Animals; Biphenyl Compounds; Disease Models, Animal; Fibrosis; Gene Expression; Gene Expression Regulation; Hypertension, Renal; Kidney; Kruppel-Like Transcription Factors; Lignans; Mice; Mice, Knockout; Models, Biological; Nephritis; Nuclear Proteins; Podocytes; Protein Binding; Signal Transduction; Sirtuin 3

Ultraviolet (UV) radiation exposure induces immunosuppression, which contributes to the development of cutaneous malignancies. We investigated the effects of honokiol, a phytochemical found in plants of the genus Magnolia, on UVB-induced immunosuppression using contact hypersensitivity (CHS) as a model in C3H/HeN mice. Topical application of honokiol (0.5 and 1.0 mg/cm

Topics: Animals; Biphenyl Compounds; Cyclooxygenase 2; Cyclooxygenase Inhibitors; Dermatitis, Contact; DNA Methylation; Female; Immunosuppression Therapy; Inflammation; Inflammation Mediators; Lignans; Methyltransferases; Mice; Models, Biological; Protective Agents; Proto-Oncogene Proteins; Skin; Ultraviolet Rays

Postoperative ileus is a common complication after intra-abdominal surgery. Nitric oxide produced by macrophages in the inflamed gastrointestinal tract plays a crucial role in the pathogeny of postoperative ileus. Honokiol, extracted from the bark of Magnolia spp., is a natural compound with a biphenolic structure. In the present study, we examined the effect of honokiol on postoperative ileus and discussed its site of action. Postoperative ileus model mice were generated by surgical intestinal manipulation. Mice were administered honokiol (10 mg kg

Topics: Animals; Biphenyl Compounds; Cytokines; Down-Regulation; Gastrointestinal Motility; Ileus; Inflammation Mediators; Leukocytes; Lignans; Macrophages, Peritoneal; Mice; Nitric Oxide Synthase Type II; Postoperative Complications; RNA, Messenger

Honokiol (HNK), one of the main medicinal components in Magnolia officinalis, possesses antimicrobial activity against a variety of pathogenic bacteria and fungi. However, little is known of the molecular mechanisms underpinning the antimicrobial activity. To explore the molecular mechanism of its antifungal activity, we determined the effects of HNK on the mRNA expression profile of Saccharomyces cerevisiae using a DNA microarray approach. HNK markedly induced the expression of genes related to iron uptake and homeostasis. Conversely, genes associated with respiratory electron transport were downregulated, mirroring the effects of iron starvation. Meanwhile, HNK-induced growth deficiency was partly rescued by iron supplementation and HNK reacted with iron, producing iron complexes that depleted iron. These results suggest that HNK treatment induced iron starvation. Additionally, HNK treatment resulted in the upregulation of genes involved in protein synthesis and drug resistance networks. Furthermore, the deletion of PDR5, a gene encoding the plasma membrane ATP binding cassette (ABC) transporter, conferred sensitivity to HNK. Overexpression of PDR5 enhanced resistance of WT and pdr5Δ strains to HNK. Taken together, these findings suggest that HNK, which can be excluded by overexpression of Pdr5, functions in multiple cellular processes in S. cerevisiae, particularly in inducing iron starvation to inhibit cell growth.

Topics: Antifungal Agents; ATP-Binding Cassette Transporters; Biphenyl Compounds; Down-Regulation; Drug Resistance, Fungal; Gene Expression Profiling; Iron; Lignans; Oligonucleotide Array Sequence Analysis; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; Signal Transduction; Up-Regulation

Topics: AMP-Activated Protein Kinase Kinases; Animals; Biphenyl Compounds; Breast Neoplasms; Cell Line, Tumor; Cell Movement; Cell Transformation, Neoplastic; Female; Humans; Lignans; Mice; Neoplastic Stem Cells; Protein Serine-Threonine Kinases; STAT3 Transcription Factor; Xenograft Model Antitumor Assays

Malignant brain glioma is the most common and aggressive type of primary intracranial neoplasm. Regular chemotherapy cannot eradicate brain glioma cells and the residual glioma cells could form vasculogenic mimicry (VM) channels under hypoxic conditions to provide nutrients for tumor cell invasion. In addition, the existence of the blood-brain barrier (BBB) restricts most antitumor drugs into brain glioma. In this study, we developed a kind of lactoferrin (Lf) modified daunorubicin plus honokiol liposomes to transport antitumor drugs across BBB, eliminate the VM channels and block tumor cell invasion. The evaluations were performed on BBB model, brain glioma cells and glioma-bearing mice. In vitro results showed that the targeting liposomes with suitable physicochemical property could enhance the drug transportation acrossing the BBB, inhibit C6 cells invasion and destroy VM channels. Action mechanism studies indicated that Lf modified daunorubicin plus honokiol liposomes could activate apoptotic enzymes caspase 3 as well as down-regulate VM protein indicators (PI3K, MMP-2, MMP-9, VE-Cadherin and FAK). In vivo results displayed the targeting liposomes improved accumulation in brain tumor tissue and exhibited obvious antitumor efficacy. Therefore, Lf modified daunorubicin plus honokiol liposomes could be used as a potential therapy for treatment of brain glioma.

Topics: Animals; Antibiotics, Antineoplastic; Apoptosis; Biphenyl Compounds; Blood-Brain Barrier; Brain Neoplasms; Cell Line, Tumor; Daunorubicin; Drug Liberation; Glioma; Lactoferrin; Lignans; Liposomes; Mice, Inbred ICR; Rats

Understanding the mechanism of action of antifungal drugs is vital for better control of mycosis, which kills >1.3 million lives every year thus remains a major health problem worldwide. In this study, we investigate the activities of three different categories of plant-derived antifungal compounds (resveratrol, honokiol and osthole) via transcriptomics and bioinformatics analysis, with the goal of discovering the common Mode-of-Action (MoA) at molecular level. The result shows that a common transcriptional response (72 gene are up-regulated while 10 genes are down-regulated, commonly) are triggered by above representative antifungal compounds in Schizosaccharomyces pombe (S. pombe) yeast. By virtue of gene set enrichment analysis (GSEA) and gene functional annotation study, we identify that the genes involved in oxidative stress response, sugar metabolism, fatty acid metabolism, amino acid metabolism and glycolysis are significantly up-regulated, while the genes involved in nucleosome assembly, transcription and RNA processing are down-regulated, by any of these antifungal compounds. These observations demonstrate that the common MoA includes a strengthened anti-oxidative cell adaptation, a faster metabolic rate and a generally suppressed gene transcriptional activity. It implies a genetically encoded common redistribution of intracellular energy flux and molecules synthesis, after the challenging of antifungal compounds.

Topics: Antifungal Agents; Biphenyl Compounds; Coumarins; Gene Expression Profiling; Gene Expression Regulation, Fungal; Lignans; Oxidative Stress; Plant Extracts; Resveratrol; Schizosaccharomyces; Stilbenes; Transcription, Genetic; Up-Regulation

The biphenyl neolignan honokiol is a neuroprotectant which has been proposed as a treatment for central nervous system disorders such as Alzheimer's disease (AD). The death of cholinergic neurons in AD is attributed to multiple factors, including accumulation and fibrillation of amyloid beta peptide (Aβ) within the brain; metal ion toxicity; and oxidative stress. In this study, we used a transgenic Caenorhabditis elegans model expressing full length Aβ

Topics: Amyloid beta-Peptides; Animals; Biphenyl Compounds; Caenorhabditis elegans; Catechin; Chelating Agents; Cholinesterase Inhibitors; Drug Stability; Free Radical Scavengers; Humans; Iron; Lignans; Molecular Docking Simulation; Molecular Structure; Neuroprotective Agents; Paralysis; Peptide Fragments; Picrates; Protein Aggregation, Pathological; Protein Multimerization; Resveratrol; Stilbenes

Curing spinal cord injury (SCI) in mammals is a daunting task because of the lack of permissive mechanisms and strong inhibitory responses at and around the lesion. The neural cell adhesion molecule L1CAM (L1) has been shown to favor axonal regrowth and enhance neuronal survival and synaptic plasticity but delivery of full-length L1 or its extracellular domain could encounter difficulties in translation to therapy in humans. We have, therefore, identified several small organic compounds that bind to L1 and stimulate neuronal survival, neuronal migration and neurite outgrowth in an L1-dependent manner. Here, we assessed the functions of two L1 mimetics, trimebutine and honokiol, in regeneration following SCI in young adult mice. Using the Basso Mouse Scale (BMS) score, we found that ground locomotion in trimebutine-treated mice recovered better than honokiol-treated or vehicle-receiving mice. Enhanced hindlimb locomotor functions in the trimebutine group were observed at 6 weeks after SCI. Immunohistology of the spinal cords rostral and caudal to the lesion site showed reduced areas and intensities of glial fibrillary acidic protein immunoreactivity in both trimebutine and honokiol groups, whereas increased regrowth of axons was observed only in the trimebutine-treated group. Both L1- and L1 mimetic-mediated intracellular signaling cascades in the spinal cord lesion sites were activated by trimebutine and honokiol, with trimebutine being more effective than honokiol. These observations suggest that trimebutine and, to a lesser extent under the present experimental conditions, honokiol have a potential for therapy in regeneration of mammalian spinal cord injuries.

Topics: Animals; Biphenyl Compounds; Cells, Cultured; Cerebellum; Female; Glial Fibrillary Acidic Protein; Gliosis; Intermediate Filaments; Lignans; Locomotion; Mice; Mice, Inbred C57BL; Neural Cell Adhesion Molecule L1; Neurons; Recovery of Function; Signal Transduction; Small Molecule Libraries; Spinal Cord Injuries; Trimebutine; Tubulin

Honokiol (HNK) is a small molecule with potent anti-inflammatory and anti-tumorigenic properties; yet the molecular targets of HNK are not well studied. Hyperactivation of the receptor tyrosine kinase c-Met and overexpression of the cytoprotective enzyme heme oxygenase-1 (HO-1) play a critical role in the growth and progression of renal cell carcinoma (RCC). Interestingly, the calcineurin inhibitor (CNI) cyclosporine A (CsA), an immunosuppressant used to prevent allograft rejection, can also increase the risk of RCC in transplant patients. We studied the potential role of c-Met signaling axis on CNI-induced renal tumor growth and tested the anti-tumor efficacy of HNK. Importantly, CNI treatment promoted c-Met induction and enhanced c-Met-induced Ras activation. We found that HNK treatment effectively down-regulated both c-Met phosphorylation and Ras activation in renal cancer cells. It inhibited the expression of both c-Met- and CNI-induced HO-1, and promoted cancer cell apoptosis. In vivo, HNK markedly inhibited CNI-induced renal tumor growth; and it decreased the expression of phospho-c-Met and HO-1 and reduced blood vessel density in tumor tissues. Our results suggest a novel mechanism(s) by which HNK exerts its anti-tumor activity through the inhibition of c-Met-Ras-HO-1 axis; and it can have significant therapeutic potential to prevent post-transplantation cancer in immunosuppressed patients.

Topics: Animals; Apoptosis; Biphenyl Compounds; Calcineurin Inhibitors; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Down-Regulation; Heme Oxygenase-1; Hepatocyte Growth Factor; Human Umbilical Vein Endothelial Cells; Humans; Kidney Neoplasms; Lignans; Mice, Nude; Neovascularization, Pathologic; Neovascularization, Physiologic; Phosphorylation; Proto-Oncogene Proteins c-met; ras Proteins

Colorectal cancer (CRC) is the second leading cause of cancer death. Hence, there is a great need to explore new efficacious drugs for the treatment of CRC. Honokiol (HNK), a natural product extracted from magnolia bark, processes various biological activities, including anticancer. In this study, we introduced cell viability assay, western blotting, real-time PCR and immunofluorescent staining to determine the anticancer effect of HNK, and the possible mechanism underlying this biological process. We found that HNK can inhibit the proliferation and induce apoptosis in HCT116 cells in a concentration- and time-dependent manner. HNK activates p53 in HCT116 and other colon cancer cells. Exogenous p53 potentiates the anticancer of HNK, while p53 inhibitor decreases this effect of HNK. Moreover, HNK upregulates the expression of bone morphogenetic protein 7 (BMP7) in colon cancer cells; Exogenous BMP7 enhances the anticancer activity of HNK and BMP7 specific antibody reduces this effect of HNK. For mechanism, we found that HNK cannot increase the level of Smad1/5/8; Exogenous BMP7 potentiates the HNK-induced activation of p53. On the contrary, BMP7 specific antibody inhibits the HNK-induced activation of p53 in colon cancer cells and partly decreases the total level of p53. Our findings suggested that HNK may be a promising anticancer drug for CRC; activation of p53 plays an important role in the anticancer activity of HNK, which may be initialized partly by the HNK-induced upregulation of BMP7.

Topics: Apoptosis; Biphenyl Compounds; Bone Morphogenetic Protein 7; Cell Proliferation; Cell Survival; Colonic Neoplasms; Gene Expression Regulation, Neoplastic; HCT116 Cells; Humans; Lignans; Signal Transduction; Smad Proteins; Tumor Suppressor Protein p53

Formyl peptide receptor 1 (FPR1) mediates bacterial and mitochondrial N-formyl peptides-induced neutrophil activation. Therefore, FPR1 is an important therapeutic target for drugs to treat septic or sterile inflammatory diseases. Honokiol, a major bioactive compound of Magnoliaceae plants, possesses several anti-inflammatory activities. Here, we show that honokiol exhibits an inhibitory effect on FPR1 binding in human neutrophils. Honokiol inhibited superoxide anion generation, reactive oxygen species formation, and elastase release in bacterial or mitochondrial N-formyl peptides (FPR1 agonists)-activated human neutrophils. Adhesion of FPR1-induced human neutrophils to cerebral endothelial cells was also reduced by honokiol. The receptor-binding results revealed that honokiol repressed FPR1-specific ligand N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein binding to FPR1 in human neutrophils, neutrophil-like THP-1 cells, and hFPR1-transfected HEK293 cells. However, honokiol did not inhibit FPR2-specific ligand binding to FPR2 in human neutrophils. Furthermore, honokiol inhibited FPR1 agonist-induced calcium mobilization as well as phosphorylation of p38 MAPK, ERK, and JNK in human neutrophils. In conclusion, our data demonstrate that honokiol may have therapeutic potential for treating FPR1-mediated inflammatory diseases.

Topics: Animals; Anti-Inflammatory Agents; Biphenyl Compounds; Brain; Cell Adhesion; Coculture Techniques; Endothelial Cells; Gene Expression Regulation; HEK293 Cells; Humans; Lignans; Magnolia; MAP Kinase Kinase 4; Mice; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Neutrophil Activation; Neutrophils; Oligopeptides; p38 Mitogen-Activated Protein Kinases; Plant Extracts; Receptors, Formyl Peptide; Receptors, Lipoxin; THP-1 Cells

To concurrently suppress multidrug resistance (MDR) and metastasis of breast cancer cells, paclitaxel (PTX) and honokiol (HNK) were coencapsulated into pH-sensitive polymeric micelles based on poly(2-ethyl-2-oxazoline)-poly(d,l-lactide) (PEOz-PLA). The physicochemical properties of dual drug-loaded PEOz-PLA micelles were characterized in size, drug loading and in vitro release. The efficiency of MDR reversal for the micelles was testified by synergetic enhancement of cytotoxicity and uptake by MCF-7/ADR cells. The flow cytometry and fluorescence polarization measurement results reinforced the conclusion that down-regulation of P-gp expression and increase of plasma membrane fluidity appeared to be possible mechanisms of MDR reversal by dual drug-loaded PEOz-PLA micelles. Further, the efficient inhibition of tumor metastasis by dual drug-loaded PEOz-PLA micelles was demonstrated by in vitro anti-invasion and anti-migration assessment in MDA-MB-231 cells and in vivo bioluminescence imaging in nude mice. The suppression of MDR and metastasis by the micelles was assigned to synergistic effects of pH-triggered drug release and HNK/PEOz-PLA-aroused P-gp inhibition, and pH-triggered drug release and PTX/HNK-aroused MMPs inhibition, respectively. In conclusion, our findings strengthen the usefulness of co-delivery of PTX and HNK by pH-responsive polymeric micelles for suppression of tumor MDR and metastasis.. Multidrug resistance (MDR) and metastasis are considered to be two of the major barriers for successful chemotherapy. The combination of a chemotherapeutic drug with a modulator has emerged as a promising strategy for efficiently treating MDR cancer and preventing tumor metastasis. Herein, a dual drug (paclitaxel and honokiol)-loaded pH-sensitive polymeric micelle system based on PEOz-PLA was successfully fabricated to ensure that tumor MDR and metastasis could be concurrently suppressed, therefore achieving distinguishing endo/lysosomal pH from physiological pH by accelerating drug release and then enhancing the cytotoxicity of paclitaxel to drug-resistant tumor cells MCF-7/ADR by increasing cellular uptake of paclitaxel, preventing in vitro invasion and migration for MDA-MB-231 cells and in vivo metastasis in nude mice. Further, the mechanism of MDR reversal by dual drug-loaded PEOz-PLA micelles was elucidated to be down-regulation of P-gp expression and increase of plasma membrane fluidity of MCF-7/ADR cells. The present findings strengthen the usefulness of co-delivery of PTX and HNK by pH-responsive polymeric micelles for suppression of tumor MDR and metastasis.

Topics: Antineoplastic Combined Chemotherapy Protocols; Biphenyl Compounds; Breast Neoplasms; Drug Delivery Systems; Female; Humans; Lignans; MCF-7 Cells; Neoplasm Metastasis; Paclitaxel

Increasing evidence indicates that amyloid β oligomer (AβO) is toxic to neurons in Alzheimer's disease (AD) brain. The aim of the present study is to evaluate the effects of honokiol on AβO-induced learning and memory dysfunction in mice.. AD mice model was established by AβO intrahippocampal injection. The cognitive function was evaluated using Morris water maze (MWM). Nissl staining was used to examine the hippocampal neuron damage. Primary hippocampal neurons were exposed to AβO. The apoptosis in the hippocampal tissues and primary neurons was assessed using terminal dexynucleotidyl transferase-mediated dUTP nick end labeling-neuronal nuclei (NeuN) and Hoechst staining, respectively. The mitochondrial membrane potential and radical oxygen species were detected using standard methods. Western blotting assay was used to check the expression levels of apoptotic and nuclear factor kappa-B (NF-κB) signaling-associated proteins and electrophoretic mobility shift assay was used to detect NF-κB-DNA binding.. Honokiol increased the time spend in the target zone of the AD mice in the MWM. In addition, honokiol dose-dependently attenuated AβO-induced hippocampal neuronal apoptosis, reactive oxygen species production and loss of mitochondrial membrane potential. Furthermore, AβO-induced NF-κB activation was inhibited by honokiol, as well as the upregulated amyloid precursor protein and β-secretase.. Honokiol attenuates AβO-induced learning and memory dysfunction in mice and it may be a potential candidate in AD therapy.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Apoptosis; Apoptosis Regulatory Proteins; Biphenyl Compounds; Cells, Cultured; Cytokines; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Hippocampus; Lignans; Male; Maze Learning; Membrane Potential, Mitochondrial; Mice; Mice, Inbred C57BL; Mitochondria; Neurons; NF-kappa B; Peptide Fragments; Reactive Oxygen Species; Signal Transduction

Cartilage degradation is the significant pathological process in osteoarthritis (OA). Inflammatory cytokines, such as interleukin-1β (IL-1β), activate various downstream mediators contributing to OA pathology. Recently, stem cell-based cartilage repair emerges as a potential therapeutic strategy that being widely studied, whereas, the outcome is still far from clinical application. In this study, we focused on an anti-inflammatory agent, honokiol, which is isolated from an herb, investigated the potential effects on human umbilical cord derived mesenchymal stem cells (hUC-MSCs) in IL-1β stimulation.. Second passage hUC-MSCs were cultured for multi-differentiation. Flow cytometry, qRT-PCR, von Kossa stain, alcian blue stain and oil red O stain were used for characterization and multi-differentiation determination. Honokiol (5, 10, 25, 50 μM) and IL-1β (10 ng/ml) were applied in hUC-MSCs during chondrogenesis. Analysis was performed by MTT, cell apoptosis evaluation, ELISA assay, qRT-PCR and western blot.. hUC-MSC was positive for CD73, CD90 and CD105, but lack of CD34 and CD45. Remarkable osteogenesis, chondrogenesis and adipogenesis were detected in hUC-MSCs. IL-1β enhanced cell apoptosis and necrosis and activated the expression of caspase-3, cyclooxygenase-2 (COX-2), interleukin-6 (IL-6) and matrix metalloproteinase (MMP)-1, -9, 13 in hUC-MSCs. Moreover, the expression of SRY-related high-mobility group box 9 (SOX-9), aggrecan and col2α1 was suppressed. Honokiol relieved these negative impacts induced by IL-1β and suppressed Nuclear factor-κB (NF-κB) pathway by downregulating expression of p-IKKα/β, p-IκBα and p-p65 in dose-dependent and time-dependent manner.. Honokiol improved cell survival and chondrogenesis of hUC-MSCs and inhibited IL-1β-induced inflammatory response, which suggested that combination of anti-inflammation and stem cell can be a novel strategy for better cartilage repair.

Topics: Anti-Inflammatory Agents; Apoptosis; Biphenyl Compounds; Caspase 3; Cell Differentiation; Cell Survival; Cells, Cultured; Chondrogenesis; Collagenases; Cyclooxygenase 2; Gene Expression Regulation; Humans; Inflammation; Interleukin-6; Lignans; Mesenchymal Stem Cells; NF-kappa B; Signal Transduction

Honokiol, a compound extracted from Magnolia officinalis, has antifungal activities by inducing mitochondrial dysfunction and triggering apoptosis in Candida albicans. However, the mechanism of honokiol-induced oxidative stress is poorly understood. The present investigation was designed to determine the specific mitochondrial reactive oxygen species (ROS)-generation component.. We found that honokiol induced mitochondrial ROS accumulation, mainly superoxide anions (O2•-) measured by fluorescent staining method. The mitochondrial respiratory chain complex I (C I) inhibitor rotenone completely blocked O2•- production and provided the protection from the killing action of honokiol. Moreover, respiratory activity and the C I enzyme activity was significantly reduced after honokiol treatment. The differential gene-expression profile also showed that genes involved in oxidoreductase activity, electron transport, and oxidative phosphorylation were upregulated.. The present work shows that honokiol may bind to mitochondrial respiratory chain C I, leading to mitochondrial dysfunction, accompanied by increased cellular superoxide anion and oxidative stress.. This work not only provides insights on the mechanism by which honokiol interferes with fungal cell, demonstrating previously unknown effects on mitochondrial physiology, but also raises a note of caution on the use of M. officinalis as a Chinese medicine due to the toxic for mitochondria and suggests the possibility of using honokiol as chemosensitizer.

Topics: Antifungal Agents; Biphenyl Compounds; Candida albicans; Candidiasis; Electron Transport Complex I; Fungal Proteins; Gene Expression Regulation, Fungal; Humans; Lignans; Magnolia; Oxidative Stress; Superoxides

Honokiol is a key component of a medicinal herb, Magnolia bark. Honokiol possesses potential pharmacological benefits for many disease conditions, especially cancer. Recent studies demonstrate that Honokiol exerts beneficial effects on cardiac hypertrophy and doxorubicin (Dox)-cardiotoxicity via deacetylation of mitochondrial proteins. However, the effects and mechanisms of Honokiol on cardiac mitochondrial respiration remain unclear. In the present study, we investigate the effect of Honokiol on cardiac mitochondrial respiration in mice subjected to Dox treatment. Oxygen consumption in freshly isolated mitochondria from mice treated with Honokiol showed enhanced mitochondrial respiration. The Dox-induced impairment of mitochondrial respiration was less pronounced in honokiol-treated than control mice. Furthermore, Luciferase reporter assay reveals that Honokiol modestly increased PPARγ transcriptional activities in cultured embryonic rat cardiomyocytes (H9c2). Honokiol upregulated the expression of PPARγ in the mouse heart. Honokiol repressed cardiac inflammatory responses and oxidative stress in mice subjected to Dox treatment. As a result, Honokiol alleviated Dox-cardiotoxicity with improved cardiac function and reduced cardiomyocyte apoptosis. We conclude that Honokiol protects the heart from Dox-cardiotoxicity via improving mitochondrial function by not only repressing mitochondrial protein acetylation but also enhancing PPARγ activity in the heart. This study further supports Honokiol as a promising therapy for cancer patients receiving Dox treatment.

Topics: Animals; Anti-Arrhythmia Agents; Antibiotics, Antineoplastic; Biphenyl Compounds; Cardiotoxicity; Cell Respiration; Doxorubicin; Heart; Lignans; Mice, Inbred C57BL; Mitochondria; Oxygen

Neuroblastoma is the most common form of childhood extracranial tumor and almost half of neuroblastoma cases occur in infants under two years old. Neuroblastoma accounts for ~6‑10% of childhood cancers and 15% of cancer‑associated childhood mortality. However, an effective treatment remains to be developed. Honokiol exhibits long‑lasting central muscle relaxation, anti‑inflammatory, antibacterial, antimicrobial, antiulcer, antioxidation, antiaging and antitumor effects. Honokiol has been previously demonstrated to kill neuroblastoma cells, however, the underlying mechanism of action remains unclear. The present study reports that honokiol inhibits the growth of neuroblastoma cells via upregulation of reactive oxygen species (ROS). MTT assays demonstrated that treatment of Neuro‑2a neuroblastoma cells with honokiol resulted in time‑ and dose‑dependent inhibition of cell proliferation, which was associated with upregulation of the protein expression of receptor‑interacting protein kinase 3 (RIP3), as demonstrated by western blot analysis. Furthermore, knockdown of RIP3 by small interfering RNA, or pharmacological inhibition of RIP3 by the RIP3 specific inhibitor necrosulfonamide, reversed honokiol‑induced loss of cell viability in Neuro‑2a cells. Importantly, honokiol significantly increased the intracellular ROS levels as determined by a 2',7'‑dichlorofluorescin diacetate assay, while ROS scavenger N‑acetyl cysteine significantly prevented the induction of ROS and RIP3 by honokiol. The results of the present study indicate that honokiol may suppress the growth of neuroblastoma Neuro‑2a cells, at least partially, through ROS‑mediated upregulation of RIP3.

Topics: Acetylcysteine; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Cell Survival; Gene Silencing; Humans; Lignans; Neuroblastoma; Reactive Oxygen Species; Receptor-Interacting Protein Serine-Threonine Kinases; RNA, Small Interfering; Time Factors; Up-Regulation

Induction of brown adipocyte-like phenotype (browning) in white adipocytes and promotion of apoptosis by dietary and pharmacological compounds is considered a novel strategy against obesity. Here, we show that honokiol exerts dual modulatory effects on adipocytes via induction of browning in 3T3-L1 white adipocytes and apoptosis as well as activation of HIB1B brown adipocytes combined with inhibition of apoptosis.. Honokiol-induced browning and apoptosis were investigated by determining expression levels of brown adipocyte-specific genes and proteins by RT-PCR and immunoblot analysis, respectively. Apoptotic data were validated by immunofluorescence and ROS levels were measured by FACS analysis.. Honokiol treatment induced browning by elevating expression levels of brown adipocyte-specific genes such as Cidea, Cox8, Fgf21, Pgc-1α, and Ucp1. Honokiol promoted apoptosis of 3T3-L1 white adipocytes and inhibited apoptosis of HIB1B brown adipocytes via opposite regulation of the pro-apoptotic protein BAX and anti-apoptotic protein Bcl-2. Honokiol also significantly increased protein expression levels of ACOX1, CPT1, p-HSL, and p-PLIN and reduced ROS levels, suggesting its possible role in fat oxidation and lipid catabolism. Honokiol-induced browning could be mediated by activation of ERK, as inhibition of ERK by FR180204 abolished expression of PGC-1α and UCP1.. Our findings suggest that honokiol exhibits a modulatory role in adipocytes via induction of browning and apoptosis in white adipocytes, promotion of catabolic lipid metabolism, as well as activation and inhibition of apoptosis in HIB1B brown adipocytes, thereby exhibiting therapeutic potential against obesity.

Topics: 3T3-L1 Cells; Adipocytes; Adipose Tissue, Brown; Adipose Tissue, White; Animals; Apoptosis; Biphenyl Compounds; Cell Line; Fluorescent Antibody Technique; Gene Expression Regulation; Lignans; Lipid Metabolism; Mice; Reactive Oxygen Species; Reverse Transcriptase Polymerase Chain Reaction

5-Fluorouracil (5-FU) is an essential chemotherapeutic agent for oral squamous cell carcinoma (OSCC). However, toxic side effects have limited its role in OSCC therapy. The aim of this study was to explore whether combination therapy with 5-FU and honokiol (HNK), a small natural organic molecule shown to induce apoptosis in OSCC cells, could enhance the anticancer activity of 5-FU without notably increasing its toxicity.. 5-FU and/or HNK were used to treat OSCC cells both in vitro and in vivo. The therapeutic effect and underlying mechanisms were evaluated by cell viability assay, flow cytometry, OSCC xenograft mouse model, and Western blot. Tumor tissue apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. Toxicity was assessed following hematoxylin and eosin staining.. Exposure to HNK + 5-FU produced a synergistic cytotoxic effect on OSCC cells. Both HNK and 5-FU could induce apoptosis through the mitochondria-mediated intrinsic pathway, and their specific signaling pathways were different. In the mouse OSCC xenograft model, treatment with 5-FU + HNK substantively retarded tumor growth, as compared to treatment with either drug individually. TUNEL analysis further confirmed that the superior in vivo antitumor efficacy of 5-FU + HNK was associated with enhanced stimulation of cell apoptosis. Notably, HNK did not increase the toxicity of 5-FU.. These findings suggest that HNK and 5-FU exert a synergistic therapeutic effect on OSCC by inducing apoptosis. HNK might thus enhance the clinical therapeutic efficacy of 5-FU without increasing its toxicity.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Biphenyl Compounds; Carcinoma, Squamous Cell; Cell Line, Tumor; Drug Synergism; Female; Fluorouracil; Humans; Lignans; Mice; Mice, Inbred BALB C; Mouth Neoplasms

A group of 36 biphenyl derivatives structurally related to honokiol were synthesized by means of Suzuki coupling reactions. Their cytotoxicities were evaluated and compared to that of honokiol. Some of the compounds were then evaluated for their ability to downregulate the secretion of the VEGF protein and the expression of the VEGF, hTERT, and c-Myc genes; the two latter involved in the activation of telomerase in tumoral cells. Some of the synthetized derivatives showed promising pharmacological features as they exhibited IC

Topics: Biphenyl Compounds; Enzyme-Linked Immunosorbent Assay; Gene Expression; HEK293 Cells; HT29 Cells; Humans; Lignans; MCF-7 Cells; Telomerase; Vascular Endothelial Growth Factor A

Lung cancer is the leading cause of cancer death in the United States. Metastasis to lymph nodes and distal organs, especially brain, leads to severe complications and death. Preventing lung cancer development and metastases is an important strategy to reduce lung cancer mortality. Honokiol (HNK), a natural compound present in the extracts of magnolia bark, has a favorable bioavailability profile and recently has been shown to readily cross the blood-brain barrier. In the current study, we evaluated the antimetastatic effects of HNK in both the lymph node and brain mouse models of lung tumor metastasis. We tested the efficacy of HNK in preventing 18 H2030-BrM3 cell (brain-seeking human lung tumor cells) migration to lymph node or brain. In an orthotopic mouse model, HNK significantly decreased lung tumor growth compared with the vehicle control group. HNK also significantly reduced the incidence of lymph node metastasis and the weight of mediastinal lymph nodes. In a brain metastasis model, HNK inhibits metastasis of lung cancer cells to the brain to approximately one third of that observed in control mice. We analyzed HNK's mechanism of action, which indicated that its effect is mediated primarily by inhibiting the STAT3 pathway. HNK specifically inhibits STAT3 phosphorylation irrespective of the mutation status of EGFR, and knockdown of STAT3 abrogated both the antiproliferative and the antimetastatic effects of HNK. These observations suggest that HNK could provide novel chemopreventive or therapeutic options for preventing both lung tumor progression and lung cancer metastasis. Cancer Prev Res; 10(2); 133-41. ©2016 AACR.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Brain Neoplasms; Cell Line, Tumor; Cell Movement; Cell Proliferation; Female; Humans; Lignans; Lung Neoplasms; Lymphatic Metastasis; Mice; Mice, Inbred NOD; Mice, SCID; Signal Transduction; STAT3 Transcription Factor; Xenograft Model Antitumor Assays

Thyroid cancer is the most common endocrine malignancy, the global incidence rate of which is rapidly rising. Surgery and radioiodine therapies are common and effective treatments only for nonmetastasized primary tumors. Therefore, effective treatment modalities are imperative for patients with radioiodine-resistant thyroid cancer. Honokiol, a biophenolic compound derived from Magnolia spp., has been shown have diverse biological and pharmacological activities, including anti-inflammatory, antioxidative, antiangiogenic, and anticancer properties. In the present study, three human thyroid cancer cell lines, namely anaplastic, follicular, and poorly differentiated thyroid cancer cells, were used to evaluate the chemotherapeutic activity of honokiol. Cell viability, cell cycle, apoptosis, and autophagy induction were determined through flow cytometry and western blot analysis. We found that honokiol treatment can suppress cell growth, induce cell cycle arrest, and enhance the induction of caspase-dependent apoptosis and autophagy in cancer cells. Moreover, honokiol treatment modulated signaling pathways including Akt/mTOR, ERK, JNK, and p38 in the studied cells. In addition, the antitumorigenic activity of honokiol was also confirmed in vitro and in vivo. Our data provide evidence that honokiol has a unique application in chemotherapy for human thyroid cancers.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Autophagy; Biphenyl Compounds; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Proliferation; Cell Survival; Humans; Lignans; Signal Transduction; Thyroid Neoplasms

AML1-ETO is the most common oncoprotein leading to acute myeloid leukemia (AML), in which 5-year survival rate is only about 30%. However, currently there are no specific therapies for AML patients with AML1-ETO. Here, we report that AML1-ETO protein is rapidly degraded by Honokiol (HNK), a natural phenolic compound isolated from the plant Magnolia officinalis. HNK induced the degradation of AML1-ETO in a concentration- and time-dependent manner in leukemic cell lines and primary AML blasts with t(8;21) translocation. Mechanistically, HNK obviously increased the expression of UbcH8, an E2-conjugase for the degradation of AML1-ETO, through triggering accumulation of acetylated histones in the promoter region of UbcH8. Knockdown of UbcH8 by small hairpin RNAs (shRNAs) prevented HNK-induced degradation of AML-ETO, suggesting that UbcH8 plays a critical role in the degradation of AML1-ETO. HNK inhibited cell proliferation and induced apoptotic death without activation of caspase-3, which was reported to cleave and degrade AML1-ETO protein. Thus, HNK-induced degradation of AML1-ETO is independent of activation of caspase-3. Finally, HNK reduced the angiogenesis and migration in Kasumi-1-injected zebrafish, decreased xenograft tumor size in a xenograft leukemia mouse model, and prolonged the survival time in mouse C1498 AML model. Collectively, HNK might be a potential treatment for t(8;21) leukemia by targeting AML1-ETO oncoprotein.

Topics: Acetylation; Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Core Binding Factor Alpha 2 Subunit; Embryo, Nonmammalian; Humans; Leukemia, Myeloid, Acute; Lignans; Male; Mice; Mice, Nude; Neoplasm Transplantation; Neovascularization, Physiologic; Oncogene Proteins, Fusion; Promoter Regions, Genetic; Proteasome Endopeptidase Complex; RUNX1 Translocation Partner 1 Protein; Ubiquitin-Conjugating Enzymes; Zebrafish

Honokiol, derived from Magnolia officinalis, has various pharmacological properties. Platelet activation plays a critical role in cardiovascular diseases. Honokiol has been reported to inhibit collagen-stimulated rabbit platelet aggregation. However, detailed further studies on the characteristics and functional activity of honokiol in platelet activation are relatively lacking. In the present study, honokiol specifically inhibited platelet aggregation and Ca

Topics: Animals; Biphenyl Compounds; Humans; Lignans; Mice; Platelet Aggregation; Protein Binding; Receptors, Collagen; Surface Plasmon Resonance; Thrombosis; Zebrafish

A major concept to sensitize cancer cells to DNA damaging agents is by inhibiting proteins in the DNA repair pathways. X-family DNA polymerases play critical roles in both base excision repair (BER) and nonhomologous end joining (NHEJ). In this study, we examined the effectiveness of honokiol to inhibit human DNA polymerase β (pol β), which is involved in BER, and DNA polymerase λ (pol λ), which is involved in NHEJ. Kinetic analysis with purified polymerases showed that honokiol inhibited DNA polymerase activity. The inhibition mode for the polymerases was a mixed-function noncompetitive inhibition with respect to the substrate, dCTP. The X-family polymerases, pol β and pol λ, were slightly more sensitive to inhibition by honokiol based on the K

Topics: Antibiotics, Antineoplastic; Biphenyl Compounds; Bleomycin; Cell Line, Tumor; DNA Polymerase beta; Enzyme Inhibitors; Humans; Kinetics; Lignans

To investigate the effects of honokiol on induction of reactive oxygen species (ROS), antioxidant defense systems, mitochondrial dysfunction, and apoptosis in Candida albicans.. To measure ROS accumulation, 2',7'-dichlorofluorescein diacetate fluorescence was used. Lipid peroxidation was assessed using both fluorescence staining and a thiobarbituric acid reactive substances (TBARS) assay. Protein oxidation was determined using dinitrophenylhydrazine derivatization. Antioxidant enzymatic activities were measured using commercially available detection kits. Superoxide dismutase (SOD) genes expression was measured using real time RT-PCR. To assess its antifungal abilities and effectiveness on ROS accumulation, honokiol and the SOD inhibitor N,N'-diethyldithiocarbamate (DDC) were used simultaneously. Mitochondrial dysfunction was assessed by measuring the mitochondrial membrane potential (mtΔψ). Honokiol-induced apoptosis was assessed using an Annexin V-FITC apoptosis detection kit.. ROS, lipid peroxidation, and protein oxidation occurred in a dose-dependent manner in C. albicans after honokiol treatment. Honokiol caused an increase in antioxidant enzymatic activity. In addition, honokiol treatment induced SOD genes expression in C. albicans cells. Moreover, addition of DDC resulted in increased endogenous ROS levels and potentiated the antifungal activity of honokiol. Mitochondrial dysfunction was confirmed by measured changes to mtΔψ. The level of apoptosis increased in a dose-dependent manner after honokiol treatment.. Collectively, these results indicate that honokiol acts as a pro-oxidant in C. albicans. Furthermore, the SOD inhibitor DDC can be used to potentiate the activity of honokiol against C. albicans.

Topics: Anti-Infective Agents; Apoptosis; Biphenyl Compounds; Candida albicans; Ditiocarb; Dose-Response Relationship, Drug; Drug Synergism; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Fungal; Lignans; Lipid Peroxidation; Membrane Potential, Mitochondrial; Microbial Sensitivity Tests; Microscopy, Fluorescence; Mitochondria; Oxidation-Reduction; Reactive Oxygen Species; Reverse Transcriptase Polymerase Chain Reaction; Superoxide Dismutase

Honokiol, a plant lignan has been shown to have antineoplastic effects against nonmelanoma skin cancer developments in mice. In this study, antineoplastic effects of honokiol were investigated in malignant melanoma models. In vitro effects of honokiol treatment on SKMEL-2 and UACC-62 melanoma cells were evaluated by measuring the cell viability, proliferation, apoptosis, cell cycle analysis, and expressions of various proteins associated with cell cycle progression and apoptosis. For the in vivo study, male nude mice inoculated with SKMEL-2 or UACC-62 cells received injections of sesame oil or honokiol for two to seven weeks. In vitro honokiol treatment caused significant decrease in cell viability, proliferation, cell cycle arrest, increased apoptosis, and modulation of apoptotic and cell cycle regulatory proteins. Honokiol caused an accumulation of cells in the G2/M phase of the cell cycle in SKMEL-2 and G0/G1 phase in UACC-62 cells. An elevated level of caspases and PARP were observed in both cell lines treated with honokiol. A decrease in the expression of various cell cycle regulatory proteins was also observed in honokiol treated cells. Honokiol caused a significant reduction of tumor growth in SKMEL-2 and UACC-62 melanoma xenografts. These findings suggest that honokiol is a good candidate for further studies as a possible treatment for malignant melanoma.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Cycle; Cell Line, Tumor; Cell Survival; Gene Expression Regulation, Neoplastic; Humans; Lignans; Male; Melanoma; Mice; Mice, Nude; Neoplasm Proteins; Xenograft Model Antitumor Assays

Pollution, especially cigarette smoke, is a major cause of skin damage.. To assess the effects of the small molecule polyphenol, honokiol, on reversing cigarette smoke-induced damage in vitro to relevant skin cells.. Keratinocytes (HaCat) cultures were exposed to cigarette smoke and, after 48 hours, IL-1α and IL-8 were measured in cell supernatants. Moreover, TIMP-2 production, apoptosis rate, and senescence β-galactosidase expression were evaluated in primary human foreskin fibroblasts (HFF-1) cultures.. Honokiol at 10 μm reduced IL-1α production by 3.4 folds (P < 0.05) and at 10 and 20 μm reduced IL-8 by 23.9% and 53.1% (P < 0.001), respectively, in HaCat keratinocytes. In HFF-1, honokiol restored TIMP-2 production by 96.9% and 91.9% (P < 0.001), respectively, at 10 and 20 μm, as well as reduced apoptosis by 47.1% (P < 0.001) and 41.3% (P < 0.01), respectively. Finally, honokiol reduced senescence-associated β-galactosidase expression in HFF-1.. Honokiol protects both HFF-1 and HaCat against cigarette smoke-induced inflammation, collagenolysis, apoptosis, and senescence.

Topics: Antioxidants; Apoptosis; beta-Galactosidase; Biphenyl Compounds; Cells, Cultured; Cellular Senescence; Fibroblasts; Humans; Inflammation; Interleukin-1alpha; Interleukin-8; Lignans; Tissue Inhibitor of Metalloproteinase-2; Tobacco Smoke Pollution

Medicinal plants have been widely used for a long history. Exploration of pharmacologically active compounds from medicinal plants present a broad prevalent of application. By examining viral mRNA expression in GCRV-infected Ctenopharyngodon idella kidney (CIK) cells treated with thirty kinds of plant extracts, we identified Magnolia officinalis Rehd et Wils. was able to preferably suppress viral replication. Further studies demonstrated that the main ingredients of magnolia bark, namely, magnolol and honokiol presented protective pharmacological function when treated GCRV-infected CIK cells with a concentration of 2.00 μg/ml and 1.25 μg/ml, respectively. Furthermore, reverse transcript quantitative polymerase chain reaction (RT-qPCR) and western blot showed that both magnolol and honokiol were efficient to restrain the replication of GCRV in CIK cells at non-toxic concentration (2.51 ± 0.51 μg/ml for magnolol, and 3.18 ± 0.61 μg/ml for honokiol). Moreover, it was found that magnolol and honokiol promoted the expression of immune-related genes. Magnolol obviously significantly increased the expression of interferon (IFN) regulatory factor (IRF)7 rather than that of IRF3 in the GCRV-infected cells, leading to the activation of type I IFN (IFN-I). Simultaneously, magnolol drastically facilitated the expression of interleukin (IL)-1β, but failed to induce the molecules in nuclear factor (NF)-κB pathways. Differently, honokiol strikingly motivated not only the expression of IL-1β, but also those of tumor necrosis factor α (TNFα) and NF-κB. Interestingly, though honokiol motivated the expression of IFN-β promoter stimulator 1 (IPS-1), IRF3 and IRF7, it failed to up-regulate the expression of IFN-I, indicating that honokiol enhanced the host innate antiviral response to GCRV infection via NF-κB pathways. Collectively, the present study revealed that magnolol and honokiol facilitated the expression of innate immune-related genes to strengthen the innate immune signaling responses to resist GCRV infection, which contributed to understanding the mechanisms by which small-molecule drugs possessed antiviral activities. In addition, these results lay a foundation for the development of broad-spectrum antiviral compounds in aquaculture industry.

Topics: Animals; Biphenyl Compounds; Carps; Cell Line; Colorimetry; Cytopathogenic Effect, Viral; Fish Diseases; Immunity, Innate; Lignans; Magnolia; Reoviridae; Reoviridae Infections; Tetrazolium Salts; Thiazoles

Combination chemotherapy is an important protocol in glioma therapy and honokiol shows synergistic anticancer effects with doxorubicin. In this paper, honokiol (HK) and doxorubicin (Dox) co-loaded Methoxy poly(ethylene glycol)-poly(ε-caprolactone) (MPEG-PCL) nanoparticles were prepared with a assembly method. The particle size (about 34 nm), morphology, X-ray Powder Diffraction (XRD), in vitro release profile, cytotoxicity and cell proliferation effects were studied in detail. The results indicated that honokiol and doxorubicin could be efficiently loaded into MPEG-PCL nanoparticles simultaneously, and could be released from the micelles in an extended period in vitro. In addition, honokiol and doxorubicin loaded in MPEG-PCL nanoparticles could efficiently suppress glioma cell proliferation and induce cell apoptosis in vitro. Furthermore, Dox-HK-MPEG-PCL micelles inhibited glioma growth more significantly than Dox-MPEG-PCL and HK-MPEG-PCL in both nude mice and zebrafish tumor models. Immunohistochemical analysis indicated that DOX-HK-MPEG-PCL micelles improved Dox's anti-tumor effect by enhancing tumor cell apoptosis, suppressing tumor cell proliferation, and inhibiting angiogenesis. Our data suggest that Dox-HK-MPEG-PCL micelles have the potential to be applied clinically in glioma therapy.

Topics: Animals; Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Disease Models, Animal; Doxorubicin; Drug Carriers; Drug Liberation; Drug Synergism; Humans; Lignans; Micelles; Nanoparticles; Neovascularization, Pathologic; Polyesters; Polyethylene Glycols; X-Ray Diffraction; Xenograft Model Antitumor Assays; Zebrafish

Topics: Administration, Oral; Animals; Antineoplastic Agents; Apoptosis; Biological Availability; Biphenyl Compounds; Caco-2 Cells; Cell Line, Tumor; Cell Proliferation; Dose-Response Relationship, Drug; Female; Humans; Lignans; Male; Mammary Neoplasms, Experimental; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Nude; Micelles; Nanoparticles; Particle Size; Structure-Activity Relationship; Surface Properties; Triple Negative Breast Neoplasms

The combination of chemotherapy drugs attracts more attention in clinical cancer trials. However, the poor water solubility of chemotherapeutic drugs restricts their anticancer application. In order to improve antitumor efficiency and reduce side effects of free drugs, we prepared paclitaxel (PTX) and honokiol (HK) combination methoxy poly(ethylene glycol)-poly(caprolactone) micelles (P-H/M) by solid dispersion method against breast cancer. The particle size of P-H/M was 28.7±2.5 nm, and transmission electron microscope image confirmed that P-H/M were spherical in shape with small particle size. After being encapsulated in micelles, the release of PTX or HK showed a sustained behavior in vitro. In addition, both the cytotoxicity and the cellular uptake of P-H/M were increased in 4T1 cells, and P-H/M induced more apoptosis than PTX-loaded micelles or HK-loaded micelles, as analyzed by flow cytometry assay and Western blot. Furthermore, the antitumor effect of P-H/M was significantly improved compared with PTX-loaded micelles or HK-loaded micelles in vivo. P-H/M were more effective in inhibiting tumor proliferation, inducing tumor apoptosis, and decreasing the density of microvasculature. Moreover, bioimaging analysis showed that drug-loaded polymeric micelles could accumulate more in tumor tissues compared with the free drug. Our results suggested that P-H/M may have potential applications in breast cancer therapy.

Topics: Angiogenesis Inhibitors; Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biocompatible Materials; Biphenyl Compounds; Blotting, Western; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Drug Carriers; Drug Liberation; Endocytosis; Female; HEK293 Cells; Humans; Immunohistochemistry; Lignans; Mice, Inbred BALB C; Micelles; Paclitaxel; Particle Size; Polymers

To conduct multiple-reaction monitoring(MRM) quantitative analysis with high-performance liquid chromatography coupled with mass spectrometry method, establish the quantification method of magnolol and honokiol in blood sample under negative ion mode with ibuprofen as internal standard, investigate the pharmacokinetic process of lignans constituents after oral administration of Weichang'an pill(WCA) at different doses, and provide theoretical basis to further reveal the material basis of WCA's anti-diarrhea effect. In the plasma samples, the linear relationship was good over the concentration range of 5.25 to 1 344.00 μg•L ⁻¹ for magnolol and 10.08 to 2 580.00 μg•L ⁻¹ for honokiol. The results of precision, stability, and extraction recovery tests showed that the determination method of plasma concentration for such compositions was stable and reliable. Dose-dependence was shown for magnolol and honokiol in the plasma concentration-time profile. The results indicated that the time to reach the maximum plasma concentration(Tmax) for lignanoids was 0.55-1.42 h, when the maximum plasma concentration(Cmax) could reach 996.36-2 330.96,189.87-1 469.43 μg•L ⁻¹ respectively for magnolol and honokiol. The lignanoids could be absorbed rapidly in the blood after oral administration of WAC pills, providing experimental basis to prove rapid and long-acting anti-diarrhea effect of WAC pills after oral administration.

Topics: Animals; Biphenyl Compounds; Drugs, Chinese Herbal; Lignans

To study the inhibition of methicillin-resistant Staphylococcus aureas (MRSA) biofilm by honokiol.. We used triphenyl tetrazolium chloride method to evaluate the inhibition of biofilm formation and mature by honokiol. We used congo red agar and spectrophotometer to detect the influence of honokiol on polysaccharide intercellular adhesion formation and extracellular DNA release. RT-PCR analysis was used to determine the effect of honokiol on expression of icaA, cidA and agrA.. Honokiol showed strong antimicrobial activity both on biofilm formation and mature biofilm of MRSA 41573. Minimum inhibitory concentration was 10 μg/mL for biofilm formation and 50 μg/mL for mature biofilm. Minimum bactericidal concentration was 20 μg/mL for biofilm formation and 100 μg/mL for mature biofilm. Honokiol showed synergy effect with vancomycin and it significantly increased the sensitivity of mature biofilm to vancomycin. Polysaccharide intercellular adhesion formation and extracellular DNA release were effectively inhibited by honokiol. Extracellular DNA release decreased by 28.3% when honokiol at 1/8 MIC. After incubated with 1/2 MIC of honokiol for 16 h, the relative expression of icaA, cidA and agrA of MRSA41573 was reduced by 59.1%, 56% and 72.3%, respectively.. Honokiol can significantly inhibit biofilm formation of MRSA41573 and its mechanism is mainly the inhibited expression of icaA and cidA to influence the synthesis of polysaccharide intercellular adhesion and extracellular DNA. Moreover, it also affect biofilm formation by QS system.

Topics: Anti-Bacterial Agents; Biofilms; Biphenyl Compounds; Drugs, Chinese Herbal; Humans; Lignans; Methicillin-Resistant Staphylococcus aureus; Microbial Sensitivity Tests; Staphylococcal Infections

Sirolimus is recognized as a P-glycoprotein (P-gp) substrate with poor water-solubility. To improve its solubility and bioabsorption, self-microemulsifying drug delivery systems (SMEDDS) containing a novel P-gp inhibitor, honokiol, were prepared. The aim of this work was to evaluate the enhanced transport of sirolimus SMEDDS as well as the roles of honokiol. In situ single-pass intestinal perfusion and in vitro human colon adenocarcinoma (Caco-2) cell models were applied to study the effects of honokiol within SMEDDS on the transport of sirolimus. The results indicated that a combination of honokiol with sirolimus in SMEDDS did not significantly alter the particle size, polydispersity index and release of drugs. In addition, the absorption rate constant (K

Topics: Administration, Oral; ATP Binding Cassette Transporter, Subfamily B, Member 1; Biological Availability; Biphenyl Compounds; Caco-2 Cells; Drug Delivery Systems; Emulsions; Humans; Intestinal Absorption; Lignans; Magnolia; Permeability; Sirolimus; Solubility

Honokiol (HK) is the main bioactive compound isolated from the bark of Magnolia officinalis. The present work is the first to report the pharmacokinetics and distribution of HK and its two metabolites of hydroxylated HK conjugated with glucuronic and sulfuric acid (M1) and HK monoglucuronide (M2) in plasma, liver, kidney and brain following oral administration of HK (40 mg/kg) to healthy Wistar rats. The results showed that only HK but not M1 or M2 was found in brain. Additionally, our work indicated that M2 not HK was the major compound in liver and plasma. The elimination of HK in liver, kidney and brain, and M2 in liver and kidney was more rapid than in plasma. The finding suggested that some of the pharmacological activity of HK might be generated by M2 but not HK.

Topics: Animals; Biphenyl Compounds; Brain; Glucuronic Acid; Kidney; Lignans; Liver; Magnolia; Male; Plasma; Rats; Rats, Wistar; Sulfuric Acids; Tissue Distribution

Zhi-Zi-Hou-Po decoction (ZZHPD) is one of the famous antidepressant Chinese formulas and is composed of Magnolia officinalis cortex (HP), Gardenia jasminoides Ellis (ZZ) and Citrus aurantium L. (ZS). Magnolol (MN) and honokiol (HN) from HP are the major active ingredients responsible for the therapeutic effects of ZZHPD. The aim of this study is to compare the pharmacokinetics and rat brain distribution of MN and HN after oral administration of HP extract and its compatibility with other herbal medicines in ZZHPD by HPLC-FLD. Compared with the HP group, Tmax (time to reach peak drug concentration in plasma) and AUC(0-τ) significantly increased in the ZZHPD and HP-ZZ groups. There was little change in the HP-ZS group in comparison with the HP group, which indicated that ZZ promotes absorption extent and defers the absorption rate of MN. The different compatibility of ZZHPD had a different degree of impact on the concentration of MN and HN in brain. The concentration of MN significantly increased in the HP-ZZ group while it decreased in the HP-ZS group compared with the HP group, which explained the concentration of compounds being slightly greater in the ZZHPD group than in the HP group. HP mixed with other medicines resulted in a decrease in HN concentration in the brain, particularly HP compatible with ZS. The results could be helpful for revealing the compatibility mechanism and providing clinical medication guidance for ZZHPD.

Topics: Administration, Oral; Animals; Biphenyl Compounds; Brain Chemistry; Chromatography, High Pressure Liquid; Drug Interactions; Drugs, Chinese Herbal; Iridoids; Lignans; Magnolia; Male; Plant Extracts; Rats; Tissue Distribution

In children, neuroblastomas are the most common and deadly solid tumor. Our previous study showed that honokiol, a small-molecule polyphenol, can traverse the blood-brain barrier and kill neuroblastoma cells. In this study, we further investigated the mechanisms of honokiol-induced insults to neuroblastoma cells. Treatment of neuroblastoma neuro-2a cells with honokiol elevated the levels of microtubule-associated protein light chain 3 (LC3)-II and induced cell autophagy in time- and concentration-dependent manners. Interestingly, pretreatment with 3-methyladenine (3-MA), an inhibitor of autophagy, led to the simultaneous attenuation of honokiol-induced cell autophagy and apoptosis but did not influence cell necrosis. As to the mechanisms, exposure of neuro-2a cells to honokiol time-dependently decreased the amount of phosphatidylinositol 3-kinase (PI3K). Sequentially, honokiol downregulated phosphorylation of protein kinase B (Akt) and mammalian target of rapamycin (mTOR) in neuro-2a cells. Furthermore, honokiol elevated the levels of glucose-regulated protein (GpR)78, an endoplasmic reticular stress (ERS)-associated protein, and amounts of intracellular reactive oxygen species (ROS). In contrast, reducing production of intracellular ROS using N-acetylcysteine, a scavenger of ROS, concurrently suppressed honokiol-induced cellular autophagy. Consequently, honokiol stimulated phosphorylation of extracellular signal-regulated kinase (ERK)1/2. However, pretreatment of neuro-2a cells with PD98059, an inhibitor of ERK1/2, lowered honokiol-induced autophagy. The effects of honokiol on inducing autophagy and apoptosis of neuroblastoma cells were further confirmed using mouse neuroblastoma NB41A3 cells as our experimental model. Fascinatingly, treatment of neuroblastoma neuro-2a and NB41A3 cells with honokiol for 12 h did not affect cell autophagy or apoptosis but caused significant suppression of cell migration. Taken together, this study showed that honokiol can induce autophagy of neuroblastoma cells and consequent apoptosis through activating the PI3K/Akt/mTOR and ERS/ROS/ERK1/2 signaling pathways and suppressing cell migration. Thus, honokiol has potential for treating neuroblastomas.

Topics: Animals; Apoptosis; Autophagy; Biphenyl Compounds; Cell Line, Tumor; Cell Movement; Endoplasmic Reticulum Stress; Extracellular Signal-Regulated MAP Kinases; Humans; Lignans; Mice; Neuroblastoma; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Reactive Oxygen Species; Signal Transduction; TOR Serine-Threonine Kinases

The orphan nuclear receptor Nur77 is implicated in the survival and apoptosis of cancer cells. The purpose of this study was to determine whether and how Nur77 serves to mediate the effect of the inflammatory cytokine TNF-α in cancer cells and to identify and characterize new agents targeting Nur77 for cancer therapy.. The effects of TNF-α on the expression and function of Nur77 were studied using in vitro and in vivo models. Nur77 expression was evaluated in tumour tissues from breast cancer patients. The anticancer effects of honokiol and its mechanism of action were assessed by in vitro, cell-based and animal studies.. TNF-α rapidly and potently induced the expression of Nur77 in breast cancer cells through activation of IκB kinase and JNK. Knocking down Nur77 resulted in TNF-α-dependent apoptosis, while ectopic Nur77 expression in MCF-7 cells promoted their growth in animals. Levels of Nur77 were higher in tumour tissues than the corresponding tissues surrounding the tumour in about 50% breast cancer patients studied. Our in vitro and animal studies also identified honokiol as an effective sensitizer of TNF-α-induced apoptosis by inhibiting TNF-α-induced Nur77 mRNA expression, which could be attributed to its interference of TNFR1's interaction with receptor-interacting protein 1 (RIPK1).. TNF-α-induced Nur77 serves as a survival factor to attenuate the death effect of TNF-α in cancer cells. With its proven human safety profile, honokiol represents a promising agent that warrants further clinical development.

Topics: Animals; Apoptosis; Biphenyl Compounds; Breast Neoplasms; Drugs, Chinese Herbal; Female; Gene Expression Regulation, Neoplastic; HeLa Cells; Hep G2 Cells; Humans; Lignans; MCF-7 Cells; Mice; Mice, Inbred BALB C; Mice, Nude; Nuclear Receptor Subfamily 4, Group A, Member 1; Tumor Necrosis Factor-alpha; Xenograft Model Antitumor Assays

Dental caries affects people of all ages and is a worldwide health concern. Streptococcus mutans is a major cariogenic bacterium because of its ability to form biofilm and induce an acidic environment. In this study, the antibacterial activities of magnolol and honokiol, the main constituents of the bark of magnolia plants, toward planktonic cell and biofilm of S. mutans were examined and compared with those of chlorhexidine. The minimal inhibitory concentrations of magnolol, honokiol and chlorhexidine for S. mutans were 10, 10 and 0.25 µg/mL, respectively. In addition, each agent showed bactericidal activity against S. mutans planktonic cells and inhibited biofilm formation in a dose- and time-dependent manner. Magnolol (50 µg/mL) had greater bactericidal activity against S. mutans biofilm than honokiol (50 µg/mL) and chlorhexidine (500 µg/mL) at 5 min after exposure, while all showed scant activity against biofilm at 30 s. Furthermore; chlorhexidine (0.5-500 µg/mL) exhibited high cellular toxicity for the gingival epithelial cell line Ca9-22 at 1 hr, whereas magnolol (50 µg/mL) and honokiol (50 µg/mL) did not. Thus; it was found that magnolol has antimicrobial activities against planktonic and biofilm cells of S. mutans. Magnolol may be a candidate for prevention and management of dental caries.

Topics: Anti-Bacterial Agents; Biofilms; Biphenyl Compounds; Cell Line; Dose-Response Relationship, Drug; Gingiva; Humans; Lignans; Magnolia; Microbial Sensitivity Tests; Microscopy, Fluorescence; Plant Bark; Plant Extracts; Streptococcus mutans

The aim of this study was to evaluate the embryo-fetal development toxicity of honokiol microemulsion. The drug was intravenously injected to pregnant SD rats at dose levels of 0, 200, 600 and 2000 μg/kg/day from day 6-15 of gestation. All the pregnant animals were observed for body weights and any abnormal changes and subjected to caesarean-section on gestation day (GD) 20; all fetuses obtained from caesarean-section were assessed by external inspection, visceral and skeletal examinations. No treatment-related external alterations as well as visceral and skeletal malformations were observed in honokiol microemulsion groups. There was no significant difference in the body weight gain of the pregnant rats, average number of corpora lutea, and the gravid uterus weight in the honokiol microemulsion groups compared with the vehicle control group. However, at a dose level of 2000 μg/kg/day, there was embryo-fetal developmental toxicity observed, including a decrease in the body length and tail length of fetuses. In conclusion, the no-observed-adverse-effect level (NOAEL) of honokiol microemulsion is 600 μg/kg/day, 75 times above the therapeutic dosage and it has embryo-fetal toxicity at a dose level of 2000 μg/kg/day, which is approximately 250 times above the therapeutic dosage.

Topics: Animals; Biphenyl Compounds; Dose-Response Relationship, Drug; Embryo, Mammalian; Emulsions; Female; Fetus; Gestational Age; Injections, Intravenous; Lignans; Maternal Exposure; Neuroprotective Agents; No-Observed-Adverse-Effect Level; Pregnancy; Rats, Sprague-Dawley; Risk Assessment

Aristolochic acids (AA) are nephrotoxic agents found in Aristolochia species whose consumption leads to the onset of a progressive tubulointerstitial fibrosis. This AA-nephropathy was first reported during the Belgian outbreak of the 1990's in which more than a hundred patients consumed slimming pills containing an Aristolochia species and Magnolia officinalis. The patients developed an end-stage kidney disease requiring dialysis or transplantation. Magnolol and honokiol are bioactive compounds from M. officinalis known for their potent antioxidant activity. As they can alleviate oxidative stress, we investigated their respective effects on AA-mediated tubulotoxicity using HK-2 cells. Magnolol and honokiol were able to reduce the oxidative stress associated with AA-treatment. Cytotoxicity alleviation was further investigated and overall cell viability measurements unexpectedly revealed that both compounds worsened the survival of AA-treated cells. Flow cytometry analyses of annexin V/PI stained cells indicated that the lignans efficiently prevented AA-induced apoptosis; but favored necrosis. Microscopy observations highlighted extensive vacuolization; other types of cell death, including autophagy, paraptosis or accelerated senescence were excluded. Ki-67 index and cell cycle analysis indicated that both magnolol and honokiol inhibited proliferation by blocking the cell cycle at the G1 phase; they also prevented the AA-induced G2/M arrest.

Topics: Aristolochic Acids; Biphenyl Compounds; Cell Cycle; Cell Line; Free Radical Scavengers; Humans; Kidney Tubules; Lignans; Molecular Structure; Oxidative Stress; Picrates

The primary objective of our study is to investigate the neuroprotective efficacy of honokiol and imipramine against restraint stress (RS)-induced cognitive impairment and depressive-like behaviour in mice. We examined whether the neuroprotective activity of honokiol and imipramine mediates through the inhibition of endoplasmic reticulum stress. Adult Swiss albino mice were restrained for 6h/day for 28 days. Honokiol (3 and 10mg/kg) and Imipramine (10 and 30mg/kg) were administered for last 7 days to the different groups. Cognitive function was assessed by Morris water maze and novel object recognition test. Forced swimming test and tail suspension test were performed to evaluate the restraint stress-induced depressive-like behaviour. Proinflammatory cytokines, brain-derived neurotrophic factor, and ER stress markers i.e. 78-kDa glucose-regulated protein (GRP78) and C/EBP homologous protein (CHOP) were quantified in the hippocampus. We observed cognitive impairment and depressive-like behaviour in RS-exposed animals. Honokiol (10mg/kg) treated group depicted marked reduction in cognitive impairment and depressive-like behaviour. However, imipramine (10 and 30mg/kg) prevented the depressive-like behaviour but failed to prevent RS-induced cognitive impairment. Moreover, proinflammatory cytokines, GRP78 and CHOP were elevated in the hippocampus of stressed mice as compared to unstressed mice. Honokiol (10mg/kg) significantly prevented the RS-induced elevated levels of proinflammatory cytokines and endoplasmic reticulum stress markers. Our results clearly suggest the beneficial potential of honokiol in restraint stress through inhibition of proinflammatory cytokines and endoplasmic reticulum stress. Honokiol could be an intriguing therapeutic approach in endoplasmic reticulum stress related neuro-pathophysiological conditions.

Topics: Animals; Behavior, Animal; Biphenyl Compounds; Brain-Derived Neurotrophic Factor; Cognition Disorders; Cytokines; Depression; Endoplasmic Reticulum Chaperone BiP; Endoplasmic Reticulum Stress; Gene Expression Regulation; Heat-Shock Proteins; Hippocampus; Imipramine; Lignans; Male; Mice; Restraint, Physical; Stress, Psychological; Transcription Factor CHOP

Honokiol is the predominant active ingredient in the commonly used traditional Chinese medicine, Magnolia, which has been confirmed in previous studies to exhibit anti-oxidation, antimicrobial, antitumor and other pharmacological effects. However, its effects on renal ischemia/reperfusion injury (IRI) remain to be elucidated. The present study aimed to examine the effects of honokiol on renal IRI, and to investigate its potential protective mechanisms in the heart. Male adult Wistar albino rats were induced into a renal IRI model. Subsequently, the levels of serum creatinine, blood urea nitrogen (BUN), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP), and the levels of serum nitrite and the kidney nitrite were examined in the IRI group. The levels of oxidative stress, inducible nitric oxide synthase (iNOS), inflammatory factors and caspase-3 were evaluated using a series of commercially available kits. The levels of phosphorylated signal transducer and activator of transcription 3 (p-STAT3) and the protein expression levels of STAT3 were determined using western blotting. Pretreatment with honokiol significantly reduced the levels of serum creatinine, BUN, ALT, AST and ALP, and the level of nitrite in the kidney of the IRI group, compared with the control group. The levels of malondialdehyde, the activity of myeloperoxidase, and the gene expression and activity of iNOS were reduced in the IRI rats, compared with the sham-operated rats, whereas the levels of superoxide dismutase and catalase were increased following treatment with honokiol in the IRI rats. In addition, the expression levels of tumor necrosis factor-α and interleukin-6 in the IRI rats were increased by honokiol. Treatment with honokiol suppressed the protein expression levels of p-STAT3 and caspase-3 in the IRI rats. These findings indicated that honokiol protects against renal IRI via the suppression of oxidative stress, iNOS, inflammation and STAT3 in the rat.

Topics: Animals; Biphenyl Compounds; Gene Expression Regulation; Humans; Inflammation; Kidney; Lignans; Nitric Oxide Synthase Type II; Oxidative Stress; Rats; Reperfusion Injury; STAT3 Transcription Factor

The high-performance liquid chromatography fingerprint method is a simple and reliable technique to evaluate the quality of leaves of Magnolia officinalis Rehd.et Wils. var. biloba Rehd.et Wils. We used the following bioactive phenolic constituents as reference compounds: rutin, afzelin, hyperoside, isoquercitrin, quercetin-3-O-α-l-rhamnoside, honokiol and magnolol. The conditions of an Agilent 1200 HPLC were: YMC-Pack-ODS-AQ column (250 × 4.6 mm id S-5 μm, 12 nm), mobile phase acetonitrile and 0.2% phosphoric acid in a gradient elute mode, flow rate 1.0 mL/min, detection wavelength 280 nm and column temperature 30°C. The analytical method was validated in terms of linearity, stability, repeatability, precision and recovery tests. While performing fingerprint analysis, we identified 11 peaks as characteristic peaks and assessed the similarities of 17 samples collected from different geological regions of China. The peak areas were used to evaluate the variation in the chemical composition of the tested samples. For this purpose, we performed hierarchical cluster analysis of the peak areas. Our results indicate that simultaneous determination of multiple ingredients could be done through chromatographic fingerprint analysis. Therefore, this high-performance liquid chromatography fingerprint method was readily utilized to evaluate the quality of leaves of M. officinalis var.biloba, which are used in several traditional herbal preparations.

Topics: Biphenyl Compounds; China; Chromatography; Chromatography, High Pressure Liquid; Glucosides; Lignans; Magnolia; Mannosides; Phenols; Plant Extracts; Plant Leaves; Proanthocyanidins; Quercetin; Reference Values; Reproducibility of Results; Rutin; Temperature

Honokiol is an active compound isolated from Magnolia officinalis that has been used without notable side effects in traditional medicine. We investigated the effects of honokiol against methylglyoxal (MG)-induced cytotoxicity in MC3T3-E1 osteoblast cells and the possible molecular mechanism(s) involved. The results showed that honokiol alleviated MG-induced cell death and the production of intracellular ROS, mitochondrial superoxide, cardiolipin peroxidation, and inflammatory cytokines. MG induction of the soluble receptor for advanced glycation end product (AGE) was reduced by pretreatment with honokiol. Furthermore, honokiol increased the levels of Nrf2 and increased the levels of glutathione and the activity of glyoxalase I. Pretreatment with honokiol prior to MG exposure reduced MG-induced mitochondrial dysfunction and alleviated MG-induced reduction of nitric oxide and PGC1α levels, suggesting that honokiol may induce mitochondrial biogenesis. It was concluded that honokiol could be useful in the attenuation of MG-induced cell damage.

Topics: Animals; Biphenyl Compounds; Cell Death; Cells, Cultured; Dose-Response Relationship, Drug; Lignans; Mice; Osteoblasts; Protective Agents; Pyruvaldehyde; Structure-Activity Relationship

Overexpression of NADPH oxidase 1 (Nox1) in melanoma cells is often associated with increased migration/metastasis rate. To develop effective treatment options, we have examined the effect of honokiol, a phytochemical from Magnolia plant, on the migratory potential of human melanoma cell lines (A375, Hs294t, SK-Mel119 and SK-Mel28) and assessed whether Nox1 is the target. Using an in vitro cell migration assay, we observed that treatment of different melanoma cell lines with honokiol for 24 h resulted in a dose-dependent inhibition of cell migration that was associated with reduction in Nox1 expression and reduced levels of oxidative stress. Treatment of cells with N-acetyl-L-cysteine, an anti-oxidant, also inhibited the migration of melanoma cells. Treatment of cells with diphenyleneiodonium chloride, an inhibitor of Nox1, significantly decreased the migration ability of Hs294t and SK-Mel28 cells. Further, we examined the effect of honokiol on the levels of core proteins (p22(phox) and p47(phox)) of the NADPH oxidase complex. Treatment of Hs294t and SK-Mel28 cells with honokiol resulted in accumulation of the cytosolic p47(phox) protein and decreased levels of the membrane-bound p22(phox) protein, thus blocking their interaction and inhibiting Nox1 activation. Our in vivo bioluminescence imaging data indicate that oral administration of honokiol inhibited the migration/extravasation and growth of intravenously injected melanoma cells in internal body organs, such as liver, lung and kidney in nude mice, and that this was associated with an inhibitory effect on Nox1 activity in these internal organs/tissues.

Topics: Animals; Apoptosis; Biphenyl Compounds; Blotting, Western; Cell Membrane; Cell Movement; Cell Proliferation; Cytosol; Enzyme Inhibitors; Female; Fluorescent Antibody Technique; Humans; Lignans; Melanoma; Mice; Mice, Nude; NADPH Oxidase 1; NADPH Oxidases; Oxidative Stress; Protein Binding; Reactive Oxygen Species; Tumor Cells, Cultured; Wound Healing; Xenograft Model Antitumor Assays

In this study we compared the effects of a diverse set of natural polyphenolics ligands on in silico interactive modelling, in vitro anti-aggregative properties and neuronal toxicity of β amyloid. The β amyloid-binding characteristics of optimised structural conformations of polyphenols with ascribed neuroprotective actions including punicalagin, myricetin, luteolin and honokiol were determined in silico. Thioflavin T and transmission electron microscopy were used to assess in vitro inhibitory effects of these polyphenols on Aβ1-42 fibril and aggregation formation. Phaeochromocytoma (PC12) cells were exposed to Aβ1-42, alone and in combination with test concentrations of each polyphenol (100 μM) and viability measured using MTT assay. Aβ1-42 evoked a concentration-dependent loss of cell viability in PC12 cells, in which all four polyphenols demonstrated significant inhibition of neurotoxicity. While all compounds variably altered the morphology of Aβ aggregation, the flavonoids luteolin and myricetin and the lignan honokiol all bound in a similar hydrophobic region of the amyloid pentamer and exerted the most pronounced inhibition of Aβ1-42 aggregation. Each of the polyphenols demonstrated neuroprotective effects in PC12 cells exposed to Aβ1-42, including punicalagin. These findings highlight some structure-activity insights that can be gleaned into the anti-aggregatory properties of bioactive polyphenols based on modelling of their binding to β-amyloid, but also serve to highlight the more general cellular neuroprotective nature of such compounds.

Topics: Amyloid beta-Peptides; Animals; Benzothiazoles; Biphenyl Compounds; Cell Survival; Flavonoids; Fluorescent Dyes; Hydrolyzable Tannins; Lignans; Luteolin; Microscopy, Electron, Transmission; Neuroprotective Agents; PC12 Cells; Peptide Fragments; Polyphenols; Rats; Thiazoles

Nearly 10-15% of women in the reproductive age were affected by endometriosis and currently facing the unmet need of effective therapeutic interventions for its management. Concerning this, the present study was intended to investigate the effect of Honokiol (HK) for the treatment of endometrial hyperplasia. The rat endometrial model was established and subsequently administered with a numerous dose of HK. The histopathology of tissues was also investigated. Results showed that, HK effectively inhibit the proliferation of rat edometeriotic tissues in a dose dependent manner. In terminal deoxynucleotidyl transferase (TdT) -mediated dUTP biotin nick end labeling (TUNEL) method, HK was able to bring apoptosis in endometrotic cells. Moreover, it also inhibits the mRNA levels of Survivin gene and Bcl-2 (B-cell lymphoma 2) in qPCR and Western blot analysis together with increases the mRNA level of apoptosis promoting factor Bax. These results clearly suggest that, HK was proficient to provoke apoptosis of rat endometriotic cells.

Topics: Animals; Apoptosis; bcl-2-Associated X Protein; Biphenyl Compounds; Endometriosis; Endometrium; Female; Lignans; Microtubule-Associated Proteins; Protective Agents; Proto-Oncogene Proteins c-bcl-2; Rats; Rats, Sprague-Dawley; RNA, Messenger; Survivin

The majority of human melanomas bears BRAF mutations and thus is treated with inhibitors of BRAF, such as vemurafenib. While patients with BRAF mutations often demonstrate an initial dramatic response to vemurafenib, relapse is extremely common. Thus, novel agents are needed for the treatment of these aggressive melanomas. Honokiol is a small molecule compound derived from Magnolia grandiflora that has activity against solid tumors and hematopoietic neoplasms. In order to increase the lipophilicity of honokiol, we have synthesized honokiol DCA, the dichloroacetate ester of honokiol. In addition, we synthesized a novel fluorinated honokiol analog, bis-trifluoromethyl-bis-(4-hydroxy-3-allylphenyl) methane (hexafluoro). Both compounds exhibited activity against A375 melanoma in vivo, but honokiol DCA was more active. Gene arrays comparing treated with vehicle control tumors demonstrated induction of the respiratory enzyme succinate dehydrogenase B (SDHB) by treatment, suggesting that our honokiol analogs induce respiration in vivo. We then examined its effect against a pair of melanomas, LM36 and LM36R, in which LM36R differs from LM36 in that LM36R has acquired vemurafenib resistance. Honokiol DCA demonstrated in vivo activity against LM36R (vemurafenib resistant) but not against parental LM36. Honokiol DCA and hexafluoro inhibited the phosphorylation of DRP1, thus stimulating a phenotype suggestive of respiration through mitochondrial normalization. Honokiol DCA may act in vemurafenib resistant melanomas to increase both respiration and reactive oxygen generation, leading to activity against aggressive melanoma in vivo.

Topics: Animals; Antineoplastic Agents; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Line, Tumor; Drug Resistance, Neoplasm; Humans; Indoles; Lignans; Melanoma; Mice; Signal Transduction; Sulfonamides; Vemurafenib; Xenograft Model Antitumor Assays

Honokiol (HNK) is a pharmacologically active small molecule that is isolated from the traditional Chinese medicinal herb, houpu. It may induce diversified types of regulated cell death, which are dependent on different cell types and varying concentrations of therapeutic agent. We previously reported that HNK triggers a cyclophilin D (CypD)-mediated regulated necrosis in various cell lines at certain concentrations (two‑fold higher than its half maximal inhibitory concentration). Subsequent study revealed that HNK induced cell death transition from early apoptosis to regulated necrosis in parallel with the increase of HNK dose. In the current study, a lower concentration of HNK (30 µg/ml) than previously reported also induced simplex CypD‑mediated mitochondrial permeability transition (MPT)‑associated regulated necrosis in the HEK‑293 human embryonic kidney cell line. HNK, at concentration of 30 µg/ml, induced necrotic cell death in HEK‑293 cells, which was demonstrated by positive staining for propidium iodide. No DNA ladder patterns or apoptotic bodies were detected in cells that underwent this type of necrotic cell death. Caspase‑8 and ‑3 were not activated during the process of HNK‑induced necrosis. In addition, pan‑caspase inhibitor, z‑VAD‑fmk and receptor‑interacting protein 1 inhibitor, necrostatin‑1 did not inhibit HNK‑induced necrosis. However, CypD inhibitor, cyclosporin A (CsA), blocked HNK‑induced necrosis. These findings indicate that 30 µg/ml HNK induced simplex CypD-mediated MPT‑associated regulated necrosis in HEK‑293 cells. Furthermore, the findings demonstrated that during HNK-triggered regulated necrosis the mammalian target of rapamycin (mTOR) signaling pathway is also inhibited. Pretreatment with CsA, therefore, inhibits HNK‑triggered regulated necrosis and reverses dephosphorylation of Akt, eIF4E‑binding protein 1 and S6 kinase. This indicated that the mTOR signaling pathway is effective downstream of the CypD‑mediated MPT and before the onset of plasma membrane breakdown during the regulated necrosis process. Therefore, it has been demonstrated for the first time, to the best of our knowledge, that the mTOR signaling pathway was inhibited downstream of the CypD-mediated MPT in the process of HNK-induced regulated necrosis.

Topics: Amino Acid Chloromethyl Ketones; Apoptosis; Biphenyl Compounds; Blotting, Western; Caspase 3; Caspase 8; Cyclophilins; Cyclosporine; HEK293 Cells; Humans; Imidazoles; Indoles; Lignans; Mitochondria; Nuclear Pore Complex Proteins; Peptidyl-Prolyl Isomerase F; Permeability; RNA-Binding Proteins; Signal Transduction; TOR Serine-Threonine Kinases

Honokiol has antitumor, antioxidative, anti-inflammatory, and antithrombotic effects. Here we aimed to identify the metabolic profile of honokiol in mouse, rat, dog, monkey, and human hepatocytes and to characterize the enzymes responsible for the glucuronidation and sulfation of honokiol. Honokiol had a high hepatic extraction ratio in all five species, indicating that it was extensively metabolized. A total of 32 metabolites, including 17 common and 15 different metabolites, produced via glucuronidation, sulfation, and oxidation of honokiol allyl groups were tentatively identified using liquid chromatography-high resolution quadrupole Orbitrap mass spectrometry. Glucuronidation of honokiol to M8 (honokiol-4-glucuronide) and M9 (honokiol-2'-glucuronide) was the predominant metabolic pathway in hepatocytes of all five species; however, interspecies differences between 4- and 2'-glucuronidation of honokiol were observed. UGT1A1, 1A8, 1A9, 2B15, and 2B17 played major roles in M8 formation, whereas UGT1A7 and 1A9 played major roles in M9 formation. Human cDNA-expressed SULT1C4 played a major role in M10 formation (honokiol-2'-sulfate), whereas SULT1A1*1, 1A1*2, and 1A2 played major roles in M11 formation (honokiol-4-sulfate). In conclusion, honokiol metabolism showed interspecies differences.

Topics: Animals; Arylsulfotransferase; Biotransformation; Biphenyl Compounds; Cells, Cultured; Chromatography, Liquid; Dogs; Glucuronides; Glucuronosyltransferase; Haplorhini; Hepatocytes; Humans; Lignans; Mice; Oxidation-Reduction; Rats; Species Specificity; Sulfuric Acid Esters; Tandem Mass Spectrometry

Eliminating cancer stem cells (CSCs) has been suggested for prevention of tumor recurrence and metastasis. Honokiol, an active compound of Magnolia officinalis, had been proposed to be a potential candidate drug for cancer treatment. We explored its effects on the elimination of oral CSCs both in vitro and in vivo.. By using the Hoechst side population (SP) technique, CSCs-like SP cells were isolated from human oral squamous cell carcinoma (OSCC) cell lines, SAS and OECM-1. Effects of honokiol on the apoptosis and signaling pathways of SP-derived spheres were examined by Annexin V/Propidium iodide staining and Western blotting, respectively. The in vivo effectiveness was examined by xenograft mouse model and immunohistochemical tissue staining.. The SP cells possessed higher stemness marker expression (ABCG2, Ep-CAM, Oct-4 and Nestin), clonogenicity, sphere formation capacity as well as tumorigenicity when compared to the parental cells. Treatment of these SP-derived spheres with honokiol resulted in apoptosis induction via Bax/Bcl-2 and caspase-3-dependent pathway. This apoptosis induction was associated with marked suppression of JAK2/STAT3, Akt and Erk signaling pathways in honokiol-treated SAS spheres. Consistent with its effect on JAK2/STAT3 suppression, honokiol also markedly inhibited IL-6-mediated migration of SAS cells. Accordingly, honokiol dose-dependently inhibited the growth of SAS SP xenograft and markedly reduced the immunohistochemical staining of PCNA and endothelial marker CD31 in the xenograft tumor.. Honokiol suppressed the sphere formation and xenograft growth of oral CSC-like cells in association with apoptosis induction and inhibition of survival/proliferation signaling pathways as well as angiogenesis. These results suggest its potential as an integrative medicine for combating oral cancer through targeting on CSCs.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Proliferation; Gene Expression Regulation, Neoplastic; Humans; Janus Kinases; Lignans; Mice; Mouth Neoplasms; Neoplasm Proteins; Neoplastic Stem Cells; Side-Population Cells; STAT Transcription Factors; Xenograft Model Antitumor Assays

The purpose of this study was to investigate the protective effect of honokiol on experimental ischemia/reperfusion injury of rat ovary.. A total of 40 female Wistar albino rats were used in this study. The rats were divided into five groups as follows: sham (Group I), torsion (Group II), torsion + detorsion (Group III), torsion + detorsion + saline (Group IV), and torsion + detorsion + honokiol (Group V). Bilateral adnexa in all the rats except for those in the sham group were exposed to torsion for 3 hours. The rats in Group IV were administered saline, whereas the rats in Group V were administered honokiol by intraperitoneal route 30 minutes before detorsion. Tissue and plasma concentrations of malondialdehyde and nitric oxide were determined. Ovarian tissue was histologically evaluated. Data analyses were performed by means of Kruskal-Wallis test and Mann-Whitney U-test (Bonferroni correction) in SPSS 15.0 (Statistical Package for Social Sciences; SPSS Inc., Chicago, IL, USA).. The torsion and detorsion groups had higher scores in vascular congestion, hemorrhage, and inflammatory cell infiltration compared with the sham group (P<0.005). In addition, total histopathological scores were significantly higher in the torsion and detorsion groups compared with the sham group (P<0.005). A significant reduction was observed in hemorrhage, inflammatory cell infiltration, and cellular degeneration scores, of all histopathological scores, in the honokiol group (P<0.005). Ovarian tissue concentrations of malondialdehyde were significantly higher in the torsion and detorsion groups compared with the sham and honokiol groups (P<0.005). Ovarian tissue concentrations of nitric oxide, on the other hand, were significantly higher in the torsion group compared with the sham, saline, and honokiol groups (P<0.005).. Honokiol has a beneficial effect on ovarian torsion-related ischemia/reperfusion injury.

Topics: Animals; Biphenyl Compounds; Female; Lignans; Ovarian Diseases; Protective Factors; Rats; Rats, Wistar; Reperfusion Injury

Our previous study showed that honokiol, a bioactive polyphenol, can traverse the blood-brain barrier and kills neuroblastoma cells.. In this study, we further evaluated the preclinical effects of honokiol on development of malignant glioma and the possible mechanisms.. Effects of honokiol on viability, caspase activities, apoptosis, and cell cycle arrest in human glioma U87 MG or U373MG cells were assayed. As to the mechanisms, levels of inactive or phosphorylated (p) p53, p21, CDK6, CDK4, cyclin D1, and E2F1 were immunodetected. Pifithrin-α (PFN-α), a p53 inhibitor, was pretreated into the cells. Finally, our in vitro findings were confirmed using intracranial nude mice implanted with U87 MG cells.. Exposure of human U87 MG glioma cells to honokiol decreased the cell viability. In parallel, honokiol induced activations of caspase-8, -9, and -3, apoptosis, and G1 cell cycle arrest. Treatment of U87 MG cells with honokiol increased p53 phosphorylation and p21 levels. Honokiol provoked signal-transducing downregulation of CDK6, CDK4, cyclin D1, phosphorylated (p)RB, and E2F1. Pretreatment of U87 MG cells with PFN-α significantly reversed honokiol-induced p53 phosphorylation and p21 augmentation. Honokiol-induced alterations in levels of CDK6, CDK4, cyclin D1, p-RB, and E2F1 were attenuated by PFN-α. Furthermore, honokiol could induce apoptotic insults to human U373MG glioma cells. In our in vivo model, administration of honokiol prolonged the survival rate of nude mice implanted with U87 MG cells and induced caspase-3 activation and chronological changes in p53, p21, CDK6, CDK4, cyclin D1, p-RB, and E2F1.. Honokiol can repress human glioma growth by inducing apoptosis and cell cycle arrest in tumor cells though activating a p53/cyclin D1/CDK6/CDK4/E2F1-dependent pathway. Our results suggest the potential of honokiol in therapies for human malignant gliomas.

Topics: Animals; Apoptosis; Benzothiazoles; Biphenyl Compounds; Caspases; Cell Cycle Checkpoints; Cell Cycle Proteins; Cell Line, Tumor; Down-Regulation; Drug Evaluation, Preclinical; Female; G1 Phase; Glioblastoma; Humans; Lignans; Mice; Mice, Inbred BALB C; Mice, Nude; Toluene; Tumor Suppressor Protein p53

Glioblastoma (GBM) is a malignant brain tumor associated with a high mortality rate. The aim of this study is to investigate the synergistic effects of honokiol (Hono) and magnolol (Mag), extracted from Magnolia officinalis, on cytotoxicity and inhibition of human GBM tumor progression in cellular and animal models. In comparison with Hono or Mag alone, co-treatment with Hono and Mag (Hono-Mag) decreased cyclin A, D1 and cyclin-dependent kinase 2, 4, 6 significantly, leading to cell cycle arrest in U87MG and LN229 human glioma cells. In addition, phosphorylated phosphoinositide 3-kinase (p-PI3K), p-Akt, and Ki67 were decreased after Hono-Mag treatment, showing proliferation inhibition. Hono-Mag treatment also reduced p-p38 and p-JNK but elevated p-ERK expression. Besides, Hono-Mag treatment induced autophagy and intrinsic and extrinsic apoptosis. Both ERK and autophagy inhibitors enhanced Hono-Mag-induced apoptosis in LN229 cells, indicating a rescuer role of ERK. In human GBM orthotopic xenograft model, the Hono-Mag treatment inhibited the tumor progression and induced apoptosis more efficiently than Temozolomide, Hono, or Mag group. In conclusion, the Hono-Mag exerts a synergistic anti-tumor effect by inhibiting cell proliferation and inducing autophagy and apoptosis in human GBM cells. The Hono-Mag may be applied as an adjuvant therapy to improve the therapeutic efficacy of GBM treatment.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Autophagy; Biphenyl Compounds; Brain Neoplasms; Cell Line, Tumor; Cell Proliferation; Cell Survival; Drug Synergism; Glioblastoma; Humans; Lignans; Mice; Mice, Nude; Xenograft Model Antitumor Assays

Honokiol, a derivative extracted from the stem and bark of Magnolia officinalis, has been reported to have anticancer effects in hepatoma cells. Recently, it was found that honokiol acted as not only a retinoid X receptor (RXR) agonist but also as a peroxisome proliferator-activated receptor gamma (PPARγ) agonist. Additionally, honokiol is capable of activating PPARγ/RXR heterodimers synergistically in the presence of rosiglitazone in 3T3-L1 adipocyte and HLE human hepatoma cells. Furthermore, synthetic PPARγ agonist thiazolidinediones exhibited growth inhibition effects in hepatoma cells through PPARγ-dependent and PPARγ-independent pathways. However, the combined effects of treatment with honokiol and PPARγ agonist are unclear in hepatoma cells.. In this study, sulforhodamine B assay, flow cytometry, and Western blot analysis were used to examine the combined effects of honokiol and PPARγ agonist (rosiglitazone) treatment on growth inhibition in SK-Hep1 and Mahlavu hepatoma cells.. Honokiol or rosiglitazone treatment in hepatoma cells induced growth inhibition at high dose by sulforhodamine B assay. Moreover, we found that combined treatment with honokiol and rosiglitazone showed more effective growth inhibition in hepatoma cells than treatment with honokiol or rosiglitazone alone. Also, treatment with honokiol and rosiglitazone induced cell cycle arrest in the G0/G1 phase; increased p21; and decreased cyclin D1, cyclin E1, and Rb expression in SK-Hep1 hepatoma cells.. Honokiol combined with rosiglitazone showed more effective growth inhibition in hepatoma cells mediated through the regulation of G0/G1 phase-related proteins p21, cyclin D1, cyclin E1, and Rb and cell cycle progression.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Proliferation; G1 Phase Cell Cycle Checkpoints; Humans; Lignans; Liver Neoplasms; PPAR gamma; Rosiglitazone; Thiazolidinediones

The stem of Magnoliae officinalis (MO) cortex is always preliminarily processed before being applied in traditional Chinese medicine. The definite bioavailability of honokiol (HO) and magnolol (MA) in processed MO (PMO) and the effect of chemical profiling change on the pharmacokinetics of HO and MA are always a greater challenge compared with those of MO. Compared with that of MO, the pharmacokinetic profiling of HO and MA in the PMO was significantly changed and the mean Tmax of HO and MA was increased by 31 and 50% (P < 0.05), respectively; the mean AUC0-t and Cmax of HO were increased by 36 and 24% (P < 0.05), respectively. Subsequently, the chemical profiling of MO and PMO was investigated by a simple and rapid LC-Q/TOF-MS coupled with multivariate analysis method. Principal component analysis and hierarchical cluster analysis of the chromatographic data demonstrated that the chemical profiling of PMO was significantly different from that of MO. Eight marker components including six alkaloids (magnocurarine, magnoflorine, roemerine and three unidentified peaks) and two lignans (obovatol and MA) were screened out by partial least-squares discriminant analysis. The results indicated that the changes of eight marker components of PMO may have an effect on the pharmacokinetic profiles of HO and MA.

Topics: Alkaloids; Animals; Biphenyl Compounds; Chromatography, Liquid; Cluster Analysis; Drugs, Chinese Herbal; Humans; Lignans; Magnolia; Male; Multivariate Analysis; Plant Stems; Principal Component Analysis; Rats; Rats, Wistar; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Beta-amyloid (Aβ) accumulation, one of the most important pathogenic traits of Alzheimer's disease (AD), has been reported to induce neurotoxicity in vitro as well as in vivo. Honokiol, isolated from the bark of Magnolia officinalis, has neuroprotective effects in different models of AD in vivo and in vitro. However, the exact mechanism for its neuroprotective effect is not well understood. The present study aimed to investigate the molecular mechanisms underlying the protective action of honokiol against Aβ1-42-induced neurotoxicity in cultured rat pheochromocytoma (PC12) cells. The results revealed that honokiol protected PC12 cells from Aβ1-42 induced cytotoxicity with increases in cell viability, GSH production and Bcl-2 expression, but decreases in the release of lactate dehydrogenase and cytochrome c, the amount of DNA fragmentation and MDA level, as well as Bax expression. Mechanistic study showed that honokiol could inhibit the activation of glycogen synthase kinase (GSK)-3β, attenuate the nuclear accumulation of β-catenin and suppress the phosphorylation of β-catenin (Ser33/Ser37/Thr41 site) in the Aβ1-42-treated PC12 cells. These results indicate that the anti-oxidative and anti-apoptotic effects of honokiol in Aβ1-42-treated PC12 cells may be mediated, at least in part, by regulation the GSK-3β and β-catenin signaling pathways.

Topics: Amyloid beta-Peptides; Animals; beta Catenin; Biphenyl Compounds; Cell Survival; Dose-Response Relationship, Drug; Glycogen Synthase Kinase 3 beta; Lignans; Neuroprotective Agents; PC12 Cells; Peptide Fragments; Rats; Signal Transduction

Adhesion molecules expressed on cerebral endothelial cells (ECs) mediate leukocyte recruitment and play a significant role in cerebral inflammation. Increased levels of adhesion molecules on the EC surface induce leukocyte infiltration into inflammatory areas and are thus hallmarkers of inflammation. Honokiol, isolated from the Chinese medicinal herb Magnolia officinalis, has various pharmacological activities, including anti-inflammatory effects, yet the nature of honokiol targeting molecules remains to be revealed. Here, we investigated the inhibitory effect of honokiol on neutrophil adhesion and vascular cell adhesion molecule-1 (VCAM-1) expression, which underlie its molecular target, and mechanisms for inactivating nuclear factor κ enhancer binding protein (NF-κB) in mouse cerebral ECs. Honokiol inhibited tumour necrosis factor-α (TNF-α)-induced neutrophil adhesion and VCAM-1 gene expression in cerebral ECs. The inflammatory transcription factor NF-κB was downregulated by honokiol. Honokiol significantly blocked TNF-α-induced NF-κB p65 nuclear translocation and degradation of the proteasome-dependent inhibitor of NF-κB α (IκBα). From docking model prediction, honokiol directly targeted the ubiquitin-ubiquitin interface of Lys48-linked polychains. Moreover, honokiol prevented the TNF-α-induced Lys48-linked polyubiquitination, including IκBα-polyubiquitin interaction. Honokiol has protective anti-inflammatory effects on TNF-α-induced neutrophil adhesion and VCAM-1 gene expression in cerebral ECs, at least in part by directly inhibiting ubiquitination-mediated IκBα degradation and then preventing NF-κB nuclear translocation.

Topics: Animals; Biphenyl Compounds; Brain; Cell Adhesion; Cells, Cultured; Endothelial Cells; Gene Expression Regulation; Humans; Lignans; Mice; Neutrophils; NF-KappaB Inhibitor alpha; Proteolysis; Tumor Necrosis Factor-alpha; Ubiquitination; Vascular Cell Adhesion Molecule-1

Understanding the link between chemoresistance and cancer progression may identify future targeted therapy for breast cancer. One of the mechanisms by which chemoresistance is attained in cancer cells is mediated through the expression of multidrug resistance proteins (MRPs). Acquiring drug resistance has been correlated to the emergence of metastasis, accounting for the progression of the disease. One of the diagnostic markers of metastatic progression is the overexpression of a transmembrane protein called Mucin 1 (MUC1) which has been implicated in reduced survival rate. The objective of this study was to understand the relationship between MUC1 and MRP1 using natural phenolic compound isolated from Magnolia grandiflora, honokiol, in mammary carcinoma cells. We provide evidence that honokiol suppresses the expression level of MUC1 and MRP1 in mammary carcinoma cells. In a time-dependent manner, honokiol-mediated reduction of MUC1 is followed by a reduction of MRP1 expression in the breast cancer cells. Additionally, silencing MUC1 suppresses the expression level of MRP1 and enhances the efficacy of doxorubicin, an MRP1 substrate. Taken together, these findings suggest MUC1 regulates the expression of MRP1 and provides a direct link between cancer progression and chemoresistance in mammary carcinoma cells.

Topics: Antineoplastic Agents, Phytogenic; Antineoplastic Combined Chemotherapy Protocols; Biphenyl Compounds; Breast Neoplasms; Down-Regulation; Doxorubicin; Drug Synergism; Female; Gene Expression; Humans; Lignans; MCF-7 Cells; Mucin-1; Multidrug Resistance-Associated Proteins; RNA, Small Interfering; Transfection

Honokiol, an active constituent extracted from the bark of Magnolia officinalis, possesses anticancer effects. Apoptosis is classified as type I programmed cell death, while autophagy is type II programmed cell death. We previously proved that honokiol induces cell cycle arrest and apoptosis of U87 MG glioma cells. Subsequently in this study, we evaluated the effect of honokiol on autophagy of glioma cells and examined the molecular mechanisms. Administration of honokiol to mice with an intracranial glioma increased expressions of cleaved caspase 3 and light chain 3 (LC3)-II. Exposure of U87 MG cells to honokiol also induced autophagy in concentration- and time-dependent manners. Results from the addition of 3-methyladenine, an autophagy inhibitor, and rapamycin, an autophagy inducer confirmed that honokiol-induced autophagy contributed to cell death. Honokiol decreased protein levels of PI3K, phosphorylated (p)-Akt, and p-mammalian target of rapamycin (mTOR) in vitro and in vivo. Pretreatment with a p53 inhibitor or transfection with p53 small interfering (si)RNA suppressed honokiol-induced autophagy by reversing downregulation of p-Akt and p-mTOR expressions. In addition, honokiol caused generation of reactive oxygen species (ROS), which was suppressed by the antioxidant, vitamin C. Vitamin C also inhibited honokiol-induced autophagic and apoptotic cell death. Concurrently, honokiol-induced alterations in levels of p-p53, p53, p-Akt, and p-mTOR were attenuated following vitamin C administration. Taken together, our data indicated that honokiol induced ROS-mediated autophagic cell death through regulating the p53/PI3K/Akt/mTOR signaling pathway.

Topics: Adenine; Animals; Apoptosis; Ascorbic Acid; Autophagy; Biphenyl Compounds; Caspase 3; Cell Line, Tumor; Dose-Response Relationship, Drug; Down-Regulation; Glioma; Lignans; Mice; Microtubule-Associated Proteins; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Reactive Oxygen Species; RNA, Small Interfering; Signal Transduction; Sirolimus; Time Factors; TOR Serine-Threonine Kinases; Tumor Suppressor Protein p53

Staphylococcus aureus (S. aureus) can attach to food, host tissues and the surfaces of medical implants and form a biofilm, which makes it difficult to eliminate. The purpose of this study was to evaluate the effect of honokiol on biofilm-grown S. aureus. In this report, honokiol showed effective antibacterial activity against S. aureus in biofilms. S. aureus isolates are capable of producing distinct types of biofilms mediated by polysaccharide intercellular adhesion (PIA) or extracellular DNA (eDNA). The biofilms' susceptibility to honokiol was evaluated using confocal laser scanning microscopy (CLSM) analysis. The transcript levels of the biofilm-related genes, the expression of PIA, and the amount of eDNA of biofilm-grown S. aureus exposed to honokiol were also investigated. The results of this study show that honokiol can detach existing biofilms, kill bacteria in biofilms, and simultaneously inhibit the transcript levels of sarA, cidA and icaA, eDNA release, and the expression of PIA.

Topics: Anti-Bacterial Agents; Anti-Infective Agents; Biofilms; Biphenyl Compounds; DNA; Lignans; Molecular Structure; Staphylococcus aureus

Honokiol (HNK), a highly promising phytochemical derived from Magnolia officinalis plant, exhibits in vitro and in vivo anticancer activity against prostate cancer but the underlying mechanism is not fully clear. This study was undertaken to delineate the role of c-Myc in anticancer effects of HNK. Exposure of prostate cancer cells to plasma achievable doses of HNK resulted in a marked decrease in levels of total and/or phosphorylated c-Myc protein as well as its mRNA expression. We also observed suppression of c-Myc protein in PC-3 xenografts upon oral HNK administration. Stable overexpression of c-Myc in PC-3 and 22Rv1 cells conferred significant protection against HNK-mediated growth inhibition and G0-G1 phase cell cycle arrest. HNK treatment decreased expression of c-Myc downstream targets including Cyclin D1 and Enhancer of Zeste Homolog 2 (EZH2), and these effects were partially restored upon c-Myc overexpression. In addition, PC-3 and DU145 cells with stable knockdown of EZH2 were relatively more sensitive to growth inhibition by HNK compared with control cells. Finally, androgen receptor overexpression abrogated HNK-mediated downregulation of c-Myc and its targets particularly EZH2. The present study indicates that c-Myc, which is often overexpressed in early and late stages of human prostate cancer, is a novel target of prostate cancer growth inhibition by HNK.

Topics: Biphenyl Compounds; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Nucleus; Cell Proliferation; Down-Regulation; Gene Expression Regulation, Neoplastic; Humans; Lignans; Male; Phosphorylation; Prostatic Neoplasms; Proto-Oncogene Proteins c-myc; Receptors, Androgen; RNA, Messenger

Since epidermal growth factor receptor (EGFR) is commonly deregulated in pre-malignant lung epithelium, targeting EGFR may arrest the development of lung cancer. Here, we showed that honokiol (2.5-7.5 μM), a bioactive compound of Magnolia officinalis, differentially suppressed proliferation (up to 93%) and induced apoptosis (up to 61%) of EGFR overexpressing tumorigenic bronchial cells and these effects were paralleled by downregulation of phospho-EGFR, phospho-Akt, phospho-STAT3 and cell cycle-related proteins as early as 6-12 h post-treatment. Autocrine secretion of EGF sensitized 1170 cells to the effects of honokiol. Molecular docking studies indicated that honokiol binds to the tyrosine kinase domain of EGFR although it was less efficient than erlotinib. However, the anti-proliferative and pro-apoptotic activities of honokiol were stronger than those of erlotinib. Upon combinatory treatment, honokiol sensitized bronchial cells and erlotinib resistant H1650 and H1975 cells to erlotinib. Furthermore, in a mouse lung tumor bioassay, intranasal instillation of liposomal honokiol (5 mg/kg) for 14 weeks reduced the size and multiplicity (49%) of lung tumors and the level of total- and phospho-EGFR, phospho-Akt and phospho-STAT3. Overall, our results indicate that honokiol is a promising candidate to suppress the development and even progression of lung tumors driven by EGFR deregulation.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biological Assay; Biphenyl Compounds; Bronchi; Carcinogenesis; Cell Cycle; Cell Proliferation; Cell Survival; Disease Progression; ErbB Receptors; Erlotinib Hydrochloride; Female; Lignans; Lung Neoplasms; Magnolia; Mice; Molecular Docking Simulation; Nitrosamines; Plant Extracts; RNA, Small Interfering

Honokiol and triphenylmethanes are small molecules with anti-tumor properties. Recently, we synthesized new honokiol analogues (HAs) that possess common features of both groups. We assessed the anti-tumor effectiveness of HAs in B-cell leukemia/lymphoma cells, namely in chronic lymphocytic leukemia (CLL) cells ex vivo and in pre-B-cell acute lymphoblastic leukemia (Nalm-6), Burkitt lymphoma (BL; Raji), diffuse large B-cell lymphoma (DLBCL; Toledo) and multiple myeloma (MM; RPMI 8226) cell lines. Four of these compounds appeared to be significantly active against the majority of cells examined, with no significant impact on healthy lymphocytes. These active HAs induced caspase-dependent apoptosis, causing significant deregulation of several apoptosis-regulating proteins. Overall, these compounds downregulated Bcl-2 and XIAP and upregulated Bax, Bak and survivin proteins. In conclusion, some of the HAs are potent tumor-selective inducers of apoptosis in ex vivo CLL and in BL, DLBCL and MM cells in vitro. Further preclinical studies of these agents are recommended.

Topics: Antineoplastic Agents; Apoptosis; Apoptosis Regulatory Proteins; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Cell Survival; Drug Screening Assays, Antitumor; Gene Expression Regulation, Neoplastic; Humans; Leukemia; Lignans; Lymphoma; Multiple Myeloma; Trityl Compounds

Honokiol (HNK) is a natural compound isolated from the magnolia plant with numerous pharmacological activities, including inhibiting epithelial-mesenchymal transition (EMT), which has been proposed as an attractive target for anti-tumor drugs to prevent tumor migration. In this study we investigated the effects of HNK on EMT in human NSCLC cells in vitro and the related signaling mechanisms.. TNF-α (25 ng/mL) in combination with TGF-β1 (5 ng/mL) was used to stimulate EMT of human NSCLC A549 and H460 cells. Cell proliferation was analyzed using a sulforhodamine B assay. A wound-healing assay and a transwell assay were performed to examine cell motility. Western blotting was used to detect the expression levels of relevant proteins. siRNAs were used to knock down the gene expression of c-FLIP and N-cadherin. Stable overexpression of c-FLIP L (H157-FLIP L) or Lac Z (H157-Lac Z) was also performed.. Treatment with TNF-α+TGF-β1 significantly enhanced the migration of A549 and H460 cells, increased c-FLIP, N-cadherin (a mesenchymal marker), snail (a transcriptional modulator) and p-Smad2/3 expression, and decreased IκB levels in the cells; these changes were abrogated by co-treatment with HNK (30 μmol/L). Further studies demonstrated that expression level of c-FLIP was highly correlated with the movement and migration of NSCLC cells, and the downstream effectors of c-FLIP signaling were NF-κB signaling and N-cadherin/snail signaling, while Smad signaling might lie upstream of c-FLIP.. HNK inhibits EMT-mediated motility and migration of human NSCLC cells in vitro by targeting c-FLIP, which can be utilized as a promising target for cancer therapy, while HNK may become a potential anti-metastasis drug or lead compound.

Topics: Animals; Antineoplastic Agents; Apoptosis; Autophagy; Biphenyl Compounds; Carcinoma, Non-Small-Cell Lung; CASP8 and FADD-Like Apoptosis Regulating Protein; Cell Line, Tumor; Cell Movement; Epithelial-Mesenchymal Transition; Humans; Lignans; Lung Neoplasms

The poor clinical outcome of pancreatic cancer (PC) is largely attributed to its aggressive nature and refractoriness to currently available therapeutic modalities. We previously reported antitumor efficacy of honokiol (HNK), a phytochemical isolated from various parts of Magnolia plant, against PC cells in short-term in vitro growth assays. Here, we report that HNK reduces plating efficiency and anchorage-independent growth of PC cells and suppresses their migration and invasiveness. Furthermore, significant inhibition of pancreatic tumor growth by HNK is observed in orthotopic mouse model along with complete-blockage of distant metastases. Histological examination suggests reduced desmoplasia in tumors from HNK-treated mice, later confirmed by immunohistochemical analyses of myofibroblast and extracellular matrix marker proteins (α-SMA and collagen I, respectively). At the molecular level, HNK treatment leads to decreased expression of sonic hedgehog (SHH) and CXCR4, two established mediators of bidirectional tumor-stromal cross-talk, both in vitro and in vivo . We also show that the conditioned media (CM) from HNK-treated PC cells have little growth-inducing effect on pancreatic stellate cells (PSCs) that could be regained by the addition of exogenous recombinant SHH. Moreover, pretreatment of CM of vehicle-treated PC cells with SHH-neutralizing antibody abolishes their growth-inducing potential on PSCs. Likewise, HNK-treated PC cells respond poorly to CM from PSCs due to decreased CXCR4 expression. Lastly, we show that the transfection of PC cells with constitutively active IKKβ mutant reverses the suppressive effect of HNK on nuclear factor-kappaB activation and partially restores CXCR4 and SHH expression. Taken together, these findings suggest that HNK interferes with tumor-stromal cross-talk via downregulation of CXCR4 and SHH and decreases pancreatic tumor growth and metastasis.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Communication; Cell Line, Tumor; Down-Regulation; Female; Gene Expression; Hedgehog Proteins; Humans; Lignans; Liver Neoplasms, Experimental; Lung Neoplasms; Mice; Pancreatic Neoplasms; Receptors, CXCR4; Tumor Microenvironment; Xenograft Model Antitumor Assays

Counter-current chromatography (CCC) has been widely used as a preparative separation method to purify natural products from plant extracts and fermentation broths. Traditionally, throughput optimization in CCC has focused on sample concentration and sample volume. In this paper sample injection was considered as consisting of three variables: injection flow rate, post-injection flow rate and sample solvent. The effects of these parameters were studied using a honokiol purification from a Magnolia officinalis bark extract as a case study aiming to achieve the highest throughput/yield ratio for greater than 99% purity of this potential anti-cancer drug obtained for submission to the Chinese FDA. An injection method was established that increased the throughput of honokiol by 46.5% (from 3.05g/h to 4.47g/h), and decreased the solvent consumption of mobile phase and stationary phase per gram of honokiol by 40.0% (from 0.68L/g to 0.41L/g) and 48.4% (from 0.40L/g to 0.21L/g) respectively. These results show the importance of understanding the whole injection process when optimizing a given CCC separation.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Countercurrent Distribution; Lignans; Magnolia; Plant Extracts; Solvents

Honokiol and magnolol, as pharmacological biphenolic compounds of Magnolia officinalis, have been reported to have antioxidant and anti-inflammatory properties. Sterol regulatory element binding protein-1 c (SREBP-1 c) plays an important role in the development and processing of steatosis in the liver. In the present study, we investigated the effects of a combination of honokiol and magnolol on SREBP-1 c-dependent lipogenesis in hepatocytes as well as in mice with fatty liver due to consumption of high-fat diet (HFD). Liver X receptor α (LXRα) agonists induced activation of SREBP-1 c and expression of lipogenic genes, which were blocked by co-treatment of honokiol and magnolol (HM). Moreover, a combination of HM potently increased mRNA of fatty acid oxidation genes. HM induced AMP-activated protein kinase (AMPK), an inhibitory kinase of the LXRα-SREBP-1 c pathway. The role of AMPK activation induced by HM was confirmed using an inhibitor of AMPK, Compound C, which reversed the ability of HM to both inhibit SREBP-1 c induction as well as induce genes for fatty acid oxidation. In mice, HM administration for four weeks ameliorated HFD-induced hepatic steatosis and liver dysfunction, as indicated by plasma parameters and Oil Red O staining. Taken together, our results demonstrated that a combination of HM has beneficial effects on inhibition of fatty liver and SREBP-1 c-mediated hepatic lipogenesis, and these events may be mediated by AMPK activation.

Topics: AMP-Activated Protein Kinases; Animals; Biphenyl Compounds; Cell Line; Diet, High-Fat; Disease Models, Animal; Drug Synergism; Drug Therapy, Combination; Enzyme Inhibitors; Fatty Liver; Hepatocytes; Humans; Lignans; Lipogenesis; Liver X Receptors; Male; Mice; Mice, Inbred C57BL; Orphan Nuclear Receptors; Signal Transduction; Sterol Regulatory Element Binding Protein 1

Honokiol, a constituent of Magnolia officinalis, has been reported to possess potent anti-cancer activity through targeting multiple signaling pathways in numerous malignancies including acute myeloid leukemia (AML). However, the underlying mechanisms remain to be defined. Here, we report that honokiol effectively decreased enzyme activity of histone deacetylases (HDACs) and reduced the protein expression of class I HDACs in leukemic cells. Moreover, treatment with proteasome inhibitor MG132 prevented honokiol-induced degradation of class I HDACs. Importantly, honokiol increased the levels of p21/waf1 and Bax via triggering acetylation of histone in the regions of p21/waf1 and Bax promoter. Honokiol induced apoptosis, decreased activity of HDACs, and significantly inhibited the clonogenic activity of hematopoietic progenitors in bone marrow mononuclear cells from patients with AML. However, honokiol did not decrease the activity of HDACs and induce apoptosis in normal hematopoietic progenitors from unbilicial cord blood. Finally, honokiol dramatically reduced tumorigenicity in a xenograft leukemia model. Collectively, our findings demonstrate that honokiol has anti-leukemia activity through inhibiting HDACs. Thus, being a relative non-toxic agent, honokiol may serve as a novel natural agent for cancer prevention and therapy in leukemia.

Topics: Acute Disease; Adult; Aged; Animals; Apoptosis; bcl-2-Associated X Protein; Biocatalysis; Biphenyl Compounds; Blotting, Western; Cell Cycle Checkpoints; Cell Line, Tumor; Cyclin-Dependent Kinase Inhibitor p21; Drugs, Chinese Herbal; Female; Histone Deacetylases; Humans; K562 Cells; Leukemia, Myeloid; Lignans; Male; Mice, Nude; Middle Aged; Proteasome Endopeptidase Complex; Reverse Transcriptase Polymerase Chain Reaction; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

Bioassay-guided fractionation of the MeOH extract of Magnolia grandiflora seeds resulted in the isolation of a new dimeric neolignan, named bishonokiol A (1), as well as two known neolignans magnolol (2) and honokiol (3). The structures of the compounds were determined on the basis of data obtained using NMR and MS. Bishonokiol A (1) showed potent anti-proliferative activities in four human cancer cell lines, with IC50 values ranging from 5.1 to 7.5 µM. Additionally, bishonokiol A (1) induced apoptosis, as well as down-regulated the expression of the anti-apoptotic protein Bcl-2 and caspase-3 cleavage in HepG2 cell line.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Caspase 3; Cell Line, Tumor; Cell Proliferation; Cell Survival; Down-Regulation; Drug Screening Assays, Antitumor; Humans; Lignans; Magnolia; Molecular Conformation; Plant Extracts; Proto-Oncogene Proteins c-bcl-2; Seeds

In this study, a kind of green solvent named polyethylene glycol (PEG) was developed for the ultrasound-assisted extraction (UAE) of magnolol and honokiol from Cortex Magnoliae Officinalis. The effects of PEG molecular weight, PEG concentration, sample size, pH, ultrasonic power and extraction time on the extraction of magnolol and honokiol were investigated to optimise the extraction conditions. Under the optimal extraction conditions, the PEG-based UAE supplied higher extraction efficiencies of magnolol and honokiol than the ethanol-based UAE and traditional ethanol-reflux extraction. Furthermore, the correlation coefficient (R(2)), repeatability (relative standard deviation, n = 6) and recovery confirmed the validation of the proposed extraction method, which were 0.9993-0.9996, 3.1-4.6% and 92.3-106.8%, respectively.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Lignans; Magnolia; Polyethylene Glycols; Ultrasonics

Application of a matrix-immobilised target enzyme for screening inhibitors is widely used in drug development, but there are few studies in insecticide discovery. In this paper, an economical and effective immobilised acetylcholinesterase (AChE) column was prepared using the sol-gel embedment method, which was further combined with high-performance liquid chromatography for screening the AChE inhibitors and insecticidal compounds from complex natural products.. AChE inhibitory constituents magnolol and honokiol were isolated from the ethanol extract of Magnolia officinalis, with IC50 values of 0.069 and 0.057 mM respectively. In an in vivo bioassay, magnolol and honokiol showed insecticidal activity against Nilaparvata lugens, with LC50 values of 0.324 and 0.137 mM, which are comparable with that of commonly used insecticide chlorpyrifos (0.233 mM). Moreover, molecular docking was carried out against a homology model of N. lugens AChE. The complexes showed that magnolol and honokiol placed themselves nicely into the active site of the enzyme and exhibited an interaction energy that was in accordance with our activity profile data.. These results demonstrate that magnolol and honokiol have great applied potential to be developed as natural insecticides, and an immobilised AChE column is very useful as a rapid screening tool for target enzymes towards potent inhibitors.

Topics: Acetylcholinesterase; Alkaloids; Animals; Biphenyl Compounds; Cholinesterase Inhibitors; Chromatography, High Pressure Liquid; Female; Hemiptera; Insecticides; Lignans; Magnolia; Plant Extracts; Sesquiterpenes

The purpose of this study was to investigate the acute and sub-chronic toxicity of honokiol microemulsion. In the acute toxicity tests, the mice were intravenously injected graded doses of honokiol microemulsion and were observed for toxic symptoms and mortality daily for 14 days. In the sub-chronic toxicity study, rats were injected honokiol microemulsion at doses of 100, 500, 2500 μg/kg body weight (BW) for 30 days. After 30 days treatment and 14 days recovery, the rats were sacrificed for hematological, biochemical and histological examination. In the acute toxicity tests, the estimated median lethal dosage (LD50) was 50.5mg/kg body weight in mice. In the sub-chronic toxicity tests, the non-toxic reaction dose was 500 μg/kg body weight. In each treatment group, degeneration or/and necrosis in vascular endothelial cells and structure change of vessel wall can be observed in the injection site (cauda vein) of a few animals while there were no changes in the vessels of other organs. The overall findings of this study indicate that the honokiol microemulsion is non-toxic up to 500 μg/kg body weight, and it has irritation to the vascular of the injection site which should be paid attention to in clinical medication.

Topics: Animals; Biphenyl Compounds; Chemistry, Pharmaceutical; Dose-Response Relationship, Drug; Emulsions; Injections, Intravenous; Lethal Dose 50; Lignans; Mice, Inbred ICR; No-Observed-Adverse-Effect Level; Rats, Sprague-Dawley; Risk Assessment; Time Factors; Toxicity Tests, Acute; Toxicity Tests, Subchronic

Melanoma is an aggressive disease with limited therapeutic options. Here, we determined the effects of honokiol (HNK), a biphenolic natural compound on melanoma cells and stemness. HNK significantly inhibited melanoma cell proliferation, viability, clonogenicity and induced autophagy. In addition, HNK significantly inhibited melanosphere formation in a dose dependent manner. Western blot analyses also demonstrated reduction in stem cell markers CD271, CD166, Jarid1b, and ABCB5. We next examined the effect of HNK on Notch signaling, a pathway involved in stem cell self-renewal. Four different Notch receptors exist in cells, which when cleaved by a series of enzymatic reactions catalyzed by Tumor Necrosis Factor-α-Converting Enzyme (TACE) and γ-secretase protein complex, results in the release of the Notch intracellular domain (NICD), which then translocates to the nucleus and induces target gene expression. Western blot analyses demonstrated that in HNK treated cells there is a significant reduction in the expression of cleaved Notch-2. In addition, there was a reduction in the expression of downstream target proteins, Hes-1 and cyclin D1. Moreover, HNK treatment suppressed the expression of TACE and γ-secretase complex proteins in melanoma cells. To confirm that suppression of Notch-2 activation is critical for HNK activity, we overexpressed NICD1, NICD2, and performed HNK treatment. NICD2, but not NICD1, partially restored the expression of Hes-1 and cyclin D1, and increased melanosphere formation. Taken together, these data suggest that HNK is a potent inhibitor of melanoma cells, in part, through the targeting of melanoma stem cells by suppressing Notch-2 signaling.

Topics: ADAM Proteins; ADAM17 Protein; Amyloid Precursor Protein Secretases; Autophagy; Basic Helix-Loop-Helix Transcription Factors; Biomarkers, Tumor; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cyclin D1; Homeodomain Proteins; Humans; Lignans; Melanoma; Neoplastic Stem Cells; Receptor, Notch2; Receptors, Notch; Signal Transduction; Transcription Factor HES-1

We present the synthesis of new derivatives of natural products magnolol (1) and honokiol (2) and their evaluation as allosteric ligands for modulation of GABAA receptor activity. New derivatives were prepared via metal assisted cross-coupling reactions in two consecutive steps. Compounds were tested by means of two-electrode voltage clamp electrophysiology at the α1β2γ2 receptor subtype at low GABA concentrations. We have identified several compounds enhancing GABA induced current (IGABA) in the range similar or even higher than the lead structures. At 3μM, compound 8g enhanced IGABA by factor of 443, compared to 162 and 338 of honokiol and magnolol, respectively. Furthermore, 8g at EC10-20 features a much bigger window of separation between the α1β2γ2 and the α1β1γ2 subtypes compared to honokiol, and thus improved subtype selectivity.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Biphenyl Compounds; GABA Modulators; Lignans; Metals; Molecular Structure; Oocytes; Patch-Clamp Techniques; Protein Subunits; Rats; Receptors, GABA-A; Recombinant Proteins; Structure-Activity Relationship; Xenopus

The emergence of antibiotic resistant strains such as methicillin-resistant Staphylococcus aureus (MRSA) reminds us an urgent need to develop a new immune-modulating agent for preventing S. aureus infection. In this study, we found that herbal medicines, honokiol and magnolol, caused a significant cellular immune modulatory effect during S. aureus infection. In mouse macrophages, these compounds drove upregulation of an antioxidant effect in response to S. aureus, resulting in a dampened total cellular reactive oxygen species (ROS) production and decreased production of inflammatory cytokines/chemokines, whereas honokiol induced increased types I and III interferon messenger RNA (mRNA) expression levels in response to MSSA infection. Moreover, the internalization of S. aureus by human alveolar epithelial cells was inhibited by these compounds. Furthermore, honokiol and magnolol treatment promoted a delay in killing during MSSA infection in Caenorhabditis elegans, suggesting antimicrobial function in vivo. In conclusion, honokiol and magnolol may be considered as attractive immune-modulating treatment for S. aureus infection.

Topics: Animals; Anti-Bacterial Agents; Biphenyl Compounds; Caenorhabditis elegans; Cytokines; Humans; Lignans; Macrophages; Methicillin; Methicillin-Resistant Staphylococcus aureus; Mice; Plant Extracts; Plants, Medicinal; Staphylococcal Infections; Staphylococcus aureus

Peritoneal dissemination is a major clinical obstacle in gastrointestinal cancer therapy, and it accounts for the majority of cancer-related mortality. Calreticulin (CRT) is over-expressed in gastric tumors and has been linked to poor prognosis. In this study, immunohistochemistry studies revealed that the up-regulation of CRT was associated with lymph node and distant metastasis in patients with gastric cancer specimens. CRT was significantly down-regulated in highly metastatic gastric cancer cell lines and metastatic animal by Honokiol-treated. Small RNA interference blocking CRT by siRNA-CRT was translocated to the cells in the early immunogenic response to Honokiol. Honokiol activated endoplasmic reticulum (ER) stress and down-regulated peroxisome proliferator-activated receptor-γ (PPARγ) activity resulting in PPARγ and CRT degradation through calpain-II activity, which could be reversed by siRNA-calpain-II. The Calpain-II/PPARγ/CRT axis and interaction evoked by Honokiol could be blocked by gene silencing or pharmacological agents. Both transforming growth factor (TGF)-β1 and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) induced cell migration, invasion and reciprocal down-regulation of epithelial marker E-cadherin, which could be abrogated by siRNA-CRT. Moreover, Honokiol significantly suppressed MNNG-induced gastrointestinal tumor growth and over-expression of CRT in mice. Knockdown CRT in gastric cancer cells was found to effectively reduce growth ability and metastasis in vivo. The present study provides insight into the specific biological behavior of CRT in epithelial-to-mesenchymal transition (EMT) and metastasis. Taken together, our results suggest that the therapeutic inhibition of CRT by Honokiol suppresses both gastric tumor growth and peritoneal dissemination by dictating early translocation of CRT in immunogenic cell death, activating ER stress, and blocking EMT.

Topics: Adult; Animals; Biomarkers, Tumor; Biphenyl Compounds; Calpain; Calreticulin; Cell Death; Cell Line, Tumor; Down-Regulation; Endoplasmic Reticulum Stress; Epithelial-Mesenchymal Transition; Female; Gene Knockdown Techniques; Humans; Lignans; Macrophages; Male; Methylnitronitrosoguanidine; Mice, Inbred BALB C; Mice, Inbred C57BL; Middle Aged; Neoplasm Invasiveness; Phagocytosis; PPAR gamma; Promoter Regions, Genetic; Protein Binding; Stomach Neoplasms; Up-Regulation

The purpose of this study was to investigate the effectiveness of honokiol, a natural molecule that was shown to have antioxidant effects, in prevention of intra-abdominal adhesion formation in a rat model.. This study was conducted on a total of 40 non-pregnant Sprague-Dawley rats, which were divided into 4 groups as follows: sham, control, saline, and honokiol groups. Both uterine horns of the rats in control, saline, and honokiol groups were exposed and a 2-cm segment of the anti-mesenteric surface of both uterine horns was traumatized by a scalpel. The saline group was administered 2 ml of saline/day intraperitoneally for 5 days after the operation. The honokiol group, on the other hand, was administered honokiol intraperitoneally at a dose of 1 mg/kg/day for 5 days after the operation. On postoperative day 14, 3 ml of intracardiac blood sample was taken from the rats for biochemical analyses, and the rats were sacrificed this way.. Adhesion and inflammation scores were significantly lower in the honokiol group compared with the saline and control groups (p < 0.008). Similarly, fibrosis score was significantly lower in the honokiol group compared with the saline group (p < 0.008).. Honokiol was found to be effective in prevention of intra-abdominal adhesion formation in a rat model. However, larger studies are needed to shed light on the exact role of honokiol in intra-abdominal adhesion formation and to determine the molecular aspects of the promising results found in this study.

Topics: Animals; Antioxidants; Biphenyl Compounds; Disease Models, Animal; Female; Lignans; Postoperative Complications; Rats; Rats, Sprague-Dawley; Tissue Adhesions; Uterus

The first step in infection by Candida albicans is adhesion to host cells or implanted medical devices and this followed by hyphal growth and biofilm formation. Yeast-to-hyphal transition has long been identified as a key factor in fungal virulence. Following biofilm formation, C. albicans is usually less sensitive or insensitive to antifungals. Therefore, development of new antifungals with inhibitory action on adhesion, yeast-hyphal transition and biofilm formation by C. albicans is very necessary.. The effects of magnolol and honokiol on hypha growth were investigated using different induction media. Their inhibitory effects were determined using the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5- carboxanilide assay, and biofilm thickness and viability were observed by a confocal scanning laser microscope. Mammalian cells were used in adhesion assays. Genes related to hyphae development and cell adhesions were analyzed by real-time reverse transcription-polymerase chain reaction. The exogenous cyclic adenosine monophosphate was used to determine the mechanisms of action of magnolol and honokiol. Caenorhabditis elegans was used as an in vivo model to estimate the antifungal activities of magnolol and honokiol.. Magnolol and honokiol inhibited adhesion, the transition from yeast to hypha, and biofilm formation by C. albicans through the Ras1-cAMP-Efg1 pathway. Moreover, magnolol and honokiol prolonged the survival of nematodes infected by C. albicans. Magnolol and honokiol have potential inhibitory effects against biofilm formation by C. albicans.. This study provides useful information towards the development of new strategies to reduce the incidence of C. albicans biofilm-associated infection.

Topics: Animals; Antifungal Agents; Biofilms; Biphenyl Compounds; Caenorhabditis elegans; Candida albicans; Cell Adhesion; Cell Line; Drugs, Chinese Herbal; Fungal Proteins; Hyphae; Lignans; Microscopy, Confocal; Models, Animal; Rats; Signal Transduction; Titanium; Virulence

Honokiol is a bioactive neolignan compound isolated from the species of Magnolia. This study was designed to elucidate the cellular mechanism by which honokiol alleviates the development of non-alcoholic steatosis. HepG2 cells were treated with honokiol for 1h, and then exposed to 1mM free fatty acid (FFA) for 24h to simulate non-alcoholic steatosis in vitro. C57BL/6 mice were fed with a high-fat diet for 28days, and honokiol (10mg/kg/day) was daily treated. Honokiol concentration-dependently attenuated intracellular fat overloading and triglyceride (TG) accumulation in FFA-exposed HepG2 cells. These effects were blocked by pretreatment with an AMP-activated protein kinase (AMPK) inhibitor. Honokiol significantly inhibited sterol regulatory element-binding protein-1c (SREBP-1c) maturation and the induction of lipogenic proteins, stearoyl-CoA desaturase-1 (SCD-1) and fatty acid synthase (FAS) in FFA-exposed HepG2 cells, but these effects were blocked by pretreatment of an AMPK inhibitor. Honokiol induced AMPK phosphorylation and subsequent acetyl-CoA carboxylase (ACC) phosphorylation, which were inhibited by genetic deletion of liver kinase B1 (LKB1). Honokiol stimulated LKB1 phosphorylation, and genetic deletion of LKB1 blocked the effect of honokiol on SREBP-1c maturation and the induction of SCD-1 and FAS proteins in FFA-exposed HepG2 cells. Honokiol attenuated the increases in hepatic TG and lipogenic protein levels and fat accumulation in the mice fed with high-fat diet, while significantly induced LKB1 and AMPK phosphorylation. Taken together, our findings suggest that honokiol has an anti-lipogenic effect in hepatocytes, and this effect may be mediated by the LKB1-AMPK signaling pathway, which induces ACC phosphorylation and inhibits SREBP-1c maturation in hepatocytes.

Topics: Acetyl-CoA Carboxylase; AMP-Activated Protein Kinases; Animals; Biphenyl Compounds; Cell Line, Tumor; Diet, High-Fat; Fatty Acid Synthases; Fatty Acids, Nonesterified; Hep G2 Cells; Hepatocytes; Humans; Lignans; Lipid Metabolism; Lipogenesis; Liver; Male; Mice; Mice, Inbred C57BL; Phosphorylation; Protein Serine-Threonine Kinases; Signal Transduction; Stearoyl-CoA Desaturase; Sterol Regulatory Element Binding Protein 1; Triglycerides

Glioblastoma multiforme (GBM) is the most common adult malignant glioma with poor prognosis due to the resistance to radiotherapy and chemotherapy, which might be critically involved in the repopulation of cancer stem cells (CSCs) after treatment. We had investigated the characteristics of cancer stem-like side population (SP) cells sorted from GBM cells, and studied the effect of Honokiol targeting on CSCs. GBM8401 SP cells possessed the stem cell markers, such as nestin, CD133 and Oct4, and the expressions of self-renewal related stemness genes, such as SMO, Notch3 and IHH (Indian Hedgehog). Honokiol inhibited the proliferation of both GBM8401 parental cells and SP cells in a dose-dependent manner, the IC50 were 5.3±0.72 and 11±1.1 μM, respectively. The proportions of SP in GBM8401 cells were diminished by Honokiol from 1.5±0.22% down to 0.3±0.02% and 0.2±0.01% at doses of 2.5 μM and 5 μM, respectively. The SP cells appeared to have higher expression of O6-methylguanine-DNA methyltransferase (MGMT) and be more resistant to Temozolomide (TMZ). The resistance to TMZ could be only slightly reversed by MGMT inhibitor O6-benzylguanine (O6-BG), but markedly further enhanced by Honokiol addition. Such significant enhancement was accompanied with the higher induction of apoptosis, greater down-regulation of Notch3 as well as its downstream Hes1 expressions in SP cells. Our data indicate that Honokiol might have clinical benefits for the GBM patients who are refractory to TMZ treatment.

Topics: Antineoplastic Agents; Apoptosis; Basic Helix-Loop-Helix Transcription Factors; Biphenyl Compounds; Brain Neoplasms; Cell Proliferation; Dacarbazine; Dose-Response Relationship, Drug; Down-Regulation; Drug Synergism; Drug Therapy, Combination; Glioblastoma; Homeodomain Proteins; Humans; Lignans; Neoplastic Stem Cells; Receptor, Notch3; Receptors, Notch; Temozolomide; Transcription Factor HES-1; Tumor Cells, Cultured

To explore the regulatory mechanisms of magnolol and honokiol on calcium-activated potassium channels signaling pathway in Enterotoxigenic Escherichia coli (ETEC)-induced diarrhea mice, the concentrations of serum chloride ion (Cl(-)), sodium ion (Na(+)), potassium ion (K(+)) and calcium ion (Ca(2+)) were measured. Additionally, the mRNA expressions of calmodulin 1 (CaM), calcium/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) and beta subunit (CaMKIIβ), ryanodine receptor 1, inositol 1,4,5-trisphosphate receptors (IP3 receptors), protein kinases C (PKC), potassium intermediate/small conductance calcium-activated channels (SK) and potassium large conductance calcium-activated channels(BK)were determined. A diarrhea mouse model was established using ETEC suspensions (3.29×10(9)CFU/ml) at a dosage of 0.02ml/g live body weight (BW). Magnolol or honokiol was intragastrically administered at dosages of 100 (M100 or H100), 300 (M300 or H300) and 500 (M500 or H500) mg/kg BW according to a 3×3 factorial arrangement. Magnolol and honokiol increased the Cl(-) and K(+) concentrations, further, upregulated the CaM, BKα1 and BKβ3 mRNA levels but downregulated the IP3 receptors 1, PKC, SK1, SK2, SK3, SK4 and BKβ4 mRNA expressions. Magnolol and honokiol did not alter the CaMKIIα, CaMKIIβ, ryanodine receptor 1, IP3 receptor 2, IP3 receptor 3, BKβ1 and BKβ2 mRNA expressions. These results clarify that magnolol and honokiol, acting through Ca(2+) channel blockade, inhibit the activation of IP3 receptor 1 to regulate the IP3-Ca(2+) store release, activate CaM to inhibit SK channels, and effectively suppress PKC kinases to promote BKα1 and BKβ3 channels opening and BKβ4 channel closing, which modulates the intestinal ion secretion.

Topics: Animals; Biphenyl Compounds; Calcium; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Calmodulin; Chlorides; Diarrhea; Enterotoxigenic Escherichia coli; Escherichia coli Infections; Ileum; Inositol 1,4,5-Trisphosphate Receptors; Lignans; Male; Mice; Potassium; Potassium Channels, Calcium-Activated; Protein Kinase C; Ryanodine Receptor Calcium Release Channel; Signal Transduction; Sodium

Randomized experimental study.. To investigate the neuroprotective effect of honokiol (HNK) on rats subjected to traumatic spinal cord injury (SCI) and the molecular mechanisms.. Inflammation contributes to the secondary injury to the spinal cord. Honokiol has been used as a neuroprotective agent because of its strong antioxidant and anti-inflammatory properties. Kruppel-like factor 4 (Klf4) is a newly identified critical target for the anti-inflammatory effect of HNK. Whether HNK can inhibit inflammatory response in rat model of SCI through mediating the expression of Klf4 has yet to be elucidated.. Eighty-four adult female Sprague-Dawley rats were randomly divided into 4 groups as sham, SCI, SCI + Vehicle (0.1% propylene glycol in saline, intraperitoneally), and SCI + HNK (20 mg/kg, intraperitoneally) groups. The influences of HNK on the proinflammatory cytokines, microglial activation, neutrophil infiltration, histological changes, and improvement in motor function were assessed.. In the SCI group, proinflammatory cytokines, microglial activation, neutrophil infiltration, and Klf4 expression levels were increased compared with the sham group (P < 0.001). HNK intervention downregulated the expression of Klf4, reduced the production of proinflammatory cytokines, inhibited microglial activation, and neutrophil infiltration (P < 0.05). Furthermore, HNK also reduced histopathology and improved functional outcome after traumatic SCI.. HNK reduces secondary tissue damage and improves locomotor function recovery after SCI through suppressing inflammatory response, and can be used as a potential therapeutic agent for SCI.. NA.

Topics: Animals; Antioxidants; Biphenyl Compounds; Cytokines; Down-Regulation; Female; Inflammation; Kruppel-Like Factor 4; Kruppel-Like Transcription Factors; Lignans; Neuroprotective Agents; Rats; Rats, Sprague-Dawley; Spinal Cord; Spinal Cord Injuries

Renal cell carcinoma (RCC) is a common urological cancer worldwide and is known to have a high risk of metastasis, which is considered responsible for more than 90% of cancer associated deaths. Honokiol is a small-molecule biphenol isolated from Magnolia spp. bark and has been shown to be a potential anticancer agent involved in multiple facets of signal transduction. In this study, we demonstrated that honokiol inhibited the invasion and colony formation of highly metastatic RCC cell line 786-0 in a dose-dependent manner. DNA-microarray data showed the significant upregulation of metastasis-suppressor gene KISS1 and its receptor, KISS1R. The upregulation was confirmed by qRT-PCR analysis. Overexpression of KISS1 and KISS1R was detected by western blotting at the translation level as well. Of note, the decreased invasive and colonized capacities were reversed by KISS1 knockdown. Taken together, the results first indicate that activation of KISS1/KISS1R signaling by honokiol suppresses multistep process of metastasis, including invasion and colony formation, in RCC cells 786-0. Honokiol may be considered as a natural agent against RCC metastasis.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Carcinoma, Renal Cell; Cell Line, Tumor; Cell Movement; Dose-Response Relationship, Drug; Gene Expression Regulation, Neoplastic; Humans; Kidney Neoplasms; Kisspeptins; Lignans; Neoplasm Invasiveness; Receptors, G-Protein-Coupled; Receptors, Kisspeptin-1; Signal Transduction

Honokiol (HKL) is a natural biphenolic compound derived from the bark of magnolia trees with anti-inflammatory, anti-oxidative, anti-tumour and neuroprotective properties. Here we show that HKL blocks agonist-induced and pressure overload-mediated, cardiac hypertrophic responses, and ameliorates pre-existing cardiac hypertrophy, in mice. Our data suggest that the anti-hypertrophic effects of HKL depend on activation of the deacetylase Sirt3. We demonstrate that HKL is present in mitochondria, enhances Sirt3 expression nearly twofold and suggest that HKL may bind to Sirt3 to further increase its activity. Increased Sirt3 activity is associated with reduced acetylation of mitochondrial Sirt3 substrates, MnSOD and oligomycin-sensitivity conferring protein (OSCP). HKL-treatment increases mitochondrial rate of oxygen consumption and reduces ROS synthesis in wild type, but not in Sirt3-KO cells. Moreover, HKL-treatment blocks cardiac fibroblast proliferation and differentiation to myofibroblasts in a Sirt3-dependent manner. These results suggest that HKL is a pharmacological activator of Sirt3 capable of blocking, and even reversing, the cardiac hypertrophic response.

Topics: Acetylation; Adenosine Triphosphatases; Animals; Biphenyl Compounds; Cardiomegaly; Cardiotonic Agents; Carrier Proteins; Cell Differentiation; Cell Proliferation; Enzyme Activation; Fibroblasts; Gene Expression Regulation; Isoproterenol; Lignans; Membrane Proteins; Mice; Mitochondria; Mitochondrial Proton-Translocating ATPases; Myocardium; Myocytes, Cardiac; Myofibroblasts; Phenylephrine; Primary Cell Culture; Reactive Oxygen Species; Signal Transduction; Sirtuin 3; Superoxide Dismutase

A series of novel benzoxazole compounds derived from a naturally occurring neolignan honokiol were prepared. Their insecticidal activity was tested against the pre-third-instar larvae of oriental armyworm (Mythimna separata Walker) in vivo. Most of the tested derivatives exhibited more potential insecticidal activity than their precursor honokiol at the concentration of 1mg/mL. Especially compound 6e, containing 4-acetyloxyphenyl groups at the C8 and C8' positions, displayed the most potent insecticidal activity with the final mortality rate of 62.1%.

Topics: Animals; Benzoxazoles; Biphenyl Compounds; Insecticides; Larva; Lignans; Molecular Structure; Moths

The aim of this study was to investigate anti-inflammatory and anti-cancer effects of honokiol (HK) in two oral squamous cancer cell carcinoma (OSCC) cell lines, HN22 and HSC4, through the regulation of inducible nitric oxide synthase (iNOS) and endoplasmic reticulum resident protein 44 (ERp44). Griess assay, zymography, and quantitative PCR were performed to study iNOS expression and subsequent nitric oxide (NO) production in OSCC cell lines. Liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteomic analysis was used to elucidate the proteins associated with ER stress and cellular cytotoxic response induced by HK. Pull-down assay and molecular modeling were performed to better understand how HK interacts with ERp44. In vitro and in vivo experiments in which ERp44 expression was knocked down were performed to better understand the effects of ERp44 on a cellular level and anti-cancer effects of HK. Expression levels of iNOS and subsequent NO secretion were reduced in OSCC cell lines treated with HK. ERp44 was significantly decreased in OSCC cell lines by HK treatment. HK directly bound to ERp44, and ERp44 knock-down significantly inhibited oral cancer cell proliferation and colony formation. Moreover, HK treatment effectively inhibited tumor growth and ERp44 levels in BALB/c nude mice bearing HN22 cell xenografts. Our findings suggest that HK inhibited inflammation and induced apoptosis by suppressing both iNOS/NO and ERp44 expression in HN22 and HSC4 cells and xenograft tumors, and thus could be a potent anti-inflammatory and anti-cancer drug candidate for human oral cancer treatment.

Topics: Animals; Anti-Inflammatory Agents; Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Calcium; Carcinoma, Squamous Cell; Chromatography, Liquid; Humans; Lignans; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Membrane Potential, Mitochondrial; Mice; Mice, Nude; Mouth Neoplasms; NF-kappa B; Nitric Oxide Synthase Type II; Oxidation-Reduction; Tandem Mass Spectrometry; Xenograft Model Antitumor Assays

Honokiol, a lignan isolated from the bark of Magnolia officinalis, has been reported to ameliorate the learning and memory impairments in senesed (SAMP8) mice. However, whether honokiol could improve scopolamine (SCOP)-induced learning and memory deficits in mice is still unknown. In this study, we aimed to investigate whether honokiol could reverse the SCOP-induced learning and memory impairments in mice and to elucidate its underlying mechanisms of action. Mice were given daily intraperitoneal injection of honokiol (10 and 20mg/kg) for 21 consecutive days. The results showed that honokiol significantly improved spatial learning and memory function (as assessed by the Morris water maze test) in the SCOP-treated mice. In addition, treatment with honokiol significantly decreased the protein and mRNA levels of interleukin (IL)-1β and the activity of acetylcholinesterase (AChE), while significantly increased the protein and mRNA levels of IL-10, and the level of acetylcholine (Ach) in the brain of the SCOP-treated mice. Moreover, honokiol also significantly suppressed the production of prostaglandin E 2 (PGE2) and mRNA expression of cyclooxygenase-2 (COX-2) in the brain of the SCOP-treated mice. Mechanistic investigations revealed that honokiol could markedly reverse the amount of phosphorylated Akt and extracellular regulated kinases 1/2 (ERK1/2) changes in the brain of the SCOP-treated mice. These results amply demonstrated that honokiol could improve learning and memory impairments induced by SCOP in mice, and the protective action may be mediated, at least in part, by inhibition of AChE activity, and amelioration of the neuroinflammatory processes in the SCOP-treated mice.

Topics: Animals; Biphenyl Compounds; Brain; Cholinesterase Inhibitors; Drugs, Chinese Herbal; Lignans; Male; Maze Learning; Memory Disorders; Mice; Mice, Inbred ICR; Scopolamine

The undesirable respiratory burst in neutrophils can lead to inflammation and tissue damage. This study investigates the effect and the underlying mechanism of 2-(4-hydroxy-3-prop-2-enyl-phenyl)-4-prop-2-enyl-phenol (honokiol), a lignan extracted from the stem bark of Magnolia officinalis Rehd. et Wils (Magnoliaceae), on N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP)-induced respiratory burst in human neutrophils. Signaling pathways regulated by honokiol which modulate fMLP-induced respiratory burst and cathepsin G release were evaluated by phosphorylation of Src family kinase induced by fMLP, Src family kinases activities and by immunoblotting analysis of the downstream targets of Src kinase. Briefly, honokiol inhibited fMLP-induced superoxide anion production (IC50 = 9.80 ± 0.21 μM, n = 4), cathepsin G release (IC50 = 14.23 ± 1.43 μM, n = 4) and migration (IC50 = 5.69 ± 1.51 μM, n = 4) in a concentration dependent manner. Further, honokiol specifically suppresses fMLP-induced Lyn (a member of the Src kinase family) phosphorylation, by inhibiting Lyn kinase activity. Consequently, honokiol attenuated the downstream targets of Lyn kinase, such as Tec translocation from the cytosol to the inner leaflet of the plasma membrane, phosphorylation of AKT, P38, PLCγ2, protein kinase C and membrane localization of p47(phox). On the other hand, fMLP-induced phosphorylation of Hck, Fgr kinase activity (other members of Src kinase), downstream phosphorylation of Vav1 and extracellular signal-regulated kinase remained unaffected. In addition, honokiol neither inhibited NADPH oxidase activity nor increased cyclic AMP levels. Honokiol is not a competitive or allosteric antagonist of fMLP. In conclusion, honokiol specifically modulates fMLP-mediated neutrophil activation by inhibiting Lyn activation which subsequently interferes with the activation of PLCγ2, AKT, p38, protein kinase C, and p47(phox).

Topics: Adult; Anti-Inflammatory Agents, Non-Steroidal; Biphenyl Compounds; Calcium; Cathepsin G; Cell Movement; Cyclic AMP; Humans; Lignans; N-Formylmethionine Leucyl-Phenylalanine; NADPH Oxidases; Neutrophils; Phospholipase C gamma; Phosphorylation; Proto-Oncogene Proteins c-akt; Receptors, Formyl Peptide; Respiratory Burst; src-Family Kinases; Superoxides; Young Adult

Counter-current chromatography (CCC) is an efficient liquid-liquid chromatography technique for separation and purification of complex mixtures like natural products extracts and synthetic chemicals. However, CCC is still a challenging process requiring some special technical knowledge especially in the selection of appropriated solvent systems. In this work, we introduced a new 9 × 9 map-based solvent selection strategy for CCC isolation of targets, which permit more than 60 hexane-ethyl acetate-methanol-water (HEMWat) solvent systems as the start candidates for the selection of solvent systems. Among these solvent systems, there are clear linear correlations between partition coefficient (K) and the system numbers. Thus, an appropriate CCC solvent system (i.e., sweet spot for K = 1) may be hit by measurement of k values of the target only in two random solvent systems. Besides this, surprisingly, we found that through two sweet spots, we could get a line ("Sweet line") where there are infinite sweet solvent systems being suitable for CCC separation. In these sweet solvent systems, the target has the same partition coefficient (K) but different solubilities. Thus, the better sweet solvent system with higher sample solubility can be obtained for high capacity CCC preparation. Furthermore, we found that there is a zone ("Sweet zone") where all solvent systems have their own sweet partition coefficients values for the target in range of 0.4 < K< 2.5 or extended range of 0.25 < K < 16. All results were validated by using 14 pure GUESSmix mimic natural products as standards and further confirmed by isolation of several targets including honokiol and magnolol from the extracts of Magnolia officinalis Rehd. Et Wils and tanshinone IIA from Salvia miltiorrhiza Bunge. In practice, it is much easier to get a suitable solvent system only by making a simple screening two to four HEMWat two-phase solvent systems to obtain the sweet line or sweet zone without special knowledge or comprehensive standards as references. This is an important advancement for solvent system selection and also will be very useful for isolation of current natural products including Traditional Chinese Medicines.

Topics: Abietanes; Biological Products; Biphenyl Compounds; Chemistry Techniques, Analytical; Chromatography, High Pressure Liquid; Countercurrent Distribution; Hexanes; Lignans; Magnolia; Methanol; Reproducibility of Results; Salvia miltiorrhiza; Solvents; Water

Here, we report the chemotherapeutic effect of honokiol, a phytochemical from Magnolia plant, on human head and neck squamous cell carcinoma (HNSCC). Treatment of HNSCC cell lines from different sub-sites, SCC-1 (oral cavity), SCC-5 (larynx), OSC-19 (tongue) and FaDu (pharynx) with honokiol inhibited their cell viability, which was associated with the: (i) induction of apoptosis, (ii) correction of dysregulatory cell cycle proteins of G0/G1 phase. Honokiol decreased the expression levels of epidermal growth factor receptor (EGFR), mTOR and their downstream signaling molecules. Treatment of FaDu and SCC-1 cell lines with rapamycin, an inhibitor of mTOR pathway, also reduced cell viability of HNSCC cells. Administration of honokiol by oral gavage (100 mg/kg body weight) significantly (P < 0.01-0.001) inhibited the growth of SCC-1 and FaDu xenografts in athymic nude mice, which was associated with: (i) inhibition of tumor cell proliferation, (ii) induction of apoptosis, (iii) reduced expressions of cyclins and Cdks, and (iv) inhibition of EGFR signaling pathway. Molecular docking analysis of honokiol in EGFR binding site indicated that the chemotherapeutic effect of honokiol against HNSCC is mediated through its firm binding with EGFR, which is better than that of gefitinib, a commonly used drug for HNSCC treatment.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Binding Sites; Biphenyl Compounds; Carcinoma, Squamous Cell; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Separation; Cell Survival; Dose-Response Relationship, Drug; ErbB Receptors; Female; Flow Cytometry; Head and Neck Neoplasms; Humans; Lignans; Magnolia; Mice; Mice, Nude; Molecular Docking Simulation; Phytotherapy; Plant Extracts; Signal Transduction; Sirolimus

A series of novel arylmethylamine derivatives of honokiol (5a-m) was prepared. Their insecticidal activity was tested against the pre-third-instar larvae of the oriental armyworm (Mythimna separata Walker), a typical lepidopteran pest. Compounds 5a, 5b, 5e, 5h, and 5k exhibited insecticidal activity equal to, or higher than, that of the positive control toosendanin.

Topics: Animals; Biphenyl Compounds; Drugs, Chinese Herbal; Insecticides; Larva; Lepidoptera; Lignans

Obesity greatly influences risk, progression and prognosis of breast cancer. As molecular effects of obesity are largely mediated by adipocytokine leptin, finding effective novel strategies to antagonize neoplastic effects of leptin is desirable to disrupt obesity-cancer axis. Present study is designed to test the efficacy of honokiol (HNK), a bioactive polyphenol from Magnolia grandiflora, against oncogenic actions of leptin and systematically elucidate the underlying mechanisms. Our results show that HNK significantly inhibits leptin-induced breast-cancer cell-growth, invasion, migration and leptin-induced breast-tumor-xenograft growth. Using a phospho-kinase screening array, we discover that HNK inhibits phosphorylation and activation of key molecules of leptin-signaling-network. Specifically, HNK inhibits leptin-induced Wnt1-MTA1-β-catenin signaling in vitro and in vivo. Finally, an integral role of miR-34a in HNK-mediated inhibition of Wnt1-MTA1-β-catenin axis was discovered. HNK inhibits Stat3 phosphorylation, abrogates its recruitment to miR-34a promoter and this release of repressor-Stat3 results in miR-34a activation leading to Wnt1-MTA1-β-catenin inhibition. Accordingly, HNK treatment inhibited breast tumor growth in diet-induced-obese mouse model (exhibiting high leptin levels) in a manner associated with activation of miR-34a and inhibition of MTA1-β-catenin. These data provide first in vitro and in vivo evidence for the leptin-antagonist potential of HNK revealing a crosstalk between HNK and miR34a and Wnt1-MTA1-β-catenin axis.

Topics: Animals; Antineoplastic Agents, Phytogenic; beta Catenin; Biphenyl Compounds; Breast Neoplasms; Cell Movement; Cell Proliferation; Cell Transformation, Neoplastic; Cyclin D1; Drugs, Chinese Herbal; Female; Histone Deacetylases; Humans; Leptin; Lignans; Magnolia; MCF-7 Cells; Mice; Mice, Nude; Mice, Obese; MicroRNAs; Neoplasm Invasiveness; Obesity; Phosphorylation; Plant Extracts; Promoter Regions, Genetic; Repressor Proteins; RNA Interference; RNA, Small Interfering; Signal Transduction; Spheroids, Cellular; STAT3 Transcription Factor; Trans-Activators; Tumor Cells, Cultured; Wnt1 Protein; Xenograft Model Antitumor Assays

Honokiol is the most important active pharmaceutical ingredient in Magnolia officinalis, which is a famous traditional Chinese medicine and commonly used in clinical practice. In order to control the quality of honokiol and related pharmaceuticals, a new certified reference material (CRM) of honokiol was developed. The studies of sample preparation, homogeneity, stability, value assignment, and uncertainty evaluation were accomplished in this paper. Three different methods, including differential scanning calorimetry (DSC), mass balance method (MB), and coulometric titration (CT), were employed to determine the purity of honokiol. Specifically, the DSC and CT methods for purity determination of honokiol were established for the first time. The purity of honokiol CRM, after validation and evaluation, was found to be 99.3%, with an expanded uncertainty of 0.5% (k = 2).

Topics: Biphenyl Compounds; Certification; Chromatography, High Pressure Liquid; Lignans; Reference Standards

Honokiol (HNK), a potential antitumor compound, has been widely studied in recent years. It induces apoptosis and affects autophagy in cancer cells, yet the mechanism of its antitumor efficacy remains obscure. Chloroquine (CQ), an autophagy inhibitor, is often applied to sensitize antitumor drugs in clinical trials. Here, we investigated the antitumor effect of HNK or CQ alone or in combination in non-small cell lung cancer (NSCLC) cells. Using an experimental approach, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) or sulforhodamine B (SRB) was used to determine the cytotoxicity of the agents. The expression levels of proteins were detected by western blotting. Apoptosis was examined via Annexin V-FITC and PI staining. H460 cell xenografts in nude mice were used to study the effects of HNK and/or CQ in vivo. Transfection with siRNA was applied to knock down cathepsin D. The results demonstrated the enhanced effects of HNK combined with CQ on the inhibition of proliferation, induction of apoptosis in vitro and the reduction in growth in vivo. It was confirmed that HNK and/or CQ triggered apoptosis via a caspase-dependent manner. Furthermore, HNK significantly increased the expression of p62 and LC3-Ⅱ in the A549 and H460 cells and inhibited autophagy and induced apoptosis in a cathepsin D-involved manner. In conclusion, an enhanced antitumor effect was demonstrated following treatment with HNK combined with CQ by inhibiting autophagy and inducing apoptosis via a caspase-dependent and cathepsin D-involved manner. This combination may be a novel and useful antitumor approach for chemotherapy in NSCLC.

Topics: Animals; Apoptosis; Autophagy; Biphenyl Compounds; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Proliferation; Chloroquine; Drug Synergism; Gene Expression Regulation, Neoplastic; Humans; Lignans; Mice; Neoplasm Proteins; Xenograft Model Antitumor Assays

Pentraxin 3 (PTX3) was identified as a marker of the inflammatory response and overexpressed in various tissues and cells related to cardiovascular disease. Honokiol, an active component isolated from the Chinese medicinal herb Magnolia officinalis, was shown to have a variety of pharmacological activities. In the present study, we aimed to investigate the effects of honokiol on palmitic acid (PA)-induced dysfunction of human umbilical vein endothelial cells (HUVECs) and to elucidate potential regulatory mechanisms in this atherosclerotic cell model. Our results showed that PA significantly accelerated the expression of PTX3 in HUVECs through the IκB kinase (IKK)/IκB/nuclear factor-κB (NF-κB) pathway, reduced cell viability, induced cell apoptosis and triggered the inflammatory response. Knockdown of PTX3 supported cell growth and prevented apoptosis by blocking PA-inducted nitric oxide (NO) overproduction. Honokiol significantly suppressed the overexpression of PTX3 in PA-inducted HUVECs by inhibiting IκB phosphorylation and the expression of two NF-κB subunits (p50 and p65) in the IKK/IκB/NF-κB signaling pathway. Furthermore, honokiol reduced endothelial cell injury and apoptosis by regulating the expression of inducible NO synthase and endothelial NO synthase, as well as the generation of NO. Honokiol showed an anti-inflammatory effect in PA-inducted HUVECs by significantly inhibiting the generation of interleukin-6 (IL-6), IL-8 and monocyte chemoattractant protein-1. In summary, honokiol repaired endothelial dysfunction by suppressing PTX3 overexpression in an atherosclerotic cell model. PTX3 may be a potential therapeutic target for atherosclerosis.

Topics: Apoptosis; Atherosclerosis; Biphenyl Compounds; C-Reactive Protein; Down-Regulation; Drugs, Chinese Herbal; Human Umbilical Vein Endothelial Cells; Humans; I-kappa B Kinase; Lignans; Magnolia; NF-kappaB-Inducing Kinase; Palmitic Acid; Protein Serine-Threonine Kinases; Serum Amyloid P-Component; Signal Transduction

Honokiol (HK), a biphenolic neolignan isolated from Magnolia officinalis, has been reported to possess anti-inflammatory and anti-angiogenic activaties. In this study, our aim was to investigate anti-psoriatic activities of HK and the involved mechanisms. In vitro, the effects of HK on the regulation of Th1/Th2 and TNF-α-induced NF-κB (p65) activation were analyzed by respective FCS and immunofluorescence. Additionally, the K14-VEGF transgenic model was used for the in vivo study. ELISA and Q-PCR were performed to evaluate serum levels of Th1/Th2 cytokines and their corresponding mRNA expressions. Effects on VEGFR-2 and p65 activation, as well as other angiogenic and inflammatory parameters were studied by immunostainings. Importantly, we found that HK significantly decreased the ratio of Th1/Th2-expression CD4(+) T cells and inhibited TNF-α-induced activation of NF-κB. The morphology and histological features of psoriasis were effectively improved by HK treatment. The expression of TNF-α and IFN-γ, and their corresponding mRNA levels were down-regulated and the expression of nuclear p65, VEGFR-2, as well as related phosphorylated proteins (p-VEGFR-2, p-ERK1/2, p-AKT and p-p38) were also suppressed. Overall, these results in our study suggested that HK exhibits anti-psoriatic effects through the inhibition of NF-κB and VEGFR-2.

Topics: Angiogenesis Inhibitors; Animals; Anti-Inflammatory Agents; Biphenyl Compounds; CD4-Positive T-Lymphocytes; Cells, Cultured; Disease Models, Animal; Down-Regulation; Gene Expression; Human Umbilical Vein Endothelial Cells; Humans; Lignans; Magnolia; Mice, Transgenic; Molecular Targeted Therapy; NF-kappa B; Phosphorylation; Phytotherapy; Psoriasis; Th1-Th2 Balance; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-2

Ma-Zi-Ren-Wan (MZRW) is a classic Chinese formula which has been used to treat human constipation in China for over 2000 years. In order to make good and rational use of this formula in the future, this paper presents the first attempt to track the pharmacokinetic features of MZRW in rat using rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Ten chemical components of MZRW, namely, rhein, emodin, aloe emodin, hesperidin, naringin, amygdalin, albiflorin, paeoniflorin, magnolol and honokiol, were simultaneously determined in rat plasma after a single oral administration (10g/kg body weight) of MZRW to rats. Geniposide and liquiritin were used as internal standards. The separation was performed on a Waters ACQUITY BEH C18 column (100mm×2.1mm, 1.7μm). The detection was conducted by multiple-reaction monitoring (MRM) in negative ionization mode. Two highest abundant MRM transitions without interference were optimized for each analyte. This method was well validated in terms of linearity, precision, accuracy, recovery, matrix effect and stability. All calibration curves had good linearity (r(2)>0.995) over the concentration range from 3.9 to 125.0ng/mL for emodin, 3.9-500.0ng/mL for amygdalin, 2.0-4000.0ng/mL for naringin and hesperidin, 3.9-2000.0ng/mL for magnolol, 7.8-2000.0ng/mL for rhein and 3.9-4000.0ng/mL for albiflorin, paeoniflorin, aloe emodin and honokiol. The intra-day and inter-day precision (relative standard deviation) was within 15%, the accuracy (relative error) ranged from -13.6% to 15.1%, and the lower limit of quantification in plasma ranged between 2.0ng/mL and 7.8ng/mL. Extraction recovery, matrix effect and stability were satisfactory. The validated method was successfully applied to a pharmacokinetic study of these ten compounds after oral administration of MZRW to rats. The pharmacokinetic parameters of each compound can facilitate clinical studies in the future.

Topics: Animals; Anthraquinones; Biphenyl Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavonoids; Glycosides; Lignans; Linear Models; Male; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Sensitivity and Specificity; Tandem Mass Spectrometry

The purpose of this study was to establish a drug screening method for small molecules extracted from traditional Chinese medicines (TCM) that have neuronal differentiation promoting effects, using P19 embryonic carcinoma cell as a cell-based model. First, the constructed plasmid (pTα1-Luc) was transfected into P19 cells to establish a screening model. Second, several TCMs were screened using the established model and all-trans-retinoic acid as a positive control. Finally, the underlying molecular mechanism was explored using immunofluorescence staining, qT-PCR, and Western blot analysis. Our results indicated that the drug screen model was established successfully and that both honokiol and hyperoside induced P19 differentiation into neurons, with the possible molecular mechanism being modulating the Wnt signaling pathway. In conclusion, the drug screening model developed in the present study provides a rapid, cell-based screening platform for identifying natural compounds with neuronal differentiation effects.

Topics: Animals; Biphenyl Compounds; Cell Differentiation; Cell Line, Tumor; Cell Proliferation; Drug Evaluation, Preclinical; Drugs, Chinese Herbal; Embryonal Carcinoma Stem Cells; Lignans; Mice; Neurons; Quercetin; Tretinoin; Wnt Signaling Pathway

Amyloid formation is associated with multiple amyloidosis diseases. Human calcitonin (hCT) is a typical amyloidogenic peptide, its aggregation is associated with medullary carcinoma of the thyroid (MTC), and also limits its clinical application. Magnolia officinalis is a traditional Chinese herbal medicine; its two major polyphenol components, magnolol (Mag) and honokiol (Hon), have displayed multiple functions. Polyphenols like flavonoids and their derivatives have been extensively studied as amyloid inhibitors. However, the anti-amyloidogenic property of a biphenyl backbone containing polyphenols such as Mag and Hon has not been reported. In this study, these two compounds were tested for their effects on hCT aggregation. We found that Mag and Hon both inhibited the amyloid formation of hCT, whereas Mag showed a stronger inhibitory effect; moreover, they both dose-dependently disassembled preformed hCT aggregates. Further immuno-dot blot and dynamic light scattering studies suggested Mag and Hon suppressed the aggregation of hCT both at the oligomerization and the fibrillation stages, while MTT-based and dye-leakage assays demonstrated that Mag and Hon effectively reduced cytotoxicity caused by hCT aggregates. Furthermore, isothermal titration calorimetry indicated Mag and Hon both interact with hCT. Together, our study suggested a potential anti-amyloidogenic property of these two compounds and their structure related derivatives.

Topics: Biphenyl Compounds; Calcitonin; Calorimetry; Cell Line, Tumor; Dynamic Light Scattering; Humans; Lignans; Magnolia; Medicine, Chinese Traditional; Microscopy, Electron, Transmission; Polyphenols; Protein Binding

To observe the anti-inflammatory effects of honokiol in primary cultures of peripheral blood mononuclear cells of rheumatoid arthritis patients, the pro-inflammatory cytokines and potential targets were investigated.. The levels of GM-CSF, IL-1β, TNF-α and IL-8 were determined by ELISA assay. The genes and proteins expression were analyzed by real-time PCR and Western blotting respectively.. The serum IL-1β, TNF-α and GM-CSF levels were 1.76-, 2.16- and 3.57-fold increased in patients with RA as compared to those of control group. Honokiol inhibited the expression levels of IL-1β, TNF-α, GM-CSF and IL-8 in PBMCs with a dose-dependent manner. Measurements obtained from supernatants were positively correlated between TNF-α and IL-1β, moreover, similar results found TNF-α levels positively correlated with GM-CSF and IL-8 activity in the supernatants of PBMCs isolated from RA patients. Furthermore, the mRNA and protein expression of IL-1β, GM-CSF and IL-8 were up-regulated when the PBMCs exposure to TNF-α, however, honokiol treatment significantly reversed the expression of IL-1β, TNF-α and GM-CSF in response to TNF-α with a dose-dependent manner.. This study demonstrates that honokiol could possess potential anti-inflammatory effects and inhibits TNF-α-induced IL-1β, GM-CSF and IL-8 production in PBMCs from rheumatoid arthritis patients.

Topics: Adult; Aged; Anti-Inflammatory Agents; Antirheumatic Agents; Arthritis, Rheumatoid; Biphenyl Compounds; Case-Control Studies; Cells, Cultured; Dose-Response Relationship, Drug; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Inflammation Mediators; Interleukin-1beta; Interleukin-8; Leukocytes, Mononuclear; Lignans; Male; Middle Aged; Primary Cell Culture; Signal Transduction; Tumor Necrosis Factor-alpha

Self-assembled pectin nanoparticles was prepared and evaluated for delivering the hydrophobic drug, honokiol (HK), to HepG2 cells. These hydrophobic drug-loaded nanoparticles were developed without using any surfactant and organic solvent. Hydroxypropyl-β-cyclodextrin (HCD) was used to fabricate an inclusion complex with HK (HKHCD) to increase the solubility of the drug and thus facilitate its encapsulation and dispersion in the pectin nanoparticles. Investigation of the in vitro release indicated that the drug-loaded nanoparticles exhibited a higher drug release rate than free honokiol and an effective sustained-release. Cytotoxicity, cell apoptosis and cellular uptake studies further confirmed that the pectin nanoparticles with galactose residues generated higher cytotoxicity than free honokiol on HepG2 cells which highly expressed asialoglycoprotein receptors (ASGR). Nevertheless, these findings were not observed in ASGR-negative A549 cells under similar condition. Therefore, pectin nanoparticles demonstrated a specific active targeting ability to ASGR-positive HepG2 cells and could be used as a potential drug carrier for treatment of liver-related tumors.

Topics: Apoptosis; Binding, Competitive; Biological Transport; Biphenyl Compounds; Capsules; Cell Survival; Drug Carriers; Drug Liberation; Hep G2 Cells; Humans; Lignans; Nanoparticles; Pectins

The antimicrobial killing activity toward methicillin-resistant Staphylococcus aureus (MRSA) has been a serious emerging global issue. In a continuing search for compounds with antibacterial activity against several microorganisms including S. aureus and MRSA, an n-hexane extract of Magnolia officinalis was found to contain magnolol. This compound exhibited potent activity against S. aureus, standard methicillin-susceptible S. aureus (MSSA), and MRSA as well as clinical MRSA isolates. When combined with oxacillin, the antibacterial activities of magnolol and honokiol against the MRSA strain were increased compared to single treatment without antibiotics at 10 µg/mL and 25 µg/mL, respectively. These activities of magnolol and honokiol were dose dependent. Also, magnolol showed synergistic effects with oxacillin against 13 clinical isolates of MRSA. It was determined that magnolol and honokiol had a synergistic effect with oxacillin against MRSA strain. Furthermore, the magnolol inhibited the expression of the resistant genes, mecA, mecI, femA, and femB, in mRNA. We concluded that the antibacterial activity of magnolol against MRSA strain is more related to the mecI's pathway and components of the cell wall than mecR1. Therefore, the results obtained in this study suggest that the combination of magnolol and antibiotics could lead to the development of new combination antibiotics against MRSA infection.

Topics: Anti-Infective Agents; Biphenyl Compounds; Hexanes; Lignans; Methicillin-Resistant Staphylococcus aureus; Microbial Sensitivity Tests; Oxacillin; Plant Extracts

Leptin, a major adipocytokine produced by adipocytes, is emerging as a key molecule linking obesity with breast cancer therefore, it is important to find effective strategies to antagonize oncogenic effects of leptin to disrupt obesity-cancer axis. Here, we examine the potential of honokiol (HNK), a bioactive polyphenol from Magnolia grandiflora, as a leptin-antagonist and systematically elucidate the underlying mechanisms. HNK inhibits leptin-induced epithelial-mesenchymal-transition (EMT), and mammosphere-formation along with a reduction in the expression of stemness factors, Oct4 and Nanog. Investigating the downstream mediator(s), that direct leptin-antagonist actions of HNK; we discovered functional interactions between HNK, LKB1 and miR-34a. HNK increases the expression and cytoplasmic-localization of LKB1 while HNK-induced SIRT1/3 accentuates the cytoplasmic-localization of LKB1. We found that HNK increases miR-34a in LKB1-dependent manner as LKB1-silencing impedes HNK-induced miR-34a which can be rescued by LKB1-overexpression. Finally, an integral role of miR-34a is discovered as miR-34a mimic potentiates HNK-mediated inhibition of EMT, Zeb1 expression and nuclear-localization, mammosphere-formation, and expression of stemness factors. Leptin-antagonist actions of HNK are further enhanced by miR-34a mimic whereas miR-34a inhibitor results in inhibiting HNK's effect on leptin. These data provide evidence for the leptin-antagonist potential of HNK and reveal the involvement of LKB1 and miR-34a.

Topics: AMP-Activated Protein Kinase Kinases; Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Blotting, Western; Breast Neoplasms; Cell Line; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Humans; Leptin; Lignans; MCF-7 Cells; Mice; MicroRNAs; Microscopy, Confocal; Protein Serine-Threonine Kinases; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; Xenograft Model Antitumor Assays

Dengue is the most widespread arbovirus infection and poses a serious health and economic issue in tropical and subtropical countries. Currently no licensed vaccine or compounds can be used to prevent or manage the severity of dengue virus (DENV) infection. Honokiol, a lignan biphenol derived from the Magnolia tree, is commonly used in Eastern medicine. Here we report that honokiol has profound antiviral activity against serotype 2 DENV (DENV-2). In addition to inhibiting the intracellular DENV-2 replicon, honokiol was shown to suppress the replication of DENV-2 in baby hamster kidney (BHK) and human hepatocarcinoma Huh7 cells. At the maximum non-toxic dose of honokiol treatment, the production of infectious DENV particles was reduced >90% in BHK and Huh7 cells. The underlying mechanisms revealed that the expression of DENV-2 nonstructural protein NS1/NS3 and its replicating intermediate, double-strand RNA, was dramatically reduced by honokiol treatment. Honokiol has no effect on the expression of DENV putative receptors, but may interfere with the endocytosis of DENV-2 by abrogating the co-localization of DENV envelope glycoprotein and the early endosomes. These results indicate that honokiol inhibits the replication, viral gene expression, and endocytotic process of DENV-2, making it a promising agent for chemotherapy of DENV infection.

Topics: Animals; Antiviral Agents; Biphenyl Compounds; Cells, Cultured; Cricetinae; Dengue Virus; Gene Expression Regulation, Viral; Humans; Lignans; Magnolia; Virus Internalization; Virus Replication

In traditional Asian medicinal systems, preparations of the root and stem bark of Magnolia species are widely used to treat anxiety and other nervous disturbances. The biphenyl-type neolignan honokiol together with its isomer magnolol are the main constituents of Magnolia bark extracts. We have previously identified a nitrogen-containing honokiol derivative (3-acetylamino-4'-O-methylhonokiol, AMH) as a high efficient modulator of GABAA receptors. Here we further elucidate the structure-activity relation of a series of nitrogenated biphenyl-neolignan derivatives by analysing allosteric modulation and agonistic effects on α1β2γ2S GABAA receptors. The strongest IGABA enhancement was induced by compound 5 (3-acetamido-4'-ethoxy-3',5-dipropylbiphenyl-2-ol, Emax: 123.4±9.4% of IGABA-max) and 6 (5'-amino-2-ethoxy-3',5-dipropylbiphenyl-4'-ol, Emax: 117.7±13.5% of IGABA-max). Compound 5 displayed, however, a significantly higher potency (EC50=1.8±1.1 μM) than compound 6 (EC50=20.4±4.3 μM). Honokiol, AMH and four of the derivatives induced significant inward currents in the absence of GABA. Strong partial agonists were honokiol (inducing 78±6% of IGABA-max), AMH (63±6%), 5'-amino-2-O-methylhonokiol (1) (59±1%) and 2-methoxy-5'-nitro-3',5-dipropylbiphenyl-4'-ol (3) (52±1%). 3-N-Acetylamino-4'-ethoxy-3',5-dipropyl-biphenyl-4'-ol (5) and 3-amino-4'-ethoxy-3',5-dipropyl-biphenyl-4'-ol (7) were less efficacious but even more potent (5: EC50=6.9±1.0 μM; 7: EC50=33.2±5.1 μM) than the full agonist GABA.

Topics: Allosteric Regulation; Animals; Biphenyl Compounds; Dose-Response Relationship, Drug; Drug Partial Agonism; Female; GABA-A Receptor Agonists; Lignans; Magnolia; Molecular Structure; Nitrogen; Oocytes; Receptors, GABA-A; Structure-Activity Relationship; Xenopus laevis

Listeria monocytogenes is considered one of the most important foodborne pathogens. The virulence-related proteins listeriolysin O (LLO) and p60 are critical factors involved in Listeria pathogenesis. In the present study, we investigated the effect of honokiol on LLO and p60 secreted from L. monocytogenes. A listeriolysin assay was used to investigate the haemolytic activities of L. monocytogenes exposed to honokiol, and the secretion of LLO and p60 was detected by immunoblot analysis. Additionally, the influence of honokiol on the transcription of LLO and p60 genes (hly and iap, respectively) was analysed by real-time reverse transcription PCR. TNF-α release assays were performed to elucidate the biological relevance of changes in LLO and p60 secretion induced by honokiol. According to the data, honokiol showed good anti-L. monocytogenes activity, with MICs of 8-16 μg ml(-1), and the secretion of LLO and p60 was decreased by honokiol. In addition, the transcription of hly and iap was inhibited by honokiol. Our results indicate that TNF-α production by RAW264.7 cells stimulated with L. monocytogenes supernatants was inhibited by honokiol. Based on these data, we propose that honokiol could be used as a promising natural compound against L. monocytogenes and its virulence factors.

Topics: Animals; Anti-Infective Agents; Bacterial Proteins; Bacterial Toxins; Biphenyl Compounds; Cell Line; Gene Expression Regulation, Bacterial; Heat-Shock Proteins; Hemolysin Proteins; Lignans; Listeria monocytogenes; Macrophages; Mice; Microbial Sensitivity Tests; Milk; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Tumor Necrosis Factor-alpha

Magnolol and honokiol, the bioactive phytochemicals contained in Magnolia officinalis, are uncommon antioxidants bearing isomeric bisphenol cores substituted with allyl functions. We have elucidated the chemistry behind their antioxidant activity by experimental and computational methods. In the inhibited autoxidation of cumene and styrene at 303 K, magnolol trapped four peroxyl radicals, with a kinh of 6.1 × 10(4) M(-1) s(-1) in chlorobenzene and 6.0 × 10(3) M(-1) s(-1) in acetonitrile, and honokiol trapped two peroxyl radicals in chlorobenzene (kinh = 3.8 × 10(4) M(-1) s(-1)) and four peroxyl radicals in acetonitrile (kinh = 9.5 × 10(3) M(-1) s(-1)). Their different behavior arises from a combination of intramolecular hydrogen bonding among the reactive OH groups (in magnolol) and of the OH groups with the aromatic and allyl π-systems, as confirmed by FT-IR spectroscopy and DFT calculations. Comparison with structurally related 3,3',5,5'-tetramethylbiphenyl-4,4'-diol, 2-allylphenol, and 2-allylanisole allowed us to exclude that the antioxidant behavior of magnolol and honokiol is due to the allyl groups. The reaction of the allyl group with a peroxyl radical (C-H hydrogen abstraction) proceeds with rate constant of 1.1 M(-1) s(-1) at 303 K. Magnolol and honokiol radicals do not react with molecular oxygen and produce no superoxide radical under the typical settings of inhibited autoxidations.

Topics: Acetonitriles; Anisoles; Antioxidants; Biphenyl Compounds; Hydrogen Bonding; Kinetics; Lignans; Molecular Structure; Oxidation-Reduction; Quantum Theory; Spectroscopy, Fourier Transform Infrared; Superoxides

The aims of the present study were to design polymeric micelles loading sirolimus with honokiol to increase drug solubility and to gain an insight into the effect of honokiol on oral transport of P-glycoprotein substrate (P-gp).. Particle size distribution, encapsulation efficiency, drug-loading content and in-vitro release of sirolimus-loaded micelles with honokiol were determined. Transport of sirolimus-loaded micelles across Caco-2 cell monolayers and jejunum segment of rats were investigated. In-vitro cytotoxicity experiments and the cellular uptake study were carried out via sulforhodamine B assay and flow cytometry, respectively.. A coadministration of honokiol with sirolimus in micelles did not significantly modify the particle size, polydispersity index and release of drugs demonstrating successful loading within the micelles. The apparent transport coefficients (Papp ) and effective permeability (Peff ) of sirolimus were increased with more amount of honokiol loaded in micelles. Cellular uptake study demonstrated that rhodamine123 uptake rate was enhanced by honokiol-loaded micelles, indicating substantial P-gp inhibition action by honokiol and mPEG-PLA-based micelles.. Oral transport of sirolimus was significantly improved by coadministration with honokiol, an inhibitor of the P-gp, in polymeric micelles formulation.

Topics: Administration, Oral; Animals; ATP Binding Cassette Transporter, Subfamily B; Biphenyl Compounds; Caco-2 Cells; Cell Survival; Chemistry, Pharmaceutical; Dose-Response Relationship, Drug; Drug Carriers; Drug Stability; Humans; Intestinal Absorption; Intestinal Mucosa; Jejunum; Kinetics; Lignans; Male; Micelles; Particle Size; Permeability; Polyesters; Polyethylene Glycols; Rats, Sprague-Dawley; Sirolimus; Solubility

Constitutive and inducible activation of signal transducer and activator of transcription 3 (STAT3) signaling facilitates the carcinogenesis in most human cancers including acute myeloid leukemia (AML). Negative regulators, such as protein tyrosine phosphatases SHP1, inhibit the activated STAT3 signaling. In this study, we investigated the effect of honokiol (HNK), a constituent of Magnolia officinalis, on the STAT3 signaling. STAT3 signaling and SHP1 expression were measured by quantitative real-time PCR and western blotting in leukemic cell lines and primary AML blasts treated with HNK. HNK decreased the phosphorylated STAT3 but not the total STAT3 through increasing the expression of SHP1. In addition, HNK inhibited transcription activity of STAT3, reduced nuclear translocation of STAT3, and decreased the expression of STAT3 target genes. Knockdown of SHP1 by small hairpin RNA (shRNA) or treatment with vanadate, a protein tyrosine phosphatases inhibitor, abolished HNK-induced STAT3 inhibition, suggesting that SHP1 plays an important role in the inhibition of STAT3 signaling by HNK. Further, HNK increased the expression of transcript factor PU.1, which had been reported to activate the expression of SHP1 via binding SHP1 promoter region. Knockdown of PU.1 reversed HNK-induced upregulation of SHP1 and inactivation of STAT3 signaling. Finally, HNK increased the expression of PU.1 and SHP1 in hematopoietic progenitors isolated from patients with AML. In conclusion, our data have shown a regulatory mechanism underlying the inhibition of STAT3 signaling by HNK. Therefore, as a relative non-toxic compound, HNK may offer a therapeutic advantage in the clinical treatment for AML.

Topics: Adult; Aged; Biphenyl Compounds; Blast Crisis; Cell Line, Tumor; Female; Gene Knockdown Techniques; Humans; Leukemia, Myeloid, Acute; Lignans; Male; Middle Aged; Phosphorylation; Protein Tyrosine Phosphatase, Non-Receptor Type 6; Proto-Oncogene Proteins; Real-Time Polymerase Chain Reaction; Signal Transduction; STAT3 Transcription Factor; Trans-Activators; Up-Regulation; Vanadates

The oncoprotein EZH2, as a histone H3K27 methyltransferase, is frequently overexpressed in various cancer types. However, the mechanisms underlying its role in urinary bladder cancer (UBC) cells have not yet fully understood. Herein, we reported that honokiol, a biologically active biphenolic compound isolated from the Magnolia officinalis inhibited human UBC cell proliferation, survival, cancer stemness, migration, and invasion, through downregulation of EZH2 expression level, along with the reductions of MMP9, CD44, Sox2 and the induction of tumor suppressor miR-143. Either EZH2 overexpression or miR-143 inhibition could partially reverse honokiol-induced cell growth arrest and impaired clonogenicity. Importantly, it was first revealed that EZH2 could directly bind to the transcriptional regulatory region of miR-143 and repress its expression. Furthermore, honokiol treatment on T24 tumor xenografts confirmed its anticancer effects in vivo, including suppression tumor growth and tumor stemness, accompanied by the dysregulation of EZH2 and miR-143 expressions. Our data suggest a promising therapeutic option to develop drugs targeting EZH2/miR-143 axis, such as honokiol, for bladder cancer treatment.

Topics: Animals; Antineoplastic Agents, Phytogenic; Binding Sites; Biphenyl Compounds; Cell Line, Tumor; Cell Movement; Cell Proliferation; Dose-Response Relationship, Drug; Down-Regulation; Enhancer of Zeste Homolog 2 Protein; Gene Expression Regulation, Neoplastic; Humans; Hyaluronan Receptors; Lignans; Male; Matrix Metalloproteinase 9; Mice, Nude; MicroRNAs; Neoplasm Invasiveness; Neoplastic Stem Cells; Polycomb Repressive Complex 2; Promoter Regions, Genetic; Signal Transduction; SOXB1 Transcription Factors; Time Factors; Transcription, Genetic; Tumor Burden; Urinary Bladder Neoplasms; Xenograft Model Antitumor Assays

This study examined the short- and long-term neuroprotective and analgesic activity of honokiol (a naturally occurring lignan isolated from Magnolia) on developing brains in neonates exposed to inflammatory pain, known to cause neuronal cell death. Postnatal day 4 (P4) neonatal rat pups were subjected to intraplantar formalin injection to four paws as a model of severe neonatal pain. Intraperitoneal honokiol (10 mg/kg) or corn oil vehicle control was administered 1 h prior to formalin insult, and animals were maintained on honokiol through postnatal day 21 (P21). Behavioral tests for stress and pain were performed after the painful insult, followed by morphological examinations of the brain sections at P7 and P21. Honokiol significantly attenuated acute pain responses 30 min following formalin insult and decreased chronic thermal hyperalgesia later in life. Honokiol-treated rats performed better on tests of exploratory behavior and performed significantly better in tests of memory. Honokiol treatment normalized hippocampal and thalamic c-Fos and hippocampal alveus substance P receptor expression relative to controls at P21. Together, these findings support that (1) neonatal pain experiences predispose rats to the development of chronic behavioral changes and (2) honokiol prevents and reduces both acute and chronic pathological pain-induced deteriorations in neonatal rats.

Topics: Analgesics; Animals; Biphenyl Compounds; Formaldehyde; Lignans; Magnolia; Male; Molecular Structure; Neuroprotective Agents; Pain; Rats

To improve the water solubility and biological activity of neoligans (magnolol and honokiol) and test the antitumor activity of the modified compounds.. The glycosylated products of magnolol and honokiol were obtained by enzymatic synthesis using a UDP-glycosyltransferase (YjiC) from Bacillus. The products were characterized by high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), and nuclear magnetic resonance (NMR) analysis. MTT assay was used to detect the growth inhibition of 4 human cancer cell lines induced by the compounds.. We obtained two glucosides of neolignans (magnolol and honokiol) for the first time by enzymatic synthesis using a UDP-glycosyltransferase. Based on the spectroscopic data, the glucosides were identified as magnolol-2- O-β-D-glucopyranoside (1) and honokiol-4'-O-β-D-glucopyranoside (2). Compounds 1-4 exhibited moderate anti-proliferative activities against the 4 human cancer cell lines, with IC50 values ranging from 9.41 to 111.21 µmol/L.. The glycoslated products show enhanced water solubility and drug sensitivity against SMMC7721 cells, suggesting their value as potential therapeutic drugs.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Line, Tumor; Chromatography, High Pressure Liquid; Glucosides; Glycosylation; Humans; Lignans; Magnetic Resonance Spectroscopy; Mass Spectrometry

Methicillin-resistant Staphylococcus aureus (MRSA) is a problematic pathogen posing a serious therapeutic challenge in the clinic. It is often multidrug-resistant (MDR) to conventional classes of antibacterial agents and there is an urgent need to develop new agents or strategies for treatment. Magnolol (ML) and honokiol (HL) are two naturally occurring diallylbiphenols which have been reported to show inhibition of MRSA. In this study their synergistic effects with antibacterial agents were further evaluated via checkerboard and time-kill assays.. The susceptibility spectrum of clinical MRSA strains was tested by the disk diffusion method. The minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs) of ML and HL were assayed by broth microdilution. The synergy was evaluated through checkerboard microdilution and time-killing experiments.. ML and HL showed similar activity against both MSSA and MRSA with MIC/MBC at 16 ~ 64 mg/L, with potency similar to amikacin (AMK) and gentamicin (GEN). When they were used in combination with conventional antibacterial agents, they showed bacteriostatic synergy with FICIs between 0.25 ~ 0.5, leading to the combined MICs decreasing to as low as 1 ~ 2 and 1 ~ 16 mg/L for ML (HL) and the agents, respectively. MIC50 of the combinations decreased from 16 mg/L to 1 ~ 4 mg/L for ML (HL) and 8 ~ 128 mg/L to 2 ~ 64 mg/L for the antibacterial agents, which exhibited a broad spectrum of synergistic action with aminoglycosides (AMK, etilmicin (ETM) and GEN), floroquinolones (levofloxacin (LEV), ciprofloxacin and norfloxacin), fosfomycin (FOS) and piperacillin. The times of dilution (TOD, the extent of decreasing in MIC value) were determined up to 16 for the combined MIC. A more significant synergy after combining was determined as ML (HL) with AMK, ETM, GEN and FOS. ML (HL) combined with antibacterial agents did not show antagonistic effects on any of the ten MRSA strains. Reversal effects of MRSA resistance to AMK and GEN by ML and HL were also observed, respectively. All the combinations also showed better dynamic bactericidal activity against MRSA than any of single ML (HL) or the agents at 24 h incubation. The more significant synergy of combinations were determined as HL (ML) + ETM, HL + LEV and HL + AMK (GEN or FOS), with △LC24 of 2.02 ~ 2.25.. ML and HL showed synergistic potentiation of antibacterial agents against clinical isolates of MRSA and warrant further pharmacological investigation.

Topics: Anti-Bacterial Agents; Biphenyl Compounds; Drug Synergism; Lignans; Methicillin; Methicillin-Resistant Staphylococcus aureus; Microbial Sensitivity Tests

Honokiol is a potential candidate for the treatment of intervertebral disc (IVD) degeneration. In this study, we develop in vitro and in vivo methods to detect the distribution of honokiol in intervertebral discs using high-performance liquid chromatography. A rat tail disc was used for both experimental models. For the in vivo animal experiment, blood samples and tail discs were collected at 15, 30, 60, 120 and 240 min after honokiol administration (30 mg/kg, i.v.). The analyte was separated by a mobile phase of methanol and 10 mM NaH2PO4 buffer at pH 2.8 (78:22, v/v) and pumped through a reversed-phase analytical column (250 × 4.6 mm, particle size 5 μm) at room temperature. The in vitro experimental results demonstrated that honokiol diffused into the intervertebral disc and was concentration-dependent. The active concentration is obtained for the therapeutic level at 15 and 30 min after honokiol administration in the in vivo model.

Topics: Acetophenones; Animals; Anti-Inflammatory Agents; Biphenyl Compounds; Chromatography, High Pressure Liquid; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; In Vitro Techniques; Injections, Intravenous; Intervertebral Disc; Lignans; Male; Molecular Structure; Permeability; Rats, Sprague-Dawley; Time Factors; Tissue Distribution

To study the quality suitability rank dividing of Magnolia officinalis on the basis of investigation on the correlation between the ratio of magnolol and honokiol in Magnoliae Officinalis Cortex and ecological factors, in order to provide scientific basis for its planting area of high-quality medicinal materials.. Based on the samples' quality analysis of 43 sampling points of Magnolia officinalis,the relationship between the ratio of magnolol and honokiol in Magnoliae officinalis Cortex and ecological factors was analyzed by statistical analysis. The geographic information system(GIS) was applied to assess the quality suitability rank dividing of Magnolia officinalis in China.. There were 12 ecological factors mainly affecting the quality of Magnoliae Officinalis Cortex; The ratio of magnolol and honokiol had obvious characteristics of regional quality. Conclusion: Magnoliae Officinalis Cortex which produced in Hubei and Chongqing is dao-di herbs.

Topics: Biphenyl Compounds; China; Geographic Information Systems; Lignans; Magnolia

Honokiol is one of the main bioactive constituents of the traditional Chinese herbal drug Magnolia bark (Cortex Magnoliae officinalis, Hou Po). The aim of this study was to probe its anti-type 2 diabetes mellitus effects and the underlying mechanism.. Type 2 diabetic mouse model was established by intraperitoneally injecting with streptozotocin. Fasting blood glucose, body weight, and lipid profile were measured. The subcutaneous adipose tissue, skeletal muscle, and liver were isolated as well as homogenized. The phospho-insulin receptor β-subunit (IRβ), IRβ, phospho-AKT, AKT, phospho-ERK1/2, ERK1/2, phosphotyrosine, and actin were examined by Western blot assay. Cell viability or cytotoxicity was analyzed by using MTT method. The inhibitory potencies of honokiol on the protein tyrosine phosphatase 1B (PTP1B) activity were performed in reaction buffer. Molecular docking and dynamic simulation were also analyzed.. In in vivo studies, oral treatment with 200 mg/kg honokiol for 8 weeks significantly decreases the fasting blood glucose in type 2 diabetes mellitus mice. The phosphorylations of the IRβ and the downstream insulin signaling factors including AKT and ERK1/2 significantly increase in adipose, skeletal muscle, and liver tissue of the honokiol-treated mice. Moreover, honokiol enhanced the insulin-stimulated phosphorylations of IRβ, AKT, and ERK1/2 in a dose-dependent manner in C2C12 myotube cells. Meanwhile, honokiol enhanced insulin-stimulated GLUT4 translocation. Importantly, honokiol exhibited reversible competitive inhibitory activity against PTP1B with good selectivity in vitro and in vivo. Furthermore, using molecular docking and dynamic simulation approaches, we determined the potential binding mode of honokiol to PTP1B at an atomic level.. These findings indicated the hypoglycemic effects of honokiol and its mechanism that honokiol improved the insulin sensitivity by targeting PTP1B. Therefore, our study may highlight honokiol as a promising insulin sensitizer for the therapy of type 2 diabetes.

Topics: Administration, Oral; Animals; Biphenyl Compounds; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Disease Models, Animal; Dose-Response Relationship, Drug; Enzyme Inhibitors; Hypoglycemic Agents; Injections, Intraperitoneal; Lignans; Mice; Molecular Docking Simulation; Molecular Dynamics Simulation; Protein Tyrosine Phosphatase, Non-Receptor Type 1; Streptozocin; Structure-Activity Relationship

This study aims to investigate the effects of honokiol on proliferation, cell cycle, and apoptosis in tumor necrosis factor (TNF)-α-induced rat aortic smooth muscle cells (RASMCs). We found that honokiol treatment showed potent inhibitory effects on TNF-α-induced RASMC proliferation, which were associated with G0/G1 cell cycle arrest and downregulation of cell cycle-related proteins, including cyclin D1, cyclin E, cyclin-dependent kinase (CDK)2 and CDK4. Furthermore, honokiol treatment led to the release of cytochrome c into cytosol and a loss of mitochondrial membrane potential (ΔΨm), as well as a decrease in the expression of Bcl-2 and an increase in the expression of Bax. Treatment with honokiol also reduced TNF-α-induced phosphorylation of p38, extracellular signal-regulated kinase 1/2, and c-Jun N-terminal kinase. Taken together, our results suggest that honokiol suppresses TNF-α-stimulated RASMC proliferation via caspase- and mitochondria-dependent apoptosis and highlight the therapeutic potential of honokiol in the prevention of cardiovascular diseases.

Topics: Animals; Aorta; Apoptosis; bcl-2-Associated X Protein; Biphenyl Compounds; Caspases; Cell Proliferation; Cell Survival; Cells, Cultured; Cyclin D1; Cyclin E; Cyclin-Dependent Kinase 2; Cyclin-Dependent Kinase 4; Cytochromes c; Drugs, Chinese Herbal; Enzyme Inhibitors; Extracellular Signal-Regulated MAP Kinases; G1 Phase Cell Cycle Checkpoints; JNK Mitogen-Activated Protein Kinases; Lignans; Male; Membrane Potential, Mitochondrial; Mitochondria; Muscle, Smooth, Vascular; Nitric Oxide Synthase; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Proto-Oncogene Proteins c-bcl-2; Rats; Rats, Sprague-Dawley; Signal Transduction; Tumor Necrosis Factor-alpha

Glioblastoma is one of the most lethal and prevalent malignant human brain tumors, with aggressive proliferation and highly invasive properties. There is still no effective cure for patients with glioblastoma. Honokiol, derived from Magnolia officinalis, can cross the blood-brain barrier (BBB) and the blood-cerebrospinal fluid barrier (BCSFB), making it a strong candidate for an effective drug for the treatment of brain tumors, including glioblastoma. In our previous study, we demonstrated that honokiol effectively induced apoptotic cell death in glioblastoma. Metastasis poses the largest problem to cancer treatment and is the primary cause of death in cancer patients. Thus, in this study, we investigated the effect of honokiol on the cell invasion process of U87MG human glioblastoma cells through brain microvascular endothelial cells (BMECs) and its possible mechanisms. Honokiol dose-dependently inhibited TNF-α-induced VCAM-1 expression in BMECs and adhesion of U87MG to BMECs. Moreover, honokiol effectively blocked U87MG invasion through BMEC-Matrigel-coated transwell membranes. Increased phosphorylation of VE-cadherin and membrane permeability by TNF-α were suppressed by honokiol in BMECs. Furthermore, we investigated the effect of honokiol on the epithelial-mesenchymal transition (EMT) in U87MG cells. Honokiol reduced the expression levels of Snail, N-cadherin and β-catenin, which are mesenchymal markers, but increased E-cadherin, an epithelial marker. In conclusion, these results suggest that honokiol inhibits metastasis by targeting the interaction between U87MG and BMECs, regulating the adhesion of U87MG to BMECs by inhibiting VCAM-1, and regulating the invasion of U87MG through BMECs by reducing membrane permeability and EMT processes of U87MG cells.

Topics: Apoptosis; Biphenyl Compounds; Brain Neoplasms; Cell Line, Tumor; Cell Membrane Permeability; Endothelial Cells; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Glioblastoma; Humans; Lignans; Neoplasm Invasiveness; Vascular Cell Adhesion Molecule-1

With the goal of developing soluble epoxide hydrolase (sEH) inhibitors with novel chemical structures, the sEH inhibitory activities of 30 natural compounds were evaluated using both a fluorescent substrate, 3-phenyl-cyano(6-methoxy-2-naphthalenyl)methyl ester- 2-oxiraneacetic acid, and a physiological substrate, 14,15-epoxyeicosatrienoic acid. To evaluate the selectivity of sEH inhibition, the inhibition of microsomal epoxide hydrolase (mEH), which plays a critical role in detoxification of toxic epoxides, was determined using human liver microsomes. Honokiol and β-amyrin acetate, isolated from Magnolia officinalis and Acer mandshuricum, respectively, displayed strong inhibition of sEH activity, with respective IC50 values of 0.57 μM and 3.4 μM determined using the fluorescent substrate, and 1.7 μM and 6.1 μM determined using 14,15-epoxyeicosatrienoic acid. mEH activity was decreased to 49% or 61% of control activity by 25 μM honokiol or β-amyrin acetate, respectively. These results suggest that β-amyrin acetate and honokiol exhibit sEH inhibitory activity, although their sEH selectivity should be improved.

Topics: Biological Products; Biphenyl Compounds; Chromatography, High Pressure Liquid; Enzyme Inhibitors; Epoxide Hydrolases; Fluorometry; Lignans; Oleanolic Acid; Spectrophotometry, Ultraviolet; Tandem Mass Spectrometry

We have shown previously that honokiol (HNK), a bioactive component of the medicinal plant Magnolia officinalis, inhibits growth of human prostate cancer cells in vitro and in vivo. However, the effect of HNK on androgen receptor (AR) signaling has not been studied.. LNCaP, C4-2, and TRAMP-C1 cells were used for various assays. Trypan blue dye exclusion assay or clonogenic assay was performed for determination of cell viability. The effects of HNK and/or its analogs on protein levels of AR and its target gene product prostate specific antigen (PSA) were determined by western blotting. RNA interference of p53 was achieved by transient transfection. Reverse transcription-polymerase chain reaction was performed for mRNA expression of AR. Nuclear level of AR was visualized by microscopy. Apoptosis was quantified by DNA fragmentation assay or flow cytometry after Annexin V-propidium iodide staining.. HNK and its dichloroacetate analog (HDCA) were relatively more effective in suppressing cell viability and AR protein level than honokiol epoxide or biseugenol. Nuclear translocation of AR stimulated by a synthetic androgen (R1881) was markedly suppressed in the presence of HNK. Downregulation of AR protein resulting from HNK exposure was attributable to transcriptional repression as well as proteasomal degradation. HNK-mediated suppression of AR protein was maintained in LNCaP cells after knockdown of p53 protein. HNK-induced apoptosis was not affected by R1881 treatment.. The present study demonstrates, for the first time, that HNK inhibits activity of AR in prostate cancer cells regardless of the p53 status.

Topics: Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Survival; Down-Regulation; Humans; Lignans; Male; Prostatic Neoplasms; Receptors, Androgen; Signal Transduction

Ten honokiol oligomers (1-10), including four novel trimers (1-4) and four novel dimers (5-8), were obtained by means of biotransformation of honokiol catalyzed by Momordica charantia peroxidase (MCP) for the first time. Their structures were established on the basis of spectroscopic methods. The biological results demonstrated that most of the oligomers were capable of inhibiting α-glucosidase with significant abilities, which were one to two orders of magnitude more potent than the substrate, honokiol. In particular, compound 2, the honokiol trimer, displayed the greatest inhibitory activity against α-glucosidase with an IC50 value of 1.38μM. Kinetic and CD studies indicated that 2 inhibited α-glucosidase in a reversible, mixed-type manner and caused conformational changes in the secondary structure of the enzyme protein. These findings suggested that 2 might be exploited as a promising drug candidate for the treatment of diabetes.

Topics: alpha-Glucosidases; Biocatalysis; Biphenyl Compounds; Dimerization; Dose-Response Relationship, Drug; Enzyme Inhibitors; Glycoside Hydrolase Inhibitors; Lignans; Molecular Structure; Momordica charantia; Peroxidase; Saccharomyces cerevisiae; Structure-Activity Relationship

Honokiol, a small-molecule polyphenol derived and isolated from the Chinese medicinal herb Magnolia officinalis, has been shown to possess a wide range of pharmacological activities. In the present study, we aimed to investigate the effects of honokiol on tumor necrosis factor-α (TNF-α)-induced migration in rat aortic smooth muscle cells (RASMCs). We found that honokiol inhibited TNF-α-induced RASMC proliferation and migration in a dose-dependent manner. At the molecular level, pretreatment with honokiol blocked TNF-α-induced protein expression of matrix metalloproteinase (MMP)-2 and MMP-9, nuclear factor (NF)-κB activation, and extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation. Moreover, NF-κB inhibitor (BAY 11-7028) and ERK inhibitor (U0126) also mimicked the inhibitory effects of honokiol in TNF-α-treated RASMCs. In conclusion, these results indicate that honokiol suppresses TNF-α-induced migration and MMP expression by blocking NF-κB activation via the ERK signaling pathway in RASMCs. Our findings support honokiol as a promising novel agent for the prevention and treatment of atherosclerosis.

Topics: Animals; Aorta; Biphenyl Compounds; Lignans; Male; MAP Kinase Signaling System; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; NF-kappa B; Rats, Sprague-Dawley; Signal Transduction; Tumor Necrosis Factor-alpha

Proinflammatory cytokine interleukin-1β (IL-1β) stimulates several mediators of cartilage degradation and plays an important role in the pathogenesis of osteoarthritis (OA). Honokiol, a low molecular weight natural product isolated from the Magnolia officinalis, has been shown to possess anti-inflammatory effect. Here, we used an in vitro model of cartilage inflammation to investigate the therapeutic potential of honokiol in OA. Human OA chondrocytes were cultured and pretreated with honokiol (2.5-10 µM) with or without IL-1β (10 ng/ml). Nitric oxide (NO) production was quantified by Griess reagent. Prostaglandin (PG)E2 , metalloproteinase-13 (MMP-13), and interleukin-6 (IL-6) productions were quantified by enzyme-linked immunosorbent assay. The expressions of collagen II, cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and nuclear factor κB (NF-κB)-related signaling molecules were determined by Western blotting. Our data showed that IL-1β markedly stimulated the expressions of iNOS and COX-2 and the productions of NO, PGE2 , and IL-6, which could be significantly reversed by honokiol. Honokiol could also suppress the IL-1β-triggered activation of IKK/IκBα/NF-κB signaling pathway. Moreover, honokiol significantly inhibited the IL-1β-induced MMP-13 production and collagen II reduction. Taken together, the present study suggests that honokiol may have a chondroprotective effect and may be a potential therapeutic choice in the treatment of OA patients.

Topics: Aged; Anti-Inflammatory Agents; Biphenyl Compounds; Cell Survival; Chondrocytes; Collagen Type II; Drugs, Chinese Herbal; Humans; I-kappa B Kinase; Lignans; Magnolia; Matrix Metalloproteinase 13; Middle Aged; NF-kappa B; Osteoarthritis; Signal Transduction

Cancer prevention remains a high priority for the scientific world. Magnolia dealbata Zucc (Magnoliaceae), a Mexican endemic species, is used for the empirical treatment of cancer.. To evaluate the cytotoxic and cancer chemopreventive effects of an ethanol extract of Magnolia dealbata seeds (MDE).. The cytotoxic effect of MDE, at concentrations ranging from 1 to 200 µg/ml, on human cancer cells and human nontumorigenic cells was evaluated using the MTT assay for 48 h. The apoptotic activities of MDE 25 μg/ml on MDA-MB231 breast cancer cells were evaluated using the TUNEL assay and the detection of caspase 3 using immunofluorescence analysis for 48 h, each. The chemopreventive effect was evaluated by administrating different doses of MDE, between 1 and 50 mg/kg, injected intraperitoneally daily into athymic mice which were implanted with MDA-MB231 cells during 28 days. The growth and weight of tumors were measured.. MDE showed cytotoxic effects on MDA-MB231 cells (IC₅₀ = 25 µg/ml) and exerted pro-apoptotic activities as determined by DNA fragmentation in MDA-MB231 cells. MDE 25 µg/ml also induces the activation of caspase 3 in MDA-MB231 cells. These results suggest that Magnolia dealbata may be an optimal source of the bioactive compounds: honokiol (HK) and magnolol (MG). MDE 50 mg/kg i.p. exerted chemopreventive effects by inhibiting the growth of MDA-MB231 tumor by 75% in athymic mice, compared to the control group.. MDE exerts cytotoxic, apoptotic and chemopreventive activities on MDA-MB231 human cancer cells.

Topics: Animals; Anticarcinogenic Agents; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Dose-Response Relationship, Drug; Humans; In Situ Nick-End Labeling; Inhibitory Concentration 50; Lignans; Magnolia; Mammary Neoplasms, Experimental; Mice, Nude; Seeds

Biphenyl neolignans such as honokiol and magnolol, which are the major active constituents of the Asian medicinal plant Magnolia officinalis, are known to exert a multitude of pharmacological and biological activities. Among these, cytotoxic and tumor growth inhibitory activity against various tumour cell lines are well-documented. To further elucidate the cytotoxic effects of honokiol derivatives, derivatizations were performed using tetrahydrohonokiol as a scaffold. The derivatizations comprised the introduction of functional groups, e.g., nitro and amino groups, as well as alkylation. This way, 18 derivatives, of which 13 were previously undescribed compounds, were evaluated against CCRF-CEM leukemia cells, U251 glioblastoma and HCT-116 colon cancer cells. The results revealed no significant cytotoxic effects in any of the three tested cell lines at a test concentration of 10 µM.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Survival; Colonic Neoplasms; Drug Screening Assays, Antitumor; Glioblastoma; HCT116 Cells; Humans; Inhibitory Concentration 50; Leukemia; Lignans; Methylation; Microwaves

Honokiol is a phytochemical component with multiple pharmacological activities, but Honokiol's wider use has been restricted by its poor solubility. Using bovine serum albumin and polyvinylpyrrolidone as stabilisers in a solvent precipitation-ultrasonication method, Honokiol nanosuspensions were prepared with a mean particle size of 116.2 nm (±2 nm), a zeta potential of -44.7 mV (±1.7 mV) and a high drug payload of 50.4 ± 0.6% (w/w). X-ray powder diffraction and differential scanning calorimetry indicated that Honokiol was in an amorphous state in the nanosuspensions, in contrast with bulk Honokiol powder. Honokiol was released faster in vitro from nanosuspensions with no burst release, and the highest 98% cumulative release was after 60 h. Honokiol nanosuspensions improved the oral bioavailability of Honokiol in in vivo studies in rats with a 3.94-fold Cmax and a 2.2-fold AUC(0-t). Remarkably, in contrast to oral administration, intraperitoneal administration of Honokiol nanosuspensions could dramatically alter the biodistribution of Honokiol, resulting in a much higher drug level and tissue bioavailability in the blood, heart and brain, benefitting the treatment of cardio-cerebro-vascular diseases.

Topics: Administration, Oral; Animals; Biological Availability; Biphenyl Compounds; Calorimetry, Differential Scanning; Cell Line, Tumor; Cerebrovascular Circulation; Coronary Circulation; Kidney; Lignans; Liver; Lung; Male; Mice; Mice, Inbred ICR; Nanoparticles; Particle Size; Rats; Rats, Sprague-Dawley; Surface Properties; Suspensions

To investigate the inhibiting effect of Honokiol (HNK) on the invasion and angiogenesis in U937 leukemia cells and the molecular mechanism.. After treated with different concentrations of HNK, the growth inhibition rate of U937 cells was determined by MTT assay, and for the adhesion and invasion abilities were assessed using cell matrix adhesion technique and Transwell(TM); assay, respectively. VEGF, VEGFR1 and MMP-9 mRNA expression levels were detected by real-time quantitative RT-PCR (qRT-PCR). VEGF protein levels were determined by ELISA.. HNK could significantly inhibit the proliferation of U937 cells in a time- and dose-dependent manner. The adhesion and invasion abilities of U937 cells were suppressed after treated with a low concentration of HNK. The expressions of VEGF, VEGFR1 and MMP-9 were down-regulated by HNK in a dose-dependent manner.. HNK can inhibit the invasion and angiogenesis of U937 cells via down-regulating VEGF, VEGFR1 and MMP-9 expressions.

Topics: Antineoplastic Agents; Biphenyl Compounds; Cell Adhesion; Cell Line, Tumor; Cell Proliferation; Dose-Response Relationship, Drug; Gene Expression Regulation, Neoplastic; Humans; Leukemia; Lignans; Matrix Metalloproteinase 9; Neoplasm Invasiveness; Neovascularization, Pathologic; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-1

Epithelial-mesenchymal transition (EMT), a critical step in the acquisition of metastatic state, is an attractive target for therapeutic interventions directed against tumor metastasis. Honokiol (HNK) is a natural phenolic compound isolated from an extract of seed cones from Magnolia grandiflora. Recent studies from our lab show that HNK impedes breast carcinogenesis. Here, we provide molecular evidence that HNK inhibits EMT in breast cancer cells resulting in significant downregulation of mesenchymal marker proteins and concurrent upregulation of epithelial markers. Experimental EMT induced by exposure to TGFβ and TNFα in spontaneously immortalized nontumorigenic human mammary epithelial cells is also completely reversed by HNK as evidenced by morphological as well as molecular changes. Investigating the downstream mediator(s) that may direct EMT inhibition by HNK, we found functional interactions between HNK, Stat3, and EMT-signaling components. In vitro and in vivo analyses show that HNK inhibits Stat3 activation in breast cancer cells and tumors. Constitutive activation of Stat3 abrogates HNK-mediated activation of epithelial markers whereas inhibition of Stat3 using small molecule inhibitor, Stattic, potentiates HNK-mediated inhibition of EMT markers, invasion and migration of breast cancer cells. Mechanistically, HNK inhibits recruitment of Stat3 on mesenchymal transcription factor Zeb1 promoter resulting in decreased Zeb1 expression and nuclear translocation. We also discover that HNK increases E-cadherin expression via Stat3-mediated release of Zeb1 from E-cadherin promoter. Collectively, this study reports that HNK effectively inhibits EMT in breast cancer cells and provide evidence for a previously unrecognized cross-talk between HNK and Stat3/Zeb1/E-cadherin axis.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Breast; Breast Neoplasms; Cadherins; Cell Line, Tumor; Epithelial-Mesenchymal Transition; Female; Homeodomain Proteins; Humans; Lignans; Magnolia; Signal Transduction; STAT3 Transcription Factor; Transcription Factors; Zinc Finger E-box-Binding Homeobox 1

Acute kidney injury (AKI) is a severe complication of sepsis, which largely contributes to the high mortality rate of sepsis. Honokiol, a natural product isolated from Magnolia officinalis (Houpo), has been shown to exhibit anti-inflammatory and antioxidant properties. Here, we investigated the effects of honokiol on sepsis-associated AKI in rats subjected to cecal ligation and puncture (CLP). We found that the administration of honokiol improved the survival of septic rats. Periodic acid-Schiff stain revealed that the morphological changes of kidney tissues in CLP rats were restored after honokiol treatment. Furthermore, honokiol reduced CLP-induced oxidative stress and inflammatory cytokine production. The levels of nitric oxide (NO) and inducible NO synthetase (iNOS) were attenuated by honokiol in septic rats. Finally, honokiol inhibited CLP-induced activation of NF-κB signaling in CLP rats. Our findings suggest that honokiol might be used as a potential therapeutic agent for complications of sepsis, especially for sepsis-induced AKI.

Topics: Acute Kidney Injury; Animals; Antioxidants; Biphenyl Compounds; Cytokines; Disease Models, Animal; Inflammation; Kidney; Lignans; Male; NF-kappa B; Nitric Oxide; Nitric Oxide Synthase Type II; Oxidative Stress; Rats; Rats, Sprague-Dawley; Sepsis; Signal Transduction

As fecal excretion is one of important routes of elimination of drugs and their metabolites, it is indispensable to investigate the metabolites in feces for more comprehensive information on biotransformation in vivo. In this study, a sensitive and reliable approach based on ultra-performance liquid chromatography/quadrupole-time-of-flight-mass spectrometry (UHPLC-Q-TOF-MS) was applied to characterize the metabolic profile of honokiol in rat feces after the administration of an equimolar mixture of honokiol and [(13)C6]-labeled honokiol. Totally 42 metabolites were discovered and tentatively identified in rat feces samples, 26 metabolites were first reported, including two novel classes of metabolites, methylated and dimeric metabolites of honokiol. Moreover, this study provided basic comparative data on the metabolites in rat plasma, feces and urine, which gave better understanding of the metabolic fate of honokiol in vivo.

Topics: Animals; Biomarkers; Biphenyl Compounds; Carbon Isotopes; Chromatography, Liquid; Feces; Isotope Labeling; Lignans; Male; Metabolome; Rats; Rats, Sprague-Dawley; Tandem Mass Spectrometry

Resistance to chemotherapy remains a major obstacle in cancer therapy. This study aimed to evaluate the molecular mechanism and efficacy of honokiol in inducing apoptosis and enhancing paclitaxel chemotherapy in pre-clinical multi-drug resistant (MDR) cancer models, including lineage-derived human MDR (KB-8-5, KB-C1, KB-V1) and their parental drug sensitive KB-3-1 cancer cell lines. In vitro analyses demonstrated that honokiol effectively inhibited proliferation in KB-3-1 cells and the MDR derivatives (IC50 ranging 3.35 ± 0.13 µg/ml to 2.77 ± 0.22 µg/ml), despite their significant differences in response to paclitaxel (IC50 ranging 1.66 ± 0.09 ng/ml to 6560.9 ± 439.52 ng/ml). Honokiol induced mitochondria-dependent and death receptor-mediated apoptosis in MDR KB cells, which was associated with inhibition of EGFR-STAT3 signaling and downregulation of STAT3 target genes. Combined treatment with honokiol and paclitaxel synergistically augmented cytotoxicity in MDR KB cells, compared with treatment with either agent alone in vitro. Importantly, the combined treatment significantly inhibited in vivo growth of KB-8-5 tumors in a subcutaneous model. Tumor tissues from the combination group displayed a significant inhibition of Ki-67 expression and an increase in TUNEL-positive cells compared with the control group. These results suggest that targeting multidrug resistance using honokiol in combination with chemotherapy drugs may provide novel therapeutic opportunities.

Topics: Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Down-Regulation; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Drug Synergism; ErbB Receptors; Humans; Ki-67 Antigen; Lignans; Mitochondria; Neoplasms; Paclitaxel; Receptors, Death Domain; Signal Transduction; STAT3 Transcription Factor

This study was aimed to investigate the effect of Honokiol (HNK) combined with Gemcitabine (GEM) on the proliferation and apoptosis of human Burkitt lymphoma Raji cells. Cell proliferation was detected by CCK-8 method to study the role of Honokiol and Gemcitabine in Raji cells. The cell apoptosis and cell cycle status were analyzed by flow cytometry. The level of apoptosis-related protein BCL-2 was measured with Western blot. The results showed that compared with cells treated with mentioned above drugs alone, the proliferative potential of cells in combination group was significantly inhibited (P < 0.01) and the inhibition rate was related to the concentration and action time of HNK; and apoptosis rate markedly increased (P < 0.01), while most Raji cells were arrested at G0/G1 phase and decreased in S phase after treatment with combination of two drugs; the expression of BCL-2 protein decreased (P < 0.01). It is concluded that Honokiol combined Gemcitabine can synergistically inhibit the proliferation, induce cell apoptosis, and down-regulate the expression of BCL-2 in Raji cells. The possible mechanism of synergistic effect may be related with arrest of cell cycle at G0/G1 phase and downregulation of the expression of BCL-2.

Topics: Apoptosis; Biphenyl Compounds; Burkitt Lymphoma; Cell Line, Tumor; Cell Proliferation; Deoxycytidine; Drug Synergism; Gemcitabine; Humans; Lignans; Proto-Oncogene Proteins c-bcl-2

This study was designed to investigate the effect of w007B, a newly synthesized derivative of honokiol, on MCAO reperfusion, and its therapeutic time window and related mechanisms in rats. Neurological deficit scores, infarct size and brain water content were measured after 24 h reperfusion following 2 h ischemia. The results showed that w007B (10 and 50 μg/kg, IV immediately after reperfusion) markedly decreased neurological deficit scores, reduced infarct size and alleviated brain water content, and then 50 μg/kg w007B given within 3 h after reperfusion (5 h after ischemia) significantly attenuated ischemia-induced brain injury. Additionally, no sign of toxicity was observed when a single dose of 50mg/kg w007B (1000 times of the highest effective dose, IP) was administered. To explore the underlying mechanisms, the expression level of apoptosis, inflammation and autophagy-related markers in brain tissue were detected with kits or by western blot. It was observed that w007B rapidly and significantly reduced caspase-3 activity and NO production in the injured semi-brain, and also lowered the level of the p65 subunit of NF-κB in the nucleus. Besides, it also reduced the expression of Beclin-1 and LC3B-II, and increased the level of p62, the autophagy-related proteins in I/R-injured hemisphere. In conclusion, w007B exerts neuroprotective effect on cerebral ischemia-reperfusion injury with wider therapeutic time window and better safety; its mechanisms may be associated with its anti-inflammation, anti-apoptosis and anti-autophagy action. These results suggest that w007B shows strong potential as a clinical neuroprotective candidate for the treatment of ischemic stroke.

Topics: Animals; Apoptosis; Autophagy; Biphenyl Compounds; Body Weight; Cerebrovascular Circulation; Disease Models, Animal; Female; Inflammation; Lignans; Male; Mice; Mice, Inbred ICR; Neuroprotective Agents; NF-kappa B; Nitric Oxide; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Sex Factors; Time Factors

In this paper, multiwalled-carbon-nanotube-based matrix solid-phase dispersion coupled to HPLC with diode array detection was used to extract and determine honokiol and magnolol from Magnoliae Cortex. The extraction efficiency of the multiwalled-carbon-nanotube-based matrix solid-phase dispersion was studied and optimized as a function of the amount of dispersing sorbent, volume of elution solvent, and flow rate of elution solvent, with the aid of response surface methodology. An amount of 0.06 g of carboxyl-modified multiwalled carbon nanotubes and 1.5 mL of methanol at a flow rate of 1.1 mL/min were selected. The method obtained good linearity (r(2) > 0.9992) and precision (RSD < 4.7%) for honokiol and magnolol, with limits of detection of 0.045 and 0.087 μg/mL, respectively. The recoveries obtained from analyzing in triplicate spiked samples were determined to be from 90.23 to 101.10% and the RSDs from 3.5 to 4.8%. The proposed method that required less samples and reagents was simpler and faster than Soxhlet and maceration extraction methods. The optimized method was applied for analyzing five real samples collected from different cultivated areas.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Lignans; Magnolia; Nanotubes, Carbon; Plant Extracts; Solid Phase Extraction

A simple, rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry method was developed and validated for the determination of honokiol in beagle dog plasma after intravenous guttae. With addition of the internal standard magnolol, plasma samples were precipitated with methanol and separated on a Shim-pack XR-ODS II (2.0 × 100 mm, 2.2 µm) with isocratic elution of methanol and water (80:20) solution at a flow rate of 0.2 mL/min. A good separation of honokiol was achieved within 3.5 min. Quantification was performed on a Waters Quattro Premier XE triple quadrupole mass spectrometer with electrospray ionization inlet in the negative multiple reaction monitoring mode. Good linearity was obtained over the concentration range of 5.12-15580 ng/mL (r(2) > 0.998). Intra- and inter-day precisions were <13.10%, and accuracy ranged from 89.21 to 99.92%. The lower limit of quantification for honokiol was 5.12 ng/mL, and honokiol was stable under various conditions (three freeze-thaw cycles, short-term temperature, post-preparative and long-term temperature conditions.). This validated method was successfully applied to the pharmacokinetic study of honokiol in dogs by intravenous guttae.

Topics: Administration, Intravenous; Animals; Biphenyl Compounds; Chromatography, High Pressure Liquid; Dogs; Female; Lignans; Linear Models; Male; Reproducibility of Results; Sensitivity and Specificity; Tandem Mass Spectrometry

Diarrhea is a big problem in piglets. Cangpo Oral Liquid (COL) is a compound of Chinese herbal medicine. The preparation was fed to piglets had diarrheal disease in order to determine its anti-diarrhea activity and potential applications in vivo.. The contents of Berberine hydrochloride, Magnolol and Honokiol in COL were performed on HPLC analysis. Organ bath was used to investigate the effect of COL on peristaltic reflexes and peristaltic waves in vitro. And anti-diarrhea activity of COL was evaluated in clinical.. Thin layer chromatography (TLC) and HPLC analyses showed that the contents of Berberine hydrochloride, Magnolol and Honokiol in COL were 970µg/mL, 130µg/mL and 300µg/mL, respectively. Administration of the COL to the organ bath caused a concentration-dependent inhibition of intestinal peristalsis. When the COL concentration in the bath was cumulatively increased, the amplitude and frequency of the peristaltic waves was lowered. The result of clinical efficacy of COL was very effective to diarrheic piglets. COL can possibly inhibit the curve of peristaltic waves in vitro; and clinical trial showed a statistically significant therapeutic effect in vivo.. In conclusion, COL can be used as an effective therapeutic agent. However, the ingredients, pharmacokinetics and specific signaling pathways of COL need to be further studied.

Topics: Animals; Antidiarrheals; Berberine; Biphenyl Compounds; Coptis; Diarrhea; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Female; Intestines; Lignans; Magnolia; Male; Peristalsis; Phytotherapy; Swine

Drug impurity profiling and identification are carried out along with the drug discovery process. Due to its inherent low concentration in drug products, the isolation and purification of impurities present a challenge to drug development processes. In our development of honokiol and quercetin as anticancer drug candidates, counter-current chromatography (CCC) and preparative HPLC were used for the impurity profiling and identification of honokiol and quercetin. Several performance parameters such as separation column volume, maximum sample loading, separation time, solvent consumption and sample throughput were investigated in order to compare the separation efficiency. We found that the sample loading capacity and therefore the throughput of preparative HPLC were not satisfactory, while CCC provided larger sample loading (especially for a sample with poor solubility), consumed less solvent and produced higher throughput than preparative HPLC. Six impurities of honokiol including one new compound were isolated in the present work.

Topics: Antineoplastic Agents; Biphenyl Compounds; Chromatography, High Pressure Liquid; Countercurrent Distribution; Drug Compounding; Drug Contamination; Lignans; Quercetin; Solvents

Renal cell carcinoma (RCC) is associated with a high frequency of metastasis and only few therapies substantially prolong survival. Honokiol, isolated from Magnolia spp. bark, has been shown to exhibit pleiotropic anticancer effects in many cancer types. However, whether honokiol could suppress RCC metastasis has not been fully elucidated. In the present study, we found that honokiol suppressed renal cancer cells' metastasis via dual-blocking epithelial-mesenchymal transition (EMT) and cancer stem cell (CSC) properties. In addition, honokiol inhibited tumor growth in vivo. It was found that honokiol could upregulate miR-141, which targeted ZEB2 and modulated ZEB2 expression. Honokiol reversed EMT and suppressed CSC properties partly through the miR-141/ZEB2 axis. Our study suggested that honokiol may be a suitable therapeutic strategy for RCC treatment.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Proliferation; Epithelial-Mesenchymal Transition; Homeodomain Proteins; Humans; Kidney Neoplasms; Lignans; Mice, Inbred BALB C; Mice, Nude; MicroRNAs; Neoplastic Stem Cells; Repressor Proteins; Signal Transduction; Tumor Burden; Zinc Finger E-box Binding Homeobox 2

Small ligands are a powerful way to control the function of protein complexes via dynamic binding interfaces. The classic example is found in gene transcription where small ligands regulate nuclear receptor binding to coactivator proteins via the dynamic activation function 2 (AF2) interface. Current ligands target the ligand-binding pocket side of the AF2. Few ligands are known, which selectively target the coactivator side of the AF2, or which can be selectively switched from one side of the interface to the other. We use NMR spectroscopy and modeling to identify a natural product, which targets the retinoid X receptor (RXR) at both sides of the AF2. We then use chemical synthesis, cellular screening and X-ray co-crystallography to split this dual activity, leading to a potent and molecularly efficient RXR agonist, and a first-of-kind inhibitor selective for the RXR/coactivator interaction. Our findings justify future exploration of natural products at dynamic protein interfaces.

Topics: Binding Sites; Biological Products; Biphenyl Compounds; Crystallography, X-Ray; Ligands; Lignans; Models, Biological; Receptors, Cytoplasmic and Nuclear; Retinoid X Receptors

Honokiol is a small biphenolic compound, which exerts antitumor activities; however, the precise mechanism of honokiol-induced apoptosis in the human colorectal cancer cells remains unclear. Here, we show that survivin and p53 display the opposite role on the regulation of honokiol-induced apoptosis in the human colorectal cancer cells. Honokiol induced the cell death and apoptosis in various colorectal cancer cell lines. Moreover, honokiol elicited the extrinsic death receptor pathway of DR5 and caspase 8 and the intrinsic pathway of caspase 9. The common intrinsic and extrinsic downstream targets of activated caspase 3 and PARP protein cleavage were induced by honokiol. Interestingly, honokiol reduced anti-apoptotic survivin protein and gene expression. Transfection with a green fluorescent protein (GFP)-survivin-expressed vector increased the colorectal cancer cell viability and resisted the honokiol-induced apoptosis. Meantime, honokiol increased total p53 and the phosphorylated p53 proteins at Ser15 and Ser46. The p53-wild type colorectal cancer cells were exhibited greater cytotoxicity, apoptosis and survivin reduction than the p53-null cancer cells after treatment with honokiol. Together, these findings demonstrate that the existence of survivin and p53 can modulate the honokiol-induced apoptosis in the human colorectal cancer cells.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Colorectal Neoplasms; Drug Resistance, Neoplasm; HCT116 Cells; Humans; Inhibitor of Apoptosis Proteins; Lignans; Signal Transduction; Survivin; Tumor Suppressor Protein p53

Magnolia bark preparations from Magnolia officinalis of Asian medicinal systems are known for their muscle relaxant effect and anticonvulsant activity. These CNS related effects are ascribed to the presence of the biphenyl-type neolignans honokiol and magnolol that exert a potentiating effect on GABAA receptors. 4-O-methylhonokiol isolated from seeds of the North-American M. grandiflora was compared to honokiol for its activity to potentiate GABAA receptors and its GABAA receptor subtype-specificity was established.. Different recombinant GABAA receptors were functionally expressed in Xenopus oocytes and electrophysiological techniques were used determine to their modulation by 4-O-methylhonokiol.. 3μM 4-O-methylhonokiol is shown here to potentiate responses of the α₁β₂γ₂ GABAA receptor about 20-fold stronger than the same concentration of honokiol. In the present study potentiation by 4-O-methylhonokiol is also detailed for 12 GABAA receptor subtypes to assess GABAA receptor subunits that are responsible for the potentiating effect.. The much higher potentiation of GABAA receptors at identical concentrations of 4-O-methylhonokiol as compared to honokiol parallels previous observations made in other systems of potentiated pharmacological activity of 4-O-methylhonokiol over honokiol.. The results point to the use of 4-O-methylhonokiol as a lead for GABAA receptor potentiation and corroborate the use of M. grandiflora seeds against convulsions in Mexican folk medicine.

Topics: Animals; Biphenyl Compounds; GABA-A Receptor Agonists; Gastrointestinal Agents; Humans; Lignans; Magnolia; Membrane Potentials; Plant Bark; Receptors, GABA-A; Xenopus laevis

Recently, increasing evidence has shown that cell cycle activation is a key factor of neuronal death and neurological dysfunction after traumatic brain injury (TBI). This study aims to investigate the effects of Honokiol, a cell cycle inhibitor, on attenuating the neuronal damage and facilitating functional recovery after TBI in rats, in an attempt to unveil its underlying molecular mechanisms in TBI. This study suggested that delayed intravenous administration of Honokiol could effectively ameliorate TBI-induced sensorimotor and cognitive dysfunctions. Meanwhile, Honokiol treatment could also reduce the lesion volume and increase the neuronal survival in the cortex and hippocampus. The neuronal degeneration and apoptosis in the cortex and hippocampus were further significantly attenuated by Honokiol treatment. In addition, the expression of cell cycle-related proteins, including cyclin D1, CDK4, pRb and E2F1, was significantly increased and endogenous cell cycle inhibitor p27 was markedly decreased at different time points after TBI. And these changes were significantly reversed by post-injury Honokiol treatment. Furthermore, the expression of some of the key cell cycle proteins such as cyclin D1 and E2F1 and the associated apoptosis in neurons were both remarkably attenuated by Honokiol treatment. These results show that delayed intravenous administration of Honokiol could effectively improve the functional recovery and attenuate the neuronal cell death, which is probably, at least in part, attributed to its role as a cell cycle inhibitior. This might give clues to developing attractive therapies for future clinical trials.

Topics: Administration, Intravenous; Animals; Apoptosis; Biphenyl Compounds; Brain Injuries; Cell Cycle; Cell Survival; Cerebral Cortex; Cognition; Disease Models, Animal; Hippocampus; Lignans; Male; Motor Activity; Neurons; Neuroprotective Agents; Random Allocation; Rats, Sprague-Dawley; Recovery of Function

Honokiol (HNK), derived from the bark of an oriental medicinal plant (Magnolia officinalis), is a promising anticancer agent with preclinical in vitro (PC-3 and LNCaP cells) and in vivo (PC-3 xenografts) efficacy against prostate cancer. However, the mechanisms affecting anticancer response to HNK are not fully understood.. Human (androgen-independent PC-3 and androgen-responsive LNCaP) and murine (Myc-CaP) prostate cancer cells, and PC-3 tumor xenografts were used for various assays. Autophagy was assessed by transmission electron microscopy, immunofluorescence (LC3 puncta), and immunoblotting (LC3BII detection). Cell viability was determined by trypan blue assay. Apoptosis was quantitated by DNA fragmentation detection and Annexin V/propidium iodide assay. Reactive oxygen species (ROS) were detected by electron paramagnetic resonance spectrometry and flow cytometric/microscopic analysis of MitoSOX red fluorescence.. Exposure of PC-3, LNCaP, and Myc-CaP cells to pharmacologic doses of HNK resulted in autophagy induction. The PC-3 tumor xenografts from HNK-treated mice contained higher levels of LC3BII protein compared with control tumors. Cell viability inhibition and apoptosis induction resulting from HNK exposure were significantly augmented by pharmacological inhibition of autophagy using 3-methyladenine as well as RNA interference of autophagy regulator ATG5. HNK-mediated increase in levels of LC3BII protein was partially but markedly diminished in the presence of antioxidants, including N-acetylcysteine, polyethylene glycol-conjugated (PEG)-superoxide dismutase, and PEG-catalase. On the other hand, antioxidants had no impact on HNK-induced apoptosis.. In conclusion, the present study demonstrates, for the first time, that HNK induces ROS-mediated cytoprotective autophagy in prostate cancer cells.

Topics: Animals; Autophagy; Biphenyl Compounds; Cell Line, Tumor; Cytoprotection; Drugs, Chinese Herbal; Humans; Lignans; Male; Mice; Mice, Transgenic; Prostatic Neoplasms; Reactive Oxygen Species

Magnolia officinalis is one of the commonly used in traditional Chinese medicine for the treatment of fever, chronic bronchitis and stomach ailments. Magnolol and honokiol are isomers with hydroxylated biphenol compound in the extract of Magnolia officinalis. This study aims to determine the isomers in rat plasma and evaluate their pharmacokinetic pattern after administration emulsion.. Sprague Dawley male rats received either an intravenous (i.v.25, mg/kg) or oral (50mg/kg) dose of the emulsion of the isomer. A sensitive and specific ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) method was developed for the investigation of the pharmacokinetics of magnolol and honokiol in rats. Kaempferol was employed as an internal standard.. The plasma samples were deproteinized with acetonitrile, the post-treatment samples were analyzed on an Agela C18 column interfaced with a triple quadrupole tandem mass spectrometer in negative electrospray ionization mode. Acetonitrile and 5 mmol/L ammonium acetate buffer solution (65: 35, v/v) was used as the mobile phase at a flow rate of 0.2 mL/min. Following oral administration of emulsion to rats, magnolol attained mean peak plasma concentrations of 426.4 ± 273.8 ng/mL at 1.20 h, whereas honokiol reached peak plasma concentrations of 40.3 ± 30.8 ng/mL at 0.45 h. The absolute bioavailability of magnolol and honokiol is 17.5 ± 9.7% and 5.3 ± 11.7%. By comparison, the AUC0-∞ of magnolol was 5.4 times higher than that of honokiol after intravenous administration, but AUC0-∞ of magnolol was about 18-fold higher than honokiol after oral administration.

Topics: Administration, Intravenous; Administration, Oral; Animals; Biphenyl Compounds; Chromatography, Liquid; Emulsions; Lignans; Male; Rats; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

Honokiol, an active constituent of oriental medicinal herb Magnolia officinalis, caused Ca(2+) mobilization and apoptosis in different cancer cells. In vivo, honokiol crossed the blood-brain or -cerebrospinal fluid barrier, suggesting that it may be an effective drug for the treatment of brain tumors, including glioblastoma. This study examined the effect of honokiol on intracellular Ca(2+) concentration ([Ca(2+)]i) and apoptosis in DBTRG-05MG human glioblastoma cells. Honokiol concentration-dependently induced a [Ca(2+)]i rise. The signal was decreased partially by removal of extracellular Ca(2+). Honokiol-triggered [Ca(2+)]i rise was not suppressed by store-operated Ca(2+) channel blockers (nifedipine, econazole, SK&F96365) and the protein kinase C (PKC) activator phorbol 12-myristate 13 acetate (PMA), but was inhibited by the PKC inhibitor GF109203X. GF109203X-induced inhibition was not altered by removal of extracellular Ca(2+). In Ca(2+)-free medium, pretreatment with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin (TG) or 2,5-di-tert-butylhydroquinone (BHQ) abolished honokiol-induced [Ca(2+)]i rise. Conversely, incubation with honokiol abolished TG or BHQ-induced [Ca(2+)]i rise. Inhibition of phospholipase C (PLC) with U73122 abolished honokiol-induced [Ca(2+)]i rise. Honokiol (20-80μM) reduced the cell viability, which was not reversed by prechelating cytosolic Ca(2+) with BAPTA-AM (1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester). Honokiol (20-60μM) enhanced reactive oxygen species (ROS) production, decreased mitochondrial membrane potential, released cytochrome c, and activated caspase-9/caspase-3. Together, honokiol induced a [Ca(2+)]i rise by inducing PLC-dependent Ca(2+) release from the endoplasmic reticulum and Ca(2+) entry via PKC-dependent, non store-operated Ca(2+) channels. Moreover, honokiol activated the mitochondrial pathway of apoptosis in DBTRG-05MG human glioblastoma cells.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Calcium; Calcium Signaling; Cell Line, Tumor; Cell Survival; Estrenes; Glioblastoma; Homeostasis; Humans; Lignans; Phosphodiesterase Inhibitors; Pyrrolidinones; Up-Regulation

Honokiol [3,5-di-(2-propenyl)-1,1-biphenyl-2,2-diol; HNK], a natural bioactive molecular compound isolated from the Magnolia officinalis, exhibits potent antitumor activity against a variety of human cancer cell lines. However, few studies have reported the antineoplastic effects of HNK on glioblastoma cells. It remains unknown how apoptosis is induced by HNK in glioblastoma cells and through which associated pathway this compound acts. The present study confirmed that HNK inhibited proliferation of glioblastoma cells by inducing a slight G0/G1 phase cell cycle arrest and apoptosis. We demonstrated for the first time that HNK triggered apoptosis of glioblastoma cells through both caspase-independent and caspase-dependent pathways, the latter including the extrinsic pathway and intrinsic pathway. Moreover, the inhibition of STAT3 signaling, ERK1/2 as well as activation of the p38 MAPK signaling pathway may be involved in apoptosis induced by HNK in U87 cells. Our findings suggest that HNK treatment could be a promising therapeutic strategy in human glioblastoma.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Gene Expression Regulation, Neoplastic; Glioblastoma; Humans; Lignans; MAP Kinase Signaling System; STAT3 Transcription Factor

Honokiol is an important bioactive compound found in the bark of Magnolia tree. It is a nonadipogenic PPARγ agonist and capable of inhibiting the growth of a variety of tumor types both in vitro and in xenograft models. However, to fully appreciate the potential chemopreventive activity of honokiol, a less artificial model system is required. To that end, this study examined the chemopreventive efficacy of honokiol in an initiation model of lung squamous cell carcinoma (SCC). This model system uses the carcinogen N-nitroso-trischloroethylurea (NTCU), which is applied topically, reliably triggering the development of SCC within 24 to 26 weeks. Administration of honokiol significantly reduced the percentage of bronchial that exhibit abnormal lung SCC histology from 24.4% bronchial in control to 11.0% bronchial in honokiol-treated group (P = 0.01) while protecting normal bronchial histology (present in 20.5% of bronchial in control group and 38.5% of bronchial in honokiol-treated group. P = 0.004). P63 staining at the SCC site confirmed the lung SCCs phenotype. In vitro studies revealed that honokiol inhibited lung SCC cells proliferation, arrested cells at the G1-S cell-cycle checkpoint, while also leading to increased apoptosis. Our study showed that interfering with mitochondrial respiration is a novel mechanism by which honokiol changed redox status in the mitochondria, triggered apoptosis, and finally leads to the inhibition of lung SCC. This novel mechanism of targeting mitochondrial suggests honokiol as a potential lung SCC chemopreventive agent.

Topics: Adenosine Triphosphate; Animals; Anticarcinogenic Agents; Apoptosis; Biphenyl Compounds; Bronchi; Carcinogens; Carcinoma, Squamous Cell; Carmustine; Caspase 3; Caspase 7; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Female; Gene Expression Regulation, Neoplastic; Green Fluorescent Proteins; Humans; Lignans; Lung Neoplasms; Mice; Mitochondria; Neoplasm Transplantation; Oxidation-Reduction; Reactive Oxygen Species

This study aimed to prepare and characterize an inclusion complex of honokiol (HNK) with sulfobutyl ether-β-cyclodextrin (SB-β-CD). The inclusion complex (HNK/CD COMP) was prepared utilizing a freeze-drying method. Phase-solubility curves were employed to obtain stability constants and thermodynamic parameters. The phase-solubility diagram showed a typical A(L)-type, indicating that the 1:1 (HNK:SB-β-CD) inclusion complex was formed. The solid inclusion complex was then characterized by differential scanning calorimetry and Fourier transform infrared spectroscopy. Results showed that HNK/CD COMP exhibited a higher drug release rate than free HNK in vitro. A comparative study of the pharmacokinetics between HNK/CD COMP and free HNK was also performed in rats. In vivo results indicated that AUC0-t and Cmax of HNK/CD COMP increased by approximately 158% and 123% compared with those of the free HNK, respectively. These results suggest that SB-β-CD will be potentially useful in the delivery of poorly soluble drugs, such as HNK.

Topics: Animals; beta-Cyclodextrins; Biological Availability; Biphenyl Compounds; Calorimetry, Differential Scanning; Drug Delivery Systems; Drug Liberation; Freeze Drying; Lignans; Male; Rats; Solubility; Spectroscopy, Fourier Transform Infrared

Honokiol, 2-(4-hydroxy-3-prop-2-enyl-phenyl)-4-prop-2-enyl-phenol, an active component of Magnolia officinalis and Magnolia grandiflora, exerts various pharmacological activities such as antitumorigenic, antioxidative, anti-inflammatory, neurotrophic, and antithrombotic effects. To investigate whether honokiol acts as a perpetrator in drug interactions, messenger ribonucleic acid (mRNA) levels of phase I and II drug-metabolizing enzymes, including cytochrome P450 (CYP), UDP-glucuronosyltransferase (UGT), and sulfotransferase 2A1 (SULT2A1), were analyzed by real-time reverse transcription polymerase chain reaction following 48-hour honokiol exposure in three independent cryopreserved human hepatocyte cultures. Honokiol treatment at the highest concentration tested (50 μM) increased the CYP2B6 mRNA level and CYP2B6-catalyzed bupropion hydroxylase activity more than two-fold in three different hepatocyte cultures, indicating that honokiol induces CYP2B6 at higher concentrations. However, honokiol treatment (0.5-50 μM) did not significantly alter the mRNA levels of phase I enzymes (CYP1A2, CYP3A4, CYP2C8, CYP2C9, and CYP2C19) or phase II enzymes (UGT1A1, UGT1A4, UGT1A9, UGT2B7, and SULT2A1) in cryopreserved human hepatocyte cultures. CYP1A2-catalyzed phenacetin O-deethylase and CYP3A4-catalyzed midazolam 1'-hydroxylase activities were not affected by 48-hour honokiol treatment in cryopreserved human hepatocytes. These results indicate that honokiol is a weak CYP2B6 inducer and is unlikely to increase the metabolism of concomitant CYP2B6 substrates and cause pharmacokinetic-based drug interactions in humans.

Topics: Biphenyl Compounds; Cells, Cultured; Cytochrome P-450 Enzyme System; Enzyme Induction; Glucuronosyltransferase; Hepatocytes; Humans; Lignans; Pharmaceutical Preparations; RNA, Messenger; Sulfotransferases

Pathological activation of the hypoxia-inducible-factor (HIF) pathway leading to expression of pro-angiogenic genes, such as vascular endothelial growth factor (VEGF), is the fundamental cause of neovascularization in ocular ischemic diseases and cancers. We have shown that pure honokiol inhibits the HIF pathway and hypoxia-mediated expression of pro-angiogenic genes in a number of cancer and retinal pigment epithelial (RPE) cell lines. The crude extracts, containing honokiol, from Magnolia plants have been used for thousands of years in the traditional oriental medicine for a number of health benefits. We have recently demonstrated that daily intraperitoneal injection of honokiol starting at postnatal day (P) 12 in an oxygen induced retinopathy mouse model significantly reduced retinal neovascularization at P17. Here, we evaluate the mechanism of HIF inhibition by honokiol in RPE cells. Using chromatin immunoprecipitation experiments, we demonstrate that honokiol inhibits binding of HIF to hypoxia-response elements present on VEGF promoter. We further show using a number of in vitro angiogenesis assays that, in addition to anti-HIF effect, honokiol manifests potent anti-angiogenic effect on human retinal micro vascular endothelial cells. Our results suggest that honokiol possesses potent anti-HIF and anti-angiogenic properties. These properties of honokiol make it an ideal therapeutic agent for the treatment of ocular neovascular diseases and solid tumors.

Topics: Angiogenesis Inhibitors; Biphenyl Compounds; Cell Line; Cell Movement; Cell Proliferation; Endothelium, Vascular; Eye Diseases; HeLa Cells; Humans; Hypoxia-Inducible Factor 1; Lignans; Luciferases; Neovascularization, Pathologic; Retinal Pigment Epithelium

A main problem in oncology is the development of drug-resistance. Some plant-derived lignans are established in cancer therapy, e.g. the semisynthetic epipodophyllotoxins etoposide and teniposide. Their activity is, unfortunately, hampered by the ATP-binding cassette (ABC) efflux transporter, P-glycoprotein. Here, we investigated the bisphenolic honokiol derived from Magnolia officinalis. P-glycoprotein-overexpressing CEM/ADR5000 cells were not cross-resistant to honokiol, but MDA-MB-231 BRCP cells transfected with another ABC-transporter, BCRP, revealed 3-fold resistance. Further drug resistance mechanisms analyzed study was the tumor suppressor TP53 and the epidermal growth factor receptor (EGFR). HCT116 p53(-/-) did not reveal resistance to honokiol, and EGFR-transfected U87.MG EGFR cells were collateral sensitive compared to wild-type cells (degree of resistance: 0.34). To gain insight into possible modes of collateral sensitivity, we performed in silico molecular docking studies of honokiol to EGFR and EGFR-related downstream signal proteins. Honokiol bound with comparable binding energies to EGFR (-7.30 ± 0.01 kcal/mol) as the control drugs erlotinib (-7.50 ± 0.30 kcal/mol) and gefitinib (-8.30 ± 0.10 kcal/mol). Similar binding affinities of AKT, MEK1, MEK2, STAT3 and mTOR were calculated for honokiol (range from -9.0 ± 0.01 to 7.40 ± 0.01 kcal/mol) compared to corresponding control inhibitor compounds for these signal transducers. This indicates that collateral sensitivity of EGFR-transfectant cells towards honokiol may be due to binding to EGFR and downstream signal transducers. COMPARE and hierarchical cluster analyses of microarray-based transcriptomic mRNA expression data of 59 tumor cell lines revealed a specific gene expression profile predicting sensitivity or resistance towards honokiol.

Topics: Antineoplastic Agents, Phytogenic; ATP Binding Cassette Transporter, Subfamily B; ATP Binding Cassette Transporter, Subfamily G, Member 2; ATP-Binding Cassette Transporters; Biphenyl Compounds; Cell Line, Tumor; Drug Resistance, Multiple; Drug Resistance, Neoplasm; ErbB Receptors; Humans; Lignans; Magnolia; Molecular Docking Simulation; Neoplasm Proteins; Oligonucleotide Array Sequence Analysis; Pharmacogenetics; Signal Transduction; Transcriptome; Tumor Suppressor Protein p53

Depression is an inflammatory, commonly occurring and lethal psychiatric disorder having high lifetime prevalence. Preclinical and clinical studies suggest that activation of immuno-inflammatory and oxido-nitrosative stress pathways play major role in the pathophysiology of depression. Honokiol (HNK) is a biphenolic neolignan possessing multiple biological activities including antioxidant, anti-inflammatory, anxiolytic, antidepressant and neuroprotective. The present study investigated the effect of HNK (2.5 and 5 mg/kg, i.p.) pretreatment (30 min prior to LPS) on lipopolysaccharide (LPS) (0.83 mg/kg, i.p.) induced depressive like behavior, neuroinflammation, and oxido-nitrosative stress in mice. HNK pretreatment at both the doses significantly attenuated LPS induced depressive-like behavior by reducing the immobility time in forced swim and tail suspension test, and by improving the anhedonic behavior observed in sucrose preference test. HNK pretreatment ameliorated LPS induced neuroinflammation by reducing IL-1β, IL-6 and TNF-α level in hippocampus (HC) and prefrontal cortex (PFC). HNK pretreatment prevented LPS evoked oxidative/nitrosative stress via improving reduced glutathione level along with reduction in the lipid peroxidation and nitrite level in HC and PFC. Pretreatment with HNK also prevented the increase in plasma corticosterone (CORT) and decrease in hippocampal BDNF level in LPS challenged mice. In conclusion, current investigation suggested that HNK pretreatment provided protection against LPS-induced depressive like behavior which may be mediated by repression of pro-inflammatory cytokines as well as oxido-nitrosative stress in HC and PFC. Our results strongly speculated that HNK could be a therapeutic approach for the treatment of depression and other pathophysiological conditions which are closely associated with neuroinflammation and oxido-nitrosative stress.

Topics: Animals; Antidepressive Agents; Biphenyl Compounds; Brain-Derived Neurotrophic Factor; Corticosterone; Depression; Disease Models, Animal; Glutathione; Hippocampus; Inflammation; Interleukin-1beta; Interleukin-6; Lignans; Lipid Peroxidation; Lipopolysaccharides; Mice; Nitrites; Oxidative Stress; Prefrontal Cortex; Tumor Necrosis Factor-alpha

Malignant melanoma is an aggressive form of skin cancer with limited effective therapeutic options. Melanoma research concentrates on maximizing the effect on cancer cells with minimal toxicity to normal cells. AMP-activated protein kinase (AMPK) is an important regulator of cellular energy homeostasis and has been shown to control tumor progression regulating the cell cycle, protein synthesis, and cell growth and/or survival. Honokiol (HNK) is a biphenolic compound derived from Magnolia officinalis, a plant that has been used in traditional Chinese and Japanese medicine for the treatment of various pathological conditions. Recent studies have shown that HNK has antitumor activity with relatively low toxicity. In this study, we demonstrated that the growth inhibitory effects of HNK on melanoma and melanoma cancer stem cells were mediated through the activation of AMPK and hence AMPK signaling in melanoma cells.. We determined the effects of HNK treatment on various melanoma cell lines. HNK-induced cell growth inhibitory effects were determined using hexosaminidase assay. Protein expression studies were done by immunoblotting. Primary spheroid assay was used to assess stemness by growing single suspension cells in ultralow attachment plates.. HNK is highly effective in inhibiting melanoma cells by attenuating protein kinase B/mammalian target of rapamycin and AMPK signaling. HNK showed significant inhibition of the spheroid-forming capacity of melanoma cells and, hence, stemness. HNK significantly decreased the number and size of melanospheres in a dose-dependent manner. Western blot analyses showed enhanced phosphorylation of AMPK in melanoma cells. Furthermore, HNK decreased the cellular adenosine triphosphate pool in a dose-dependent manner with maximum effects observed at 48 hours.. The results suggest that HNK can target melanoma cells and mark them for cell death through AMPK signaling. Further studies are warranted for developing HNK as an effective chemopreventive/therapeutic agent in melanoma.

Topics: AMP-Activated Protein Kinases; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Survival; Humans; Immunoblotting; Lignans; Melanoma; Melanoma, Cutaneous Malignant; Microscopy, Electron, Scanning; Signal Transduction; Skin Neoplasms

Biphenylic compounds related to the natural products magnolol and 4'-O-methylhonokiol were synthesized, evaluated and optimized as positive allosteric modulators (PAMs) of GABA(A) receptors. The most efficacious compounds were the magnolol analog 5-ethyl-5'-hexylbiphenyl-2,2'-diol (45) and the honokiol analogs 4'-methoxy-5-propylbiphenyl-2-ol (61), 5-butyl-4'-methoxybiphenyl-2-ol (62) and 5-hexyl-4'-methoxybiphenyl-2-ol (64), which showed a most powerful potentiation of GABA-induced currents (up to 20-fold at a GABA concentration of 3μM). They were found not to interfere with the allosteric sites occupied by known allosteric modulators, such as benzodiazepines and N-arachidonoylglycerol. These new PAMs will be useful as pharmacological tools and may have therapeutic potential for mono-therapy, or in combination, for example, with GABA(A) receptor agonists.

Topics: Allosteric Regulation; Animals; Biological Products; Biphenyl Compounds; Lignans; Oocytes; Patch-Clamp Techniques; Protein Binding; Protein Isoforms; Receptors, GABA-A; Structure-Activity Relationship; Xenopus

In recent years, epidemiological data confirm that cannabis-related emergencies, cannabis-use disorders and dependence are significantly increased. Cannabis is generally considered a little dangerous substances of abuse, however, chronic consumption has been associated to the development of mental disorders, cognitive deficits, chronic bronchitis, emphysema, increased risk of myocardial infarction in the hour after use, increased mortality after myocardial infarction, liver inflammation and steatosis in patients affected by hepatitis C. In this article we described the pharmacological characteristics of Magnolia officinalis bark active principles suggesting a potential application in the treatment of both cannabis dependence and cannabis-related disorders.

Topics: Animals; Biphenyl Compounds; Cannabis; Humans; Inflammation; Lignans; Magnolia; Marijuana Abuse; Plant Bark; Plant Extracts; Polyphenols; Rats; Receptor, Cannabinoid, CB1; Receptor, Cannabinoid, CB2; Receptors, Cannabinoid; Receptors, G-Protein-Coupled

This study was aimed to investigate the effect of Honokiol (HNK) on proliferation and apoptosis of acute myeloid leukemia HL-60 cells and its potential mechanism. Inhibitory effect of HNK on the HL-60 cell proliferation was detected by MTT assay. Flow cytometry was used to detect the change of cell cycle and AnnexinV/PI staining was used to detect apoptosis. Western blot was applied to analyze the cell cycle protein (cyclins), cyclin-dependent kinase (CDK), P53, P21, P27, BCL-2, BCL-XL, Bax, caspase-3/9 and proteins for MAPK signal pathway. The results showed that HNK could inhibit the proliferation of HL-60 cells in time- and dose dependent ways. HNK arrested HL-60 cells in G0/G1 phase, and S phase cells decreased significantly (P < 0.05). The expression of cyclin D1, cyclin A, cyclin E and CDK2/4/6 were significantly down-regulated (P < 0.05), the expression of P53 and P21 was significantly upregulated after treating for 24 h with HNK (P < 0.05). After 24 h treatment with HNK, HL-60 cell apoptosis increased significantly with the upregulation of activated caspase-3, -9, BAX expression and the downregulation of BCL-2, BCL-XL expression. The MAPK subfamily, P38 and JNK were not significantly changed, but the expression of MEK1/2-ERK1/2 was significantly downregulated (P < 0.05). It is concluded that HNK arrestes the cells at G0/G1 phase and induces HL-60 cell apoptosis through the intervention of MEK1/2-ERK1/2 signaling pathway.

Topics: Apoptosis; bcl-2-Associated X Protein; Biphenyl Compounds; Caspase 3; Cell Cycle; Cell Proliferation; Cyclin D1; Cyclin E; Cyclin-Dependent Kinase 2; HL-60 Cells; Humans; Lignans; Oncogene Proteins; Signal Transduction

Cell division protein, FtsZ, has been identified as a new potential antimicrobial target against multidrug-resistant (MDR) and methicillin-resistant Staphylococcus aureus (MRSA). By using computer-aided simulation, the phenolic compounds magnolol and honokiol from Magnolia officinalis were shown to have high anchor energies to FtsZ of S. aureus. The calculated binding energies of magnolol and honokiol for this FtsZ (PDB Code: 4DXD) were established to be -7.6 kcal/mol and -8.2 kcal/mol, respectively. Both of them showed polymerization inhibition efficacy for this FtsZ at 100 ppm, which confirmed the simulation results. Their antibacterial activity against S. aureus including multidrug-resistant (MDR) and methicillin-resistant S. aureus (MRSA) with minimum inhibitory concentration (MIC) values in the range of 8-16 ppm. These findings support the use of computer-aided simulation to screen natural compounds for this cell division protein, FtsZ, and this method can be a quick and promising approach for the development of antimicrobial agents against multi-drug resistant S. aureus.

Topics: Anti-Bacterial Agents; Biphenyl Compounds; Humans; Lignans; Magnolia; Methicillin-Resistant Staphylococcus aureus; Microbial Sensitivity Tests; Plant Exudates; Staphylococcal Infections; Staphylococcus aureus

Honokiol, a cell-permeable phenolic compound derived from the bark of magnolia trees and present in Asian herbal teas, has a unique array of pharmacological actions, including the inhibition of multiple autonomic responses. We determined the effects of honokiol on calcium signaling underlying transmission mediated by human M3 muscarinic receptors expressed in Chinese hamster ovary (CHO) cells. Receptor binding was determined in radiolabelled ligand binding assays; changes in intracellular calcium concentrations were determined using a fura-2 ratiometric imaging protocol; cytotoxicity was determined using a dye reduction assay.. Honokiol had a potent (EC50 ≈ 5 μmol/l) inhibitory effect on store operated calcium entry (SOCE) that was induced by activation of the M3 receptors. This effect was specific, rapid and partially reversible, and was seen at concentrations not associated with cytotoxicity, inhibition of IP3 receptor-mediated calcium release, depletion of ER calcium stores, or disruption of M3 receptor binding.. It is likely that an inhibition of SOCE contributes to honokiol disruption of parasympathetic motor functions, as well as many of its beneficial pharmacological properties.

Topics: Animals; Biphenyl Compounds; Calcium; Calcium Signaling; CHO Cells; Cricetinae; Cytoplasm; Gene Expression Regulation; Humans; Inositol 1,4,5-Trisphosphate; Ion Transport; Lignans; Receptor, Muscarinic M3

Honokiol, isolated from the Chinese traditional herb magnolia, is a poorly water-soluble component and has been found to have anti-tumor properties. In the current study, honokiol submicron lipid emulsions (HK-SLEs) were prepared by high pressure homogenization technology. After HK-SLEs were physically characterized, their pharmacokinetics, tissue distribution and antitumor activity after intravenous (i.v.) administration to tumor-burdened mice were examined, using honokiol solution (HK-SOL) as the control. The results showed that the mean particle size, zeta potential, pH value, osmolality, drug loading (DL)% and entrapment efficiency (EE)% of HK-SLEs were 186.6 +/- 1.7 nm, -35.65 +/- 0.67 mV, 7.22 +/- 0.26, 298 +/- 2.3 mOsm/L, 7.1 +/- 0.2% and 95.5 +/- 0.2%, respectively. HK-SLEs were stable for at least 12 months when stored at 4 +/- 2 degrees C. The pharmacokinetic results showed that the drug concentration-time curves of HK-SLEs and HK-SOL could both be described by an open two-compartment model. The half-life of HK-SLEs (t1/2(alpha) = 8.014 min, t1/2(beta) = 35.784 min) was remarkably prolonged compared to that of HK-SOL (t1/2(alpha) = 4.318 min, t1/2(beta) = 15.522 min). HK-SLEs exhibited a greater AUC and reduced plasma clearance. The tissue distribution results indicated that HK-SLEs have better targeting properties to lung and tumor tissues compared with those of HK-SOL. Both HK-SLEs and HK-SOL tended to accumulate in brain tissue. In vivo study showed that HK-SLEs treatment caused significant inhibition of mouse sarcoma S180 tumor growth compared to HK-SOL. These results suggest that HK-SLEs might be an effective parenteral carrier for honokiol delivery in cancer treatment.

Topics: Animals; Antineoplastic Agents, Phytogenic; Area Under Curve; Biphenyl Compounds; Cell Line, Tumor; Chromatography, High Pressure Liquid; Drug Compounding; Drug Stability; Electrochemistry; Emulsions; Hydrogen-Ion Concentration; Injections, Intravenous; Lignans; Mice; Neoplasms, Experimental; Osmolar Concentration; Particle Size; Reproducibility of Results; Soybean Oil; Tissue Distribution

In the course of screening for neuroprotective natural products, Magnoliae Cortex showed potent inhibition of hippocampal neuronal HT22 cell death. Obovatol, honokiol, and magnolol were isolated from the ethanolic extract of Magnoliae Cortex. Isolated compounds obovatol, honokiol, and magnolol were protective against 5mM glutamate-induced cell death. When cells were stressed using glutamate, cell viability decreased to 16.98±4.58% over the control (100.00±10.15%). In contrast, 10 μM obovatol, 10 μM honokiol, and 50 μM magnolol increased cell viability to 91.80±1.70%, 93.59±1.93%, and 85.36±7.40%, respectively. The neuroprotective effects of obovatol and honokiol were attributable to the inhibition of intracellular reactive oxygen species production, followed by protection of the mitochondrial membrane potential (ΔΨm), recovery of Bcl-2 and Bid levels, inhibition of apoptosis-inducing factor expression, and phosphorylation of mitogen-activated protein kinases such as p38 kinases, extracellular signal-regulated kinases, and c-Jun N-terminal kinases. On the contrary, magnolol did not show any significant effect on the ΔΨm and apoptotic factors. Among three compounds, obovatol most strongly scavenged 2,2-diphenyl-1-picrylhydrazyl radicals and inhibited the elevation of intracellular reactive oxygen species levels in glutamate-stressed HT22 cells. These data suggest that obovatol and honokiol may have clinical applications for preventing neurodegenerative disorders.

Topics: Animals; Apoptosis; Apoptosis Inducing Factor; BH3 Interacting Domain Death Agonist Protein; Biphenyl Compounds; Cell Death; Cell Line; Cell Survival; Extracellular Signal-Regulated MAP Kinases; Glutamic Acid; JNK Mitogen-Activated Protein Kinases; Lignans; Magnoliaceae; Membrane Potential, Mitochondrial; Mice; Neurons; Neuroprotective Agents; Oxidative Stress; p38 Mitogen-Activated Protein Kinases; Phenyl Ethers; Phosphorylation; Picrates; Plant Extracts; Reactive Oxygen Species; Signal Transduction

Honokiol is a pharmacologically active small molecule with multifunctional antitumor effects. Although plenty of literature is available on honokiol-triggered apoptosis and programmed necrosis, few studies have investigated the potential existence of death mode transition from apoptosis to programmed necrosis. In the current study, we demonstrated that the necrotic cell population (PI-positive) gradually increased and the early-stage apoptotic cell population (PI-negative and AV-positive) decreased in a dose- and time-dependent manner following honokiol treatment. Furthermore, we demonstrated that these PI-positive cells were under necrotic cell death, since no late-apoptosis characteristics including conspicuous chromatin condensation or DNA ladder patterns were detected. These results demonstrated that cells suffered death mode transition from early-stage apoptosis to programmed necrosis with the increase of honokiol dose or treatment time. The protein expression of RIP3 markedly increased in parallel with HNK-triggered death mode transition, while the expression of RIP1 decreased. Cyclophilin D expression increased during cell death mode transition, and inhibition of cyclophilin D by cyclosporin A clearly blocked HNK-triggered programmed necrosis. These data indicated that honokiol-induced programmed necrosis and death mode transition are potentially RIP3‑dependent, cyclophilin D-regulated. Further results showed that blocked cyclophilin D by cyclosporin A inhibited HNK-induced necrosis, but did not affect HNK-induced RIP3 overexpression. This indicated that cyclophilin D was a potential modulator at downstream of RIP3. In conclusion, honokiol triggers a potential RIP3-dependent cell death mode transition from early-stage apoptosis to programmed necrosis, which is highly regulated by cyclophilin D.

Topics: Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cyclophilins; Humans; Lignans; Necrosis; Peptidyl-Prolyl Isomerase F; Receptor-Interacting Protein Serine-Threonine Kinases

Malignant pleural mesothelioma (MPM) is an extremely aggressive type of cancer and is associated with a poor patient prognosis due to its rapid progression. Novel therapeutic agents such as honokiol (HNK) improve the clinical outcomes of cancer therapy, yet the mechanisms involved have not been fully elucidated. The present study examined the regulatory effects of HNK on the growth and apoptosis of MSTO-211H mesothelioma cells and investigated its anticancer mechanism. The results revealed that HNK significantly reduced the cell viability and increased the sub-G1 population in MSTO-211H cells and suppressed the expression of the specificity protein 1 protein (Sp1). HNK reduced the transcriptional activity of Sp1 regulatory proteins, including cyclin D1, Mcl-1 and survivin, and, thus, induced apoptosis signaling pathways by increasing Bax, reducing Bid and Bcl-xl and activating caspase-3 and PARP in mesothelioma cells. The results suggest that Sp1, a novel molecular target of HNK, may be related to cell cycle arrest and apoptosis induction through the modulation of signal transduction pathways in MPM.

Topics: Antineoplastic Agents; Apoptosis; Apoptosis Regulatory Proteins; Biphenyl Compounds; Cell Cycle Proteins; Cell Line, Tumor; Cell Proliferation; Cell Survival; Down-Regulation; Drug Screening Assays, Antitumor; G1 Phase Cell Cycle Checkpoints; Humans; Lignans; Mesothelioma; Pleural Neoplasms; Signal Transduction; Sp1 Transcription Factor

Lung cancer remains a leading cause of death due to its metastasis to distant organs. We have examined the effect of honokiol, a bioactive constituent from the Magnolia plant, on human non-small cell lung cancer (NSCLC) cell migration and the molecular mechanisms underlying this effect. Using an in vitro cell migration assay, we found that treatment of A549, H1299, H460 and H226 NSCLC cells with honokiol resulted in inhibition of migration of these cells in a dose-dependent manner, which was associated with a reduction in the levels of cyclooxygenase-2 (COX-2) and prostaglandin E2 (PGE2). Celecoxib, a COX-2 inhibitor, also inhibited cell migration. Honokiol inhibited PGE2-enhanced migration of NSCLC cells, inhibited the activation of NF-κB/p65, an upstream regulator of COX-2, in A549 and H1299 cells, and treatment of cells with caffeic acid phenethyl ester, an inhibitor of NF-κB, also inhibited migration of NSCLC cells. PGE2 has been shown to activate β-catenin signaling, which contributes to cancer cell migration. Therefore, we checked the effect of honokiol on β-catenin signaling. It was observed that treatment of NSCLC cells with honokiol degraded cytosolic β-catenin, reduced nuclear accumulation of β-catenin and down-regulated matrix metalloproteinase (MMP)-2 and MMP-9, which are the down-stream targets of β-catenin and play a crucial role in cancer cell metastasis. Honokiol enhanced: (i) the levels of casein kinase-1α, glycogen synthase kinase-3β, and (ii) phosphorylation of β-catenin on critical residues Ser(45), Ser(33/37) and Thr(41). These events play important roles in degradation or inactivation of β-catenin. Treatment of celecoxib also reduced nuclear accumulation of β-catenin in NSCLC cells. FH535, an inhibitor of Wnt/β-catenin pathway, inhibited PGE2-enhanced cell migration of A549 and H1299 cells. These results indicate that honokiol inhibits non-small cell lung cancer cells migration by targeting PGE2-mediated activation of β-catenin signaling.

Topics: beta Catenin; Biphenyl Compounds; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Movement; Cell Nucleus; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Dinoprostone; Drugs, Chinese Herbal; Gene Knockdown Techniques; Humans; Lignans; Lung Neoplasms; Neoplasm Invasiveness; RNA Interference; Signal Transduction; Sulfonamides; Transcription Factor RelA

Honokiol has shown the ability to induce the apoptosis of several different cancer cell lines. Considering that mitochondria are involved in apoptosis, the aim of the present work was to investigate the effects of honokiol on mitochondria. The effects of honokiol on the permeability of H⁺ and K⁺, membrane potential, membrane fluidity, respiration and swelling of mitochondria isolated from the rat liver were assessed. The results show that honokiol can significantly induce mitochondrial swelling, decrease membrane potential and affect the respiration of mitochondria. Meanwhile, honokiol does not have a direct effect on the mitochondrial permeability transition pore.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Line, Tumor; Lignans; Membrane Potential, Mitochondrial; Mitochondria, Liver; Mitochondrial Membranes; Oxygen Consumption; Permeability; Rats; Rats, Wistar

A general approach based on stable isotope labeling and UPLC/Q-TOF-MS analysis of in vivo novel metabolites of honokiol has been developed in our study. In this method, urine samples were collected after intravenous administration of mixture of regular and [(13)C6]-labeled honokiol at 1:1 ratio to healthy rats. The metabolites could be easily recognized by the determination of a chromatographically co-eluted pair of isotopomers (MS doublet peaks) with similar peak intensities and mass difference corresponding to that between isotope-labeled and non-isotope-labeled honokiol. A total of 51 metabolites were detected, 37 of which were tentatively identified based on mass accuracy (<5 ppm). Among them, 33 of honokiol metabolites were first reported with 5 metabolites belonging to phase I and other 32 metabolites belonging to phase II metabolites. Our results highlighted that the main phase I metabolic pathways of honokiol in rats were oxidation, and the phase II metabolic pathways were sulfation, glucuronidation, acetylation as well as amino acids conjugation. This was the first research focused on the biotransformation of honokiol in rats, and the identification of these metabolites might provide us essential information for further pharmacological and clinical studies of honokiol.

Topics: Animals; Biotransformation; Biphenyl Compounds; Chromatography, Liquid; Drugs, Chinese Herbal; Isotope Labeling; Lignans; Magnolia; Male; Rats; Rats, Wistar; Tandem Mass Spectrometry

Two honokiol dimers, houpulins A and B (1 and 2), and two magnolol derivatives, houpulins C and D (3 and 4), were isolated and characterized from an ethanol extract obtained from the roots of Magnolia officinalis. The chemical structures were determined based on spectroscopic and physicochemical analyses, which included 1D and 2D NMR, as well as mass spectrometry data. These four oligomers possess new carbon skeletons postulated to be biosynthesized from the coupling of three or four C6-C3 subunits. In addition, the new oligomers were evaluated for inhibition of superoxide anion generation and elastase release, and houpulin B (2) was identified as a new anti-inflammatory lead compound.

Topics: Adult; Anti-Inflammatory Agents, Non-Steroidal; Biphenyl Compounds; Carbon; Dimerization; Ethanol; Humans; Inhibitory Concentration 50; Lignans; Magnolia; Neutrophils; Pancreatic Elastase; Plant Roots; Superoxides; Young Adult

Honokiol ((3,5-di-(2-propenyl)-1,1-biphenyl-2,2-diol), a component of Magnolia officinalis, stimulates apoptosis and is thus considered for the treatment of malignancy. In analogy to apoptosis of nucleated cells, erythrocytes may enter eryptosis, a suicidal death characterized by cell shrinkage and by breakdown of cell membrane phosphatidylserine asymmetry with phosphatidylserine-exposure at the erythrocyte surface. Eryptosis may be triggered following increase of cytosolic Ca(2+)-activity ([Ca(2+)]i). The present study explored, whether honokiol elicits eryptosis. Cell volume has been estimated from forward scatter, phosphatidylserine-exposure from annexin V binding, hemolysis from hemoglobin release, [Ca(2+)]i from Fluo3-fluorescence, and ceramide from fluorescent antibodies. As a result, a 48 h exposure to honokiol was followed by a slight but significant increase of [Ca(2+)]i (15 μM), significant decrease of forward scatter (5 μM), significant increase of annexin-V-binding (5 μM) and significant increase of ceramide formation (15 μM). Honokiol further induced slight, but significant hemolysis. Honokiol (15 μM) induced annexin-V-binding was significantly blunted but not abrogated in the nominal absence of extracellular Ca(2+). In conclusion, honokiol triggers suicidal erythrocyte death or eryptosis, an effect at least in part due to stimulation of Ca(2+) entry and ceramide formation.

Topics: Biphenyl Compounds; Calcium; Cell Death; Cell Size; Cells, Cultured; Erythrocytes; Hemolysis; Humans; Lignans; Phosphatidylserines

Although calcineurin inhibitors (CNIs) are very useful in preventing allograft rejection, they can mediate a rapid progression of post-transplantation malignancies. The CNI cyclosporine A (CsA) can promote renal tumor growth through activation of the proto-oncogene ras and over-expression of the angiogenic cytokine VEGF; the ras activation also induces over-expression of the cytoprotective enzyme HO-1, which promotes survival of renal cancer cells. Here, we show that the natural product honokiol significantly inhibited CsA-induced and Ras-mediated survival of renal cancer cells through the down-regulations of VEGF and HO-1. Thus, honokiol treatment may help to prevent tumor-promoting effects of CsA in transplant patients.

Topics: Apoptosis; Biphenyl Compounds; Calcineurin; Calcineurin Inhibitors; Cell Line, Tumor; Cell Proliferation; Cyclosporine; Down-Regulation; Gene Expression Regulation; Genes, ras; Heme Oxygenase-1; Humans; Kidney Neoplasms; Lignans; Phosphorylation; Proto-Oncogene Mas; raf Kinases; ras Proteins; Signal Transduction; Transcriptional Activation; Transfection; Vascular Endothelial Growth Factor A

Oxaliplatin is an important drug in the chemotherapy of colorectal carcinoma, but its toxicity, especially dose-related neurosensory toxicity, is not well tolerated. In this study, we investigated whether honokiol could augment the anti-tumor effect of oxaliplatin in colon cancer HT-29 cells in vitro and whether honokiol could be used with oxaliplatin to decrease oxaliplatin dose. We used the normal colon cells, human colonic epithelial cells (HCoEpiCs) as control cells. Cell proliferation, apoptosis, prostaglandin E2 (PGE2) and vascular endothelial growth factor (VEGF) levels were also investigated. Expression levels of cyclo-oxygenase 2 (COX-2), VEGF, AKT/p-AKT, extracellular signal-related kinase (ERK)1/2/p-ERK1/2, nuclear factor kappa B (NF-κB) P65/p-P65, and caspase-3 were measured. Honokiol or oxaliplatin suppressed the proliferation of HT-29 cells in a concentration-dependent manner, but only high concentrations of honokiol would suppress the proliferation of HCoEpiCs. HT-29 cells were more sensitive to oxaliplatin treatment in the presence of honokiol. Oxaliplatin combined with honokiol improved the apoptosis rate of HT-29 cell and reduced PGE2 and VEGF secretion levels. Expression levels of COX-2 and VEGF protein and phosphorylation of AKT, ERK1/2, and NF-κB P65 were also inhibited. Caspase-3 levels were upregulated after honokiol treatment. Therefore, honokiol can be used in combination with oxaliplatin in the chemotherapy of colon cancer. This combination allows a reduction in oxaliplatin dose, and thereby reduces its adverse effects. It may also enhance the chemotherapeutic effect of oxaliplatin for this disease.

Topics: Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Blotting, Western; Caspase 3; Cell Line; Cell Proliferation; Cell Survival; Colonic Neoplasms; Cyclooxygenase 2; Dinoprostone; Dose-Response Relationship, Drug; Drug Synergism; Drugs, Chinese Herbal; Enzyme-Linked Immunosorbent Assay; Extracellular Signal-Regulated MAP Kinases; HT29 Cells; Humans; Lignans; NF-kappa B; Organoplatinum Compounds; Oxaliplatin; Proto-Oncogene Proteins c-akt; Vascular Endothelial Growth Factor A

Honokiol, a natural molecule isolated from Magnolia officinalis Rehd. et Wils., is widely known as an antitumor agent. In present work, an analysis of in vivo biotransformation and metabolites of honokiol has been performed by a combined method based on stable isotope cluster technique with honokiol-[(13)C6]-labeled and ultra-high performance liquid chromatography/quadrupole-time-of-flight-mass spectrometry (UHPLC/Q-TOF-MS). The metabolites could be easily identified by the determination of a chromatographically co-eluted pair of isotopomers (MS doublet peaks) with similar peak intensities and mass difference corresponding to that between isotope-labeled and non-isotope-labeled honokiol. A total of eighteen metabolites were detected and tentatively identified, fourteen of which were reported for the first time. The results indicated that the main metabolic pathways of honokiol in rats were hydroxylation, methylation, sulfation and glucuronidation. This study provided the first essential information on biotransformation and metabolites of honokiol in rats, which was very useful for further pharmacological and clinical studies of honokiol as a potent drug candidate.

Topics: Animals; Biphenyl Compounds; Chromatography, High Pressure Liquid; Glucuronides; Glutamates; Isotope Labeling; Lignans; Male; Mass Spectrometry; Metabolic Networks and Pathways; Rats; Rats, Sprague-Dawley

Glucose regulated protein 78 (GRP78) functions as a sensor of endoplasmic reticulum (ER) stress. The aim of this study was to test the hypothesis that molecules that bind to GRP78 induce the unfolded protein response (UPR) and enhance cell death in combination with ER stress inducers.. Differential scanning calorimetry (DSC), measurement of cell death by flow cytometry and the induction of ER stress markers using western blotting.. Epigallocatechin gallate (EGCG), a flavonoid component of Green Tea Camellia sinensis, and honokiol (HNK), a Magnolia grandiflora derivative, bind to unfolded conformations of the GRP78 ATPase domain. Epigallocatechin gallate and HNK induced death in six neuroectodermal tumour cell lines tested. Levels of death to HNK were twice that for EGCG; half-maximal effective doses were similar but EGCG sensitivity varied more widely between cell types. Honokiol induced ER stress and UPR as predicted from its ability to interact with GRP78, but EGCG was less effective. With respect to cell death, HNK had synergistic effects on melanoma and glioblastoma cells with the ER stress inducers fenretinide or bortezomib, but only additive (fenretinide) or inhibitory (bortezomib) effects on neuroblastoma cells.. Honokiol induces apoptosis due to ER stress from an interaction with GRP78. The data are consistent with DSC results that suggest that HNK binds to GRP78 more effectively than EGCG. Therefore, HNK may warrant development as an antitumour drug.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Catechin; Cell Line, Tumor; Cell Survival; Endoplasmic Reticulum Chaperone BiP; Endoplasmic Reticulum Stress; Heat-Shock Proteins; Humans; Lignans; Molecular Targeted Therapy; Molecular Weight; Neoplasms; Protein Binding

Peroxisome proliferator-activated receptor gamma (PPARγ) agonists are clinically used to counteract hyperglycemia. However, so far experienced unwanted side effects, such as weight gain, promote the search for new PPARγ activators.. We used a combination of in silico, in vitro, cell-based and in vivo models to identify and validate natural products as promising leads for partial novel PPARγ agonists.. The natural product honokiol from the traditional Chinese herbal drug Magnolia bark was in silico predicted to bind into the PPARγ ligand binding pocket as dimer. Honokiol indeed directly bound to purified PPARγ ligand-binding domain (LBD) and acted as partial agonist in a PPARγ-mediated luciferase reporter assay. Honokiol was then directly compared to the clinically used full agonist pioglitazone with regard to stimulation of glucose uptake in adipocytes as well as adipogenic differentiation in 3T3-L1 pre-adipocytes and mouse embryonic fibroblasts. While honokiol stimulated basal glucose uptake to a similar extent as pioglitazone, it did not induce adipogenesis in contrast to pioglitazone. In diabetic KKAy mice oral application of honokiol prevented hyperglycemia and suppressed weight gain.. We identified honokiol as a partial non-adipogenic PPARγ agonist in vitro which prevented hyperglycemia and weight gain in vivo.. This observed activity profile suggests honokiol as promising new pharmaceutical lead or dietary supplement to combat metabolic disease, and provides a molecular explanation for the use of Magnolia in traditional medicine.

Topics: 3T3-L1 Cells; Adipose Tissue; Animals; Biological Products; Biphenyl Compounds; Cell Differentiation; Diabetes Mellitus, Experimental; HEK293 Cells; Humans; Lignans; Mice; Molecular Docking Simulation; PPAR gamma

Honokiol is known to suppress the growth of cancer cells; however, to date, its antiperitoneal dissemination effects have not been studied in an orthotopic mouse model. In the present study, we evaluated the antiperitoneal dissemination potential of Honokiol in an orthotopic mouse model and assessed associations with tumor growth factor-β1 (TGFβ1) and cells stimulated by a carcinogen, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Our results demonstrate that tumor growth, peritoneal dissemination and peritoneum or organ metastasis of orthotopically implanted MKN45 cells were significantly decreased in Honokiol-treated mice and that endoplasmic reticulum (ER) stress was induced. Honokiol-treated tumors showed increased epithelial signatures such as E-cadherin, cytokeratin-18 and ER stress marker. In contrast, decreased expression of vimentin, Snail and tumor progression locus 2 (Tpl2) was also noted. TGFβ1 and MNNG-induced downregulation of E-cadherin and upregulation of Tpl2 were abrogated by Honokiol treatment. The effect of Tpl2 inhibition in cancer cells or endothelial cells was associated with inactivation of CCAAT/enhancer binding protein B, nuclear factor kappa-light-chain-enhancer of activated B cell and activator protein-1 and suppression of vascular endothelial growth factor. Inhibition of Tpl2 in gastric cancer cells by small interfering RNA or pharmacological inhibitor was found to effectively reduce growth ability and vessel density in vivo. Honokiol-induced reversal of epithelial-to-mesenchymal transition (EMT) and ER stress-induced apoptosis via Tp12 may involve the paralleling processes. Taken together, our results suggest that the therapeutic inhibition of Tpl2 by Honokiol thwarts both gastric tumor growth and peritoneal dissemination by inducing ER stress and inhibiting EMT.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Blotting, Western; Cell Adhesion; Cell Movement; Cell Proliferation; Fluorescent Antibody Technique; Humans; Immunoenzyme Techniques; Immunoprecipitation; Lignans; Luciferases; Male; MAP Kinase Kinase Kinases; Mice; Mice, Inbred BALB C; Mice, Nude; Peritoneal Neoplasms; Phytotherapy; Proto-Oncogene Proteins; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Stomach Neoplasms; Tumor Cells, Cultured

Honokiol (HK), a novel plant-derived natural product, is a physiologically activated compound with polyphenolic structure, and has been identified to function as an anticancer agent. It has been widely used in several diseases as a traditional medicine for a long time. We investigated whether HK could show anticancer effects on two oral squamous cell lines (OSCCs), HN-22 and HSC-4. We demonstrated that HK-treated cells showed dramatic reduction in cell growth and apoptotic cell morphologies. Intriguingly, the transcription factor specificity protein 1 (Sp1) was significantly inhibited by HK in a dose-dependent manner. Furthermore, we checked changes in cell cycle regulatory proteins and anti-apoptotic proteins at the molecular level, which are known as Sp1 target genes. The important key regulators in the cell cycle such as p27 and p21 were up-regulated by HK-mediated down-regulation of Sp1, whereas anti-apoptotic proteins including Mcl-1 and survivin were decreased, resulting in caspase-dependent apoptosis. Taken together, results from this study suggest that HK could modulate Sp1 transactivation and induce apoptotic cell death through the regulation of cell cycle and suppression of anti‑apoptotic proteins. In addition, HK may be used in cancer prevention and therapies to improve the clinical outcome as an anticancer drug.

Topics: Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Down-Regulation; Gene Expression Regulation, Neoplastic; Humans; Lignans; Mouth Neoplasms; Plants; Sp1 Transcription Factor

This study investigated the effect of honokiol (HON) and magnolol (MAG), phenolic compounds in Magnolia plants, on adiposity and adiposity-related metabolic disturbances in mice fed high-fat diet (HFD), and the potential underlying mechanisms focusing on the lipid metabolism and inflammatory response.. C57BL/6J mice were fed HFD (45 kcal% fat) with or without HON (0.02%, w/w) or MAG (0.02%, w/w) for 16 wk. Despite no changes in body weight, food intake, and hepatic fat accumulation, HON and MAG significantly lowered the weight of white adipose tissue (WAT) as well as adipocyte size and protected against insulin resistance induced by HFD. These effects were associated with increases in energy expenditure and adipose fatty acid oxidation and decreases in fatty acid synthase activity and expression of genes related to fatty acid synthesis, desaturation, and uptake, as well as adipocyte differentiation in WAT. Moreover, HON and MAG significantly lowered the expression of proinflammatory genes in WAT and elevated the plasma IL-10 level. Particularly, HON significantly decreased the plasma resistin level and increased the plasma adiponectin level compared to the control group.. HON and MAG have potential as novel agents for amelioration of adiposity and associated insulin resistance and inflammation.

Topics: Adipogenesis; Adiponectin; Adipose Tissue, White; Adiposity; Animals; Biphenyl Compounds; Blood Glucose; Body Weight; Chemokine CCL2; Cholesterol; Diet, High-Fat; Dietary Supplements; Energy Metabolism; Glucose Tolerance Test; Inflammation; Insulin Resistance; Interleukin-10; Interleukin-6; Lignans; Liver; Male; Mice; Mice, Inbred C57BL; Triglycerides; Tumor Necrosis Factor-alpha

α-Santalol is active component of sandalwood oil and has been shown to have chemopreventive effects against chemically and UVB-induced skin cancer development in mice. α-Santalol is also shown to have skin permeation enhancing effects. Honokiol and magnolol isolated from Magnolia officinalis bark extract have also been shown to have chemopreventive effects against chemically and UVB-induced skin cancer in mice. This study was conducted to investigate the combination effects of α-santalol, honokiol and magnolol to study any additive/synergistic effects to lower the doses required for chemoprevention. Pretreatment of combinations of α-santalol with honokiol and magnolol significantly decreased tumor multiplicity upto 75% than control, α-santalol, honokiol and magnolol alone in SKH-1 mice. Combination of α-santalol with honokiol and magnolol also decreased cell viability, proliferation, and enhanced apotosis in comparison to α-santalol, honokiol and magnolol alone in Human epidrmoid carcinoma A431 cells. Overall, the results of present study indicated combinations of α-santalol with honokiol and magnolol could provide chemoprevention of skin cancer at lower doses than given alone.

Topics: Animals; Anticarcinogenic Agents; Apoptosis; Biphenyl Compounds; Cell Proliferation; Drug Therapy, Combination; Female; Lignans; Mice; Neoplasms, Radiation-Induced; Polycyclic Sesquiterpenes; Sesquiterpenes; Skin Neoplasms; Ultraviolet Rays

Aberrant activation of the hypoxia inducible factor (HIF) pathway is the underlying cause of retinal neovascularization, one of the most common causes of blindness worldwide. The HIF pathway also plays critical roles during tumor angiogenesis and cancer stem cell transformation. We have recently shown that honokiol is a potent inhibitor of the HIF pathway in a number of cancer and retinal pigment epithelial cell lines. Here we evaluate the safety and efficacy of honokiol, digoxin, and doxorubicin, three recently identified HIF inhibitors from natural sources. Our studies show that honokiol has a better safety to efficacy profile as a HIF inhibitor than digoxin and doxorubicin. Further, we show for the first time that daily intraperitoneal injection of honokiol starting at postnatal day (P) 12 in an oxygen-induced retinopathy (OIR) mouse model significantly reduced retinal neovascularization at P17. Administration of honokiol also prevents the oxygen-induced central retinal vaso-obliteration, characteristic feature of the OIR model. Additionally, honokiol enhanced physiological revascularization of the retinal vascular plexuses. Since honokiol suppresses multiple pathways activated by HIF, in addition to the VEGF signaling, it may provide advantages over current treatments utilizing specific VEGF antagonists for ocular neovascular diseases and cancers.

Topics: Animals; Antibiotics, Antineoplastic; Biphenyl Compounds; Cell Line; Digoxin; Doxorubicin; Drugs, Chinese Herbal; Enzyme Inhibitors; Humans; Hypoxia-Inducible Factor 1; Lignans; Mice; Mice, Inbred C57BL; Oxygen; Retina; Retinal Neovascularization; Transcriptional Activation

Honokiol is a bioactive component isolated from the medicinal herbs Magnolia officinalis and Magnolia grandiflora that has antioxidative, anti-inflammatory, antithrombotic, and antitumor activities. The inhibitory potentials of honokiol on eight major human cytochrome P450 (CYP) enzymes 1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, and 3A4, and four UDP-glucuronosyltransferases (UGTs) 1A1, 1A4, 1A9, and 2B7 in human liver microsomes were investigated using liquid chromatography-tandem mass spectrometry. Honokiol strongly inhibited CYP1A2-mediated phenacetin O-deethylation, CYP2C8-mediated amodiaquine N-deethylation, CYP2C9-mediated diclofenac 4-hydroxylation, CYP2C19-mediated [S]-mephenytoin 4-hydroxylation, and UGT1A9-mediated propofol glucuronidation with K(i) values of 1.2, 4.9, 0.54, 0.57, and 0.3 μM, respectively. Honokiol also moderately inhibited CYP2B6-mediated bupropion hydroxylation and CYP2D6-mediated bufuralol 1'-hydroxylation with K(i) values of 17.5 and 12.0 μM, respectively. These in vitro results indicate that honokiol has the potential to cause pharmacokinetic drug interactions with other co-administered drugs metabolized by CYP1A2, CYP2C8, CYP2C9, CYP2C19, and UGT1A9.

Topics: Aryl Hydrocarbon Hydroxylases; Biphenyl Compounds; Bupropion; Drugs, Chinese Herbal; Enzyme Inhibitors; Ethanolamines; Glucuronosyltransferase; Herb-Drug Interactions; Humans; Hydroxylation; Inactivation, Metabolic; Inhibitory Concentration 50; Lignans; Liver; Microsomes, Liver; Phenacetin

To develop a UPLC method for the simultaneous determination of liquiritin, narirutin, hesperidin, ammonium glycyrrhetate, honokiol and magnolol in Huoxiang Zhengqi oral liquid.. A Zorbax Eclipse C18 column was used with the mobile phase of acetonitrile and 0. 05% phosphate acid by gradient elution at the detection wavelength of 220 nm. The flow rate was 0.42 mL x min(-1) and the column temperature was 30 degrees C.. The calibration curves were linear in the ranges of 0.001 7-0.034, 0.003 4-0.068, 0.006 4-0.128, 0.012 8-0.256, 0.003 2-0.064, 0.006 4-0.128 microg, respectively. The average recoveries were 103.3%, 98.39%, 98.29%, 102.1%, 98.45%, 102.2% with RSDs of 2.1%,1.0%, 0.50%, 2.3%, 0.9%, 2.0%, respectively.. The UPLC method was simple, rapid and accurate, it could be used for quality control of Huoxiang Zhengqi oral liquid.

Topics: Administration, Oral; Biphenyl Compounds; Chromatography, High Pressure Liquid; Disaccharides; Drugs, Chinese Herbal; Flavanones; Glucosides; Hesperidin; Lignans; Pharmaceutical Solutions

Honokiol, a major bioactive constituent of the bark of Magnolia officinalis has been confirmed to have the neuroprotective effect on ischemic stroke in rats. This study was designed to observe the therapeutic time window of honokiol microemulsion on cerebral ischemia-reperfusion injury to support its potential for future clinical trials and further explore the underlying mechanisms. Honokiol microemulsion (50μg/kg, i.v. at 0, 1 or 3h after reperfusion) significantly reduced neurological deficit, infarct volume and brain water content in rats subjected to cerebral ischemia-reperfusion, and honokiol (0.1-10μM) significantly attenuated oxygen-glucose deprivation- or glutamate-induced injury of fetal rat cortical neurons. In co-immunoprecipitation and western blot test, honokiol decreased the intensity of nNOS related to PSD95 but failed to affect that of PSD95 related to NR2B in NR2B-PSD95-nNOS complex, and it also inhibited the translocation of nNOS from cytosol to membrane without affecting total nNOS expression, and then markedly decreased NO production in cortical neurons. Besides, the results of whole-cell patch-clamp recordings showed that honokiol reversibly inhibited the NMDA current by about 64%. In conclusion, honokiol has a therapeutic window of at least 5h after the onset of cerebral ischemia or 3h after reperfusion in rats, which may be in part ascribed to the disruption of the PSD95-nNOS interaction leading to the inhibition of neurotoxic NO production.

Topics: Animals; Biphenyl Compounds; Bisbenzimidazole; Blotting, Western; Cell Hypoxia; Cells, Cultured; Central Nervous System Depressants; Cerebral Cortex; Coloring Agents; Cytosol; Disks Large Homolog 4 Protein; Glucose; Glutamic Acid; Immunoprecipitation; In Vitro Techniques; Infarction, Middle Cerebral Artery; Intracellular Signaling Peptides and Proteins; L-Lactate Dehydrogenase; Lignans; Male; Membrane Proteins; Nitric Oxide; Nitric Oxide Synthase Type I; Rats; Rats, Sprague-Dawley; Receptors, N-Methyl-D-Aspartate; Reperfusion Injury; Translocation, Genetic

Non-small-cell lung cancer (NSCLC) represents approximately 80% of all types of lung cancer. Here, we report the chemotherapeutic effect of honokiol, a phytochemical from Magnolia grandiflora, on NSCLC cells and the molecular mechanisms underlying these effects using in vitro and in vivo models. Treatment of NSCLC cells (A549, H1299, H460 and H226) with honokiol (20, 40 and 60 µM) inhibited histone deacetylase (HDAC) activity, reduced the levels of class I HDAC proteins and enhanced histone acetyltransferase activity in a dose-dependent manner. These effects of honokiol were associated with a significant reduction in the viability of NSCLC cells. Concomitant treatment of cells with a proteasome inhibitor, MG132, prevented honokiol-induced degradation of class I HDACs, suggesting that honokiol reduced the levels of HDACs in NSCLC cells through proteasomal degradation. Valproic acid, an inhibitor of HDACs, exhibited a similar pattern of reduced viability and induction of death of NSCLC cells. Treatment of A549 and H1299 cells with honokiol resulted in an increase in G 1 phase arrest, and a decrease in the levels of cyclin D1, D2 and cyclin dependent kinases. Further, administration of honokiol by oral gavage significantly inhibited the growth of subcutaneous A549 and H1299 tumor xenografts in athymic nude mice, which was associated with the induction of apoptotic cell death and marked inhibition of class I HDACs proteins and HDAC activity in the tumor xenograft tissues. Together, our study provides new insights into the role of class I HDACs in the chemotherapeutic effects of honokiol on lung cancer cells.

Topics: Acetylation; Animals; Biphenyl Compounds; Bronchi; Carcinoma, Non-Small-Cell Lung; Cell Death; Cell Line, Tumor; Cell Proliferation; Cell Survival; Epithelial Cells; G1 Phase Cell Cycle Checkpoints; Histone Acetyltransferases; Histone Deacetylase Inhibitors; Histone Deacetylases; Histones; Hydroxamic Acids; Lignans; Liver Neoplasms; Mice; Mice, Nude; Proteasome Endopeptidase Complex; Proteolysis; Valproic Acid; Xenograft Model Antitumor Assays

Multidrug resistance (MDR) is a major challenge in cancer therapy. Apoptosis tolerance is one of the key mechanisms of MDR. Honokiol, a small-molecule pharmacologically active component, exhibits competent cytotoxicity in a variety of human cancer cells through apoptosis and other forms of programmed cell death (such as programmed necrosis). Although much work has been done on its antitumor effects, little attention has been paid on systemic evaluation of efficacy of honokiol combined with other chemotherapeutic agents, especially in drug‑resistant cell lines. Here, we systematically and quantitatively assess its combinational effect with different chemotherapeutic agents using the combination index (CI) equation. We found that honokiol synergized with chemotherapeutic agents both in sensitive and resistant, solid and non-solid (MCF-7, HL-60, MCF-7/ADR and HL-60/ADR) cell lines. Honokiol (40 µg/ml) induced necrotic cell death in MCF-7/ADR cells with characterized morphological and biochemical features. Co-incubation with honokiol and etoposide (VP-16) activated a complex death modality, which was composed of necrotic cell death and apoptosis. This dual-death pathway was shut down when pretreated with pan-caspase inhibitor (z-VAD-fmk) and cyclophilin D inhibitor (cyclosporin A). Western blot analysis results proved that honokiol also enhanced VP-16-induced apoptosis potentially via blocking nuclear factor‑κB (NF-κB) activation. Our data for the first time quantitatively demonstrate that honokiol synergizes frequently-used chemotherapeutic agents via enhanced apoptosis and additional programmed necrotic death. These findings indicate a promising way to circumvent MDR and apoptosis tolerance.

Topics: Apoptosis; Biphenyl Compounds; Breast Neoplasms; Cyclophilins; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Drug Synergism; Etoposide; Female; Humans; Lignans; MCF-7 Cells; Peptidyl-Prolyl Isomerase F

This study was designed to investigate the effects of honokiol, a neuroprotective agent, on cerebral edema in cerebral ischemia reperfusion (IR) mice and its mechanism of anti-inflammation. Honokiol (0.7-70μg/kg) significantly reduced brain water contents and decreased the exudation of Evans blue dye from brain capillaries in cerebral IR mice. Honokiol (0.1-10μM) significantly reduced the p65 subunit level of NF-κB in the nucleus of primary culture-microglia. It (0.01-10μM) evidently reduced nitric oxide (NO) level in the microglia culture medium and in the microglia and astrocytes coculture medium. Honokiol (0.01-10μM) significantly decreased the level of TNF-α in the microglia medium or coculture cell medium. Honokiol (10μM) decreased the level of Regulated upon Activation Normal T-cell Expressed and Secreted (RANTES/CCL5) protein in medium of microglia or astrocytes. In conclusion, Honokiol has a potent anti-inflammatory effect in cerebral ischemia-reperfusion mice and this effect might be attributed to its inhibition ability on the NF-κB activation, consequently blocking the production of inflammatory factors including: NO, tumor necrosis factor-α (TNF-α) and RANTES/CCL5 in glial cells. These results provide evidence for the anti-inflammatory effect of honokiol for the potential treatment of ischemic stroke.

Topics: Animals; Anti-Inflammatory Agents; Astrocytes; Biphenyl Compounds; Brain Edema; Brain Ischemia; Capillary Permeability; Chemokine CCL5; Coculture Techniques; Cytokines; Drugs, Chinese Herbal; Female; Inflammation; Lignans; Male; Mice; Microglia; Neuroglia; NF-kappa B; Nitric Oxide; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Tumor Necrosis Factor-alpha

Our previous report has shown that honokiol (HNK), a constituent of Magnolia officinalis, induces a novel form of non-apoptotic programmed cell death in human leukemia NB4 and K562 cells. In this study, we further explored the relationship between the cell death pathway and cytoplasmic vacuolization and studied the underlying mechanism of leukemia cell death mediated by honokiol. The results showed that low concentrations of honokiol activated an novel alternative cell death fitted the criteria of paraptosis, such as cytoplasmic vacuolization derived from endoplasmic reticulum swelling, lack of caspase activation, and lack of apoptotic morphology. Results further indicated that the cell death was time- and concentration-dependent. In addition, honokiol-induced paraptosis did not involve membrane blebbing, chromatin condensation and phosphatidylserine exposure at the outer of the plasma membrane. The mechanism of the cell death may be associated, at least in part, with the increased generation of reactive oxygen species. Further analysis showed that honokiol induces cell death predominantly via paraptosis and to a certain extent via apoptosis in NB4 cells, and predominantly via apoptosis and to a certain extent via paraptosis in K562 cells. These observations suggest that cell death occurs via more than one pathway in leukemia cells and targeting paraptosis may be an alternative and promising avenue for honokiol in leukemia therapy.

Topics: Apoptosis; Biphenyl Compounds; Caspases; Cell Line, Tumor; Cell Membrane; Cytoplasm; Endoplasmic Reticulum; Humans; Leukemia; Lignans; Metabolic Networks and Pathways; Necrosis; Phosphatidylserines; Reactive Oxygen Species; Vacuoles

The bark of magnolia has been used in Oriental medicine to treat a variety of remedies, including some neurological disorders. Magnolol (Mag) and honokiol (Hon) are isomers of polyphenolic compounds from the bark of Magnolia officinalis, and have been identified as major active components exhibiting anti-oxidative, anti-inflammatory, and neuroprotective effects. In this study, we investigate the ability of these isomers to suppress oxidative stress in neurons stimulated by the ionotropic glutamate receptor agonist N-methyl-D-aspartate (NMDA) and oxidative and inflammatory responses in microglial cells activated by interferon-γ (IFNγ) and lipopolysaccharide (LPS). We also attempt to elucidate the mechanism and signaling pathways involved in cytokine-induced production of reactive oxygen species (ROS) in microglial cells.. Dihydroethidium (DHE) was used to assay superoxide production in neurons, while CM-H2DCF-DA was used to test for ROS production in murine (BV-2) and rat (HAPI) immortalized microglial cells. NADPH oxidase inhibitors (for example, diphenyleneiodonium (DPI), AEBSF, and apocynin) and immunocytochemistry targeting p47phox and gp91phox were used to assess the involvement of NADPH oxidase. Western blotting was used to assess iNOS and ERK1/2 expression, and the Griess reaction protocol was employed to determine nitric oxide (NO) concentration.. Exposure of Hon and Mag (1-10 μM) to neurons for 24 h did not alter neuronal viability, but both compounds (10 μM) inhibited NMDA-stimulated superoxide production, a pathway known to involve NADPH oxidase. In microglial cells, Hon and Mag inhibited IFNγ±LPS-induced iNOS expression, NO, and ROS production. Studies with inhibitors and immunocytochemical assay further demonstrated the important role of IFNγ activating the NADPH oxidase through the p-ERK-dependent pathway. Hon and, to a lesser extent, Mag inhibited IFNγ-induced p-ERK1/2 and its downstream pathway for ROS and NO production.. This study highlights the important role of NADPH oxidase in mediating oxidative stress in neurons and microglial cells and has unveiled the role of IFNγ in stimulating the MAPK/ERK1/2 signaling pathway for activation of NADPH oxidase in microglial cells. Hon and Mag offer anti-oxidative or anti-inflammatory effects, at least in part, through suppressing IFNγ-induced p-ERK1/2 and its downstream pathway.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Antioxidants; Biphenyl Compounds; Cell Line, Transformed; Cells, Cultured; Inflammation; Inflammation Mediators; Lignans; Magnolia; Mice; Microglia; Neurons; Oxidative Stress; Polyphenols; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species

Magnolol and honokiol, predominant active compounds in the family Magnoliaceae, are known to exhibit strong anti-inflammatory activities against dermal disorders. We attempted to modify the structures of magnolol and honokiol by methoxylation to optimize the skin delivery ability. Absorption of these permeants into and through the skin was performed at both an infinite dose and saturated solubility. Superoxide anion and elastase released from human neutrophils were the biomarkers used to examine anti-inflammatory potencies of these permeants. The safety of the permeants was evaluated by keratinocyte viability and in vivo bioengineering techniques. Topical magnolol and honokiol at an infinite dose (7.5 mM) showed skin accumulations of 0.22 and 0.16 nmol/mg, respectively. Methoxylation significantly enhanced their skin absorption. Deposition amounts of dimethylmagnolol and dimethylhonokiol were respectively 15- and 7-fold greater than those of magnolol and honokiol. Contrary to the skin accumulation results, the transdermal penetration across skin decreased following methoxylation. No transdermal delivery occurred for dimethylhonokiol. Skin uptake of 4'-O-methylhonokiol was 2-fold higher than that of 2-O-methylhonokiol, although they are isomers. Methoxylated permeants demonstrated selective absorption into follicles, which showed 3-5-fold higher follicular amounts compared to magnolol and honokiol. The relative order of anti-inflammatory activities was honokiol>2-O-methylmagnolol>dimethylhonokiol>magnolol. The other compounds exhibited negligible or negative responses in activated neutrophils. Magnolol and honokiol induced slight but significant keratinocyte cytotoxicity and stratum corneum disruption. Daily administration of methoxylated permeants, especially dimethylhonokiol, produced no skin irritation for up to 7 days. Methoxylated magnolol and honokiol can be efficient and safe candidates for treating inflammatory skin disorders.

Topics: Adult; Animals; Anti-Inflammatory Agents; Biphenyl Compounds; Cell Survival; Cells, Cultured; Female; Humans; In Vitro Techniques; Keratinocytes; Lignans; Mice; Mice, Nude; Neutrophils; Pancreatic Elastase; Skin; Skin Absorption; Skin Irritancy Tests; Superoxides; Swine; Young Adult

Most of the anti-breast cancer drugs are often limited owing to drug resistance and serious adverse reactions. Therefore, development of more targeted and low toxic drugs from traditional Chinese medicines for breast cancer are needed. At the same time, establishment of fast and effective drug screening methods are urgently required. We describe here a 2D LC method of MDA-MB-231 cell membrane chromatography combined with HPLC/MS for recognition, separation, and identification of target components from traditional Chinese medicine Cortex Magnolia officinalis. The MDA-MB-231 cells membrane was used to prepare the chromatographic stationary phase in the first dimension. The active compounds had a retention characteristic on the cell membrane chromatography model (10 × 2.0 mm, 5 μm). The retention fractions were enriched using an online C(18) column (10 × 1.0 mm, 5 μm) and were analyzed by the second dimension RP chromatography. Finally, the activity of the retention fractions was tested through in vitro experiments. Results showed that the retention fractions were honokiol and magnolol and the inhibition rate on MDA-MB-231 cell growth were 23 and 64 μM, respectively. These results support the conclusion that this coupled analytical technique could be an efficient method in drug discovery.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Chromatography, Liquid; Drug Screening Assays, Antitumor; Drugs, Chinese Herbal; Female; Humans; Lignans; Magnolia; Mass Spectrometry

To investigate the chemical constituents of Elephantopus mollis.. Compounds were separated and purified by various column chromatographies including macroporous resin, ODS, Sephadex LH-20 and silica gel columns. The structures were identified by their physicochemical properties and spectroscopic data.. Nine compounds were identified as 2beta-deethoxy-2-hydroxyphantomolin (1), 2beta-methoxy-2-deethoxyphantomolin (2), 2beta-methoxy-2-deethoxy-8-O-deacylphantomolin-8-O-tigli-nate (3), molephantinin (4), betulinic acid (5), magnolol (6), honokiol (7), dibutly phthalate (8) and tricin (9).. Compounds 5-9 are isolated from this plant for the first time.

Topics: Asteraceae; Betulinic Acid; Biphenyl Compounds; Dibutyl Phthalate; Flavonoids; Lignans; Magnetic Resonance Spectroscopy; Molecular Structure; Pentacyclic Triterpenes; Triterpenes

The activation of signal transducers and activators of transcription 3 (STAT3) has been closely linked with the proliferation, survival, invasion, and angiogenesis of hepatocellular carcinoma (HCC) and represents an attractive target for therapy. In the present report, we investigated whether honokiol mediates its effect through interference with the STAT3 activation pathway. The effect of honokiol on STAT3 activation, associated protein kinases, and phosphatase, STAT3-regulated gene products and apoptosis was investigated using both functional proteomics tumor pathway technology platform and different HCC cell lines. We found that honokiol inhibited both constitutive and inducible STAT3 activation in a dose- and time-dependent manner in HCC cells. The suppression was mediated through the inhibition of activation of upstream kinases c-Src, Janus-activated kinase 1, and Janus-activated kinase 2. Vanadate treatment reversed honokiol-induced down-regulation of STAT3, suggesting the involvement of a tyrosine phosphatase. Indeed, we found that honokiol induced the expression of tyrosine phosphatase SHP-1 that correlated with the down-regulation of constitutive STAT3 activation. Moreover, deletion of SHP-1 gene by siRNA abolished the ability of honokiol to inhibit STAT3 activation. The inhibition of STAT3 activation by honokiol led to the suppression of various gene products involved in proliferation, survival, and angiogenesis. Finally, honokiol inhibited proliferation and significantly potentiated the apoptotic effects of paclitaxel and doxorubicin in HCC cells. Overall, the results suggest that honokiol is a novel blocker of STAT3 activation and may have a great potential for the treatment of HCC and other cancers.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Carcinoma, Hepatocellular; Caspase 3; Cell Line, Tumor; Cell Proliferation; Cell Survival; CSK Tyrosine-Protein Kinase; G1 Phase; Genes, Reporter; Humans; Interleukin-6; Janus Kinase 1; Janus Kinase 2; Lignans; Liver Neoplasms; Models, Biological; Poly(ADP-ribose) Polymerases; Protein Tyrosine Phosphatase, Non-Receptor Type 6; Protein-Tyrosine Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction; src-Family Kinases; STAT3 Transcription Factor

Metastasizing osteosarcoma has a mean 5-year survival rate of only 20% to 30%. Therefore, novel chemotherapeutics for more effective treatment of this disease are required.. The antineoplastic activity of honokiol, which was demonstrated previously in numerous malignancies, was studied in vivo in C3H mice subcutaneously injected with syngeneic β-galactosidase bacterial gene (lacZ)-expressing LM8 osteosarcoma (LM8-lacZ) cells. In vitro cytotoxic effects of honokiol were investigated in 8 human and 2 murine osteosarcoma cell lines with different in vivo metastatic potential.. Seven days after subcutaneous flank injection of LM8-lacZ cells, daily intraperitoneal treatment of mice with 150 mg/kg honokiol reduced the number of micrometastases in the lung by 41% and reduced the number of macrometastases in the lung and liver by 69% and 80%, respectively, compared with control. Primary tumor growth was not inhibited. In osteosarcoma cell lines, honokiol inhibited the metabolic activity with a half-maximal concentration (IC(50) ) between 8.0 μg/mL and 16 μg/mL. Cyclosporin A partially reversed the inhibition of metabolic activity in LM8-lacZ cells. Cell proliferation and wound healing migration of LM8-lacZ cells were inhibited by honokiol with an IC(50) between 5.0 μg/mL and 10 μg/mL. Higher concentrations caused rapid cell death, which was distinct from necrosis, apoptosis, or autophagy but was associated with swelling of the endoplasmic reticulum, cytoplasmic vacuolation, and morphologically altered mitochondria.. Honokiol exhibited prominent antimetastatic activity in experimental osteosarcoma and caused rapid cell death in vitro that was unrelated to necrosis, apoptosis, or autophagy. The authors concluded that honokiol has considerable potential for the treatment of metastasizing osteosarcoma.

Topics: Animals; Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Bone Neoplasms; Cell Death; Cell Line, Tumor; Cell Proliferation; Drug Screening Assays, Antitumor; Female; Humans; Lignans; Liver Neoplasms; Lung Neoplasms; Mice; Mice, Inbred C3H; Osteosarcoma

Interleukin-8 (IL-8) and tumour necrosis factor-alpha (TNF-α) from keratinocyte play important roles in initiating the inflammatory process of acne. They are used as major elements to evaluate the anti-inflammatory activity of drugs. In this study, various active constituents extracted from Chinese medicinal herbs were tested for their anti-inflammatory effects against P. acnes using ELISA. Among the constituents, matrine, baicalin, ursolic acid, sodium danshensu, magnolol, honokiol, hesperidin and andrographolide significantly reduced IL-8 and TNF-α by human HaCaT keratinocyte cells pretreated with heat-killed P. acnes. Excepting hesperidin, these active constituents presented dose-dependent inhibitory effects. Our studies showed that all of them exhibited low cytotoxicity at 5 µg mL⁻¹ in tested cell lines, and even at 50 µg mL⁻¹, in the cases of matrine, baicalin, ursolic acid and sodium danshensu. Based on the obtained results, it can be suggested that these active constituents are potential acne-mitigating candidates for cosmetic applications.

Topics: Anti-Inflammatory Agents; Biphenyl Compounds; Cell Line; Drugs, Chinese Herbal; Flavonoids; Humans; Interleukin-8; Keratinocytes; Lignans; Propionibacterium acnes; Triterpenes; Ursolic Acid

As the molecular biology of vestibular schwannoma (VS) is better understood, new means of targeting the pathways involved for intervention in schwannoma cells are being developed. Honokiol, a bioactive constituent of Magnolia officinalis, has attracted attention due to its diverse biological effects. This study was conducted to determine the inhibitory effect of honokiol on schwannoma cell proliferation.. HEI 193 cells were used to investigate the growth-inhibitory effects of honokiol. Cell proliferation was assessed by MTT assays. Apoptosis was measured by flow cytometry analysis and immunofluorescence staining including Hoechst 33342 and TUNEL. Western blot analysis was used to assess the potential inhibition of extracellular signal-regulated kinase (ERK) and AKT signaling by honokiol.. Honokiol exhibited significant antiproliferative activity in a dose-dependent manner on HEI 193 cells. Significant apoptosis was detected on schwannoma cells with 7 mg/mL(IC50) honokiol. Western blot analysis showed significant inhibition of ERK phosphorylation.. Honokiol, a low molecular weight natural product, inhibits cell proliferation and promotes apoptosis in schwannoma cells by targeting the ERK pathway. Our data suggest that honokiol can be evaluated as a chemotherapeutic agent for VS.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Proliferation; Humans; Lignans; Neuroma, Acoustic; Tumor Cells, Cultured

Honokiol, a small active molecular compound extracted from magnolia, has recently been shown to inhibit hepatitis C virus (HCV) infection in vitro.. This study further characterized aspects of the HCV lifecycle affected by the antiviral functions of honokiol.. The influence of honokiol on HCV infection, entry, translation and replication was assessed in Huh-7.5.1 cells using cell culture-derived HCV (HCVcc), HCV pseudo-type (HCVpp) and sub-genomic replicons.. Honokiol had strong antiviral effect against HCVcc infection at non-toxic concentrations. Combined with interferon-α, its inhibitory effect on HCVcc was more profound than that of ribavirin. Honokiol inhibited the cell entry of lentiviral particles pseudo-typed with glycoproteins from HCV genotypes 1a, 1b, and 2a, but not of the vesicular stomatitis virus. It had inefficient activity on HCV internal ribosome entry site (IRES)-translation at concentrations with significant anti-HCVcc effects. The expression levels of components of replication complex, NS3, NS5A and NS5B, were down-regulated by honokiol in a dose-dependent manner. It also inhibited HCV replication dose dependently in both genotypes 1b and 2a sub-genomic replicons.. Honokiol inhibits HCV infection by targeting cell entry and replication and, only at a concentration >30 μM, IRES-mediated translation of HCV life cycle. Based on its high therapeutic index (LD(50) /EC(90)  = 5.4), honokiol may be a promising drug for the treatment of HCV infection.

Topics: Antiviral Agents; Biphenyl Compounds; Cell Line, Tumor; Dose-Response Relationship, Drug; Drug Therapy, Combination; Drugs, Chinese Herbal; Gene Expression Regulation, Viral; Genes, Reporter; Genotype; HEK293 Cells; Hepacivirus; Humans; Interferon-alpha; Lignans; Magnolia; Phenotype; Plants, Medicinal; Protein Biosynthesis; Transfection; Viral Proteins; Virus Internalization; Virus Replication

An efficient synthesis of honokiol with Suzuki-Miyaura cross coupling obtained an overall yield of 45%. The proposed approach successfully synthesized several structurally similar alkyl, alkenyl and alkynyl analogues, seven of which showed potential neuropreventive activity against MPP(+)-induced and CHP/TBHP oxidative stress induced neuroblastoma cell death.

Topics: Biphenyl Compounds; Catalysis; Cell Death; Cell Line, Tumor; Cell Survival; Chemistry, Pharmaceutical; Drug Design; Humans; Lignans; Models, Chemical; Neuroblastoma; Neuroprotective Agents; Oxidative Stress; Parkinson Disease; Reactive Oxygen Species; Time Factors

A versatile synthetic route is reported towards the preparation of new analogues for potent neurotrophic agent biaryl-type lignan honokiol. A focused 24-membered library of derivatives containing five different groups at 5'-position of honokiol has been prepared in fair to good overall yields. Compared to the natural product, or to analogues with a short alkyl chain in this position, these new derivatives have lost most of the neurotrophic activity.

Topics: Animals; Biphenyl Compounds; Cells, Cultured; Humans; Immunohistochemistry; Lignans; Molecular Structure; Nerve Growth Factors; Neurites; Neurogenesis; Small Molecule Libraries; Structure-Activity Relationship

We investigated the properties of honokiol, a natural rexinoid, as a regulator of retinoid X receptor (RXR) heterodimers with various partner nuclear receptors (NRs), in comparison with those of the synthetic rexinoid bexarotene. Honokiol alone was hardly capable of activating peroxisome proliferator-activated receptor (PPAR) γ/RXR, RXR/liver X receptor (LXR), and RXR/vitamin D receptor (VDR) heterodimers, whereas it effectively potentiated their activation by agonists for the partner NRs of the RXR heterodimers. These findings were further supported by increased mRNA and protein levels for the respective NR target genes. Bexarotene alone activated PPARγ/RXR and RXR/LXR heterodimers, but not RXR/VDR heterodimers, and facilitated the activation of all three RXR heterodimers by the respective PPARγ, LXR, and VDR agonists. When the potencies of honokiol and bexarotene were compared, honokiol was able to serve as a subsidiary agonist in the activation of RXR heterodimers in a similar manner to bexarotene. However, it seemed to potentiate the activation of PPARγ/RXR heterodimers by the PPARγ agonist rosiglitazone more efficiently than bexarotene, and was a less potent RXR agonist than bexarotene. In conclusion, we have demonstrated that honokiol is a rexinoid that possesses distinct properties from bexarotene, and mainly has subsidiary roles in the activation of RXR heterodimers by potentiating the activation of RXR heterodimers by agonists for the partner NRs.

Topics: 3T3-L1 Cells; Animals; Anticarcinogenic Agents; Bexarotene; Biphenyl Compounds; Caco-2 Cells; Dimerization; Humans; Lignans; Liver X Receptors; Magnolia; Mice; Orphan Nuclear Receptors; Plant Extracts; PPAR gamma; Receptors, Calcitriol; Receptors, Cytoplasmic and Nuclear; Retinoid X Receptors; RNA, Messenger; Rosiglitazone; Signal Transduction; Tetrahydronaphthalenes; Thiazolidinediones

Severe acute pancreatitis remains a life-threatening disease with a high mortality rate among a defined proportion of those affected. Apoptosis has been hypothesized to be a beneficial form of cell death in acute pancreatitis. Honokiol, a low-molecular-weight natural product, possesses the ability of anti-inflammation and apoptosis induction. Here, we investigate whether honokiol can ameliorate severe acute pancreatitis and the associated acute lung injury in a mouse model. Mice received six injections of cerulein at 1-h intervals, then given one intraperitoneal injection of bacterial lipopolysaccharide for the induction of severe acute pancreatitis. Moreover, mice were intraperitoneally given vehicle or honokiol 10 min after the first cerulein injection. Honokiol protected against the severity of acute pancreatitis in terms of increased serum amylase and lipase levels, pancreas pathological injury, and associated acute lung injury. Honokiol significantly reduced the increases in serum tumor necrosis factor-α, interleukin 1, and nitric oxide levels 3 h and serum high-mobility group box 1 24 h after acute pancreatitis induction. Honokiol also significantly decreased myeloperoxidase activities in the pancreas and the lungs. Endoplasmic reticulum stress-related molecules eIF2α (phosphorylated) and CHOP protein expressions, apoptosis, and caspase-3 activity were increased in the pancreas of mice with severe acute pancreatitis, which was unexpectedly enhanced by honokiol treatment. These results suggest that honokiol protects against acute pancreatitis and limits the spread of inflammatory damage to the lung in a severe acute pancreatitis mouse model. The acceleration of pancreatic cell apoptosis by honokiol may play a pivotal role.

Topics: Acinar Cells; Acute Disease; Acute Lung Injury; Amylases; Animals; Apoptosis; Biphenyl Compounds; Caspase 3; Disease Models, Animal; Endoplasmic Reticulum; Enzyme Inhibitors; HMGB1 Protein; Interleukin-1beta; Lignans; Lipase; Lung; Male; Mice; Mice, Inbred BALB C; Pancreas; Pancreatitis; Time Factors; Transcription Factor CHOP; Tumor Necrosis Factor-alpha

Neuroblastomas, an embryonic cancer of the sympathetic nervous system, often occur in young children. Honokiol, a small-molecule polyphenol, has multiple therapeutic effects and pharmacological activities. This study was designed to evaluate whether honokiol could pass through the blood-brain barrier (BBB) and induce death of neuroblastoma cells and its possible mechanisms. Primary cerebral endothelial cells (CECs) prepared from mouse brain capillaries were cultured at a high density for 4 days, and these cells formed compact morphologies and expressed the ZO-1 tight-junction protein. A permeability assay showed that the CEC-constructed barrier obstructed the passing of FITC-dextran. Analyses by high-performance liquid chromatography and the UV spectrum revealed that honokiol could traverse the CEC-built junction barrier and the BBB of ICR mice. Exposure of neuroblastoma neuro-2a cells and NB41A3 cells to honokiolinduced cell shrinkage and decreased cell viability. In parallel, honokiol selectively induced DNA fragmentation and cell apoptosis rather than cell necrosis. Sequential treatment of neuro-2a cells with honokiol increased the expression of the proapoptotic Bax protein and its translocation from the cytoplasm to mitochondria. Honokiol successively decreased the mitochondrial membrane potential but increased the release of cytochrome c from mitochondria. Consequently, honokiol induced cascade activation of caspases-9, -3, and -6. In comparison, reducing caspase-6 activity by Z-VEID-FMK, an inhibitor of caspase-6, simultaneously attenuated honokiol-induced DNA fragmentation and cell apoptosis. Taken together, this study showed that honokiol can pass through the BBB and induce apoptotic insults to neuroblastoma cells through a Bax-mitochondrion-cytochrome c-caspase protease pathway. Therefore, honokiol may be a potential candidate drug for treating brain tumors.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; bcl-2-Associated X Protein; Biphenyl Compounds; Blood-Brain Barrier; Brain Neoplasms; Caspases; Cell Line, Tumor; Cell Survival; Cells, Cultured; Cytochromes c; DNA Fragmentation; Endothelial Cells; Humans; Lignans; Membrane Potential, Mitochondrial; Mice; Mice, Inbred ICR; Mitochondria; Neuroblastoma; Proto-Oncogene Proteins c-bcl-2; Tight Junctions

Honokiol, a constituent of Magnolia obovata, has various pharmacological effects, including protection against cerebral ischemia. However, few studies have been conducted to evaluate the possible neuroprotective effects of honokiol against cerebral ischemia. We recently reported that cerebral ischemic neuronal damage could be triggered by glucose intolerance that develops after the onset of ischemic stress (i.e., post-ischemic glucose intolerance). In addition, suppression of post-ischemic glucose intolerance significantly ameliorated ischemic neuronal damage. Here, we investigated the effects of honokiol on the development of post-ischemic glucose intolerance and neuronal damage. Mice were subjected to middle cerebral artery occlusion (MCAO) for 2 h. The development of post-ischemic glucose intolerance on day 1 and neuronal damage on day 3 after MCAO were significantly reduced by intraperitoneal administration of honokiol (10 mg/kg) compared with the vehicle-treated group. Honokiol did not affect serum insulin or adiponectin levels. However, honokiol significantly decreased the expression of phosphoenolpyruvate carboxykinase and increased the expression of 5'-AMP-activated protein kinase (AMPK) on day 1 after MCAO, compared with the vehicle-treated MCAO group. The results of this study suggest that honokiol could prevent post-ischemic glucose intolerance in an AMPK-dependent manner, which may be involved in the neuroprotective effects of honokiol against cerebral ischemia.

Topics: Adiponectin; AMP-Activated Protein Kinases; Animals; Biphenyl Compounds; Brain Ischemia; Glucose Intolerance; Infarction, Middle Cerebral Artery; Insulin; Lignans; Male; Mice; Phosphoenolpyruvate Carboxykinase (ATP)

To investigate the protective effect of honokiol, a phytochemical used in traditional medicine, on testicular injury after torsion/detorsion (T/D) in a rat model. Testicular torsion is a medical emergency that can cause impairment of semen quality and permanent testicular atrophy or loss.. Male Wistar rats were randomized to each time point of each group (n = 6/time point/group). After 2 hours of torsion, the testes were counter-rotated to the natural position. The rats in each group underwent a sham operation, T/D, or T/D with honokiol treatment (5 mg/kg and 10 mg/kg intraperitoneally, immediately before detorsion). Bilateral orchiectomy was performed at 6 and 24 hours and 3 months after detorsion. The testes were examined histologically. Apoptosis and endoplasmic reticulum stress were detected by Western blot.. Histologic examination revealed that testicular T/D induced acute injury after 6 and 24 hours, and spermatogenesis was decreased at 3 months of follow-up. At 24 hours after T/D, increases were found in the activation of apoptosis-related molecules [poly (ADP-ribose) polymerase and caspases 3 and 7], and the expression levels of endoplasmic reticulum stress-associated molecules (phosphorylated-eukaryotic translation initiation factor 2 subunit α and CCAAT/enhancer-binding protein homologous protein). These increases were significantly reversed with honokiol treatment. Furthermore, honokiol effectively reversed the inhibition of spermatogenesis in testes treated with T/D for 3 months.. The results of our study have shown that the endoplasmic reticulum stress-related apoptotic pathway is involved in testicular injury after testicular T/D. It remains to be determined whether alterations in this pathway would have a protective affect against reperfusion damage.

Topics: Animals; Apoptosis; Biphenyl Compounds; Drugs, Chinese Herbal; Endoplasmic Reticulum; Lignans; Male; Rats; Rats, Wistar; Reperfusion Injury; Spermatic Cord Torsion; Spermatogenesis

A method for the separation and determination of honokiol and magnolol in Magnolia officinalis and its medicinal preparation is developed by capillary zone electrophoresis and response surface methodology. The concentration of borate, content of organic modifier, and applied voltage are selected as variables. The optimized conditions (i.e., 16 mmol/L sodium tetraborate at pH 10.0, 11% methanol, applied voltage of 25 kV and UV detection at 210 nm) are obtained and successfully applied to the analysis of honokiol and magnolol in Magnolia officinalis and Huoxiang Zhengqi Liquid. Good separation is achieved within 6 min. The limits of detection are 1.67 µg/mL for honokiol and 0.83 µg/mL for magnolol, respectively. In addition, an artificial neural network with "3-7-1" structure based on the ratio of peak resolution to the migration time of the later component (R(s)/t) given by Box-Behnken design is also reported, and the predicted results are in good agreement with the values given by the mathematic software and the experimental results.

Topics: Biphenyl Compounds; Computational Biology; Drugs, Chinese Herbal; Electrophoresis, Capillary; Lignans; Magnolia; Models, Statistical; Neural Networks, Computer; Plant Bark; Reproducibility of Results

The extracts from leaves, heartwood, barks and roots of the Streblus asper were investigated for anti-HBV activities, separately. The results suggested that the MeOH extracts of the heartwood, barks, and roots exhibited good anti-HBV activities. Further investigations displayed that ethyl acetate and n-butanol soluble parts of their MeOH extracts showed more significant anti-HBV activities. Moreover, a new lignan, together with 11 known compounds, was isolated from n-butanol-soluble part of MeOH extract of the roots of S. asper. The structures were elucidated by spectroscopic methods, including 1D NMR ((1)H NMR, (13)C NMR), 2D NMR (HMQC, HMBC) and HR-EI-MS experiments. Compounds 1-3 were evaluated for their anti-HBV activities. Honokiol showed significant anti-HBV activity with IC(50) values of 3.14μM and 4.74μM for HBsAg and HBeAg with no cytotoxicity respectively, while lamivudine (3TC, positive control) exhibited weak anti-HBV activity with IC(50) values of 11.81μM and 25.80μM for HBsAg and HBeAg respectively.

Topics: Antiviral Agents; Biphenyl Compounds; Hep G2 Cells; Hepatitis B; Hepatitis B virus; Humans; Inhibitory Concentration 50; Lamivudine; Lignans; Molecular Structure; Moraceae; Phytotherapy; Plant Extracts; Plant Roots; Plant Structures

Cancer stem cells are implicated in resistance to ionizing radiation (IR) and chemotherapy. Honokiol, a biphenolic compound has been used in traditional Chinese medicine for treating various ailments. In this study, we determined the ability of honokiol to enhance the sensitivity of colon cancer stem cells to IR. The combination of honokiol and IR suppressed proliferation and colony formation while inducing apoptosis of colon cancer cells in culture. There were also reduced numbers and size of spheroids, which was coupled with reduced expression of cancer stem cell marker protein DCLK1. Flow cytometry studies confirmed that the honokiol-IR combination reduced the number of DCLK1+ cells. In addition, there were reduced levels of activated Notch-1, its ligand Jagged-1, and the downstream target gene Hes-1. Furthermore, expression of components of the Notch-1 activating γ-secretase complex, presenilin 1, nicastrin, Pen2, and APH-1 was also suppressed. On the other hand, the honokiol effects were mitigated when the Notch intracellular domain was expressed. To determine the effect of honokiol-IR combination on tumor growth in vivo, nude mice tumor xenografts were administered honokiol intraperitoneally and exposed to IR. The honokiol-IR combination significantly inhibited tumor xenograft growth. In addition, there were reduced levels of DCLK1 and the Notch signaling-related proteins in the xenograft tissues. Together, these data suggest that honokiol is a potent inhibitor of colon cancer growth that targets the stem cells by inhibiting the γ-secretase complex and the Notch signaling pathway. These studies warrant further clinical evaluation for the combination of honokiol and IR for treating colon cancers.

Topics: Animals; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Colonic Neoplasms; Combined Modality Therapy; Down-Regulation; Drugs, Chinese Herbal; HCT116 Cells; Humans; Immunohistochemistry; Lignans; Male; Mice; Mice, Nude; Neoplastic Stem Cells; Nitric Oxide Synthase; Signal Transduction; Transfection; Xenograft Model Antitumor Assays

Honokiol, a small-molecule polyphenol isolated from magnolia species, is widely known for its therapeutic potential as an antiinflammatory, antithrombosis, and antioxidant agent, and more recently, for its protective function in the pathogenesis of carcinogenesis. In the present study, we sought to examine the effectiveness of honokiol in inhibiting migration and invasion of breast cancer cells and to elucidate the underlying molecular mechanisms.. Clonogenicity and three-dimensional colony-formation assays were used to examine breast cancer cell growth with honokiol treatment. The effect of honokiol on invasion and migration of breast cancer cells was evaluated by using Matrigel invasion, scratch-migration, spheroid-migration, and electric cell-substrate impedance sensing (ECIS)-based migration assays. Western blot and immunofluorescence analysis were used to examine activation of the liver kinase B1 (LKB1)-AMP-activated protein kinase (AMPK) axis. Isogenic LKB1-knockdown breast cancer cell line pairs were developed. Functional importance of AMPK activation and LKB1 overexpression in the biologic effects of honokiol was examined by using AMPK-null and AMPK-wild type (WT) immortalized mouse embryonic fibroblasts (MEFs) and isogenic LKB1-knockdown cell line pairs. Finally, mouse xenografts, immunohistochemical and Western blot analysis of tumors were used.. Analysis of the underlying molecular mechanisms revealed that honokiol treatment increases AMP-activated protein kinase (AMPK) phosphorylation and activity, as evidenced by increased phosphorylation of the downstream target of AMPK, acetyl-coenzyme A carboxylase (ACC) and inhibition of phosphorylation of p70S6kinase (pS6K) and eukaryotic translation initiation factor 4E binding protein 1 (4EBP1). By using AMPK-null and AMPK-WT (MEFs), we found that AMPK is required for honokiol-mediated modulation of pACC-pS6K. Intriguingly, we discovered that honokiol treatment increased the expression and cytoplasmic translocation of tumor-suppressor LKB1 in breast cancer cells. LKB1 knockdown inhibited honokiol-mediated activation of AMPK and, more important, inhibition of migration and invasion of breast cancer cells. Furthermore, honokiol treatment resulted in inhibition of breast tumorigenesis in vivo. Analysis of tumors showed significant increases in the levels of cytoplasmic LKB1 and phospho-AMPK in honokiol-treated tumors.. Taken together, these data provide the first in vitro and in vivo evidence of the integral role of the LKB1-AMPK axis in honokiol-mediated inhibition of the invasion and migration of breast cancer cells. In conclusion, honokiol treatment could potentially be a rational therapeutic strategy for breast carcinoma.

Topics: Adaptor Proteins, Signal Transducing; AMP-Activated Protein Kinase Kinases; AMP-Activated Protein Kinases; Animals; Antineoplastic Agents; Biphenyl Compounds; Breast Neoplasms; Cell Cycle Proteins; Cell Line, Tumor; Cell Movement; Cell Transformation, Neoplastic; Enzyme Activation; Female; Gene Expression; Humans; Lignans; Mice; Mice, Nude; Neoplasm Invasiveness; Phosphoproteins; Phosphorylation; Protein Processing, Post-Translational; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins c-akt; Ribosomal Protein S6 Kinases; Spheroids, Cellular; TOR Serine-Threonine Kinases; Tumor Burden; Xenograft Model Antitumor Assays

The National Center for Complementary and Alternative Medicine (NCCAM) estimates that nearly 40% of adults in the United States use alternative medicines, often in the form of an herbal supplement. Extracts from the tree bark of magnolia species have been used for centuries in traditional Chinese and Japanese medicines to treat a variety of neurological diseases, including anxiety, depression, and seizures. The active ingredients in the extracts have been identified as the bi-phenolic isomers magnolol and honokiol. These compounds were shown to enhance the activity of GABA(A) receptors, consistent with their biological effects. The GABA(A) receptors exhibit substantial subunit heterogeneity, which influences both their functional and pharmacological properties. We examined the activity of magnolol and honokiol at different populations of both neuronal and recombinant GABA(A) receptors to characterize their mechanism of action and to determine whether sensitivity to modulation was dependent upon the receptor's subunit composition. We found that magnolol and honokiol enhanced both phasic and tonic GABAergic neurotransmission in hippocampal dentate granule neurons. In addition, all recombinant receptors examined were sensitive to modulation, regardless of the identity of the α, β, or γ subunit subtype, although the compounds showed particularly high efficacy at δ-containing receptors. This direct positive modulation of both synaptic and extra-synaptic populations of GABA(A) receptors suggests that supplements containing magnolol and/or honokiol would be effective anxiolytics, sedatives, and anti-convulsants. However, significant side-effects and risk of drug interactions would also be expected.

Topics: Animals; Biphenyl Compounds; Drugs, Chinese Herbal; GABA Agents; HEK293 Cells; Hippocampus; Humans; Inhibitory Postsynaptic Potentials; Lignans; Neurons; Rats; Receptors, GABA-A; Synaptic Transmission

Honokiol, a naturally occurring biphenyl, possesses anti-neoplastic properties. We investigated activities of honokiol against adult T-cell leukemia (ATL) associated with human T-cell leukemia virus type 1 (HTLV-1).. Cell viability was assessed using colorimetric assay. Propidium iodide staining was performed to determine cell cycle phase. Apoptotic effects were evaluated by 7A6 detection and caspases activity. Expressions of cell cycle- and apoptosis-associated proteins were analyzed by Western blot. We investigated the efficacy of honokiol in mice harboring tumors of HTLV-1-infected T-cell origin.. Honokiol exhibited cytotoxic activity against HTLV-1-infected T-cell lines and ATL cells. We identified two different effects of honokiol on HTLV-1-infected T-cell lines: cell cycle inhibition and induction of apoptosis. Honokiol induced G1 cell cycle arrest by reducing the expression of cyclins D1, D2, E, CDK2, CDK4, CDK6 and c-Myc, while apoptosis was induced via reduced expression of cIAP-2, XIAP and survivin. The induced apoptosis was also associated with activation of caspases-3 and -9. In addition, honokiol suppressed the phosphorylation of IκBα, IKKα, IKKβ, STAT3, STAT5 and Akt, down-regulated JunB and JunD, and inhibited DNA binding of NF-κB, AP-1, STAT3 and STAT5. These effects resulted in the inactivation of survival signals including NF-κB, AP-1, STATs and Akt. Honokiol was highly effective against ATL in mice. Our data suggested that honokiol is a systemically available, non-toxic inhibitor of ATL cell growth that should be examined for potential clinical application.. Our findings provide a rationale for clinical evaluation of honokiol for the management of ATL.

Topics: Adult; Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Blotting, Western; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Proliferation; Cell Survival; Electrophoretic Mobility Shift Assay; Female; Humans; Leukemia-Lymphoma, Adult T-Cell; Lignans; Mice; Mice, Inbred ICR; Mice, SCID; NF-kappa B; Signal Transduction; Tumor Cells, Cultured

In this study, the binding modes of honokiol (HK) and magnolol (MG) with human serum albumin (HSA) have been established under imitated physiological condition, which was very important to understand the pharmacokinetics and toxicity of HK or MG. The experimental results proved that the fluorescence of HSA was quenched by HK or MG through a static quenching procedure. The binding constants of HK-HSA and MG-HSA complexes were 5.304 and 263.755×10(4) L mol(-1) at 298 K, respectively. The binding process was a spontaneous molecular interaction procedure, in which the hydrophobic interaction played a major role in the formation of the HK-HSA complex, whereas, the binding interaction between MG and HSA might involve the hydrophobic interaction strongly and electrostatic interaction. In addition, the effect of HK/MG on the secondary structure of HSA was analyzed using CD, UV-vis absorption, Fourier transform infrared (FT-IR), synchronous fluorescence and three-dimensional fluorescence spectra. According to Förster no-radiation energy transfer theory, the binding distance of HSA to HK or MG was calculated to be 1.842 or 1.238 nm. Besides, the effects of common ions on the binding constants of HSA-HK/MG systems were also discussed.

Topics: Biphenyl Compounds; Circular Dichroism; Humans; Hydrophobic and Hydrophilic Interactions; Lignans; Protein Binding; Serum Albumin; Spectrometry, Fluorescence; Spectroscopy, Fourier Transform Infrared; Static Electricity

Honokiol possesses many pharmacological activities including anti-cancer properties. Here in, we designed and synthesized honokiol analogs that block major honokiol metabolic pathway which may enhance their effectiveness. We studied their cytotoxicity in human cancer cells and evaluated possible mechanism of cell cycle arrest. Two analogs, namely 2 and 4, showed much higher growth inhibitory activity in A549 human lung cancer cells and significant increase of cell population in the G0-G1 phase. Further elucidation of the inhibition mechanism on cell cycle showed that analogs 2 and 4 inhibit both CDK1 and cyclin B1 protien levels in A549 cells.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Blotting, Western; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Humans; Inhibitory Concentration 50; Lignans; Lung Neoplasms; Molecular Structure; Structure-Activity Relationship

Decoctions of the Chinese herb houpu contain honokiol and are used to treat a variety of mental disorders, including depression. Depression commonly presents alongside sleep disorders and sleep disturbances, which appear to be a major risk factor for depression. Here, we have evaluated the somnogenic effect of honokiol and the mechanisms involved.. Honokiol was administered i.p. at 20:00 h in mice. Flumazenil, an antagonist at the benzodiazepine site of the GABA(A) receptor, was administered i.p. 15 min before honokiol. The effects of honokiol were measured by EEG and electromyogram (EMG), c-Fos expression and in vitro electrophysiology.. Honokiol (10 and 20 mg·kg⁻¹) significantly shortened the sleep latency to non-rapid eye movement (non-REM, NREM) sleep and increased the amount of NREM sleep. Honokiol increased the number of state transitions from wakefulness to NREM sleep and, subsequently, from NREM sleep to wakefulness. However, honokiol had no effect on either the amount of REM sleep or EEG power density of both NREM and REM sleep. Honokiol increased c-Fos expression in ventrolateral preoptic area (VLPO) neurons, as examined by immunostaining, and excited sleep-promoting neurons in the VLPO by whole-cell patch clamping in the brain slice. Pretreatment with flumazenil abolished the somnogenic effects and activation of the VLPO neurons by honokiol.. Honokiol promoted NREM sleep by modulating the benzodiazepine site of the GABA(A) receptor, suggesting potential applications in the treatment of insomnia, especially for patients who experience difficulty in falling and staying asleep.

Topics: Animals; Biphenyl Compounds; Dose-Response Relationship, Drug; Electroencephalography; Electromyography; Flumazenil; GABA Modulators; Gene Expression; Injections, Intraperitoneal; Lignans; Male; Mice; Mice, Inbred C57BL; Patch-Clamp Techniques; Proto-Oncogene Proteins c-fos; Receptors, GABA-A; Sleep; Sleep Initiation and Maintenance Disorders; Sleep, REM; Time Factors

The MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide) assay is a classical method for screening cytotoxic anti-cancer agents. Candidate drugs from the MTT assay need in vivo models to test their efficiency and to assess the absorption, distribution, metabolism, excretion, and toxicity of the drugs. An in vivo screening model could increase the rate of development of anti-cancer drugs. Here, we used zebrafish to screen a library of 502 natural compounds and compared the results with those from an MTT assay of the MCF7 breast cancer cell line. We identified 59 toxic compounds in the zebrafish screen, 21 of which were also identified by the MTT assay, and 28 of which were already known for their anti-cancer and apoptosis-inducing effects. These compounds induced apoptosis and activated the p53 pathway in zebrafish within 3h treatment. Our results indicate that zebrafish is a simple, reliable and highly efficient in vivo tool for cancer drug screening, and could complement the MTT assay.

Topics: Animals; Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Camptothecin; Cell Line, Tumor; Coloring Agents; Dimethyl Sulfoxide; Drug Screening Assays, Antitumor; Emodin; Gene Expression; Humans; Lignans; Naphthoquinones; Rotenone; Tetrazolium Salts; Thiazoles; Transcriptional Activation; Tumor Suppressor Protein p53; Zebrafish

Hypoxia-inducible-factor (HIF)-mediated expression of pro-angiogenic genes under hypoxic conditions is the fundamental cause of pathological neovascularization in retinal ischemic diseases and cancers. Recent studies have shown that histone lysine demethylases (KDMs) play a key role in the amplification of HIF signaling and expression of pro-angiogenic genes. Thus, the inhibitors of the HIF pathway or KDMs can have profound therapeutic value for diseases caused by pathological neovascularization. Here, we show that hypoxia-mediated expression of KDMs is a conserved process across multiple cell lines. Moreover, we report that honokiol, a biphenolic phytochemical extracted from Magnolia genus which has been used for thousands of years in the traditional Japanese and Chinese medicine, is a potent inhibitor of the HIF pathway as well as hypoxia-induced expression of KDMs in a number of cancer and retinal pigment epithelial cell lines. Further, treating the cells with honokiol leads to inhibition of KDM-mediated induction of pro-angiogenic genes (adrenomedullin and growth differentiation factor 15) under hypoxic conditions. Our results provide an evidence-based scientific explanation for therapeutic benefits observed with honokiol and warrant its further clinical evaluation for the treatment of pathological neovascularization in retinal ischemic diseases and cancers.

Topics: Angiogenesis Inhibitors; Biphenyl Compounds; Cell Hypoxia; Cell Line; Cell Line, Tumor; Gene Expression; Histone Demethylases; Humans; Hypoxia; Hypoxia-Inducible Factor 1; Lignans; Neovascularization, Pathologic; Oxygen

The cannabinoid type-2 G protein-coupled (CB₂) receptor is an emerging therapeutic target for pain management and immune system modulation. In a mouse model of Alzheimer's disease (AD) the orally administered natural product 4'-O-methylhonokiol (MH) has been shown to prevent amyloidogenesis and progression of AD by inhibiting neuroinflammation. In this commentary we discuss an intriguing link between the recently found CB₂ receptor-mediated molecular mechanisms of MH and its anti-inflammatory and protective effects in AD animal models. We argue that the novel cannabimimetic MH may exert its beneficial effects via modulation of CB₂ receptors expressed in microglial cells and astrocytes. The recent findings provide further evidence for a potential role of CB₂ receptors in the pathophysiology of AD, spurring target validation and drug discovery.

Topics: Alzheimer Disease; Animals; Biphenyl Compounds; Humans; Inflammation; Lignans; Plants, Medicinal; Receptor, Cannabinoid, CB2; Receptors, Cannabinoid

One way to link chronic inflammation with cancer is through the intrinsic inflammatory pathway, in which genetic alterations that induce malignant transformation also produce a cancer-promoting, inflammatory microenvironment. Signal transducer and activator of transcription 3 (STAT3) contributes to the intrinsic inflammatory pathway in Barrett's esophagus. In human tumors, honokiol (a polyphenol in herbal teas) has growth-inhibitory and proapoptotic effects associated with suppressed activation of STAT3. We used human Barrett's epithelial and esophageal adenocarcinoma cell lines to determine effects of honokiol on cell number, necrosis, apoptosis, and anchorage-independent growth and to explore STAT3's role in those effects. We determined Ras activity and expression of phosphorylated ERK1/2, phosphorylated Akt, and phosphorylated STAT3 in the presence or absence of honokiol. Cells were infected with constitutively active Stat3-C to assess effects of honokiol-induced STAT3 inhibition on apoptosis. Honokiol decreased cell number and increased necrosis and apoptosis in transformed Barrett's cells, but not in nontransformed cells. In adenocarcinoma cells, honokiol also increased necrosis and apoptosis and decreased anchorage-independent growth. Within 30 min of honokiol treatment, transformed Barrett's cells decreased expression of phosphorylated STAT3; decreases in Ras activity and phosphorylated ERK1/2 expression were detected at 24 h. Infection with Stat3-C significantly reduced apoptosis after honokiol treatment. Honokiol causes necrosis and apoptosis in transformed Barrett's and esophageal adenocarcinoma cells, but not in nontransformed Barrett's cells, and the proapoptotic effects of honokiol are mediated by its inhibition of STAT3 signaling. These findings suggest a potential role for targeting the intrinsic inflammatory pathways as a therapeutic strategy to prevent Barrett's carcinogenesis.

Topics: Adenocarcinoma; Antineoplastic Agents, Phytogenic; Apoptosis; Barrett Esophagus; Biphenyl Compounds; Cell Line, Tumor; Cell Transformation, Neoplastic; Esophageal Neoplasms; Humans; Lignans; Signal Transduction; STAT3 Transcription Factor

WO(3) nanowires (nano-WO(3)) were synthesized using a simple hydrothermal technique, and then used to modify the carbon paste electrode (CPE). The electrochemical behavior of honokiol on the unmodified CPE and nano-WO(3) modified CPE was compared. It was found that nano-WO(3) remarkably increased the oxidation signal of honokiol. The influences of supporting electrolyte, amount of nano-WO(3), accumulation potential and time were studied on the oxidation peak current of honokiol. Based on the strong enhancement effect of nano-WO(3), a sensitive, simple and rapid electrochemical method was developed for the detection of honokiol. The linear range was from 3×10(-8) to 2.0×10(-5)M, and the detection limit was as low as 1×10(-8)M after 2-min accumulation. Finally, it was used to determine honokiol in traditional Chinese medicines, and the recovery was over the range between 96.5% and 98.8%.

Topics: Biphenyl Compounds; Carbon; Drugs, Chinese Herbal; Electrochemical Techniques; Electrodes; Humans; Hydrogen-Ion Concentration; Lignans; Limit of Detection; Medicine, Chinese Traditional; Microscopy, Electron, Scanning; Nanowires; Oxidation-Reduction; Oxides; Surface Properties; Tungsten

To study the anti-tumor effect of tanshinon II A, tetrandrine, honokiol, curcumin, oridonin and paeonol on leukemia cell lines SUP-B15, K562, CEM, HL-60 and NB4.. To study the anti-tumor effect of tanshinone II A, tetrandrine, honokiol, curcumin, The leukemia cell lines were exposed to the six Chinese herbal components for 96 hours. The proliferative inhibitory effects were detected with MTT and described by IC50 value.. Tanshinone II A inhibited the proliferations of SUP-B15, K562, CEM, HL-60 and NB4 cell lines, with HL-60 showing the least impact. Tetrandrine, honokiol, curcumin and oridonin inhibited the proliferations of SUP-B15, K562, CEM, HL-60 and NB4 cell lines and there was no significant difference between the cell lines. Paeonol did not have significant inhibitory effect on leukemia cell lines.. Tetrandrine, honokiol, curcumin and oridonin inhibit the proliferation of five cell lines SUP-B15, K562, CEM, HL-60, NB4, and the effects are similar, which means that their anticancer effects are quite broad. Tanshinone II A has better anti-leukemia effects on SUP-B15, K562, CEM, NB4 than on HL-60. The effect of paeonol against leukemia cell lines is poor.

Topics: Abietanes; Acetophenones; Antineoplastic Agents, Phytogenic; Benzylisoquinolines; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Curcumin; Diterpenes, Kaurane; Drugs, Chinese Herbal; HL-60 Cells; Humans; K562 Cells; Leukemia; Lignans; Plants, Medicinal

Embryonic stem cells, which are characterized by pluripotency and self-renewal, have recently been highlighted in drug discovery. In particular, the potential of ES cells to differentiate into specific-cell types make them an extremely useful tool in the evaluation of the biological activity of test compounds. Honokiol, a major neolignan derived from the bark of Magnolia obovata, has been shown an anti-tumor activity. However, the precise mechanism of action in the anti-tumor activity of honokiol is still poorly understood. Here, we evaluated the antiangiogenic activity of honokiol using mouse ES cell-derived embryoid bodies. mES-derived EBs were formed using hanging drop cultures and vascular formation was induced on gelatincoated plates in EGM-2 medium. The growth inhibition of honokiol was found to be more sensitive in the differentiated EB-derived endothelial cells compared to the undifferentiated EB-derived cells. Honokiol also inhibited the vascular formation of mES cells on 3-D collagen gel and decreased the expression of endothelial biomarkers VEGFR2 and PECAM in the differentiated EB-derived endothelial cells. In addition, honokiol suppressed the MAPK and mTOR signaling pathways in the EB-derived endothelial cells. Therefore, the anti-angiogenic activity of honokiol is associated in part with the suppression of PECAM and MAPK/mTOR pathways in EB-derived endothelial cells.

Topics: Angiogenesis Inhibitors; Animals; Biphenyl Compounds; Cell Differentiation; Cell Line; Embryoid Bodies; Embryonic Stem Cells; Endothelial Cells; Lignans; Mice; Mitogen-Activated Protein Kinase Kinases; Platelet Endothelial Cell Adhesion Molecule-1; Signal Transduction; TOR Serine-Threonine Kinases; Vascular Endothelial Growth Factor Receptor-2

Honokiol has shown chemopreventive effects in chemically-induced and UVB-induced skin cancer in mice. In this investigation, we assessed the time-effects of a topical low dose of honokiol (30 μg), and then the effects of different honokiol doses (30, 45, and 60 μg) on a UVB-induced skin cancer model to find an optimal dose and time for desirable chemopreventive effects. UVB radiation (30 mJ/cm(2), 5 days/week for 25 or 27 weeks) was used to induce skin carcinogenesis in SKH-1 mice. For the time-response experiment 30 μg honokiol in acetone was applied topically to the animals before the UVB exposure (30 min, 1 h, and 2 h) and after the UVB exposure (immediately, 30 min, and 1 h). Control groups were treated with acetone. For the dose-response study, animals were treated topically with acetone or honokiol (30, 45, and 60 μg) one hour before the UVB exposure. In the time-response experiment, honokiol inhibited skin tumor multiplicity by 49-58% while reducing tumor volumes by 70-89%. In the dose-response study, honokiol (30, 45, and 60 μg) significantly decreased skin tumor multiplicity by 36-78% in a dose-dependent manner, while tumor area was reduced by 76-94%. Honokiol (60 μg) significantly reduced tumor incidence by 40% as compared to control group. Honokiol applied in very low doses (30 μg) either before or after UVB radiation shows chemopreventive effects. Honokiol (30, 45, and 60 μg) prevents UVB-induced skin cancer in a dose-dependent manner. Honokiol can be an effective chemopreventive agent against skin cancer.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Female; Lignans; Mice; Mice, Hairless; Neoplasms, Radiation-Induced; Skin Neoplasms; Tumor Burden; Ultraviolet Rays; Weight Gain

Glioblastoma is one of the most lethal and common malignant human brain tumors, with aggressive proliferation and highly invasive properties. Honokiol derived from Magnolia officinalis is able to cross the blood-brain barrier (BBB) and the blood-cerebrospinal fluid barrier (BCSFB), suggesting a strong possibility that it could be an effective drug for the treatment of brain tumors, including glioblastoma. Thus, we investigated the effects of honokiol on the expression of adhesion molecules in TNF-α-stimulated endothelial cells, and cancer growth and invasion were determined in T98G human glioblastoma cells. Honokiol dose-dependently inhibited the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in human umbilical vein endothelial cells (HUVECs) stimulated with TNF-α for 6 h. Pretreatment with honokiol significantly reduced the adhesion of T98G cells to HUVECs. Moreover, honokiol inhibited the invasion of T98G cells, suggesting that honokiol has an anti-metastatic effect. In addition, honokiol increased the cytotoxicity of T98G cells in a dose- and time-dependent manner as assayed by MTT. TUNEL assay showed that honokiol significantly induced apoptosis in T98G cells at doses of 10 µM or more. The induction of apoptotic cell death was mediated by the downregulation of the anti-apoptotic protein Bcl-2 and the upregulation of the pro-apoptotic protein Bax. Taken together, the results of this study suggest that honokiol exerts an anticancer effect by preventing metastasis and inducing apoptotic cell death of brain tumor cells.

Topics: Apoptosis; Biphenyl Compounds; Blood-Brain Barrier; Brain Neoplasms; Cell Adhesion; Cell Adhesion Molecules; Cell Line, Tumor; Gene Expression Regulation, Neoplastic; Glioblastoma; Human Umbilical Vein Endothelial Cells; Humans; Intercellular Adhesion Molecule-1; Lignans; Magnolia; Plant Extracts; Tumor Necrosis Factor-alpha; Vascular Cell Adhesion Molecule-1

To investigate the effect of prescription compatibility on the pharmacokinetics of components from Dachengqi Decoction (DCQD, ) in rats.. Twenty-four male rats were randomly and equally divided into the DCQD group, Dahuang (Radix et Rhizoma Rhei, Polygonaceae) group, Houpo (Magnolia officinalis Rehd., Magnoliaceae) group, and Zhishi (Fructus Aurantii Immaturus, Rutaceae) group. The blood samples were collected before dosing and subsequently at 10, 15, 20, 30, 45 min, 1, 2, 4, 8, and 12 h following gavage. The levels of aloe-emodin, rhein, emodin, chrysophanol, honokiol, magnolol, hesperidin, and naringin in rat serum were quantified using a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for pharmacokinetic study.. The area under the curve (AUC), mean retention time (MRT), the peak concentration (C(max)) of aloe-emodin, rhein, emodin, and chrysophanol in the DCQD group were significantly different compared with the Dahuang group (P <0.05, respectively). The mean plasma concentration, C(max), and the absorption of Dahuang's component in the DCQD group were obviously lower at each time point than those in the Dahuang group, while the elimination process of Dahuang's component was obviously delayed (P <0.05). Half-lives of aloe-emodin, chrysophanol, and rhein were also extended in the DCQD group (P <0.05, respectively). In the DCQD group, the mean plasma concentration, AUC, C(max) and absorption of honokiol, and magnolol were significantly lower (P <0.01, respectively) at each time point than those in the Houpo group, while the drug distribution half-life time (T(1/2α)), the drug eliminated half-life time (T(1/2β)), MRT, and time of peak concentration (T(max)) were significantly delayed (P <0.05, respectively). Pharmacokinetic parameters of hesperidin and naringin in the Zhishi group were not significantly different as compared with the DCQD group (P >0.05, respectively), while the MRT of naringin was significantly longer.. The compatibility in Chinese medicine could affect the drug's pharmacokinetics in DCQD, which proves that the prescription compatibility principle of Chinese medicine formulations has its own pharmacokinetic basis.

Topics: Administration, Oral; Animals; Anthraquinones; Biphenyl Compounds; Drug Incompatibility; Emodin; Flavanones; Hesperidin; Lignans; Male; Plant Extracts; Rats; Rats, Sprague-Dawley

Honokiol, a small-molecular weight natural product, has previously been reported to activate apoptosis and inhibit gastric tumorigenesis. Whether honokiol inhibits the angiogenesis and metastasis of gastric cancer cells remains unknown.. We tested the effects of honokiol on angiogenic activity and peritoneal dissemination using in vivo, ex vivo and in vitro assay systems. The signaling responses in human gastric cancer cells, human umbilical vascular endothelial cells (HUVECs), and isolated tumors were detected and analyzed. In a xenograft gastric tumor mouse model, honokiol significantly inhibited the peritoneal dissemination detected by PET/CT technique. Honokiol also effectively attenuated the angiogenesis detected by chick chorioallantoic membrane assay, mouse matrigel plug assay, rat aortic ring endothelial cell sprouting assay, and endothelial cell tube formation assay. Furthermore, honokiol effectively enhanced signal transducer and activator of transcription (STAT-3) dephosphorylation and inhibited STAT-3 DNA binding activity in human gastric cancer cells and HUVECs, which was correlated with the up-regulation of the activity and protein expression of Src homology 2 (SH2)-containing tyrosine phosphatase-1 (SHP-1). Calpain-II inhibitor and siRNA transfection significantly reversed the honokiol-induced SHP-1 activity. The decreased STAT-3 phosphorylation and increased SHP-1 expression were also shown in isolated peritoneal metastatic tumors. Honokiol was also capable of inhibiting VEGF generation, which could be reversed by SHP-1 siRNA transfection.. Honokiol increases expression and activity of SPH-1 that further deactivates STAT3 pathway. These findings also suggest that honokiol is a novel and potent inhibitor of angiogenesis and peritoneal dissemination of gastric cancer cells, providing support for the application potential of honokiol in gastric cancer therapy.

Topics: Animals; Apoptosis; Biphenyl Compounds; Calpain; Cell Line, Tumor; Cell Proliferation; Cells, Cultured; Human Umbilical Vein Endothelial Cells; Humans; Lignans; Mice; Neovascularization, Pathologic; Phosphorylation; Protein Tyrosine Phosphatase, Non-Receptor Type 6; RNA, Small Interfering; Signal Transduction; STAT3 Transcription Factor; Stomach Neoplasms; Up-Regulation; Xenograft Model Antitumor Assays

This study aimed to evaluate the effect of honokiol and its structural analogs on the functional activity and gene expression of P-glycoprotein (P-gp) in order to identify effective P-gp inhibitors from natural products which have additional health-promoting effects.. The interaction characteristics of honokiol, magnolol and 4-O-methylhonokiol with P-gp were determined in NCI/ADR-RES cells overexpressing P-gp.. All three compounds down-regulated the expression of P-gp in a concentration- and time-dependent manner, leading to 2.5- to 4.1-fold reductions of P-gp expression in NCI/ADR-RES cells. Accordingly, down-regulation of P-gp resulted in the significant enhancement of the intracellular accumulation of calcein, a P-gp substrate. Furthermore, pre-treatment with honokiol, magnolol or 4-O-methylhonokiol significantly increased the susceptibility of cancer cells to daunorubicin-induced cytotoxicity in NCI/ADR-RES cells.. The present study suggests that honokiol, magnolol and 4-O-methylhonokiol could be promising agents for reducing the multidrug resistance of cancer cells to anticancer drugs via the down-regulation of P-gp expression.

Topics: Antineoplastic Agents, Phytogenic; Antineoplastic Combined Chemotherapy Protocols; ATP Binding Cassette Transporter, Subfamily B, Member 1; Biphenyl Compounds; Cell Line, Tumor; Daunorubicin; Down-Regulation; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Humans; Lignans; Magnolia; Plant Extracts

Melanomas are aggressive neoplasms with limited therapeutic options. Therefore, developing new therapies with low toxicity is of utmost importance. Honokiol is a natural compound that recently has shown promise as an effective anticancer agent.. The effect of honokiol on melanoma cancer cells was assessed in vitro. Proliferation and physiologic changes were determined using hexosaminidase assay and transmission electron microscopy. Protein expression was assessed by immunoblotting.. Honokiol treatment inhibited cell proliferation and induced death. Electron microscopy showed autophagosome formation. Reduced levels of cyclin D1 accompanied cell-cycle arrest. Honokiol also decreased phosphorylation of AKT (known as protein kinase B) and mammalian target of rapamycin, and inhibited γ-secretase activity by down-regulating the expression of γ-secretase complex proteins, especially anterior pharynx-defective 1.. Honokiol is highly effective in inhibiting melanoma cancer cells by attenuating AKT/mammalian target of rapamycin and Notch signaling. These studies warrant further clinical evaluation for honokiol alone or with present chemotherapeutic regimens for the treatment of melanomas.

Topics: Amyloid Precursor Protein Secretases; Animals; Antineoplastic Agents, Phytogenic; Autophagy; Biomarkers, Tumor; Biphenyl Compounds; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Dose-Response Relationship, Drug; Down-Regulation; Immunoblotting; Lignans; Melanoma; Mice; Proto-Oncogene Proteins c-akt; Signal Transduction; TOR Serine-Threonine Kinases

To reveal the relationship between the quality and the leaf shape of Magnolia officinalis produced in Hubei Enshi.. Determined the content of magnolol and honokiol by HPLC with methanol-water (78:22) and the detective wavelength was 294 nm.. The content of magnolol and honokiol in the leaf of Magnolia officinalis was usually higher if the leaf apex was convex and the leaves were short. In certain years, the content of magnolol and honokiol in Cortex Magnoliae officinalis was positively correlated with the bark thickness.. The leaf shape of Magnolia officinalis produced in Hubei Enshi is directly related to the phenolic content of leaf itself. But it has nothing to do with the content of magnolol and honokiol in Cortex Magnoliae officinalis.

Topics: Biphenyl Compounds; China; Chromatography, High Pressure Liquid; Lignans; Magnolia; Plant Bark; Plant Leaves; Plants, Medicinal; Quality Control

Honokiol, 5,5'-diallyl-2,4'-dihydroxy- biphenyl, by comparison with its isomer magnolol, 5,5'-diallyl- 2,2'-dihydroxy- biphenyl, has been characterized by steady-state and time-resolved spectroscopy as well as (1)H NMR. Honokiol shows more complex pH dependence of absorption and fluorescence characteristics compared with magnolol. Honokiol possesses much weaker acidity than magnolol both in the ground and excited states. Its weak photoacidity is similar to that of 4-hydroxy- biphenyl or 4, 4'-dihydroxy- biphenyl rather than 2-hydroxy- biphenyl or 2, 2'-hydroxy- biphenyl. The electron effect and geometry configuration of substitution has been discussed.

Topics: Acids; Biphenyl Compounds; Hydrogen-Ion Concentration; Lignans; Magnetic Resonance Spectroscopy; Photochemistry; Spectrometry, Fluorescence

Oxidative stress-induced apoptosis in lens epithelial cells plays an important role in cataract formation, and its prevention may be of therapeutic interest. This study was performed to investigate the protective effect and mechanisms of honokiol on H(2)O(2)-induced apoptosis in human lens epithelial (HLE) cells. HLE cells (SRA01-04) were pretreated with honokiol at concentrations of 5μM, 10μM and 20μM before 50μM H(2)O(2) treatment. The results demonstrated that pretreatment of honokiol inhibited the activation of caspase-3 and caspase-9 and downregulated the expression of Bcl-2. Mechanistically, honokiol suppressed H(2)O(2)-induced phosphorylation of ERK1/2, p38 mitogen-activated protein kinase (MAPK), JNK and Akt. Honokiol also inhibited H(2)O(2)-induced nuclear factor-κB (NF-κB)/p65 phosphorylation and translocation in HLE cells. These results demonstrate that honokiol suppresses H(2)O(2)-induced HLE cell apoptosis via interference with the MAPKs, Akt and NF-κB signaling, suggesting that honokiol might have a potential effect against cataract formation.

Topics: Apoptosis; Biphenyl Compounds; Caspase 3; Caspase 9; Cell Line; Down-Regulation; Enzyme Activation; Epithelial Cells; Humans; Hydrogen Peroxide; Lens, Crystalline; Lignans; Mitogen-Activated Protein Kinases; Protein Transport; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; Signal Transduction; Transcription Factor RelA; Up-Regulation

Honokiol has been shown to possess a lot of pharmacologic benefits, including antioxidative, antiangiogenic and antineoplastic effects. In the present study, we investigated the anti-inflammatory effects of honokiol and the signaling mechanisms involved in lipopolysaccharide (LPS)-induced conditions in human renal mesangial cells (HRMCs). Honokiol did not significantly change HRMC viability when used at a concentration of <20 μmol/l but markedly altered cell viability at concentrations of >40 μmol/l. In this study, LPS treatment led to a marked upregulation of the levels of IL-1β, IL-18, TNF-α, TGF-β1, CCL2, CCL3, and CCL5 in HRMCs. The expression of COX-2, iNOS, and their products PGE(2) and NO also increased. The upregulation of these molecules was significantly abolished by honokiol in a dose-dependent manner. Moreover, honokiol almost completely reversed IL-1β, CCL3, and NO expression at 10 μmol/l, and IL-18, TNF-α, TGF-β1, and COX-2 expression at 20 μmol/l. In addition, phospho-NF-κB p65 at Ser536, phospho-Akt, and phospho-p42/44 were dramatically suppressed by honokiol in LPS-treated HRMCs. These results indicate that honokiol can inhibit the LPS-induced expression of inflammatory cytokines and mediators in HRMCs. The anti-inflammatory mechanisms of honokiol are partly due to the suppression of the phospho-NF-κB p65, phospho-Akt and phospho-p42/44 pathways.

Topics: Anti-Inflammatory Agents; Biphenyl Compounds; Cell Survival; Cyclooxygenase 2; Cytokines; Dose-Response Relationship, Drug; Gene Expression Regulation; Humans; Inflammation Mediators; Lignans; Lipopolysaccharides; Mesangial Cells; Nitric Oxide Synthase Type II; Phosphorylation; Signal Transduction

In the present study, we report the effects of honokiol, a phytochemical from Magnolia spp., on cancer cell migration capacity and the molecular mechanisms underlying these effects using breast cancer cell lines as an in vitro model. Using cell migration assays, we found that the treatment of human breast cancer cells (MCF-7) and murine mammary cancer cells (4T1) with honokiol resulted in a dose-dependent inhibition of migration of these cells, which was associated with a reduction in nitric oxide (NO) levels. The cell migration capacity was decreased in the presence of NG-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NO synthase. Honokiol reduced the elevated levels of cyclic guanosine monophosphate (cGMP) in the cells, while the treatment of 4T1 cells with guanylate cyclase (GC) inhibitor 1-H-[1,2,4]oxadiaxolo[4,3-a]quinolalin-1-one (ODQ) reduced the migration of cells and the levels of cGMP. The presence of 8-bromoguanosine 3'5'-cyclic monophosphate, an analogue of cGMP, enhanced the migration of these cells, suggesting a role for GC in the migration of 4T1 cells. Honokiol also inhibited the levels of cyclooxygenase-2 (COX-2) and prostaglandin (PG) E2 in 4T1 cells. The transfection of 4T1 cells with COX-2 siRNA resulted in a reduction in cell migration. ODQ and L-NAME also decreased the levels of PGE2 in 4T1 cells suggesting a role for COX-2/PGE2 in cell migration. Moreover, honokiol inhibited the activation of nuclear factor κB (NF-κB), an upstream regulator of COX-2 and iNOS, in 4T1 cells. These results indicate that NO and COX-2 are the key targets of honokiol in the inhibition of breast cancer cell migration, an essential step in invasion and metastasis.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Breast Neoplasms; Carcinoma; Cell Line, Tumor; Cell Movement; Cyclooxygenase 2; Drug Evaluation, Preclinical; Drugs, Chinese Herbal; Female; Humans; Lignans; Magnolia; Mice; Models, Biological; Molecular Targeted Therapy; Nitric Oxide; Phytotherapy

Renal fibrosis acts as the common pathway leading to the development of end-stage renal disease. The present study investigated, in vivo and in vitro, the anti-fibrotic and anti-inflammatory effects, particularly on the epithelial to mesenchymal transition of renal tubular cells, exerted by honokiol, a phytochemical used in traditional medicine, and mechanisms underlying these effects.. Anti-fibrotic effects in vivo were assayed in a rat model of renal fibrosis [the unilateral ureteral obstruction (UUO) model]. A rat tubular epithelial cell line (NRK-52E) was stimulated by transforming growth factor-β1 (TGF-β1) and treated with honokiol to explore possible mechanisms of these anti-fibrotic effects. Gene or protein expression was analysed by Northern or Western blotting. Transcriptional regulation was investigated using luciferase activity driven by a connective tissue growth factor (CTGF) promoter.. Honokiol slowed development of renal fibrosis both in vivo and in vitro. Honokiol treatment attenuated tubulointerstitial fibrosis and expression of pro-fibrotic factors in the UUO model. Honokiol also decreased expression of the mRNA for the chemokine CCL2 and for the intracellular adhesion molecule-1, as well as accumulation of type I (α1) collagen and fibronectin in UUO kidneys. Phosphorylation of Smad-2/3 induced by TGF-β1 and CTGF luciferase activity in renal tubular cells were also inhibited by honokiol.. Honokiol suppressed expression of pro-fibrotic and pro-inflammatory factors and of extracellular matrix proteins. Honokiol may become a therapeutic agent to prevent renal fibrosis.

Topics: Actins; Animals; Biphenyl Compounds; Cell Line; Chemokine CCL2; Connective Tissue Growth Factor; Extracellular Matrix Proteins; Fibrosis; Genes, Reporter; Intercellular Adhesion Molecule-1; Kidney; Kidney Tubules; Lignans; Luciferases; Male; Phosphorylation; Rats; Rats, Wistar; RNA, Messenger; Signal Transduction; Smad2 Protein; Smad3 Protein; Transforming Growth Factor beta1; Ureteral Obstruction

Sepsis has a high mortality rate despite the recent advances in intensive care medicine and antibiotics. Honokiol, a low molecular weight natural product, is known to possess anti-inflammatory activity. Here, we investigate whether honokiol can ameliorate acute lung injury and lethal response in murine models of sepsis.. Mice were intraperitoneally given vehicle or honokiol 30 min after the induction of sepsis by cecal ligation and puncture (CLP) and endotoxemia by administration of E. coli lipopolysaccharide (LPS).. The productions of serum tumor necrosis factor-α (TNF-α), nitric oxide (NO), and high mobility group box 1 (HMGB 1) were increased in mice during sepsis, which could be reversed by honokiol. Honokiol could also effectively reduce the increased blood lipid peroxidation and nitrotyrosine in septic mice. Honokiol significantly reversed the inductions of inducible NO synthase and nuclear factor-κB (NF-κB) activation in the lungs of mice during sepsis. Honokiol also effectively rescued the lung edema, lung pathological changes, and lethality in septic mice.. These findings suggest that honokiol is capable of suppressing the lethal response and acute lung injury associated with sepsis, and support the potential use of honokiol as a therapeutic agent for the conditions associated with septic shock.

Topics: Acute Lung Injury; Animals; Anti-Infective Agents; Biphenyl Compounds; Endotoxemia; Inflammation; Lignans; Lipopolysaccharides; Male; Mice; Mice, Inbred ICR; Oxidative Stress; Sepsis

Our previous report has demonstrated that 5-formylhonokiol (FH), a derivative of honokiol (HK), exerts more potent anti-proliferative activities than honokiol in several tumor cell lines. In present study, we first explored the antiangiogenic activities of 5-formylhonokiol on proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVECs) for the first time in vitro. Then we investigated the in vivo antiangiogenic effect of 5-formylhonokiol on zebrafish angiogenesis model. In order to clarify the underlying molecular mechanism of 5-formylhonokiol, we investigated the signaling pathway involved in controlling the angiogenesis process by western blotting assay. Wound-healing results showed that 5-formylhonokiol significantly and dose-dependently inhibited migration of cultured human umbilical vein enthothelial cells. The invasiveness of HUVEC cells was also effectively suppressed at a low concentration of 5-formylhonokiol in the transwell assay. Further F-actin imaging revealed that inhibitory effect of 5-formylhonokiol on invasion may partly contribute to the disruption of assembling stress fiber. Tube formation assay, which is associated with endothelial cells migration, further confirmed the anti-angiogenesis effect of 5-formylhonokiol. In in vivo zebrafish angiogenesis model, we found that 5-formylhonokiol dose-dependently inhibited angiogenesis. Furthermore, western blotting showed that 5-formylhonokiol significantly down-regulated extracellular signal-regulated kinase (ERK) expression and inhibited the phosphorylation of ERK but not affecting the total protein kinase B (Akt) expression and related phosphorylation, suggesting that 5-formylhonokiol might exert anti-angiogenesis capacity via down-regulation of the ERK signal pathway. Taken together, these data suggested that 5-formylhonokiol might be a viable drug candidate in antiangiogenesis and anticancer therapies.

Topics: Actins; Angiogenesis Inhibitors; Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Blotting, Western; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cells, Cultured; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Embryo, Nonmammalian; Endothelium, Vascular; Extracellular Signal-Regulated MAP Kinases; Humans; Lignans; Neovascularization, Physiologic; Signal Transduction; Umbilical Veins; Wound Healing; Zebrafish

Adipose tissue plays an essential role in energy homeostasis as a metabolic and endocrine organ. Accordingly, adipocytes are emerging as a major drug target for obesity and obesity-mediated metabolic syndrome. Dysfunction of enlarged adipocytes in obesity is involved in obesity-mediated metabolic syndrome. Adipocytokines, such as adiponectin released from small adipocytes, are able to prevent these disorders. In this study, we found that honokiol, an ingredient of Magnolia officinalis used in traditional Chinese and Japanese medicines, enhanced adipocyte differentiation in 3T3-L1 preadipocytes. Oil Red O staining showed that treatment with honokiol in the presence of insulin dose-dependently increased lipid accumulation in 3T3-L1 preadipoyctes although its activity was weak compared with rosiglitazone. During adipocyte differentiation, the expression of peroxisome proliferator-activated receptor γ2 (PPARγ2) mRNA and PPARγ target genes such as adipocyte protein 2 (aP2), adiponectin, and GLUT4 was induced by treatment with 10 μM honokiol. However, honokiol failed to show direct binding to the PPARγ ligand-binding domain in vitro. In preadipocytes, treatment with honokiol in the presence of insulin increased the phosphorylation of extracellular signal-regulated kinase (ERK) 1/2 protein and Akt protein, early insulin signaling pathways related to adipocyte differentiation, compared with insulin-only treatment. Taken together, our results suggest that honokiol promotes adipocyte differentiation through increased expression of PPARγ2 mRNA and potentiation of insulin signaling pathways such as the Ras/ERK1/2 and phosphoinositide-3-kinase (PI3K)/Akt signaling pathways.

Topics: 3T3-L1 Cells; Adipocytes; Animals; Biphenyl Compounds; Cell Differentiation; Immunoblotting; Insulin; Lignans; Magnetic Resonance Spectroscopy; Magnolia; Mice; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Proto-Oncogene Proteins c-akt; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction

Entrapping inclusion complexes in liposomes has been proposed to increase the entrapment efficiency (EE) and stability of liposomes compared with conventional liposomes. In the present study, a stable honokiol-in-hydroxypropyl-β-cyclodextrin-in-liposome (honokiol-in-HP-β-CD-in-liposome) was developed as honokiol delivery system by a novel method. The final molar ratio of honokiol/HP-β-CD/lipid was selected as 1:2:2. The mean particle size was 123.5 nm, the zeta potential was -25.6 mV, and the EE was 91.09 ± 2.76%. The release profile in vitro demonstrated that honokiol is released from honokiol-in-HP-β-CD-in-liposome with a sustained and slow speed. Crystallographic study indicated that honokiol was first bound within HP-β-CD and then the inclusion complex was encapsulated within liposomes. Honokiol-in-HP-β-CD-in-liposome without freeze dry kept stable for at least 6 months at 4°C. Pharmacokinetic study revealed that honokiol-in-HP-β-CD-in-liposome significantly retarded the elimination and prolonged the residence time in circulating system. The data of bioactivity showed that honokiol-in-HP-β-CD-in-liposome remained similar antiproliferative activity in A549 and HepG2 tumor cells compared to free honokiol. These results suggested that we had successfully prepared honokiol-in-HP-β-CD-in-liposome. The novel honokiol formulation was easy to push industrialization forward and might be a potential carrier for honokiol delivery in tumor chemotherapy.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Animals; Antineoplastic Agents, Phytogenic; beta-Cyclodextrins; Biphenyl Compounds; Cell Survival; Chemistry, Pharmaceutical; Chromatography, High Pressure Liquid; Crystallization; Drug Carriers; Drug Compounding; Drug Stability; Hep G2 Cells; Humans; Injections, Intravenous; Lignans; Liposomes; Male; Microscopy, Electron, Transmission; Particle Size; Rats; Rats, Sprague-Dawley; Solubility; Surface Properties; X-Ray Diffraction

PEGylated liposomal honokiol had been developed with the purpose of improving the solubility and pharmacokinetics compared with free honokiol. Human plasma protein binding ability of honokiol was also investigated. PEGylated liposomal honokiol was prepared by thin film evaporation-sonication method. Its mean particle size was 98.68 nm, mean zeta potential was -20.6 mV and encapsulation efficiency were 87.68±1.56%. The pharmacokinetics of PEGylated liposomal honokiol was studied after intravenous administration in Balb/c mice. There were significant differences of parameters T(1/2β) and AUC(0→∞) between them and liposome lengthened T(1/2β) and AUC(0→∞) values. The mean T(1/2β) value of PEGylated liposomal honokiol and free honokiol were 26.09 min and 13.46 min, respectively. The AUC(0→∞) ratio of PEGylated liposomal honokiol to free honokiol was about 1.85-fold (219.24 μg/mL min/118.68 μg/mL min) (P=0.000). Examination of protein binding ability showed that honokiol with 0.5, 8.0 and 20 μg/mL concentrations in human plasma achieved the percent of bound between 60% and 65%. The results suggested that PEGylated liposomal honokiol improved the solubility, increased the drug concentration in plasma, and withstanded the clearance. Besides, the percent of protein bound of honokiol in human plasma was between 60% and 65%.

Topics: Animals; Area Under Curve; Biphenyl Compounds; Blood Proteins; Female; Half-Life; Humans; Injections, Intravenous; Lignans; Liposomes; Male; Mice; Mice, Inbred BALB C; Particle Size; Polyethylene Glycols; Protein Binding; Solubility

The biotransformation of honokiol, a major constituent of the bark of Magnolia officinalis, was investigated in rat and human livers. When isolated, rat livers were perfused with 10 µM honokiol and two metabolites, namely hydroxylated honokiol conjugated with glucuronic and sulfuric acid (M1) and honokiol monoglucuronide (M2), were quantified in bile and perfusate by high-performance liquid chromatography. The hepatic extraction ratio and clearance of honokiol was very high in rat liver (E: 0.99 ± 0.01 and 35.8 ± 0.04 mL/min, respectively) leading to very low bioavailability (F = 0.007 ± 0.001). M2 formation was also highly efficient in human liver microsomes [V(max) /K(m) = 78.1 ± 6.73 µL/(min mg)], which appeared to be catalyzed mainly by UDP-glucuronosyltransferases 1A1, A3, 1A8, and 1A10, indicating hepatic and extrahepatic glucuronidation. Monosulfation of honokiol to the minor metabolite honokiol monosulfate [V(max) /K(m) = 27.9 ± 4.33 µL/(min mg)] by human liver cytosol was less pronounced and is mediated by sulfotransferases 1A1* 1, 1A1* 2, 1A2, 1A3, 1B1, and 1E1. P450-mediated oxidation of honokiol by liver microsomes, however, was below detection limit. In summary, this study established that glucuronidation and sulfation are the main metabolic pathways for honokiol in rat and human liver, suggesting their major contribution to clearance in vivo.

Topics: Animals; Bile; Biotransformation; Biphenyl Compounds; Chromatography, High Pressure Liquid; Cytosol; Glucuronic Acid; Humans; In Vitro Techniques; Insecta; Lignans; Liver; Magnolia; Male; Metabolic Detoxication, Phase I; Microsomes, Liver; Perfusion; Rats; Rats, Wistar; Species Specificity; Sulfuric Acids; Tissue Distribution

Honokiol (HNK) is a small organic molecule purified from magnolia species and has demonstrated anticancer activities in a variety of cancer cell lines; however, its effect on oral squamous cell carcinoma (OSCC) cells is unknown. We investigated the antitumor activities of HNK on OSCC cells in vitro for the first time. The inhibitory effects of HNK on the growth and proliferation of OSCC cells were demonstrated via in vitro 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and propidium iodide (PI) assays, and the apoptotic cells were investigated by the observation of morphological changes and detection of DNA fragmentation via PI, TdT-mediated dUTP-biotin nick end labeling (TUNEL), and DNA ladder assays, as well as flow cytometry assay. The results showed that HNK inhibited the growth and proliferation of OSCC cells in vitro in a time and dose-dependent manner. The inhibitory effect was associated with the cell apoptosis induced by HNK, evidenced by the morphological features of apoptotic cells, TUNEL-positive cells and a degradation of chromosomal DNA into small internucleosomal fragments. The study also demonstrated here that the inhibition or apoptosis mediated by 15 microg x mL(-1) or 20 microg x mL(-1) of HNK were more stronger compared with those of 20 microg x mL(-1) 5-fluorouracil (5-Fu, the control) applied to OSCC cells, when the ratio of OSCC cell numbers were measured between the treatment of different concentrations of HNK to the 5-Fu treatment for 48 h. HNK is a promising compound that can be potentially used as a novel treatment agent for human OSCC.

Topics: Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Biphenyl Compounds; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Cell Survival; Drugs, Chinese Herbal; Flow Cytometry; Fluorouracil; Humans; In Situ Nick-End Labeling; Lignans; Magnolia; Mouth Neoplasms; Phytotherapy; Plant Extracts

Melanoma continues to be a therapeutic challenge for the medical community owing to the scarcity of effective agents available to treat the disease. Honokiol, a traditional Chinese herb, has been proven to have anti-cancer effects in various cell types, therefore we hypothesized it may have similar cytotoxic capabilities against melanoma cells in vitro.. Two cell lines, SK-MEL2 and MeWo, were grown in culture and exposed to increasing doses of Honokiol. Cell proliferation, cytochrome c release into the cytosol, intra-cellular caspase activity, and mitochondrial depolarization were then evaluated after treatment with honokiol.. Melanoma cells in culture underwent cell death, had increased cytosolic cytochrome c, showed greater caspase activity, and demonstrated increased mitochondrial depolarization after treatment when compared to controls.. It appears that honokiol is an effective inhibitor of cultured human melanoma cells.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Blotting, Western; Caspases; Cell Proliferation; Cytochromes c; Drugs, Chinese Herbal; Enzyme-Linked Immunosorbent Assay; Humans; In Vitro Techniques; Lignans; Melanoma; Tumor Cells, Cultured

Hypoxia-inducible factor-1α (HIF-1α) plays a pivotal role in the reaction of a tumour to hypoxia. In this study, we examined the inhibitory effect of a natural compound, honokiol, on HIF-1α activity and tumour growth in combination with radiation.. The inhibitory effect of honokiol on hypoxia-responsive element (HRE) controlled luciferase activity and HIF-1α accumulations stimulated by CoCl(2), or hypoxia was examined. Effect of honokiol on HIF-1α levels within hypoxic tumour microenvironment was investigated by immunohistochemical and in vivo bioluminescent studies. The in vivo radiosensitising activity of honokiol was evaluated with subcutaneous murine colon carcinoma, CT26, xenografts of BALB/c mice treated with honokiol, radiation, or both.. Suppression of luciferase (luc) activity in HRE-luc stable cells by honokiol was in agreement with the results of decreased HIF-1α accumulation. In CT26-HRE-luc tumour-bearing mice, the inhibitory effect of intraperitoneally injected honokiol on HIF-1α-regulated luciferase activities induced by either CoCl(2) or radiation could be monitored non-invasively. Lastly, honokiol in combination with irradiation produced synergistic delay of CT26 tumour growth.. Our data suggest that honokiol can exert its anticancer activity as a HIF-1α inhibitor by reducing HIF-1α protein level and suppressing the hypoxia-related signaling pathway. The animal experiment indicates that honokiol improves the therapeutic efficacy of radiation.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cobalt; Enzyme-Linked Immunosorbent Assay; Female; HeLa Cells; Humans; Hypoxia; Hypoxia-Inducible Factor 1; Immunohistochemistry; Lignans; Luciferases; Melanoma, Experimental; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Neoplasm Transplantation; Neoplasms

Some Magnolia (Magnoliaceae) species are used for the empirical treatment of diabetes mellitus, but the antidiabetic properties of Magnolia dealbata have not yet been experimentally validated. Here we report that an ethanolic extract of Magnolia dealbata seeds (MDE) and its active principles honokiol (HK) and magnolol (MG) induced the concentration-dependent 2-NBDG uptake in murine 3T3-F442A and human subcutaneous adipocytes. In insulin-sensitive adipocytes, MDE 50 μg/ml induced the 2-NBDG uptake by 30% respect to insulin, while HK and MG, 30 μM each, did it by 50% (murine) and 40% (human). The simultaneous application of HK and MG stimulated 2-NBDG uptake by 70% in hormone-sensitive cells, on which Magnolia preparations exerted synergic effects with insulin. In insulin-resistant adipocytes, MDE, HK and MG induced 2-NBDG uptake by 57%, 80% and 96% respect to Rosiglitazone (RGZ), whereas HK and MG simultaneously applied stimulated 2-NBDG uptake more efficiently than RGZ (120%) in both murine and human adipocytes. Inhibitors of the insulin-signaling pathway abolished the glucose uptake induced by Magnolia dealbata preparations, suggesting that their antidiabetic effects are mediated by this signaling pathway. In addition, MDE, HK and MG exerted only mild to moderate proadipogenic effects on 3T3-F442A and human preadipocytes, although the combined application of HK and MG markedly increased the lipid accumulation in both cell types. In summary, Magnolia dealbata and its active principles HK and MG stimulate glucose uptake in insulin-sensitive and insulin-resistant murine and human adipocytes using the insulin signaling pathway.

Topics: 3T3 Cells; 4-Chloro-7-nitrobenzofurazan; Adipocytes; Adipogenesis; Animals; Biphenyl Compounds; Cell Survival; Deoxyglucose; Drug Synergism; Glucose; Humans; Hypoglycemic Agents; Insulin; Lignans; Lipid Metabolism; Magnolia; Mice; Plant Extracts; Signal Transduction

In this study, we investigated the mechanisms underlying the anti-inflammatory effects of honokiol in tumor necrosis factor (TNF)-α-stimulated rheumatoid arthritis synovial fibroblasts (RASFs). RASFs pre-treated with honokiol (0-20 μM) were stimulated with TNF-α (20 ng/ml). The levels of prostaglandin E2 (PGE2), nitric oxide (NO), soluble intercellular adhesion molecule-1 (sICAM-1), transforming growth factor-β1 (TGF-β1), monocyte chemotactic protein-1 (MCP-1), and macrophage inflammatory protein-1α (MIP-1α) in supernatants were determined by enzyme-linked immunosorbent assay (ELISA) and Griess assay. In addition, protein expression levels of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and phosphorylated Akt, nuclear factor kappa B (NFκB), and extracellular signal-regulated kinase (ERK)1/2 were determined by western blot. The expression of NFκB-p65 was assessed by immunocytochemical analysis. TNF-α treatment significantly up-regulated the levels of PGE2, NO, sICAM-1, TGF-β1, MCP-1, and MIP-1α in the supernatants of RASFs, increased the protein expression of COX-2, iNOS, and induced phosphorylation of Akt, IκB-α, NFκB, and ERK1/2 in RASFs. TNF-α-induced expression of these molecules was inhibited in a dose-dependent manner by pre-treatment with honokiol. The inhibitory effect of honokiol on NFκB-p65 activity was also confirmed by immunocytochemical analysis. In conclusion, honokiol is a potential inhibitor of TNF-α-induced expression of inflammatory factors in RASFs, which holds promise as a potential anti-inflammatory drug.

Topics: Biphenyl Compounds; Blotting, Western; Cells, Cultured; Chemokines; Cytokines; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Fluorescent Antibody Technique; Humans; Inflammation Mediators; Lignans; Synovial Membrane; Tumor Necrosis Factor-alpha; Up-Regulation

Gliosarcoma is one of the most common malignant brain tumors, and anti-angiogenesis is a promising approach for the treatment of gliosarcoma. However, chemotherapy is obstructed by the physical obstacle formed by the blood-brain barrier (BBB) and blood-cerebrospinal fluid barrier (BCSFB). Honokiol has been known to possess potent activities in the central nervous system diseases, and anti-angiogenic and anti-tumor properties. Here, we hypothesized that honokiol could cross the BBB and BCSFB for the treatment of gliosarcoma.. We first evaluated the abilities of honokiol to cross the BBB and BCSFB by measuring the penetration of honokiol into brain and blood-cerebrospinal fluid, and compared the honokiol amount taken up by brain with that by other tissues. Then we investigated the effect of honokiol on the growth inhibition of rat 9L gliosarcoma cells and human U251 glioma cells in vitro. Finally we established rat 9L intracerebral gliosarcoma model in Fisher 344 rats and human U251 xenograft glioma model in nude mice to investigate the anti-tumor activity.. We showed for the first time that honokiol could effectively cross BBB and BCSFB. The ratios of brain/plasma concentration were respectively 1.29, 2.54, 2.56 and 2.72 at 5, 30, 60 and 120 min. And about 10% of honokiol in plasma crossed BCSFB into cerebrospinal fluid (CSF). In vitro, honokiol produced dose-dependent inhibition of the growth of rat 9L gliosarcoma cells and human U251 glioma cells with IC(50) of 15.61 µg/mL and 16.38 µg/mL, respectively. In vivo, treatment with 20 mg/kg body weight of honokiol (honokiol was given twice per week for 3 weeks by intravenous injection) resulted in significant reduction of tumor volume (112.70±10.16 mm(3)) compared with vehicle group (238.63±19.69 mm(3), P = 0.000), with 52.77% inhibiting rate in rat 9L intracerebral gliosarcoma model, and (1450.83±348.36 mm(3)) compared with vehicle group (2914.17±780.52 mm(3), P = 0.002), with 50.21% inhibiting rate in human U251 xenograft glioma model. Honokiol also significantly improved the survival over vehicle group in the two models (P<0.05).. This study provided the first evidence that honokiol could effectively cross BBB and BCSFB and inhibit brain tumor growth in rat 9L intracerebral gliosarcoma model and human U251 xenograft glioma model. It suggested a significant strategy for offering a potential new therapy for the treatment of gliosarcoma.

Topics: Animals; Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Blood-Brain Barrier; Brain Neoplasms; Cell Line, Tumor; Cerebrospinal Fluid; Chromatography, High Pressure Liquid; Female; Glioma; Gliosarcoma; Humans; Lignans; Male; Mice; Mice, Nude; Neoplasm Transplantation; Rats; Rats, Inbred F344; Rats, Sprague-Dawley

Honokiol, an active agent extracted from magnolia bark, has been reported that induces anxiolytic action in a mouse elevated plus-maze test. However, the mechanism of anxiolytic action induced by honokiol remains unclear. This study was to investigate the change in two forms of glutamic acid decarboxylase (GABA synthesized enzymes) GAD(65) and GAD(67) in the cortex and hippocampus areas while the anxiolytic actions induced by chronic administration of honokiol in mice. Mice treated with 7 daily injection of honokiol (1mg/kg, p.o.) caused anxiolytic action which was similar to that was induced by 7 daily injection of diazepam (2mg/kg, p.o.) in the elevated plus-maze test. In addition, the activity of hippocampal GAD(65) of honokiol treated mice was significantly increased than that of the vehicle or diazepam treated groups. These data suggest that honokiol causes diazepam-like anxiolytic action, which may be mediated by altering the synthesis of GABA in the brain of mice.

Topics: Animals; Anti-Anxiety Agents; Anxiety; Biphenyl Compounds; Central Nervous System Depressants; Cerebral Cortex; Diazepam; Glutamate Decarboxylase; Hippocampus; Humans; Lignans; Magnolia; Male; Maze Learning; Mice; Mice, Inbred ICR; Phytotherapy; Plant Bark; Plant Extracts

There has been a strong interest in searching for natural therapies for osteoporosis. Honokiol is a phenolic compound isolated from the bark of Magnolia officinalis, a plant widely used in traditional medicine. In the present study, the effects of honokiol on the function of osteoblastic MC3T3-E1 cells were studied. Honokiol caused a significant elevation of cell growth, alkaline phosphatase activity, collagen synthesis, mineralization, glutathione content, and osteoprotegerin release in the cells (P<0.05). Moreover, honokiol significantly (P<0.05) decreased the production of osteoclast differentiation inducing factors such as TNF-α, IL-6, and receptor activator of nuclear factor-kB ligand (RANKL) in the presence of antimycin A, which inhibits mitochondrial electron transport and has been used as a ROS generator. These results demonstrate that honokiol may have positive effects on skeletal structure.

Topics: Alkaline Phosphatase; Animals; Antimycin A; Biphenyl Compounds; Bone Resorption; Calcification, Physiologic; Cell Growth Processes; Cell Line; Collagen; Glutathione; Humans; Interleukin-6; Lignans; Magnolia; Mice; Osteoblasts; Osteoporosis; Osteoprotegerin; Phytotherapy; RANK Ligand; Tumor Necrosis Factor-alpha

Honokiol (HNK) is a phenolic compound isolated from the bark of houpu (Magnolia officinalis), a plant widely used in traditional Chinese and Japanese medicine. While substantial evidence indicates that HNK possesses anti-inflammatory activity, its effect on dendritic cells (DCs) during the inflammatory reaction remains unclear. The present study investigates how HNK affects lipopolysaccharide (LPS)-stimulated human monocyte-derived DCs. Our experimental results show that HNK inhibits the inflammatory response of LPS-induced DCs by (1) suppressing the expression of CD11c, CD40, CD80, CD83, CD86, and MHC-II on LPS-activated DCs, (2) reducing the production of TNF-α, IL-1β, IL-6, and IL-12p70 but increasing the production of IL-10 and TGF-β1 by LPS-activated DCs, (3) inhibiting the LPS-induced DC-elicited allogeneic T-cell proliferation, and (4) shifting the LPS-induced DC-driven Th1 response toward a Th2 response. Further, our results show that HNK inhibits the phosphorylation levels of ERK1/2, p38, JNK1/2, IKKα, and IκBα in LPS-activated DCs. Collectively, the findings show that the anti-inflammatory actions of HNK on LPS-induced DCs are associated with the NF-κB and mitogen-activated protein kinase (MAPK) signaling pathways.

Topics: Biphenyl Compounds; Cell Differentiation; Cell Line; Cell Proliferation; Cell Survival; Cytokines; Dendritic Cells; Endocytosis; Enzyme Activation; Humans; Inflammation; Lignans; Lipopolysaccharides; MAP Kinase Signaling System; Mitogen-Activated Protein Kinases; Monocytes; NF-kappa B; Phenotype; Th1 Cells; Th2 Cells

A series of 31 analogues of the neolignan honokiol (a major constituent of Magnolia officinalis) was synthesized, and their effects on GABA(A) receptors expressed in Xenopus oocytes were investigated. Honokiol enhanced chloride currents (I(GABA)) through GABA(A) receptors of seven different subunit compositions with EC(50) values ranging from 23.4 μM (α(5)β(2)) to 59.6 μM (α(1)β(3)). Honokiol was most efficient on α(3)β(2) (maximal I(GABA) enhancement 2386%) > α(2)β(2) (1130%) > α(1)β(2) (1034%) > α(1)β(1) (260%)). On α(1)β(2)-receptors, N-substituted compounds were most active with 3-acetylamino-4'-O-methylhonokiol (31), enhancing I(GABA) by 2601% (EC(50) (α(1)β(2)) = 3.8 μM). Pharmacophore modeling gave a model with an overall classification accuracy of 91% showing three hydrophobic regions, one acceptor and one donor region. Unlike honokiol, 31 was most efficient on α(2)β(2)- (5204%) > α(3)β(2)- (3671%) > α(1)β(2)-receptors (2601%), suggesting a role of the acetamido group in subunit-dependent receptor modulation.

Topics: Animals; Biphenyl Compounds; Lignans; Models, Molecular; Oocytes; Receptors, GABA-A; Structure-Activity Relationship; Xenopus laevis

The antioxidant properties of magnolol and honokiol were evaluated in the experimental systems of reducing ONOO(-) and (1)O(2), bleaching β-carotene in linoleic acid (LH) emulsion, and trapping 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) cationic radical (ABTS(+)*) and 2,2'-diphenyl-1-picrylhydrazyl radical (DPPH), and then were applied to inhibit the oxidation of DNA induced by Cu(2+)/glutathione (GSH) and 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH). Magnolol and honokiol were active to reduce ONOO(-) and (1)O(2). Honokiol showed a little higher activity to protect LH and to inhibit Cu(2+)/GSH-induced oxidation of DNA than magnolol. In addition, honokiol exhibited higher activities to trap ABTS(+)* and DPPH than magnolol. In particular, honokiol trapped 2.5 radicals while magnolol only trapped 1.8 radicals in protecting DNA against AAPH-induced oxidation. The obtained results suggested that low antioxidant ability of magnolol may be related to the intramolecular hydrogen bond formed between di-ortho-hydroxyl groups, which hindered the hydrogen atom in hydroxyl group to be abstracted by radicals. Therefore, the antioxidant capacity of magnolol was lower than that of honokiol.

Topics: Antioxidants; Biphenyl Compounds; DNA; Free Radical Scavengers; Lignans

Survival rates for patients with pancreatic cancer are extremely poor due to its asymptomatic progression to advanced and metastatic stage for which current therapies remain largely ineffective. Therefore, novel therapeutic agents and treatment approaches are desired to improve the clinical outcome. In this study, we determined the effects of honokiol, a biologically active constituent of oriental medicinal herb Magnolia officinalis/grandiflora, on two pancreatic cancer cell lines, MiaPaCa and Panc1, alone and in combination with the standard chemotherapeutic drug, gemcitabine. Honokiol exerted growth inhibitory effects on both the pancreatic cancer cell lines by causing cell cycle arrest at G₁ phase and induction of apoptosis. At the molecular level, honokiol markedly decreased the expression of cyclins (D1 and E) and cyclin-dependent kinases (Cdk2 and Cdk4), and caused an increase in Cdk inhibitors, p21 and p27. Furthermore, honokiol treatment led to augmentation of Bax/Bcl-2 and Bax/Bcl-xL ratios to favor apoptosis in pancreatic cancer cells. These changes were accompanied by enhanced cytoplasmic accumulation of NF-κB with a concomitant decrease in nuclear fraction and reduced transcriptional activity of NF-κB responsive promoter. This was associated with decreased phosphorylation of inhibitor of kappa B alpha (IκB-α) causing its stabilization and thus increased cellular levels. Importantly, honokiol also potentiated the cytotoxic effects of gemcitabine, in part, by restricting the gemcitabine-induced nuclear accumulation of NF-κB in the treated pancreatic cancer cell lines. Altogether, these findings demonstrate, for the first time, the growth inhibitory effects of honokiol in pancreatic cancer and indicate its potential usefulness as a novel natural agent in prevention and therapy.

Topics: Apoptosis; bcl-2-Associated X Protein; bcl-X Protein; Biphenyl Compounds; Cell Cycle; Cell Cycle Proteins; Cell Line, Tumor; Cell Nucleus; Cell Proliferation; Cell Survival; Deoxycytidine; Drug Screening Assays, Antitumor; Drug Synergism; G1 Phase; Gemcitabine; Humans; Lignans; NF-kappa B; Pancreatic Neoplasms; Protein Transport

Honokiol is a phenolic compound isolated from the bark of Magnolia officinalis, a plant widely used in traditional medicine. Antimycin A, which inhibits complex III of the electron transport system, has been used as a reactive oxygen species generator in biological systems. In the present study, we investigated the protective effects of honokiol on antimycin A-induced dysfunction in osteoblastic MC3T3-E1 cells.. Osteoblastic MC3T3-E1 cells were pre-incubated with honokiol before treatment with antimycin A, and markers of mitochondrial function and oxidative damage were examined. In addition, the effects of honokiol on the activation of PI3K (phosphoinositide 3-kinase) and CREB (cAMP-responsive element-binding protein) were examined in MC3T3-E1 cells.. Honokiol significantly (P < 0.05) increased cell viability and calcium deposition and decreased the production of ROS in the presence of antimycin A. Moreover, pretreatment with honokiol prior to antimycin A exposure significantly reduced antimycin A-induced mitochondrial membrane potential (MMP) dissipation, complex IV inactivation, nitrotyrosine formation, and thioredoxin reductase inactivation. Honokiol also induced the activation of PI3K and CREB inhibited by antimycin A, which demonstrates that honokiol utilizes the PI3K and CREB pathway to augment metabolic activity inhibited by antimycin A.. Honokiol may reduce or prevent osteoblast degeneration in osteoporosis or other degenerative disorders.

Topics: Animals; Anti-Bacterial Agents; Antimycin A; Antioxidants; Biphenyl Compounds; Cell Survival; Cyclic AMP Response Element-Binding Protein; Lignans; Magnolia; Membrane Potential, Mitochondrial; Mice; Mitochondria; Osteoblasts; Oxidative Stress; Oxygen; Phosphatidylinositol 3-Kinases; Reactive Oxygen Species; Superoxides

Honokiol is a small-molecule polyphenol isolated from the species Magnolia obovata. We hypothesized that honokiol attenuated vascular contractions through the inhibition of the RhoA/Rho-kinase signalling pathway.. Rat aortic rings were denuded of endothelium, mounted in organ baths, and subjected to contraction or relaxation. Phosphorylation of 20kDa myosin light chains (MLC(20) ), myosin phosphatase targeting subunit 1 (MYPT1) and protein kinase C (PKC)-potentiated inhibitory protein for heterotrimeric myosin light chain phosphatase (MLCP) of 17kDa (CPI17) were examined by immunoblot. We also measured the amount of guanosine triphosphate RhoA as a marker for RhoA activation.. Pretreatment with honokiol dose-dependently inhibited the concentration-response curves in response to sodium fluoride (NaF) or thromboxane A(2) agonist U46619. Honokiol decreased the phosphorylation levels of MLC(20) , MYPT1(Thr855) and CPI17(Thr38) as well as the activation of RhoA induced by 8.0mm NaF or 30nm U46619.. These results demonstrated that honokiol attenuated vascular contraction through the inhibition of the RhoA/Rho-kinase signalling pathway.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Animals; Aorta; Biphenyl Compounds; Dose-Response Relationship, Drug; Lignans; Magnolia; Male; Muscle Contraction; Muscle, Smooth, Vascular; Myosin Light Chains; Myosin-Light-Chain Phosphatase; Phosphorylation; Plant Extracts; Pyridones; Rats; Rats, Sprague-Dawley; rho-Associated Kinases; rhoA GTP-Binding Protein; Signal Transduction; Sodium Fluoride; Vasoconstriction; Vasodilator Agents

DNA mismatch repair is required for correcting any mismatches that are created during replication and recombination, and a defective mismatch repair system contributes to DNA damage-induced growth arrest. The colorectal cancer cell line HCT116 is known to have a mutation in the hMLH1 mismatch repair gene resulting in microsatellite instability and defective mismatch repair. Honokiol is a biphenolic compound that has been used in traditional Chinese medicine for treating various ailments including cancer. This study was designed to test the hypothesis that honokiol enhances the radiosensitivity of cancer cells with mismatch repair defect (HCT116) compared with those that are mismatch repair proficient (HCT116-CH3). We first determined that the combination of honokiol and γ-irradiation treatment resulted in dose-dependent inhibition of proliferation and colony formation in both cell lines. However, the effects were more pronounced in HCT116 cells. Similarly, the combination induced higher levels of apoptosis (caspase 3 activation, Bax to Bcl2 ratio) in the HCT116 cells compared with HCT116-CH3 cells. Cell cycle analyses revealed higher levels of dead cells in HCT116 cells. The combination treatment reduced expression of cyclin A1 and D1 and increased phosphorylated p53 in both cell lines, although there were significantly lower amounts of phosphorylated p53 in the HCT116-CH3 cells, suggesting that high levels of hMLH1 reduce radiosensitivity. These data demonstrate that honokiol is highly effective in radiosensitizing colorectal cancer cells, especially those with a mismatch repair defect.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Colorectal Neoplasms; DNA Mismatch Repair; HCT116 Cells; Humans; Lignans; Radiation Tolerance; Radiation-Sensitizing Agents; Tumor Cells, Cultured

To study the transport mechanism of honokiol in Caco-2 cell model.. The analysis was performed on a Kromasil 100-5 C18 column (4.5 mm x 250 mm, 5 microm) eluted with acetonitrile-water (70: 30) as mobile phase. The detection wavelength was set at 203 nm. Two-way transport of honokiol was studied by using Caco-2 cell model, and the effects of time, drug concentration, inhibitor, pH, temperature on the transport of honokiol was investigated. The drug concentrations were determined by high performance liquid chromatography(HPLC) and used to calculate the apparent permeability coefficient.. The standard curve of honokiol was Y = 24 044X - 3 763.6 (r = 0.999 8), and the detection limit was 0.04 micromol x L(-1). In Caco-2 cell model, the transport amounts from the top side to the base side of were more than that from the base side to the top side under the same concentration. The transport amounts increased with time both in AP --> BL and BL --> AP directions. Verapamil could improve the transport amounts of AP --> BL. There were no effects of pH on the transport of AP --> BL. Both in AP --> BL and BL --> AP directions, the transport showed temperature dependence.. Honokiol is transported through the intestinal mucosa via a passive diffusion mechanism primarily, coexisting with a carrier-mediated transport, at the same time effected by P-gp.

Topics: Biological Transport; Biphenyl Compounds; Caco-2 Cells; Chromatography, High Pressure Liquid; Diffusion; Humans; Lignans; Permeability

Honokiol, widely known as an antitumor agent, has been used as an antiangiogenesis drug lead. In this paper, 47 honokiol analogues and derivatives were investigated for their antiangiogenic activity by application of the transgenic zebrafish screening model, antiproliferative and cytotoxic activity against HUVECs, and three tumor cell lines by MTT assay. 3',5-Diallyl-2,4'-dihydroxy-[1,1'-biphen-yl]-3,5'-dicarbaldehyde (8c) was found to suppress the newly grown segmental vessels from the dorsal aorta of zebrafish and prevent inappropriate vascularization as well as exhibit more potent inhibitory effects on the proliferation of HUVECs, A549, HepG2, and LL/2 cells (IC(50) = 15.1, 30.2, 10.7, and 21.7 μM, respectively) than honokiol (IC(50) = 52.6, 35.0, 16.5, and 65.4 μM, respectively). Analogue 8c also effectively inhibited the migration and capillary-like tube formation of HUVECs in vitro. The antiangiogenic effect and antiproliferative activity of these structurally modified honokiol analogues and derivatives have led to the establishment of a structure-activity relationship.

Topics: Allyl Compounds; Angiogenesis Inhibitors; Animals; Animals, Genetically Modified; Benzaldehydes; Biphenyl Compounds; Cell Line; Cell Line, Tumor; Cell Movement; Cell Proliferation; Drug Screening Assays, Antitumor; Embryo, Nonmammalian; Endothelial Cells; Endothelium, Vascular; Humans; In Vitro Techniques; Lignans; Magnolia; Neovascularization, Pathologic; Neovascularization, Physiologic; Structure-Activity Relationship; Zebrafish

This paper aims to investigate the metabolism and pharmacokinetics of curcumin, demethoxycurcumin and bisdemethoxycurcumin in mice tumor. To improve water solubility, nanoparticle formulations were prepared as curcuminoids-loaded solid lipid nanoparticles (curcuminoids-SLNs) and curcumin-loaded solid lipid nanoparticles (curcumin-SLNs). After intragastric administration to tumor-bearing ICR mice, the plasma and tumor samples were analyzed by liquid chromatography with ion trap mass spectrometry. We discovered that curcuminoids were mainly present as glucuronides in plasma, whereas in free form in tumor tissue. A validated LC/MS/MS method was established to determine the three free curcuminoids in tumor homogenate. Samples were separated on a Zorbax SB-C(18) column, eluted with acetonitrile-water (containing 0.1% formic acid), and detected by TSQ Quantum triple quadrupole mass spectrometer in selected reaction monitoring mode. The method showed good linearity (r(2)=0.997-0.999) over wide dynamic ranges (2-6000 ng/mL). Variations within- and between-batch never exceeded 11.2% and 13.4%, respectively. The extraction recovery rates ranged from 78.3% to 87.7%. The pharmacokinetics of curcuminoids in mice tumor fit two-compartment model and first order elimination. For curcumin-SLNs group, the dosing of 250 mg/kg of curcumin resulted in AUC((0-48 h)) of 2285 ngh/mL and C(max) of 209 ng/mL. For curcuminoids-SLNs group, the dosing equivalent to 138 mg/kg of curcumin resulted in higher tumor concentrations (AUC=2811 ngh/mL, C(max)=285 ng/mL). It appeared that co-existing curcuminoids improved the bioavailability of curcumin.

Topics: Animals; Area Under Curve; Biphenyl Compounds; Chromatography, Liquid; Curcumin; Diarylheptanoids; Drug Delivery Systems; Drug Stability; Lignans; Linear Models; Lipids; Male; Mice; Mice, Inbred ICR; Nanoparticles; Neoplasm Transplantation; Neoplasms; Reproducibility of Results; Tandem Mass Spectrometry

Magnolia officinalis, a component of Asian herbal teas, has long been employed in traditional Japanese and Chinese medicine to treat numerous maladies. Honokiol, a biphenolic compound, is now considered to be one of the major active ingredients of Magnolia extract, and is under intense investigation for its anti-angiogenic, anti-inflammatory, anti-tumor and neuroprotective properties. Biochemically, honokiol has been recognized to modulate the nuclear factor κ B (NF-κB) signal transduction pathway suggesting that it possesses anti-inflammatory properties. Inflammation is intimately associated with bone turnover and skeletal deterioration and consequently, anti-inflammatory drugs may hold significant promise as bone protective agents to stem bone loss in osteoporotic conditions. We and others have demonstrated that suppression of NF-κB blunts osteoclastic bone resorption, but promotes osteoblastic bone formation. Indeed previous studies have demonstrated the anti-osteoclastogenic effects of honokiol, however, activities on osteoblast differentiation and activity have yet to be investigated. In this study, we show that honokiol is a potent inducer of in vitro osteoblast differentiation by virtue of its capacity to suppress basal and tumor necrosis factor alpha (TNFα)-induced NF-κB activation and to alleviate the suppressive action of TNFα on bone morphogenetic protein (BMP)-2-induced Smad activation. Our data confirm that honokiol may have considerable promise as a dual anabolic/anti-catabolic agent for the amelioration of multiple osteoporotic diseases.

Topics: Animals; Anti-Inflammatory Agents; Biphenyl Compounds; Blotting, Western; Bone Morphogenetic Protein 2; Bone Resorption; Cell Differentiation; Cell Line; Drugs, Chinese Herbal; Humans; Lignans; Magnolia; Mice; NF-kappa B; Osteoclasts; Osteogenesis; Osteoporosis; Signal Transduction; Smad Proteins; Tumor Necrosis Factor-alpha

Honokiol is a plant lignan isolated from bark and seed cones of Magnolia officinalis. Recent studies from our laboratory indicated that honokiol pretreatment decreased ultraviolet B-induced skin cancer development in SKH-1 mice. The aim of the present investigation was to study the effects of honokiol on human epidermoid squamous carcinoma A431 cells and to elucidate possible mechanisms involved in preventing skin cancer. A431 cells were pretreated with different concentrations of honokiol for a specific time period and investigated for effects on apoptosis and cell cycle analysis. Treatment with honokiol significantly decreased cell viability and cell proliferation in a concentration- and time-dependent manner. Honokiol pretreatment at 50 μmol/L concentration induced G0/G1 cell cycle arrest significantly (P < 0.05) and decreased the percentage of cells in the S and G2/M phase. Honokiol down-regulated the expression of cyclin D1, cyclin D2, Cdk2, Cdk4 and Cdk6 proteins and up-regulated the expression of Cdk's inhibitor proteins p21 and p27. Pretreatment of A431 cells with honokiol leads to induction of apoptosis and DNA fragmentation. These findings indicate that honokiol provides its effects in squamous carcinoma cells by inducing cell cycle arrest at G0/G1 phase and apoptosis.

Topics: Animals; Anticarcinogenic Agents; Apoptosis; Biphenyl Compounds; Carcinoma, Squamous Cell; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Cyclin D2; Cyclin-Dependent Kinase 2; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinase 6; Cyclin-Dependent Kinase Inhibitor p21; Cyclin-Dependent Kinase Inhibitor p27; DNA Fragmentation; Gene Expression Regulation, Neoplastic; Humans; Lignans; Mice; Skin Neoplasms

Multi-drug resistance is of great concern for public health worldwide and necessitates the search for new antimicrobials from sources such as plants. Several Magnolia (Magnoliaceae) species have been reported to exert antimicrobial effects on sensitive and multidrug-resistant microorganisms. However, the antimicrobial properties of Magnolia dealbata have not been experimentally evaluated. The antimicrobial effects of an ethanol extract of Magnolia dealbata seeds (MDE) and its active compounds honokiol (HK) and magnolol (MG) were tested against the phytopathogen Clavibacter michiganensis subsp. michiganensis and several human multi-drug resistant pathogens using the disk-diffusion assay. The effects of MDE and its active compounds on the viability of human peripheral blood mononuclear cells (PBMC) were evaluated using MTT assay. MDE and its active compounds had antimicrobial activity (inhibition zone > 10 mm) against C. michiganensis, Pseudomonas aeruginosa, Acinetobacter baumannii, Acinetobacter lwoffii, Candida albicans, Candida tropicalis and Trichosporon belgeii. The results suggest that M. dealbata and its active compounds have selective antimicrobial effects against drug-resistant fungal and Gram (-) bacteria and exert minimal toxic effects on human PMBC.

Topics: Anti-Bacterial Agents; Antifungal Agents; Bacteria; Biphenyl Compounds; Fungi; Humans; Leukocytes, Mononuclear; Lignans; Magnolia; Plant Extracts

To investigate the apoptosis-inducing effect of honokiol on human non-Hodgkin lymphoma Raji cells and the possible mechanism.. Raji cells were treated with different concentrations of honokiol, and the proliferation of the cells was detected using MTT assay. Flow cytometry was employed to analyze the cell cycle changes and apoptosis of honokiol-treated cells. Caspase 8 activity in the cells was measured by caspase 8 kit, and RT-PCR was used to detect the expression of apoptosis-related genes Bcl-2, Bad, and Bax.. Honokiol significantly inhibited the growth of Raji cells in a time- and dose-dependent manner, with IC(50) concentration of 17.53, 12.61, and 7.4 µg/ml at 12, 24, 48 h, respectively. Flow cytometry revealed cell cycle arrest at G0/G1 phase following honokiol treatment. The apoptosis rates of Raji cells treated with 7.5 and 15 µg/ml honokiol were significantly higher than that of the control cells [(18.24∓2.53)%, (28.44∓2.48)% vs (4.84∓1.15)%, P<0.01]. Caspase 8 activity in Raji cells was significantly enhanced by honokiol (P<0.05). The mRNA expression of the apoptosis-promoting gene Bad was significantly increased following honokiol treatment (P<0.01), while the expressions of Bcl-2 and Bax remained unchanged.. Honokiol can induce apoptosis in Raji cells possibly in relation to enhancement of caspase 8 activity and Bad gene expression.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; bcl-Associated Death Protein; Biphenyl Compounds; Burkitt Lymphoma; Caspase 8; Cell Line, Tumor; Cell Proliferation; Humans; Lignans; Lymphoma, Non-Hodgkin

In this article, we demonstrated a novel injectable polymer matrix: honokiol (HK) loaded poly (epsilon-caprolactone)-poly(ethylene glycol)-poly(epsilon-caprolactone) (PCL-PEG-PCL, PCEC) nanoparticles in thermosensitive poly(ethylene glycol)-poly(epsilon-caprolactone)-poly(ethylene glycol) (PEG-PCL-PEG, PECE) hydrogel for the drug local delivery. First, HK, as a model hydrophobic drug, was loaded into PCL-PEG-PCL nanoparticles by emulsion solvent evaporation method to overcome its poor water solubility. Then, the HK-loaded PCEC nanoparticles (HK-PCEC) were incorporated into thermosensitive PEG-PCL-PEG hydrogel, which was sol at low temperature and could gel as a depot for sustained release of drug in situ after topical injection. The HK-PCEC incorporated PECE hydrogel (HK-PCEC-PECE) was biodegradable and could be gradually eliminated from the injection site in about 2 weeks after subcutaneously injected into mice. The in vitro release studies indicated that HK could be released from HK-PCEC and HK-PCEC-PECE in a sustained manner. Such biodegradable smart drug-delivery system might have great potential application in injectable hydrophobic drug local delivery system.

Topics: Animals; Biocompatible Materials; Biphenyl Compounds; Drug Delivery Systems; Hydrogel, Polyethylene Glycol Dimethacrylate; Hydrophobic and Hydrophilic Interactions; Lignans; Mice; Nanoparticles; Polyesters; Polyethylene Glycols; Temperature

Chondrosarcoma is a malignant primary bone tumor that responds poorly to both chemotherapy and radiation therapy. In the present study, we investigated the anti-cancer effect of a honokiol, an active component isolated and purified from the Magnolia officinalis in human chondrosarcoma cells. Honokiol-induced cell apoptosis in human chondrosarcoma cell lines (including: JJ012 and SW1353) but not primary chondrocytes. Honokiol also induces upregulation of Bax and Bak, downregulation of Bcl-XL and dysfunction of mitochondria in chondrosarcoma cells. Honokiol triggered endoplasmic reticulum (ER) stress, as indicated by changes in cytosol-calcium levels. We also found that honokiol increased the expression and activities of glucose-regulated protein 78 (GRP78) and calpain. Transfection of cells with GRP78 or calpain siRNA reduced honokiol-mediated cell apoptosis in JJ012 cells. Importantly, animal studies have revealed a dramatic 53% reduction in tumor volume after 21days of treatment. This study demonstrates that honokiol may be a novel anti-cancer agent targeting chondrosarcoma cells.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Bone Neoplasms; Calcium; Calpain; Cell Line, Tumor; Chondrosarcoma; Endoplasmic Reticulum; Endoplasmic Reticulum Chaperone BiP; Heat-Shock Proteins; Humans; Lignans; Male; Mice; Mice, Inbred BALB C; Mitochondria; Xenograft Model Antitumor Assays

Honokiol (HK) shows potential application in cancer treatment, but its poor water solubility restricts clinical application greatly. In this paper, monomethoxy poly(ethylene glycol)-poly(lactic acid) (MPEG-PLA) was synthesized by ring-opening polymerization and processed into nanoparticle for honokiol delivery. Chemical structure of the synthesized polymer was confirmed by (1)H NMR, and its molecular weight was determined by gel permeation chromatography (GPC). Honokiol loaded MPEG-PLA nanoparticles were prepared by solvent extract method. And particle size distribution, morphology, drug loading, drug release profile and anticancer activity in vitro were studied in detail. The described honokiol loaded MPEG-PLA nanoparticles in this paper might be a novel formulation for honokiol delivery.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Cell Survival; Chemistry, Pharmaceutical; Chromatography, Gel; Dose-Response Relationship, Drug; Drug Carriers; Female; Hemolysis; Humans; Kinetics; Lignans; Magnetic Resonance Spectroscopy; Molecular Structure; Molecular Weight; Nanoparticles; Nanotechnology; Ovarian Neoplasms; Particle Size; Polyesters; Polyethylene Glycols; Solubility; Solvents; Surface Properties; Technology, Pharmaceutical

A rapid expansion of supercritical solution (RESS) technology was presented for the micronization of Chinese medicinal material. Magnolia bark extract (MBE) obtained by supercritical carbon dioxide (scCO(2)) extraction technology was chosen as the experimental material. RESS process produced 4.7 microm size MBE microparticles (size distribution, 0.2-24.1 microm), which was significantly smaller than the 55.3 microm size particles (size distribution, 8.3-102.4 microm) obtained from conventional mechanical milling. Dissolution rate study showed that drug dissolution was significantly enhanced by the RESS progress. At 90 min, the amount dissolved of mechanical milling MBE was 6.37 mg x l(-1), which was significantly lower than that of micronized MBE (14.77 mg x l(-1)), according to the results of ANOVA (p<0.01). The effect of extraction temperature (30, 40, 50 degrees Celsius), extraction pressure (200, 250, 300 bar) and nozzle size (50, 100, 200 microm) on the size distribution of microparticles was investigated. The characteristics of microparticles were also studied by differential scanning calorimetry (DSC), infrared spectroscopy (IR), scanning electron microscopy (SEM), and image analysis. This study demonstrates that RESS is applicable for preparing microparticles of MBE at low operating temperature; the process is simple without residual solvent.

Topics: Biphenyl Compounds; Calorimetry, Differential Scanning; Carbon Dioxide; Drugs, Chinese Herbal; Equipment Design; Lignans; Magnolia; Molecular Structure; Particle Size; Plant Bark; Solubility; Solutions; Spectrophotometry, Infrared; Technology, Pharmaceutical; Time Factors

Honokiol (HNK), a natural small molecular product, inhibited proliferation of HepG2 cells and exhibited anti-tumor activity in nude mice. In this article, we applied a novel sensitive stable isotope labeling with amino acids in cell culture-based quantitative proteomic method and a model of nude mice to investigate the correlation between HNK and the hotspot migration molecule Ras GTPase-activating-like protein (IQGAP1). The quantitative proteomic analysis showed that IQGAP1 was 0.53-fold down-regulated under 10 microg/mL HNK exposure for 24 h on HepG2 cells. Migration ability of HepG2 cells under HNK treatment was correlated with its expression level of IQGAP1. In addition, the biochemical validation on HepG2 cells and the tumor xenograft model further demonstrated that HNK decreased the expression level of IQGAP1 and its upstream proteins Cdc42/Rac1. These data supported that HNK can modulate cell adhesion and cell migration by acting on Cdc42/Rac1 signaling via IQGAP1 interactions with its upstream Cdc42/Rac1 proteins, which is a new molecular mechanism of HNK to exert its anti-tumor activity.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; cdc42 GTP-Binding Protein; Cell Movement; Down-Regulation; Hep G2 Cells; Histocytochemistry; Humans; Isotope Labeling; Lignans; Mice; Mice, Nude; Proteome; Proteomics; rac1 GTP-Binding Protein; ras GTPase-Activating Proteins; Reproducibility of Results; Statistics, Nonparametric; Tumor Burden

This paper developed a new hydrophobic honokiol transdermal delivery system. First, Honokiol was loaded into Pluronic F127 micelles by direct dissolution method assisted by ultrasound. Then the obtained honokiol-loaded F127 micelles were incorporated into thermosensitive F127 hydrogel, which made the composite system bioadhesive. The particle size, drug loading, and encapsulation efficiency were determined. The sol-gel transitions of the copolymer, honokiol release profile in vitro, and the permeation studies in vitro were studied in detail. The lower critical solution temperature (LCST) of the composite system decreases with increase in the mass of honokiol in the system. Honokiol could be sustained released from the system in vitro. In in vitro permeation studies, honokiol could be absorbed per cutem. The described drug delivery system might have great potential application for transdermal delivery of hydrophobic drugs such as honokiol.

Topics: Administration, Cutaneous; Animals; Anti-Infective Agents; Anti-Inflammatory Agents, Non-Steroidal; Biphenyl Compounds; Cell Survival; Chemistry, Pharmaceutical; Chromatography, High Pressure Liquid; Excipients; Hydrogels; Lignans; Micelles; Particle Size; Permeability; Poloxamer; Rats; Rats, Sprague-Dawley; Skin Absorption; Temperature

The present study was performed to evaluate the antihypertensive effects of honokiol in vivo in spontaneously hypertensive rats (SHR). The effects of honokiol were investigated by determination of the blood pressure, vascular reactivity, oxidative parameters, and histologic change in the aorta. Long-term administration of honokiol (400 mg/kg/d) to SHR decreased systolic blood pressure significantly. Honokiol (200, 400 mg/kg/d) enhanced the aortic relaxation in response to acetylcholine after 49-d treatment, but had no significant effects on the relaxation to sodium nitroprusside. The oral administration of honokiol significantly increased the plasma level of NO(2(-))/NO(3(-)), but decreased the level of malondialdehyde in liver of SHR compared with the control vehicle. In addition, SHR administered honokiol showed significant reductions in the elastin bands and media thickness in the aorta. These results suggest that chronic treatment with honokiol exerts an antihypertensive effect in SHR, and its vasorelaxant action and antioxidant properties may contribute to reducing the elevated blood pressure.

Topics: Acetylcholine; Animals; Antihypertensive Agents; Antioxidants; Aorta; Biphenyl Compounds; Blood Pressure; Elastin; Hypertension; Lignans; Liver; Magnolia; Malondialdehyde; Nitrates; Nitrites; Nitroprusside; Plant Extracts; Rats; Rats, Inbred SHR; Vasodilator Agents

Honokiol, a neolignan, is a physiologically active component of kouboku (Magnolia obovata), a herb used in traditional Chinese medicine. This study investigated the effects of honokiol on the differentiation and function of osteoclasts induced by receptor activator of nuclear factor-kappaB ligand (RANKL). Honokiol markedly inhibited RANKL-induced tartrate-resistant acid phosphatase (TRAP) activity and the formation of TRAP-positive multinucleated cells in both bone marrow-derived monocytes and RAW264 cells. In experiments to elucidate its mechanism of action, honokiol was found to suppress RANKL-induced phosphorylation of p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK). The RANKL-induced expressions of c-Fos and nuclear factor of activated T cells-c1 (NFATc1), which are crucial transcriptional factors for osteoclastogenesis, were also reduced by treatment with honokiol. Furthermore, honokiol induced disruption of the actin rings in mature osteoclasts (mOCs) without affecting the cell viability and suppressed osteoclastic pit formation on dentin slices. Taken together, these results suggest that honokiol inhibits osteoclast differentiation by suppressing the activation of MAPKs (p38 MAPK, ERK and JNK), decreasing the expressions of c-Fos and NFATc1, and attenuates bone resorption by disrupting the actin rings in mOCs. Therefore, honokiol could prove useful for the treatment of bone diseases associated with excessive bone resorption.

Topics: Acid Phosphatase; Actins; Animals; Biphenyl Compounds; Bone Density Conservation Agents; Bone Marrow Cells; Bone Resorption; Cell Differentiation; Dentin; Isoenzymes; Lignans; Magnolia; Mice; Mitogen-Activated Protein Kinases; NFATC Transcription Factors; Osteoclasts; Phosphorylation; Plant Extracts; Proto-Oncogene Proteins c-fos; RANK Ligand; Tartrate-Resistant Acid Phosphatase

Honokiol, a major phenolic constituent of Magnolia sp., has various pharmacological activities. To improve the solubility and antibacterial activity of honokiol against E. coli and P. aeruginosa, new honokiol-derivatives (honokiol-acetate, honokiol-succinic acid, honokiol-glycerol, honokiol-glycine, honokiol-glucose and honokiol-mannose) were synthesized and their solubility and antimicrobial activities were investigated. Among the tested compounds, honokiol-glycine showed improved water solubility and antibacterial activities against E. coli and P. aeruginosa when compared to honokiol.

Topics: Anti-Infective Agents; Antifungal Agents; Bacteria; Biphenyl Compounds; Escherichia coli; Fungi; Lignans; Microbial Sensitivity Tests; Pseudomonas aeruginosa; Solubility

Much recent research has demonstrated that honokiol, a phenolic compound originally isolated from Magnolia officinalis, has potent anticancer activities; however, the detailed molecular mechanism of its anti-inflammatory activity has not yet been fully addressed. In this study we demonstrated that honokiol inhibited lipopolysaccharide (LPS)-induced tumor necrosis factor-alpha secretion in macrophages, without affecting the activity of the tumor necrosis factor-alpha converting enzyme. At the same time, honokiol not only inhibited nitric oxide expression in LPS-stimulated murine macrophages but also inhibited the LPS-induced phosphorylation of ERK1/2, JNK1/2, and p38. By means of confocal microscope analysis we demonstrated that phosphorylation and membrane translocation of protein kinase C-alpha, as well as NF-kappaB activation, were inhibited by honokiol in LPS-stimulated macrophages. Furthermore, it was found that honokiol neither antagonizes the binding of LPS to cells nor alters the cell surface expression of toll-like receptor 4 and CD14. Our current results have exhaustively described the anti-inflammatory properties of honokiol, which could lead to the possibility of its future pharmaceutical application in the realm of immunomodulation.

Topics: Animals; Anti-Inflammatory Agents; Biphenyl Compounds; Cell Line; Cells, Cultured; Down-Regulation; Humans; Lignans; Lipopolysaccharides; Macrophages; Mice; Mitogen-Activated Protein Kinases; NF-kappa B; Nitric Oxide; Protein Kinase C; Signal Transduction; Tumor Necrosis Factor-alpha

An efficient protocol for the in vitro propagation of Magnolia dealbata Zucc., an important medicinal plant that is the source of the anxiolytic and anticancer compounds honokiol and magnolol, was established. This plant is wild-crafted, and conservationists have expressed concerns with regard to the sustainability of production. In the present work, two factors were found to be of importance for the regeneration of M. dealbata and the production of honokiol and magnolol. These factors were the type of explants and the combination and concentration of plant-growth regulators. Green, compact, nodular organogenic callus was obtained from leaf explants in a medium fortified with Murashige and Skoog salts and supplemented with 1.5 mg/L 2,4-dicholorophenoxyacetic acid and 1.5 mg/L kinetin. Shoots multiplication from callus cultures was achieved in the Murashige and Skoog (MS) medium with 1.5 mg/L thidiazuron (TDZ). Phenol secretion was controlled by the addition of 250 mg/L of activated charcoal. For rooting, shoots were transferred to MS medium supplemented with several auxins. After root induction, the plants were hardened in earthen pots containing sand, soil, and vermiculite. The contents of honokiol (HK) and magnolol (MG) were determined in different plant materials by high-performance liquid chromatography-diode-array detection techniques. This analysis revealed that the honokiol and magnolol content in aerial and underground parts of micropropagated M. dealbata were higher than that observed in wild plants (both 6 months old). Our results suggest that conservation of M. dealbata is possible by means of in vitro multiplication of leaf-derived callus. The usefulness of M. dealbata regeneration and production of HK and MG may be attributed to the proper selection of explant sourcing and identification of the correct growth medium to support adequate growth. This careful selection of explants and growth medium leads to a very useful source of plant material for pharmacological and phytomedicinal screening applications and, above all, would safeguard this plant species from the threat of extinction.

Topics: Biphenyl Compounds; Conservation of Natural Resources; Culture Media; Lignans; Magnolia; Mexico; Plant Roots; Plant Shoots; Tissue Culture Techniques

Honokiol, an active component isolated and purified from Chinese traditional herb magnolia, has been shown to inhibit growth and induce apoptosis in different cancer cell lines. This study shows that honokiol can induce a cell death distinct from apoptosis at lower concentrations. The death was characterized by cytoplasmic vacuolization with the endoplasmic reticulum swelling and accompanied by apoptosis at higher concentrations in NB4 and K562 cells. The two death processes may be in sequence at lower concentrations and in parallel with the increase of honokiol concentration. Membrane-associated cytotoxicity was involved in honokiol-induced paraptosis and apoptosis. Furthermore, honokiol inhibited concentration-dependent cell adhesion to extracellular matrix for NB4 cells. In addition, the cytotoxicity of honokiol combined treatment with imatinib was schedule- and concentration-dependent and the sequential administration of honokiol before imatinib appeared to be more beneficial in K562 cells. Taken together, the data suggest that honokiol induced a novel cell death pathway and there was cross-talk between apoptotic and non-apoptotic programmed cell death caused by honokiol in leukemia cells. Moreover, honikiol exhibited schedule-dependent synergy in combination with imatinib and sequential administration of imatinib followed by honokiol could be the optimal sequence to combine these two drugs in K562 cells.

Topics: Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Benzamides; Biphenyl Compounds; Cell Line, Tumor; Cell Membrane; Cell Proliferation; Drug Administration Schedule; Drug Synergism; Humans; Imatinib Mesylate; Leukemia; Lignans; Piperazines; Pyrimidines; Vacuoles

This paper describes the application of high-performance counter-current chromatography (HPCCC) as a fast, useful and economic alternative for the separation and purification of seven honokiol derivatives (two of them are isomers), which were synthesized by a one-pot procedure. Five honokiol derivatives were successfully separated by n-hexane-ethyl acetate-methanol-water solvent system at three different volume ratios in a step-gradient elution. Two derivatives were obtained through a cycle elution mode. The whole separation process produced 366.3 mg, 323.6 mg, 242.8 mg, 216.2 mg, 203.5 mg, 185.8 mg and 279.3 mg of 3'-formylhonokiol (1), 2'-methoxy-3'-formylhonokiol (2), 2'-methoxyhonokiol (3), 4-methoxyhonokiol (4), 3',5-diformylhonokiol (5), 2',4-dimethoxy-3'-formylhonokiol (6) and 2',4-dimethoxyhonokiol (7) from crude sample of 3 g with purities of 98.7%, 99.3%, 98.6%, 98.2%, 99.0%, 98.4% and 99.2%, respectively. The purities and structural identification were determined by HPLC, (1)H NMR, (13)C NMR and mass spectroscopy.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Countercurrent Distribution; Lignans; Magnetic Resonance Spectroscopy

This study aimed to investigate the utility of honokiol, a naturally occurring compound, in the treatment of head and neck squamous cell carcinoma (HNSCC) as well as its ability to target the epidermal growth factor receptor (EGFR), a critical therapeutic target in HNSCC, and to enhance the effects of other EGFR-targeting therapies.. Human HNSCC cell lines and the xenograft animal model of HNSCC were used to test the effects of honokiol treatment.. Honokiol was found to inhibit growth in human HNSCC cell lines, with 50% effective concentration (EC(50)) values ranging from 3.3 to 7.4 micromol/L, and to induce apoptosis, as shown through Annexin V staining. These effects were associated with inhibition of EGFR signaling, including downstream inhibition of mitogen-activated protein kinase, Akt, and signal transducer and activator of transcription 3 (STAT3), and expression of STAT3 target genes, Bcl-X(L) and cyclin D1. Furthermore, honokiol enhanced the growth inhibitory and anti-invasion activity of the EGFR-targeting agent erlotinib. Although HNSCC xenograft models did not show significant inhibition of in vivo tumor growth with honokiol treatment alone, the combination of honokiol plus cetuximab, a Food and Drug Administration-approved EGFR inhibitor for this malignancy, significantly enhanced growth inhibition. Finally, HNSCC cells rendered resistant to erlotinib retained sensitivity to the growth inhibitory effects of honokiol.. These results suggest that honokiol may be an effective therapeutic agent in HNSCC, in which it can augment the effects of EGFR inhibitors and overcome drug resistance.

Topics: Animals; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Antineoplastic Agents, Phytogenic; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Biphenyl Compounds; Blotting, Western; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cetuximab; Dose-Response Relationship, Drug; Drug Synergism; Drugs, Chinese Herbal; ErbB Receptors; Erlotinib Hydrochloride; Head and Neck Neoplasms; Humans; Lignans; Mice; Mice, Nude; Quinazolines; Signal Transduction; Xenograft Model Antitumor Assays

Skin cancer is the most prevalent of all cancer types and its incidence is expected to increase substantially. Chemoprevention involves the administration of chemical agents to prevent initiation, promotion and/or progression that occurs during neoplastic development. Honokiol, a plant lignan isolated from bark and seed cones of Magnolia officinalis, has been shown to have chemopreventive effects on chemically induced skin cancer development.. The objective of this investigation was to study the chemopreventive effects of honokiol on UVB-induced skin tumor development in SKH-1 mice, a model relevant to humans, and to elucidate the possible role of apoptotic proteins involved in the prevention of skin tumor development.. Female SKH-1 mice were divided into two groups. Group 1 received acetone (0.2 ml, topical) and Group 2 received honokiol (30 microg in 0.2 ml acetone, topical) one hour before UVB treatment. Tumor initiation and promotion were carried out by UVB radiation (30 mJ/cm(2)/day), 5 days a week for 30 weeks. Tumor counts and mouse weights were taken weekly.. The honokiol-pretreated group exhibited a 45% reduction in tumor multiplicity as compared to the control group. Mechanistic studies showed the possible involvement of caspase-3, caspase-8, caspase-9, poly (ADP-ribose) polymerase (PARP) and p53 activation (p<0.05) leading to the induction of DNA fragmentation and apoptosis.. Pretreatment with honokiol, at concentrations in micrograms per application compared with milligram applications of other potential chemopreventive agents, prevents UVB-induced skin cancer development, possibly by activating proapoptotic proteins through both intrinsic and extrinsic pathways.

Topics: Animals; Anticarcinogenic Agents; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Caspases; Disease Models, Animal; Female; Lignans; Mice; Neoplasms, Radiation-Induced; Skin Neoplasms; Tumor Suppressor Protein p53; Ultraviolet Rays

This study aims to develop self-assembled poly(ethylene glycol)-poly(epsilon-caprolactone)-poly(ethylene glycol) (PEG-PCL-PEG, PECE) micelles to encapsulate hydrophobic honokiol (HK) in order to overcome its poor water solubility and to meet the requirement of intravenous administration. Honokiol loaded micelles (HK-micelles) were prepared by self-assembly of PECE copolymer in aqueous solution, triggered by its amphiphilic characteristic assisted by ultrasonication without any organic solvents, surfactants and vigorous stirring. The particle size of the prepared HK-micelles measured by Malvern laser particle size analyzer were 58 nm, which is small enough to be a candidate for an intravenous drug delivery system. Furthermore, the HK-micelles could be lyophilized into powder without any adjuvant, and the re-dissolved HK-micelles are stable and homogeneous with particle size about 61 nm. Furthermore, the in vitro release profile showed a significant difference between the rapid release of free HK and the much slower and sustained release of HK-micelles. Moreover, the cytotoxicity results of blank micelles and HK-micelles showed that the PECE micelle was a safe carrier and the encapsulated HK retained its potent antitumor effect. In short, the HK-micelles were successfully prepared by an improved method and might be promising carriers for intravenous delivery of HK in cancer chemotherapy, being effective, stable, safe (organic solvent and surfactant free), and easy to produce and scale up.

Topics: Adenocarcinoma; Animals; Antineoplastic Agents; Biphenyl Compounds; Cell Growth Processes; Cell Line, Tumor; Colonic Neoplasms; Drug Delivery Systems; Drug Screening Assays, Antitumor; Drug Stability; Drugs, Chinese Herbal; Humans; Lignans; Lung Neoplasms; Mice; Micelles; Microscopy, Electron, Transmission; Particle Size; Polyesters; Polyethylene Glycols; Solubility; Sonication; X-Ray Diffraction

The hydrophobic ionic liquid of [BMIM][PF(6)] was successfully used for the ultrasound-assisted extraction of hydrophobic magnolol and honokiol from cortex Magnoliae officinalis. To obtain the best extraction efficiencies, some ultrasonic parameters including the concentration of [BMIM][PF(6)], pH, ultrasonic power and ultrasonic time were evaluated. The results obtained indicated that the [BMIM][PF(6)]-based ultrasound-assisted extraction efficiencies of magnolol and honokiol were greater than those of the [BMIM][BF(4)]-based ultrasound-assisted extraction (from 48.6 to 45.9%) and the traditional ethanol reflux extraction (from 16.2 to 13.3%). Furthermore, the proposed extraction method is validated by the recovery, correlation coefficient (R(2)) and reproducibility (RSD, n=5), which were 90.8-102.6, 0.9992-0.9998, and 1.6-5.4%, respectively.

Topics: Biphenyl Compounds; Drugs, Chinese Herbal; Hydrogen-Ion Concentration; Hydrophobic and Hydrophilic Interactions; Imidazoles; Ionic Liquids; Lignans; Magnolia; Molecular Structure; Solvents; Ultrasonics

To evaluate the regulatory effects of honokiol on high-glucose (HG)-induced inflammatory responses of human renal mesangial cells (HRMCs).. We performed MTS assays to determine the non-cytotoxic concentration of honokiol for HRMCs. Enzyme-linked immunosorbent assays were performed to analyze the expressions of the proteins interleukin (IL)-1β, IL-18, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β1, monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1α, RANTES, and prostaglandin (PG) E2. The total nitric oxide (NO) concentration was determined using the Griess reaction.. Treatment with 50 mmol/L glucose markedly increased the level of IL-1β, IL-18, TNF-α, PGE2, NO, TGF-β1, MCP-1, MIP-1α, and RANTES. Honokiol (~20 μmol/L) treatment inhibited the HG-induced expression of inflammatory cytokines such as IL-1β, IL-18, TNF-α, PGE2, NO, and TGF-β1 in a dose-dependent manner. Moreover, it markedly inhibited the expression of chemokines such as MCP-1, MIP-1α, and RANTES, which are upregulated under HG conditions.. Honokiol inhibits the HG-induced expression of inflammatory factors in HRMCs. Honokiol may be considered a promising drug with potent anti-inflammatory activities in addition to its strong anti-cancer, anti-angiogenesis, and anti-neurodegenerative effects.

Topics: Animals; Anti-Allergic Agents; Biphenyl Compounds; Cell Survival; Cells, Cultured; Cytokines; Glucose; Humans; Lignans; Mesangial Cells; Up-Regulation

ABCA1, a member of the ATP-binding cassette transporter family, regulates high-density lipoprotein (HDL) metabolism and reverses cholesterol transport. Its expression is upregulated mainly by the activation of the liver X receptor (LXR), retinoid X receptor (RXR), and peroxisome proliferator-activated receptors (PPARs). To identify natural compounds that can upregulate ABCA1 expression, we developed a reporter assay using U251-MG (human glioma cell line) cells that stably express a human ABCA1 promoter-luciferase and performed a cell-based high-throughput screening of 118 natural compounds. Using this system, we identified honokiol, a compound extracted from Magnolia officinalis, as an activator of the ABCA1 promoter. We found that honokiol also increased ABCA1 mRNA and protein expression levels in a dose-dependent manner in U251-MG cells without significant cell death and also increased ABCA1, ABCG1 and apolipoprotein E (apoE) expression levels in THP-1 macrophages. PPAR antagonists did not diminish the induction of ABCA1 expression by honokiol in U251-MG cells. Cotreatment of the cells with honokiol and T0901317 (synthetic LXR ligand) further increased the ABCA1 expression level, whereas cotreatment with 9-cis retinoic acid had no additive effect compared with treatment with honokiol alone. We also found that honokiol has binding affinity to RXRbeta. In this study, we identified for the first time honokiol as an upregulator of ABCA1 expression, which is mediated by the binding of honokiol to RXRbeta as a ligand.

Topics: ATP Binding Cassette Transporter 1; ATP-Binding Cassette Transporters; Base Sequence; Biphenyl Compounds; Blotting, Western; Cell Line, Tumor; DNA Primers; Humans; In Situ Nick-End Labeling; Ligands; Lignans; Promoter Regions, Genetic; Retinoid X Receptor beta

Peroxisome proliferator-activated receptor-gamma (PPAR-gamma), COX-2 and 15-lipoxygenase (LOX)-1 have been shown to be involved in tumour growth. However, the roles of PPAR-gamma, COX-2 or 15-LOX-1 in gastric tumourigenesis remain unclear. Here, we investigate the role of 15-LOX-1 induction by honokiol, a small-molecular weight natural product, in PPAR-gamma and COX-2 signalling during gastric tumourigenesis.. Human gastric cancer cell lines (AGS, MKN45, N87 and SCM-1) were cultured with or without honokiol. Gene and protein expressions were analysed by RT-PCR and Western blotting respectively. Small interfering RNAs (siRNAs) for COX-2, PPAR-gamma and 15-LOX-1 were used to interfere with the expressions of these genes. A xenograft gastric tumour model in mouse was used for in vivo study.. PPAR-gamma and COX-2 proteins were highly expressed in gastric cancer cells. Inhibitors, or siRNA for COX-2 or PPAR-gamma, significantly decreased cell viability. Honokiol markedly inhibited PPAR-gamma and COX-2 expressions in gastric cancer cells and tumours of xenograft mice, and induced apoptosis and cell death. Honokiol markedly activated cellular 15-LOX-1 expression and 13-S-hydroxyoctadecadienoic acid (a primary product of 15-LOX-1 metabolism of linoleic acid) production. 15-LOX-1 siRNA could reverse the honokiol-induced down-regulation of PPAR-gamma and COX-2, and cell apoptosis. 15-LOX-1 was markedly induced in tumours of xenograft mice treated with honokiol.. These findings suggest that induction of 15-LOX-1-mediated down-regulation of a PPAR-gamma and COX-2 pathway by honokiol may be a promising therapeutic strategy for gastric cancer.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Arachidonate 15-Lipoxygenase; Biphenyl Compounds; Blotting, Western; Cell Line, Tumor; Cell Survival; Cyclooxygenase 2; Cyclooxygenase Inhibitors; Dose-Response Relationship, Drug; Enzyme Activation; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Humans; Lignans; Linoleic Acids; Male; Mice; Mice, Inbred BALB C; Mice, Nude; PPAR gamma; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; Signal Transduction; Stomach Neoplasms; Time Factors; Tumor Burden; Xenograft Model Antitumor Assays

In the course of a study on lignan profiles of tropical and subtropical members of the Magnoliaceae, Magnolia garrettii, an evergreen tree known from northern Thailand, Vietnam, and southern Yunnan (China), was investigated. The work resulted in the isolation of two dimeric lignans from the dichloromethane extract of the leaves of M. garrettii, garrettilignan A (1) and garrettilignan B (2), each substituted with two additional p-allylphenolic moieties. Garrettilignans A (1) and B (2) represent new skeletal types within the neolignan class. Additionally, four known neolignans, magnolol, honokiol, 4'-methylhonokiol, and obovatol, were identified.

Topics: Austria; Biphenyl Compounds; Lignans; Magnolia; Molecular Structure; Nuclear Magnetic Resonance, Biomolecular; Phenyl Ethers; Plant Leaves

To evaluate the interaction of fluconazole (FLC) and honokiol (HNK) in vitro and vivo against azole-resistant (azole-R) clinical isolates of Candida albicans.. A checkerboard microdilution method was used to study the in vitro interaction of FLC and HNK in 24 azole-R clinical isolates of C. albicans. In vivo antifungal activity was performed to further analyse the interaction between FLC and HNK. In the in vitro study, synergism was observed in all 24 FLC-resistant strains tested as determined by fractional inhibitory concentration index (FICI), and in 22 strains by Delta E models. No antagonistic activity was observed in any of the strains tested. These positive interactions were also confirmed by using the time-killing test for the selected strain C. albicans YL371, which shows strong susceptible to the combination of HNK and FLC. In the in vivo study, the mice with candidiasis were treated successfully by a combination therapy of HNK with FLC, the results showed a decrease of the colony forming unit in infected and treated animals compared to the controls, at the conditions of the treatment used in this study.. Synergistic activity of HNK and FLC against clinical isolates of FLC-resistant C. albicans was observed in vitro and in vivo.. This report might provide a potential therapeutic method to overcome the problem of drug-resistance in C. albicans.

Topics: Animals; Antifungal Agents; Biphenyl Compounds; Candida albicans; Candidiasis; Colony Count, Microbial; Drug Resistance, Fungal; Drug Synergism; Fluconazole; Humans; Lignans; Mice; Microbial Sensitivity Tests; Microbial Viability; Treatment Outcome

Screening of a total of 86 crude drugs for retinoid X receptor (RXR) ligands demonstrated that the methanol extract of the bark of Magnolia obovata markedly activated the transcriptional activity of RXRalpha in luciferase reporter assays. Thereafter, honokiol (1) was isolated as a constituent able to activate RXR selectively as a natural rexinoid, but not RARalpha. The activity of 1 was more potent than those of phytanic acid and docosahexaenoic acid, both of which are known to be natural RXR agonists. Honokiol (1) is capable of activating a RXR/LXR heterodimer, resulting in the induction of ATP-binding cassette transporter A1 mRNA and protein expression in RAW264.7 cells, as well as an increase in [(3)H]cholesterol efflux from peritoneal macrophages. These effects of 1 were enhanced synergistically in the presence of an LXR agonist, 22(R)-hydroxycholesterol. The results obtained demonstrate that 1, a newly identified natural rexinoid, regulates the functions of RXR/LXR heterodimer and abrogates foam cell formation by the induction of ABCA1 via activation of the RXR/LXR heterodimer.

Topics: Animals; ATP Binding Cassette Transporter 1; ATP-Binding Cassette Transporters; Bexarotene; Biphenyl Compounds; Cholesterol; Humans; Ligands; Lignans; Macrophages; Magnolia; Male; Mice; Mice, Inbred Strains; Molecular Structure; Plant Bark; Plants, Medicinal; Receptors, Retinoic Acid; Retinoic Acid Receptor alpha; Retinoid X Receptors; Tetrahydronaphthalenes

Excitotoxicity plays a major role in various neurological disorders. In this study, we explored the behavioral and neurotoxic effects of intraventricular NMDA administration in mice. After NMDA injection, acute seizures were followed by impairments in locomotor activity, motor performance on a rotarod, and climbing ability. Mice killed 1 day after NMDA administration showed increased synaptosomal reactive oxygen species ROS production and calcium concentration and decreased mitochondrial membrane potential, mitochondrial reductase activities, and neuronal membrane Na+, K+ -ATPase and mg2+ -ATPase activities. One and 3 days after excitotoxic injury, Golgi stains showed that dendritic length and spine density were significantly decreased in neurons of the hippocampal dentate gyrus. Some mice received honokiol, tea polyphenol plus memantine, and honokiol plus memantine prior to NMDA treatment; the occurrence of generalized seizures was attenuated, seizure scores were reduced, latency to generalized seizures was prolonged, and motor impairments were lessened. Moreover, all of the neurochemical changes of the synaptosomes were also ameliorated. In conclusion, the behavioral and neurotoxic effects of intracerebroventricular injection of NMDA were ameliorated by treatment with honokiol alone or combined treatment with either tea polyphenol plus memantine or honokiol plus memantine, but only partly by either tea polyphenol or memantine alone. The therapeutic potential of these neuroprotective regimens in treating excitotoxicity-related diseases merits for further investigation.

Topics: Analysis of Variance; Animals; Biphenyl Compounds; Ca(2+) Mg(2+)-ATPase; Calcium; Disease Models, Animal; Drug Interactions; Excitatory Amino Acid Agonists; Flavonoids; Lignans; Linear Models; Male; Membrane Potential, Mitochondrial; Mice; Motor Activity; N-Methylaspartate; Neurons; Neuroprotective Agents; Neurotoxicity Syndromes; Phenols; Phytotherapy; Polyphenols; Psychomotor Performance; Reactive Oxygen Species; Rotarod Performance Test; Silver Staining; Sodium-Potassium-Exchanging ATPase; Synaptosomes

Plant-derived compounds with potent anti-inflammatory activity have attracted a great deal of attention as a source for novel anti-arthritic agents with minimal side effects. We attempted to determine the anti-arthritic effects of orally administered honokiol isolated from Magnolia species. The oral administration of honokiol inhibited the progression and severity of type II collagen (CII)-induced arthritis (CIA) by reducing clinical arthritis scores and paw swelling. The histological analysis demonstrated preserved joint space; and the immunohistochemical data showed that the levels of interleukin (IL)-17, matrix metalloproteinase (MMP)-3, MMP-9, MMP-13, and receptor activator for nuclear factor-κB ligand, as well as nitrotyrosine formation, were substantially suppressed in the honokiol-treated CIA mice. The elevated serum levels of tumor necrosis factor-α and IL-1β in the CIA mice were also restored to control levels via honokiol treatment. In the CIA mice, honokiol inhibited CII- or lipopolysaccharide-stimulated cytokine secretion in spleen cells, as well as CII-stimulated spleen cell proliferation. Furthermore, honokiol treatment reduced CIA-induced oxidative damage in the liver and kidney tissues of CIA mice. Collectively, the oral administration of honokiol inhibited CIA development by reducing the production of pro-inflammatory cytokines, MMP expressions, and oxidative stress. Thus, honokiol is an attractive candidate for an anti-arthritic agent.

Topics: Animals; Arthritis, Experimental; Biphenyl Compounds; Cattle; Cytokines; Disease Progression; Inflammation; Inflammation Mediators; Lignans; Male; Matrix Metalloproteinase Inhibitors; Matrix Metalloproteinases; Mice; Mice, Inbred DBA; Oxidation-Reduction; Oxidative Stress

Recently, intermittent counter-current extraction (ICcE) has been developed and shown its advantage in improving resolution between targeted compounds. However, how to choose suitable parameters to increase the throughput has not been systematically studied yet. In present work, we first calculated theoretically the conditions to carry out ICcE elution mode. Then, honokiol and magnolol were separated as model compounds using ICcE elution mode to confirm our conclusion. After parameters like sample concentration and sample feed were optimized in analytical high-performance counter-current chromatography (HPCCC), the separation process was scaled up to preparative HPCCC successfully. 12.8 g honokiol and 16.1g magnolol were separated from 30 g mixture with purities of 98.6% and 93.7%. And the throughput of target isolation of ICcE elution mode was at least 3.75 x higher than isocratic elution mode with the same HPCCC instruments. Our results confirmed our theory calculation and demonstrated the enormous potential of ICcE on preparative separation of binary mixture.

Topics: Biphenyl Compounds; Chemical Fractionation; Countercurrent Distribution; Drugs, Chinese Herbal; Lignans; Models, Chemical

To develop newer and more effective chemopreventive agents for skin cancer, we assessed the effect of honokiol, a phytochemical from the Magnolia plant, on ultraviolet (UV) radiation-induced skin tumorigenesis using the SKH-1 hairless mouse model. Topical treatment of mice with honokiol in a hydrophilic cream-based topical formulation before or after UVB (180 mJ/cm(2)) irradiation resulted in a significant protection against photocarcinogenesis in terms of tumor multiplicity (28-60%, P < 0.05 to <0.001) and tumor volume per tumor-bearing mouse (33-80%, P < 0.05 to 0.001, n = 20). Honokiol also inhibited and delayed the malignant progression of papillomas to carcinomas. To investigate the in vivo molecular targets of honokiol efficacy, tumors and tumor-uninvolved skin samples from the tumor-bearing mice were analyzed for inflammatory mediators, cell cycle regulators and survival signals using immunostaining, western blotting and enzyme-linked immunosorbent assay. Treatment with honokiol significantly inhibited UVB-induced expression of cyclooxygenase-2, prostaglandin E(2) (P < 0.001), proliferating cell nuclear antigen and proinflammatory cytokines, such as tumor necrosis factor-α (P < 0.001), interleukin (IL)-1β (P < 0.01) and IL-6 (P < 0.001) in the skin as well as in skin tumors. Western blot analysis revealed that honokiol: (i) inhibited the levels of cyclins D1, D2 and E and associated cyclin-dependent kinases (CDKs)2, CDK4 and CDK6, (ii) upregulated Cip/p21 and Kip/p27 and (iii) inhibited the levels of phosphatidylinositol 3-kinase and the phosphorylation of Akt at Ser(473) in UVB-induced skin tumors. Together, our results indicate that honokiol holds promise for the prevention of UVB-induced skin cancer by targeting inflammatory mediators, cell cycle regulators and cell survival signals in UVB-exposed skin.

Topics: Administration, Topical; Animals; Anti-Infective Agents; Biphenyl Compounds; Blotting, Western; Cell Cycle; Cell Transformation, Neoplastic; Cyclin-Dependent Kinases; Cyclooxygenase 2; Drugs, Chinese Herbal; Enzyme-Linked Immunosorbent Assay; Female; Immunoenzyme Techniques; Inflammation Mediators; Lignans; Magnolia; Mice; Mice, Hairless; Papilloma; Phosphatidylinositol 3-Kinases; Phosphorylation; Proto-Oncogene Proteins c-akt; Signal Transduction; Skin Neoplasms; Ultraviolet Rays

Amyloid β peptide (Aβ) induced toxicity is a well-established pathway of neuronal cell death which might play a role in Alzheimer's disease. In this regard, the toxic effect of Aβ on a cultured Aβ-sensitive neuronal cell line was used as a primary screening tool for potential anti-Alzheimer's therapeutic agents. The effects of nine pure compounds (vitamin E, α-asarone, salidroside, baicolin, magnolol, gastrodin, bilobalide, honokiol and β-asarone) from selected Chinese herbs on neuronal cell death induced by Aβ in NGF-differentiated PC12 cells were examined. Only two of the studied compounds, honokiol and magnolol, significantly decreased Aβ-induced cell death. Further experiments indicated that their neuroprotective effects are possibly mediated through reduced ROS production as well as suppression of intracellular calcium elevation and inhibition of caspase-3 activity. The results provide for the first time a scientific rationale for the clinical use of honokiol and magnolol in the treatment of Alzheimer's disease.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Apoptosis; Biphenyl Compounds; Calcium; Caspase 3; Cell Survival; Drugs, Chinese Herbal; Lignans; Magnolia; Neuroprotective Agents; PC12 Cells; Rats; Reactive Oxygen Species

Chronic airway inflammation is a hallmark of asthma, an immune-based disease with great societal impact. Honokiol (HNK), a phenolic neurotransmitter receptor (γ-aminobutyric acid type A) agonist purified from magnolia, has anti-inflammatory properties, including stabilization of inflammation in experimentally induced arthritis. The present study tested the prediction that HNK could inhibit the chronic inflammatory component of allergic asthma. C57BL/6 mice sensitized to and challenged with OVA had increased airway hyperresponsiveness to methacholine challenge and eosinophilia compared with naive controls. HNK-treated mice showed a reduction in airway hyperresponsiveness as well as a significant decrease in lung eosinophilia. Histopathology studies revealed a marked drop in lung inflammation, goblet cell hyperplasia, and collagen deposition with HNK treatment. Ag recall responses from HNK-treated mice showed decreased proinflammatory cytokines in response to OVA, including TNF-α-, IL-6-, Th1-, and Th17-type cytokines, despite an increase in Th2-type cytokines. Regulatory cytokines IL-10 and TGF-β were also increased. Assessment of lung homogenates revealed a similar pattern of cytokines, with a noted increase in the number of FoxP3(+) cells in the lung. HNK was able to alter B and T lymphocyte cytokine secretion in a γ-aminobutyric acid type A-dependent manner. These results indicate that symptoms and pathology of asthma can be alleviated even in the presence of increased Th2 cytokines and that neurotransmitter agonists such as HNK have promise as a novel class of anti-inflammatory agents in the treatment of chronic asthma.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Biphenyl Compounds; Cytokines; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Eosinophilia; Female; Fluorescent Antibody Technique; Hypersensitivity; Lignans; Lung; Mice; Mice, Inbred C57BL; Th2 Cells

Honokiol, an active principle extracted from Magnolia officinalis, has great potential as a cancer treatment. However, its poor water solubility greatly hampers its delivery to the tumor sites at an effective concentration. In this study, an amphiphilic polymer-drug conjugate was successfully prepared by condensation of low molecular weight monomethoxy-poly(ethylene glycol) (MPEG)-2000 with honokiol (HK) through an ester linkage to increase the hydrophilicity of honokiol. The MPEG-honokiol (MPEG-HK) conjugate prepared formed nano-sized micelles, with a mean particle size of less than 20 nm (MPEG-HK, 360 µg ml(-1)) in water, which could be well dispersed in water. The nanoparticles obtained were characterized by particle size distribution, morphology and zeta potential. The stability and hydrolysis profile of the polymeric pro-drug in phosphate-buffered saline (PBS) and plasma were also studied and the results showed that only 20% of the conjugated honokiol was released in 2.0 h in beagle dog plasma, while in PBS the time required to reach 20% of honokiol release was >200 h. Meanwhile, the inhibitory activity of the honokiol conjugate was found to be retained in vitro against LL/2 cell lines with an IC50 value of 10.7 µg ml(-1). These results suggest that the polymer-drug conjugate provides a potential new approach to hydrophobic drugs, such as honokiol, in formulation design.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Colloids; Diffusion; Dogs; Drug Compounding; Lignans; Materials Testing; Micelles; Nanocapsules; Particle Size; Plasma; Polyethylene Glycols

In this paper, we prepared a novel cationic self-assembled micelle from poly(epsilon-caprolactone)-poly(ethyl glycol)-poly(epsilon-caprolactone) grafted polyethyleneimine (PCEC-g-PEI). The PCEC-g-PEI micelles, formed by self-assembly method, had mean particle size of ca. 82 nm and zeta potential of +22.5 mV at 37 degrees C, and could efficiently transfer pGFP into HEK293 cells in vitro. Meanwhile, as a model hydrophobic chemotherapeutic drug, honokiol was loaded into PCEC-g-PEI micelles by direct dissolution method assisted by ultrasonication. The honokiol loaded cationic PCEC-g-PEI micelles could effectively adsorb DNA onto its surface, while it could release honokiol in an extended period in vitro. This study demonstrated a novel DNA and hydrophobic chemotherapeutic drug co-delivery system.

Topics: Biphenyl Compounds; Cell Survival; DNA; Drug Delivery Systems; Gene Transfer Techniques; HEK293 Cells; Humans; Lignans; Micelles; Nanoconjugates; Particle Size; Polyesters; Polyethylene Glycols; Polyethyleneimine; Temperature

The combination chemotherapy is an important protocol in cancer therapy. Honokiol shows synergistic anticancer effect with doxorubicin. In this paper, honokiol and doxorubicin co-loaded MPEG-PLA nanoparticles were prepared. The particle size, morphology, in vitro release profile, cytotoxicity and cell proliferation study were studied in detail. The results indicated that honokiol and doxorubicin could be efficiently loaded into MPEG-PLA nanoparticles simultaneously, and could be released out in an extended period in vitro. Meanwhile, honokiol and doxorubicin in MPEG-PLA nanoparticle could efficiently suppress cancer cell proliferation in vitro. The described honokiol and doxorubicin co-loaded MPEG-PLA nanoparticle might be a novel anticancer agent.

Topics: Antineoplastic Agents; Biphenyl Compounds; Cell Line, Tumor; Doxorubicin; Humans; Lactic Acid; Lignans; Nanoparticles; Polyesters; Polyethylene Glycols; Polymers

Self-assembled polymeric micelles are widely applied in drug delivery system (DDS). In this study, honokiol (HK) loaded micelles were prepared from biodegradable poly(epsilon-caprolactone)-poly(ethylene glycol)-poly(epsilon-caprolactone) (PCEC) copolymers. Micelles were prepared by self-assembly of triblock copolymer PCEC in distilled water triggered by its amphiphilic character without any organic solvent. Drug loading and encapsulation efficiency were determined by adjusting the weight ratio of HK and PCEC. The particle size and zeta potential distribution of obtained micelles were determined using Malvern laser particle sizer, and spherical geometry were observed on atomic force microscope (AFM). Otherwise, the thermo-sensitivity of honokiol-loaded micelles was monitored. And the cytotoxicity results of drug loaded micelles showed that the encapsulated honokiol remained potent antitumor effect. Moreover, in vitro release profile demonstrated a significant difference between rapid release of free honokiol and much slower and sustained release of HK-loaded micelles. These results suggested that we have successfully prepared honokiol-loaded micelles in an improved method which is safer and more efficient. The prepared micelles might be potential carriers for honokiol delivery in cancer chemotherapy.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Line, Tumor; Delayed-Action Preparations; Drug Carriers; Drug Delivery Systems; Humans; Lasers; Lignans; Lung Neoplasms; Micelles; Microscopy, Atomic Force; Particle Size; Polyesters; Polyethylene Glycols; Temperature

Honokiol is a naturally occurring neolignan abundant in Magnoliae Cortex and has showed anti-proliferative and pro-apoptotic effects in a wide range of human cancer cells. However, the molecular mechanisms on the anti-proliferative activity in cancer cells have been poorly elucidated. In this study, we evaluated the growth inhibitory activity of honokiol in cultured estrogen receptor (ER)-negative MDA-MB-231 human breast cancer cells. Honokiol exerted anti-proliferative activity with the cell cycle arrest at the G0/G1 phase and sequential induction of apoptotic cell death in a concentration-dependent manner. The honokiol-induced cell cycle arrest was well correlated with the suppressive expression of CDK4, cyclin D1, CDK2, cyclin E, c-Myc, and phosphorylated retinoblastoma protein (pRb) at Ser780. Apoptosis caused by honokiol was also concomitant with the cleavage of caspases (caspase-3, -8, and -9) and Bid along with the suppressive expression of Bcl-2, but it was independent on the expression of Bax and p53. In addition, honokiol-treated cells exhibited the cleavage of poly (ADP-ribose) polymerase (PARP) and DNA fragmentation. In the analysis of signal transduction pathway, honokiol down-regulated the expression and phosphorylation of c-Src, epidermal growth factor receptor (EGFR), and Akt, and consequently led to the inactivation of mTOR and its downstream signal molecules including 4E-binding protein (4E-BP) and p70 S6 kinase. These findings suggest that honokiol-mediated inhibitory activity of cancer cell growth might be related with the cell cycle arrest and induction of apoptosis via modulating signal transduction pathways.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Breast Neoplasms; Cell Cycle; Cell Cycle Proteins; Cell Line, Tumor; CSK Tyrosine-Protein Kinase; Down-Regulation; ErbB Receptors; Female; Humans; Lignans; Phosphorylation; Protein-Tyrosine Kinases; Signal Transduction; src-Family Kinases; Tumor Suppressor Proteins

Esophageal adenocarcinoma (EAC) is a highly lethal cancer in western countries. EAC cells are believed to develop from esophageal epithelial cells through complex transformation processes involving inflammation and oxidative stress. The purpose of this study was to compare the redox status of malignant and nonmalignant esophageal epithelial cells and to test their responses to bile acid-induced oxidative stress and to treatment with honokiol (HNK), a natural product with anticancer activity. We demonstrated that esophageal adenocarcinoma cells express significantly higher levels of antioxidant molecules and were resistant to reactive oxygen species (ROS) stress induced by bile acid, but were sensitive to the cytotoxic action of HNK. Mechanistic study showed that HNK caused cancer cell death by disruption of mitochondrial transmembrane potential and was correlated with cyclophilin D (CypD) expression. Inhibition of CypD by cyclosporin A or abrogation of its expression by siRNA significantly suppressed the cytotoxicity of HNK, suggesting that CypD may be a key molecule that mediates the cytotoxicity. Our study suggests that the high antioxidant capacity in EAC cells confers on them the ability to survive the oxidative microenvironment in the reflux esophagus, and that HNK is a promising compound to kill the transformed cells preferentially.

Topics: Adenocarcinoma; Base Sequence; Bile Acids and Salts; Biphenyl Compounds; Cell Line, Transformed; Cyclophilins; Epithelial Cells; Esophageal Neoplasms; Esophagus; Humans; Lignans; Oxidation-Reduction; Peptidyl-Prolyl Isomerase F; RNA, Small Interfering

Most conventional methods for delivering chemotherapeutic agents fail to achieve therapeutic concentrations of drugs, despite reaching toxic systemic levels. Novel controlled drug delivery systems are designed to deliver drugs at predetermined rates for predefined periods at the target organ and overcome the shortcomings of conventional drug formulations therefore could diminish the side effects and improve the life quality of the patients. Thus, a suitable controlled drug delivery system is extremely important for chemotherapy.. A novel biodegradable thermosensitive composite hydrogel, based on poly(ethylene glycol)-poly(epsilon-caprolactone)-poly(ethylene glycol) (PEG-PCL-PEG, PECE) and Pluronic F127 copolymer, was successfully prepared in this work, which underwent thermosensitive sol-gel-sol transition. And it was flowing sol at ambient temperature but became non-flowing gel at body temperature. By varying the composition, sol-gel-sol transition and in vitro drug release behavior of the composite hydrogel could be adjusted. Cytotoxicity of the composite hydrogel was conducted by cell viability assay using human HEK293 cells. The 293 cell viability of composite hydrogel copolymers were yet higher than 71.4%, even when the input copolymers were 500 microg per well. Vitamin B12 (VB12), honokiol (HK), and bovine serum albumin (BSA) were used as model drugs to investigate the in vitro release behavior of hydrophilic small molecular drug, hydrophobic small molecular drug, and protein drug from the composite hydrogel respectively. All the above-mentioned drugs in this work could be released slowly from composite hydrogel in an extended period. Chemical composition of composite hydrogel, initial drug loading, and hydrogel concentration substantially affected the drug release behavior. The higher Pluronic F127 content, lower initial drug loading amount, or lower hydrogel concentration resulted in higher cumulative release rate.. The results showed that composite hydrogel prepared in this paper were biocompatible with low cell cytotoxicity, and the drugs in this work could be released slowly from composite hydrogel in an extended period, which suggested that the composite hydrogel might have great potential applications in biomedical fields.

Topics: Biocompatible Materials; Biphenyl Compounds; Cell Line; Cell Survival; Drug Delivery Systems; Humans; Hydrogels; Lignans; Poloxamer; Polyesters; Polyethylene Glycols; Temperature; Vitamin B 12

Lymph nodes metastasis of tumor could be a crucial early step in the metastatic process. Induction of tumor lymphangiogenesis by vascular endothelial growth factor-D may play an important role in promoting tumor metastasis to regional lymph nodes and these processes can be inhibited by inactivation of the VEGFR-3 signaling pathway. Honokiol has been reported to possess potent antiangiogenesis and antitumor properties in several cell lines and xenograft tumor models. However, its role in tumor-associated lymphangiogenesis and lymphatic metastasis remains unclear. Here, we established lymph node metastasis models by injecting overexpressing VEGF-D Lewis lung carcinoma cells into C57BL/6 mice to explore the effect of honokiol on tumor-associated lymphangiogenesis and related lymph node metastasis. The underlying mechanisms were systematically investigated in vitro and in vivo. In in vivo study, liposomal honokiol significantly inhibited the tumor-associated lymphangiogenesis and metastasis in Lewis lung carcinoma model. A remarkable delay of tumor growth and prolonged life span were also observed. In in vitro study, honokiol inhibited VEGF-D-induced survival, proliferation and tube-formation of both human umbilical vein endothelial cells (HUVECs) and lymphatic vascular endothelial cells (HLECs). Western blotting analysis showed that liposomal honokiol-inhibited Akt and MAPK phosphorylation in 2 endothelial cells, and downregulated expressions of VEGFR-2 of human vascular endothelial cells and VEGFR-3 of lymphatic endothelial cells. Thus, we identified for the first time that honokiol provided therapeutic benefit not only by direct effects on tumor cells and antiangiogenesis but also by inhibiting lymphangiogenesis and metastasis via the VEGFR-3 pathway. The present findings may be of importance to investigate the molecular mechanisms underlying the spread of cancer via the lymphatics and explore the therapeutical strategy of honokiol on antilymphangiogenesis and antimetastasis.

Topics: Angiogenesis Inhibitors; Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Carcinoma, Lewis Lung; Cell Line; Endothelial Cells; Female; Humans; Lignans; Liposomes; Lymphangiogenesis; Lymphatic Metastasis; Mice; Mice, Inbred C57BL; Vascular Endothelial Growth Factor D; Vascular Endothelial Growth Factor Receptor-3; Xenograft Model Antitumor Assays

To characterize antifungal principles from the methanol extract of Magnolia obovata and to evaluate their antifungal activities against various plant pathogenic fungi.. Four neolignans were isolated from stem bark of M. obovata as antifungal principles and identified as magnolol, honokiol, 4-methoxyhonokiol and obovatol. In mycelial growth inhibition assay, both magnolol and honokiol displayed more potent antifungal activity than 4-methoxyhonokiol and obovatol. Both magnolol and honokiol showed similar in vivo antifungal spectrum against seven plant diseases tested; both compounds effectively suppressed the development of rice blast, tomato late blight, wheat leaf rust and red pepper anthracnose. 4-Methoxyhonokiol and obovatol were highly active to only rice blast and wheat leaf rust respectively.. The extract of M. obovata and four neolignans had potent in vivo antifungal activities against plant pathogenic fungi.. Neolignans from Magnolia spp. can be used and suggested as a novel antifungal lead compound for the development of new fungicide and directly as a natural fungicide for the control of plant diseases such as rice blast and wheat leaf rust.

Topics: Allyl Compounds; Antifungal Agents; Biphenyl Compounds; Chromatography, Thin Layer; Fungi; Lignans; Magnolia; Microbial Sensitivity Tests; Mycelium; Phenyl Ethers; Plant Bark; Plant Diseases; Plant Extracts

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Humans; Lignans; Magnolia; Neoplasms; Plant Extracts

Honokiol, a bioactive component isolated from the Chinese herb Magnolia officinalis, is known for its potent antioxidative and anti-inflammatory effects. To study whether honokiol can protect skeletal muscle from sports injuries, we set up an eccentric exercise bout protocol for rats consisting of downhill running on a treadmill and examined the effect of oral administration of honokiol at 1 h before eccentric exercise at a dose of 5 mg/kg on day 1 (HK5 x 1) or 1 mg/kg/day for 5 consecutive days (HK1 x 5). Eccentric exercise was implemented for 3-5 consecutive days, and induced remarkable tissue damage. This damage was associated with an increase in serum creatine levels, increase in protein nitrotyrosylation, poly-ADP-ribose-polymerase (PARP) upregulation, lipid peroxidation, and leukocyte infiltration. The degree of muscle damage also paralleled dramatic gene expression for cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and inflammation-associated cytokines (interleukin (IL)-1beta, IL-6, tumor necrosis factor-alpha, and monocyte chemoattractant protein-1), possibly through activation of nuclear factor kappa-B (NF-kappaB), a crucial proinflammatory transcription factor. Both honokiol treatments (HK5 x 1 and HK1 x 5) significantly ameliorated eccentric exercise-induced muscle damage as revealed by suppression of cell fragmentation, protein nitrotyrosylation and PARP upregulation, as well as reductions in lipid peroxidation and leukocyte infiltration, possibly through downregulating gene expression for COX-2, iNOS, and the proinflammatory cytokines by modulation of NF-kappaB activation. In conclusion, the present study demonstrates for the first time that honokiol exhibits protective effects against eccentric exercise-induced skeletal muscle damage in rats, probably by modulating inflammation-mediated damage to muscle cells.

Topics: Animals; Biphenyl Compounds; Chemokine CCL2; Creatinine; Cyclooxygenase 2; Dose-Response Relationship, Drug; Gene Expression; Inflammation; Interleukin-1beta; Interleukin-6; Lignans; Lipid Peroxidation; Male; Muscle, Skeletal; Oxidative Stress; Physical Conditioning, Animal; Poly(ADP-ribose) Polymerases; Proteins; Rats; Rats, Wistar; Time Factors; Tumor Necrosis Factor-alpha; Up-Regulation

Ethanol induces hepatic steatosis via a complex mechanism that is not well understood. Among the variety of molecules that have been proposed to participate in this mechanism, the sterol regulatory element (SRE)-binding proteins (SREBPs) have been identified as attractive targets for therapeutic intervention. In the present study, we evaluated the effects of honokiol on alcoholic steatosis and investigated its possible effect on the inhibition of SREBP-1c maturation. In in vitro studies, H4IIEC3 rat hepatoma cells developed increased lipid droplets when exposed to ethanol, but co-treatment with honokiol reversed this effect. Honokiol inhibited the maturation of SREBP-1c and its translocation to the nucleus, the binding of nSREBP-1c to SRE or SRE-related sequences of its lipogenic target genes, and the expression of genes for fatty acid synthesis. In contrast, magnolol, a structural isomer of honokiol, had no effect on nSREBP-1c levels. Male Wistar rats fed with a standard Lieber-DeCarli ethanol diet for 4 weeks exhibited increased hepatic triglyceride and decreased hepatic glutathione levels, with concomitantly increased serum alanine aminotransferase and TNF-alpha levels. Daily administration of honokiol (10 mg/kg body weight) by gavage during the final 2 weeks of ethanol treatment completely reversed these effects on hepatotoxicity markers, including hepatic triglyceride, hepatic glutathione, and serum TNF-alpha, with efficacious abrogation of fat accumulation in the liver. Inhibition of SREBP-1c protein maturation and of the expression of Srebf1c and its target genes for hepatic lipogenesis were also observed in vivo. A chromatin immunoprecipitation assay demonstrated inhibition of specific binding of SREBP-1c to the Fas promoter by honokiol in vivo. These results demonstrate that honokiol has the potential to ameliorate alcoholic steatosis by blocking fatty acid synthesis regulated by SREBP-1c.

Topics: Active Transport, Cell Nucleus; Animals; Biphenyl Compounds; Cell Line, Tumor; Cytoprotection; Disease Models, Animal; Dose-Response Relationship, Drug; Ethanol; fas Receptor; Fatty Acids; Fatty Liver, Alcoholic; Genes, Reporter; Glutathione; Lignans; Lipogenesis; Liver; Male; Promoter Regions, Genetic; Protective Agents; Rats; Rats, Wistar; Sterol Regulatory Element Binding Protein 1; Transcription, Genetic; Transfection; Triglycerides; Tumor Necrosis Factor-alpha

In this article, poly(epsilon-caprolactone)-poly(ethylene glycol)-poly(epsilon-caprolactone) (PCL-PEG-PCL, PCEC) nanoparticles were successfully prepared for honokiol delivery in vitro. Blank or honokiol loaded PCL-PEG-PCL nanoparticles were prepared in moderate condition by solvent diffusion method without using any surfactants. The prepared blank PCL-PEG-PCL nanoparticles are mono-dispersed and smaller than 200 nm. The particle size increased with increase in polymer concentration and oil-water (O/W) ratio. The prepared PCL-PEG-PCL nanoparticles (40 mg/mL, ca. 106 nm) did not induce hemolysis in vitro. And the 50% inhibiting concentration (IC50) of it (48 h) on HEK293 cells was higher than 5 mg/mL. Honokiol could be efficiently loaded into PCL-PEG-PCL nanoparticles and released from these nanoparticles in an extended period in vitro. After honokiol (HK) was entrapped into PCL-PEG-PCL nanoparticles, the particle size increased with the increase in HK/PCEC mass ratio in feed, and the encapsulated honokiol retained potent anticancer activity in vitro. The PCL-PEG-PCL nanoparticle was suitable for honokiol delivery, and such honokiol loaded PCL-PEG-PCL nanoparticle was a novel honokiol formulation.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Line; Cell Line, Tumor; Drug Carriers; Drugs, Chinese Herbal; Erythrocytes; Female; Hemolysis; Humans; Lignans; Nanoparticles; Ovarian Neoplasms; Particle Size; Polyesters; Polyethylene Glycols; Rabbits; Solvents

It has been demonstrated that spray-drying is a powerful method to prepare dry powders for pulmonary delivery. This paper prepared dispersible dry powders based on chitosan and mannitol containing honokiol nanoparticles as model drug. The results showed that the prepared microparticles are almost spherical and have appropriate aerodynamic properties for pulmonary delivery (aerodynamic diameters was between 2.8-3.3 microm and tapped density ranging from 0.14-0. 18 g/cm(3)). Moreover, surface morphology and aerodynamic properties of the powders were strongly affected by the content of mannitol. Fourier transform infra-red (FTIR) spectrum of powders indicated that the honokiol nanoparticles were successfully incorporated into microparticles. In vitro drug release profile was also observed. The content of mannitol in powders significantly influenced the release rate of honokiol from matrices.

Topics: Administration, Inhalation; Administration, Intranasal; Biphenyl Compounds; Chemistry, Pharmaceutical; Chitosan; Drug Carriers; Drug Compounding; Drug Delivery Systems; Drug Stability; Excipients; Lignans; Lung; Mannitol; Nanoparticles; Powders; Respiratory Mucosa; Technology, Pharmaceutical

This study aims to develop a novel composite drug delivery system (CDDS) for hydrophobic honokiol delivery: honokiol loaded micelles in thermosensitive hydrogel (honokiol micelles/hydrogel) based on biodegradable poly(ethylene glycol)-poly(epsilon-caprolactone)-poly(ethylene glycol) (PEG-PCL-PEG, PECE) copolymers. In our work, we found that PECE copolymers with different molecular weight and PEG/PCL ratios could be administered to form micelles or thermosensitive hydrogel, respectively. Honokiol loaded PECE micelles (honokiol micelles) were prepared by self-assembly of biodegradable PECE copolymer (PEG5000-PCL5000-PEG5000) triggered by its amphiphilic characteristic assisted by ultrasonication without using any organic solvents and surfactants. Meanwhile, biodegradable and injectable thermosensitive PECE hydrogel (PEG550-PCL2400-PEG550) with a lower sol-gel transition temperature at around physiological temperature was also prepared successfully. Furthermore, the obtained honokiol micelles/hydrogel CDDS was a free-flowing sol at ambient temperature and became a nonflowing gel at body temperature. The cytotoxicity results showed that the CDDS was a safe carrier and the encapsulated honokiol retained its potent antitumor effect. In addition, the in vitro release profile demonstrated a significant difference between rapid release of free honokiol and much slower and sustained release of honokiol micelles/hydrogel. The results suggested that the CDDS might have great potential applications in cancer chemotherapy.

Topics: Animals; Biphenyl Compounds; Cell Line, Tumor; Cell Proliferation; Drug Carriers; Hydrogels; Lignans; Mice; Polyesters; Polyethylene Glycols; Temperature

PNAS-4, a novel pro-apoptotic gene activated during the early response to DNA damage, can inhibit proliferation via apoptosis when overexpressed in some tumor cells. Recent studies have indicated that honokiol can induce apoptosis, inhibit angiogenesis, and suppress tumor growth. In the present study, we investigated whether mouse PNAS-4 (mPNAS-4) could augment the apoptosis of tumor cells induced by honokiol in vitro, and whether the antiangiogenic activity of honokiol and induction of apoptosis by mPNAS-4 could work cooperatively to improve the antitumor efficacy in vivo. In vitro, mPNAS-4 inhibited proliferation of murine colorectal carcinoma CT26 and Lewis lung carcinoma LL2 cells through induction of apoptosis, and significantly augmented the apoptosis of CT26 and LL2 cells induced by honokiol. Compared with treatment with mPNAS-4 or honokiol alone, in vivo systemic administration of an expression plasmid encoding mPNAS-4 and low-dose honokiol significantly suppressed tumor growth through the enhanced induction of apoptosis and the augmented inhibition of angiogenesis. Our data suggest that the combined treatment with mPNAS-4 plus honokiol augments antitumor effects in vitro and in vivo, and that the improved antitumor activity in vivo may be associated with enhanced induction of apoptosis and augmented inhibition of angiogenesis. The present study may provide a novel and effective method for the treatment of cancer.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Apoptosis Regulatory Proteins; Biphenyl Compounds; Carcinoma, Lewis Lung; Cell Proliferation; Colonic Neoplasms; Combined Modality Therapy; Female; Genetic Therapy; Humans; In Situ Nick-End Labeling; Lignans; Liposomes; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Nitric Oxide Synthase; Tumor Cells, Cultured

A series of honokiol analogues were synthesized by modifying the 5- and/or 3'-position(s) of honokiol to assess their anti-tumor effects. Some compounds exerted more potent anti-proliferative activities than those of honokiol on K562 leukemia cells, A549 alveolar basal epithelial cells, SPC-A1 adenocarcinoma cells and A2780 human ovarian carcinoma cells in vitro. Compounds 2b, 3a, and 3c displayed most potent anti-proliferative activities against these tested cell strains and their anti-drug resistance effects were evaluated in vitro on cisplatin-resistant A2780 human ovarian carcinoma cells. The structure-activity relationship was also proposed.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Line, Tumor; Drug Resistance, Neoplasm; Drug Screening Assays, Antitumor; Humans; Lignans; Structure-Activity Relationship

The purpose of this study is to explore the feasibility and superiority of using rapid expansion of supercritical solution (RESS) technology in the field of traditional Chinese medicine. The extract of magnolia bark (EMB) was obtained by supercritical carbon dioxide (SCF-CO2) extraction technology. Microparticles of EMB were manufactured by RESS technology. The effects of operating temperature and pressure on the contents of the active ingredient in the particles were evaluated by HPLC. The effect of expansion conditions on the particle size distribution of EMB particles was investigated. The smallest sample (mean size: 4.7 microm) was obtained under the RESS condition: pressure of 25 MPa, temperature of 50 degrees C and a nozzle size of 100 microm. The characteristics of microparticles were also studied by differential scanning calorimetry (DSC), scanning electron microscopy (SEM) and image analysis. The dissolution rate study showed that microparticles had a significantly faster dissolution rate than normal material particles. After oral raw EMB suspension, the mean areas under the plasma concentration-time curves (AUC(0-t)) of honokiol and magnolol were found to be (4.23 +/- 0.36) and (5.46 +/- 0.57) mg x h x L(-1), respectively, which were increased significantly, i.e. (5.41 +/- 0.63) and (7.24 +/- 0.83) mg x h x L(-1) when micronized EMB suspension was administered orally in SD rats (P < 0.05). Similarly, the mean maximum plasma concentrations of honokiol and magnolol increased from (1.55 +/- 0.22) and (2.35 +/- 0.14) mg x L(-1) (raw EMB) to (2.31 +/- 0.17) and (2.84 +/- 0.21) mg x L(-1) (micronized EMB), respectively. The results of t-test demonstrated that AUC(0-t) and Cmax value for honokiol and magnolol was significantly increased with the micronization compared to raw EBM (P < 0.05). This study demonstrated that the RESS was applicable for preparing microparticles of EMB at low operating temperature. The process was simple, free of environment pollution and without residual solvent.

Topics: Administration, Oral; Animals; Area Under Curve; Biphenyl Compounds; Drug Compounding; Drugs, Chinese Herbal; Lignans; Magnolia; Male; Microspheres; Particle Size; Plant Bark; Plants, Medicinal; Rats; Rats, Sprague-Dawley; Solubility

Honokiol and magnolol, important anxiolytic and anti-cancer agents, have been produced in cell-suspension cultures of the endangered Mexican plant Magnolia dealbata Zucc. In vitro cultures of the plant were established, and the accumulation of honokiol and magnolol in callus and cell-suspension cultures was measured. Leaf samples were the best explants for callus establishment and metabolite production, and Murashige and Skoog medium supplemented with 1 mg/L 2, 4-dichlorophenoxyacetic acid and 1 mg/L kinetin yielded 2.3 mg/g of honokiol and 5.9 mg/g of magnolol. Bacterial and fungal contamination was inhibited with a multiple-step tissue sterilization procedure. Oxidation was inhibited with 1 g/L activated charcoal. Cell-suspension batch cultures derived from friable callus obtained from leaves of this species were grown for 30 days in shaker flasks containing Murashige and Skoog medium. Throughout the growth cycle, honokiol and magnolol levels, fresh and dry weight, and sucrose uptake were determined. The effects of carbon source concentration on biomass accumulation and the synthesis of bioactive compounds were studied. By using 3 mL of inocula supplemented with 3% (w/v) sucrose, maximum yields of honokiol (8.1 mg/g) and magnolol (13.4 mg/g) were obtained after 25 days. These yields were 300% and 382%, respectively, of the yields of honokiol and magnolol obtained from field-grown plants.

Topics: Anti-Anxiety Agents; Antineoplastic Agents, Phytogenic; Biomass; Biphenyl Compounds; Carbohydrates; Cells, Cultured; Chromatography, High Pressure Liquid; Kinetics; Lignans; Magnolia; Mexico

Magnolia bark combined with ginger rhizome is a common drug pair in traditional Chinese prescriptions for the treatment of depression. In the present study, we examined antidepressant-like effects of the mixture of honokiol and magnolol (HMM) from magnolia bark and essential oil from ginger rhizome (OGR) alone and in combination in chronic unpredictable mild stress (CUMS) of rats. Behavioral (sucrose intake, immobility time of forced swimming test) and biochemical parameters [serotonin (5-HT) in prefrontal cortex, hippocampus, and striatum, gastric mucosa cholecystokinin (CCK) and serum gastrin (GAS) levels] were simultaneously examined in the CUMS rats. 20 mg/kg HMM alone, but not OGR, significantly increased sucrose intake and reduced immobility time in the CUMS rats. Moreover, 20 mg/kg HMM and 14 mg/kg OGR in combination exhibited significant synergistic effects on sucrose intake increase and immobility time reduction in the CUMS rats. HMM elevated 5-HT levels in various brain regions, and OGR reduced gastric mucosa CCK and serum GAS levels in the CUMS rats. These results suggested that the synergistic antidepressant-like effects of compatibility of HMM with OGR might be mediated simultaneously by regulation of the serotonergic and gastroenteric system functions. These findings also provided a pharmacological basis for the clinical application of this drug pair of magnolia bark and ginger rhizome in traditional Chinese medicine.

Topics: Animals; Antidepressive Agents; Biphenyl Compounds; Brain; Cholecystokinin; Depression; Drug Synergism; Drug Therapy, Combination; Gastric Mucosa; Gastrins; Lignans; Male; Oils, Volatile; Rats; Rats, Wistar; Serotonin; Swimming; Zingiber officinale

The regulatory effects of honokiol on the cellular responses of macrophages and monocytes were evaluated. Specifically, we investigated the effects of honokiol with respect to lipopolysaccharide (LPS)-induced cytotoxicity, LPS- or phorbol-12-myristate-13-acetate (PMA)-mediated morphological changes, and relevant events (FITC-dextran-induced phagocytic uptake). Honokiol blocked the LPS-induced cytotoxicity of RAW264.7 cells in a dose-dependent manner. In addition, honokiol appeared to block the production of cytotoxic cytokines such as interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha, nitric oxide (NO), and reactive oxygen species (ROS). Moreover, honokiol strongly prevented the morphological changes in RAW 264.7 and U937 cells that were induced by LPS and PMA. The surface levels of marker proteins, which are up-regulated under the morphological changes of RAW264.7 and U937 cells, were also diminished. The data presented here strongly suggest that the honokiol modulates various cellular responses managed by macrophages and monocytes. [BMB reports 2009; 42(9): 574-579].

Topics: Anti-Allergic Agents; Biphenyl Compounds; Carcinogens; Cells, Cultured; Humans; Interleukin-1beta; Lignans; Lipopolysaccharides; Macrophages; Monocytes; Nitric Oxide; Nitric Oxide Synthase; Reactive Oxygen Species; Tetradecanoylphorbol Acetate; Tumor Necrosis Factor-alpha; U937 Cells

The antinociceptive effects of honokiol and magnolol, two major bioactive constituents of the bark of Magnolia officinalis, were investigated on animal paw licking responses and thermal hyperalgesia induced by glutamate receptor agonists including glutamate, N-methyl-D-aspartate (NMDA), and metabotropic glutamate 5 receptor (mGluR5) activator (RS)-2-chloro-5-hydroxyphenylglycine (CHPG), as well as inflammatory mediators such as substance P and prostaglandin E2 (PGE2) in mice. The actions of honokiol and magnolol on glutamate-induced c-Fos expression in the spinal cord dorsal horn were also examined. Our data showed that honokiol and magnolol blocked glutamate-, substance P- and PGE2-induced inflammatory pain with similar potency and efficacy. Consistently, honokiol and magnolol significantly decreased glutamate-induced c-Fos protein expression in superficial (I-II) laminae of the L4-L5 lumbar dorsal horn. However, honokiol was more selective than magnolol for inhibition of NMDA-induced licking behavioral and thermal hyperalgesia. In contrast, magnolol was more potent to block CHPG-mediated thermal hyperalgesia. These results demonstrate that honokiol and magnolol effectively decreased the inflammatory pain. Furthermore, their different potency on inhibition of nociception provoked by NMDA receptor and mGluR5 activation should be considered.

Topics: Analgesics; Animals; Anti-Infective Agents; Biphenyl Compounds; Dinoprostone; Excitatory Amino Acid Agents; Glycine; Immunohistochemistry; Inflammation; Lignans; Male; Mice; N-Methylaspartate; Phenylacetates; Proto-Oncogene Proteins c-fos; Receptor, Metabotropic Glutamate 5; Receptors, Metabotropic Glutamate

Honokiol (HK) can efficiently inhibit the growth of tumors. However, its clinical applications have been restricted by its extreme hydrophobicity. We hope to improve its water solubility by nanotechnology. And we wonder whether a novel honokiol nanoparticles-loaded thermosensitive poly(ethylene glycol)-poly(epsilon-caprolactone)-poly(ethylene glycol) (PEG-PCL-PEG, PECE) hydrogel (HK-hydrogel) could improve the therapeutic efficacy on malignant pleural effusion (MPE). To evaluate the therapeutic effects of HK-hydrogel on MPE, MPE-bearing mice were administered intrapleurally with HK-hydrogel, HK nanoparticles (HK-NP), blank hydrogel, or normal saline (NS) at days 4 and 11 after Lewis lung carcinoma (LLC) cells inoculation, respectively. Pleural tumor foci and survival time were observed, and antiangiogenesis of HK-hydrogel was determined by CD31. Histological analysis and assessment of apoptotic cells were also conducted in tumor tissues. HK-hydrogel reduced the number of pleural tumor foci, while prolonging the survival time of MPE-bearing mice, more effectively, as compared with control groups. In addition, HK-hydrogel successfully inhibited angiogenesis as assessed by CD31 (P < 0.05). Histological analysis of pleural tumors exhibited that HK-hydrogel led to the increased rate of apoptosis. This work is important for the further application of HK-hydrogel in the treatment of MPE.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Crystallization; Drug Carriers; Hydrogels; Lignans; Macromolecular Substances; Materials Testing; Mice; Mice, Inbred C57BL; Molecular Conformation; Nanomedicine; Nanoparticles; Particle Size; Pleural Effusion, Malignant; Radiography; Surface Properties; Temperature; Treatment Outcome

In this article, a novel composite drug delivery system, honokiol nanoparticles in biodegradable hydrogels based on chitosan (CS) and beta-glycerophosphate (beta-GP), was prepared. CS/beta-GP solution was liquid at room temperature and turned into gel as temperature increased. With increase in beta-GP concentration, the sol-gel transition temperature decreased accordingly. Honokiol nanoparticles with diameter of about 30 nm were prepared by emulsion solvent evaporation method. The sol-gel transition temperature of CS/beta-GP system decreased as F-127 presented in honokiol nanoparticles. In vitro release profiles were studied, the results showed that honokiol could be slowly released from CS/beta-GP gel over at least 2 weeks and the release rate was greatly influenced by initial drug loading. The described injectable hydrogels based on chitosan (CS) and beta-glycerophosphate (beta-GP) might have potential application as local drug delivery for honokiol.

Topics: Biphenyl Compounds; Chitosan; Chromatography, High Pressure Liquid; Drug Delivery Systems; Hydrogels; Lignans; Microscopy, Atomic Force; Nanoparticles; Temperature

Honokiol has been known to have antitumour activity. This study was conducted to evaluate the antiproliferative potential of honokiol against the hepG2 heptocellular cell line and its mechanism of action.. hepG2 cells were treated with honokiol of 0-40 microg/ml concentration. The cytotoxic effect of honokiol was determined by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The apoptosis was evaluated by flow cytometry. Western blots were used to analyse the expression of various proteins (procaspase-9, procaspase-3, cleaved caspase-3, cytochrome c, Bcl-2, Bax, Bad, Bcl-X(L) and p38).. Honokiol induced apoptosis with a decreased expression of procaspase-3 and -9 and an increased expression of active caspase-3. Exposure of hepG2 cells to honokiol resulted in the downregulation of Bcl-X(L) and Bcl-2 expression and the release of mitochondrial cytochrome c to the cytosol. In addition, honokiol activated the p38 mitogen-activated protein kinase (MAPK) pathway, and the inhibition of this pathway by SB203580 reduced honokiol-induced apoptosis and activation of caspase-3.. Honokiol induces apoptosis of hepG2 human hepatocellular carcinoma cells through activation of the p38 MAPK pathway, and, in turn, activation of caspase-3.

Topics: Apoptosis; Biphenyl Compounds; Blotting, Western; Carcinoma, Hepatocellular; Caspases; Cell Line, Tumor; Cell Proliferation; Flow Cytometry; Humans; Imidazoles; Lignans; p38 Mitogen-Activated Protein Kinases; Pyridines; Signal Transduction; Tetrazolium Salts; Thiazoles

Honokiol, a novel antitumor agent, could induce apoptosis and inhibit the growth of vascular endothelium in several tumor cell lines and xenograft models. It has been suggested that the antitumor effect of chemotherapy could be increased by combining it with an antiangiogenesis agent in anticancer strategy. The present study explored the potential to increase the antitumor effect of adriamycin by combining it with honokiol in mouse 4T1 breast cancer models, and the underlining mechanism was investigated. Honokiol was encapsulated in liposomes to improve the water insolubility. In vitro, liposomal honokiol inhibited the proliferation of 4T1 cells via apoptosis and significantly enhanced the apoptosis of 4T1 cells induced by adriamycin. In vivo, the systemic administration of liposomal honokiol and adriamycin significantly decreased tumor growth through increased tumor cell apoptosis compared with either treatment alone. Collectively, these findings suggest that liposomal honokiol may augment the induction of apoptosis in 4T1 cells in vitro and in vivo, and this combined treatment has shown synergistic suppression in tumor progression according to the analysis of isobologram. The present study may be important in future exploration of the potential application of the combined approach in the treatment of breast cancer.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Cell Survival; Dose-Response Relationship, Drug; Doxorubicin; Drug Combinations; Drug Screening Assays, Antitumor; Drug Synergism; Drugs, Chinese Herbal; Female; Lignans; Liposomes; Mice; Mice, Inbred BALB C; Neovascularization, Pathologic; Xenograft Model Antitumor Assays

Honokiol, an active component isolated and purified from Chinese traditional herb magnolia, was demonstrated to inhibit growth and induce apoptosis of different cancer cell lines such as human leukaemia, colon, and lung cancer cell lines; to attenuate the angiogenic activities of human endothelial cells in vitro; and to efficiently suppress the growth of angiosarcoma in nude mice. In this study, we have demonstrated that treatment of different human breast cancer cell lines with honokiol resulted in a time- and concentration-dependent growth inhibition in both estrogen receptor-positive and -negative breast cancer cell lines, as well as in drug-resistant breast cancer cell lines such as adriamycin-resistant and tamoxifen-resistant cell lines. The inhibition of growth was associated with a G1-phase cell cycle arrest and induction of caspase-dependent apoptosis. The effects of honokiol might be reversely related to the expression level of human epidermal growth receptor 2, (HER-2, also known as erbB2, c-erbB2) since knockdown of her-2 expression by siRNA significantly enhanced the sensitivity of the her-2 over-expressed BT-474 cells to the honokiol-induced apoptosis. Furthermore, inhibition of HER-2 signalling by specific human epidermal growth receptor 1/HER-2 (EGFR/HER-2) kinase inhibitor lapatinib synergistically enhanced the anti-cancer effects of honokiol in her-2 over-expressed breast cancer cells. Finally, we showed that honokiol was able to attenuate the PI3K/Akt/mTOR (Phosphoinositide 3-kinases/Akt/mammalian target of rapamycin) signalling by down-regulation of Akt phosphorylation and upregulation of PTEN (Phosphatase and Tensin homolog deleted on chromosome Ten) expression. Combination of honokiol with the mTOR inhibitor rapamycin presented synergistic effects on induction of apoptosis of breast cancer cells. In conclusion, honokiol, either alone or in combination with other therapeutics, could serve as a new, promising approach for breast cancer treatment.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Biphenyl Compounds; Breast Neoplasms; Cell Line, Tumor; Dose-Response Relationship, Drug; Drug Resistance, Neoplasm; Drug Screening Assays, Antitumor; Drug Synergism; Female; Gene Expression Regulation, Neoplastic; Humans; Lapatinib; Lignans; Magnolia; Medicine, Chinese Traditional; Quinazolines; Signal Transduction; Sirolimus; Time Factors

Elevated phospholipase D (PLD) activity provides a survival signal in several human cancer cell lines and suppresses apoptosis when cells are subjected to the stress of serum withdrawal. Thus, targeting PLD survival signals has potential to suppress survival in cancer cells that depend on PLD for survival. Honokiol is a compound that suppresses tumor growth in mouse models. The purpose of this study was to investigate the effect of honokiol on PLD survival signals and the Ras dependence of these signals.. The effect of honokiol upon PLD activity was examined in human cancer cell lines where PLD activity provides a survival signal. The dependence of PLD survival signals on Ras was investigated, as was the effect of honokiol on Ras activation.. We report here that honokiol suppresses PLD activity in human cancer cells where PLD has been shown to suppress apoptosis. PLD activity is commonly elevated in response to the stress of serum withdrawal, and, importantly, the stress-induced increase in PLD activity is selectively suppressed by honokiol. The stress-induced increase in PLD activity was accompanied by increased Ras activation, and the stress-induced increase in PLD activity in MDA-MB-231 breast cancer cells was dependent on a Ras. The PLD activity was also dependent on the GTPases RalA and ADP ribosylation factor. Importantly, honokiol suppressed Ras activation.. The data provided here indicate that honokiol may be a valuable therapeutic reagent for targeting a large number of human cancers that depend on Ras and PLD for their survival.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cell Survival; Enzyme Activation; Humans; Lignans; Mice; Models, Chemical; Neoplasm Transplantation; Neoplasms; Phospholipase D; ras Proteins; Signal Transduction

To study the phenols in Cortex of Magnolia officinalis of biernial seedings produced in Enshi, Hubei.. The content of magnolol and honokiol in cortexes were determined by HPLC. The chromatograms of 10 samples seedling cortexes were recorded and compared.. The content of magnolol and honokiol in Cortex of Magnolia officinalis of the seedlings from Enshi was higher than other samples. There were ten characteristic absorption bands in the HPLC chromatograms, which differed from the cortex of adult trees.. The results can be used to identify the quality of the seedlings for the breeding.

Topics: Age Factors; Biphenyl Compounds; China; Chromatography, High Pressure Liquid; Lignans; Magnolia; Plant Bark; Plant Stems; Plants, Medicinal; Seedlings

To study pharmacokinetics of honokiol in Cortex of Magnolia officinalis and its compound prescription Houpu Sanwu Decoction (HPSWD) in rat, and discuss the change on pharmacokinetic process affected by other components.. The rats were divided into two groups, one was supplied with HPSWD and the other was administrated with Cortex of Magnolia officinalis. Concentrations of honokiol in rat plasma were then determined using HPLC method and main pharmacokinetic parameters were estimated.. The plasma concentrations of honokiol of both groups were conformed to the two-compartment model with first order absorption. There existed significant differences in AUC, C(max), T(max), CL/F among Cortex of Magnolia officinalis and HPSWD.. The results indicate that Rhubarb and Immature Orange Fruit in HPSWD can influence the asorption, distribution and elimination of honokiol.

Topics: Administration, Oral; Animals; Area Under Curve; Biological Availability; Biphenyl Compounds; Chromatography, High Pressure Liquid; Drug Combinations; Drugs, Chinese Herbal; Lignans; Magnolia; Male; Rats; Rats, Wistar

Honokiol, a compound extracted from Magnolia officinalis, has antitumor and antiangiogenic properties in several tumor models in vivo. Among the downstream pathways inhibited by honokiol is nuclear factor kappa beta (NFkappabeta). A prime physiologic stimulus of NFkappabeta is reactive oxygen species. The chemical structure of honokiol suggests that it may be an effective scavenger of reactive oxygen species. In this work, we have studied the reactions of honokiol with superoxide and peroxyl radicals in cell-free and cellular systems using electron spin resonance (ESR) and high-performance liquid chromatography (HPLC) techniques. Honokiol efficiently scavenged superoxide radicals in xanthine oxidase and cytochrome P-450 cell-free systems with the rate constant 3.2x10(5)M(-1)s(-1), which is similar to reactivity of ascorbic acid but 20-times higher than reactivity of vitamin E analog trolox. Honokiol potently scavenged intracellular superoxide within melanoma cells. In addition, honokiol scavenged peroxyl radicals generated by 2,2'-azo-bis(2-amidinopropane hydrochloride) (AAPH). The rate constant of the reaction of honokiol with peroxyl radicals (1.4x10(6)M(-1)s(-1)) was calculated from the competition with spin trap 5-(ethoxycarbonyl)-5-methyl-1-pyrroline N-oxide (EMPO), and was found close to reactivity of trolox (2.5x10(6)M(-1)s(-1)). Therefore, honokiol is an effective scavenger of both superoxide and peroxyl radicals, which may be important for physiological activity of honokiol.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Line, Tumor; Chromatography, High Pressure Liquid; Electron Spin Resonance Spectroscopy; Free Radical Scavengers; Humans; Kinetics; Lignans; Peroxides; Spin Labels; Superoxides

Targeting death receptor-mediated apoptosis has emerged as an effective strategy for cancer therapy. However, certain types of cancer cells are intrinsically resistant to death receptor-mediated apoptosis. In an effort to identify agents that can sensitize cancer cells to death receptor-induced apoptosis, we have identified honokiol, a natural product with anticancer activity, as shown in various preclinical studies, as an effective sensitizer of death receptor-mediated apoptosis. Honokiol alone moderately inhibited the growth of human lung cancer cells; however, when combined with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), greater effects on decreasing cell survival and inducing apoptosis than TRAIL alone were observed, indicating that honokiol cooperates with TRAIL to enhance apoptosis. This was also true to Fas-induced apoptosis when combined with Fas ligand or an agonistic anti-Fas antibody. Among several apoptosis-associated proteins tested, cellular FLICE-inhibitory protein (c-FLIP) was the only one that was rapidly down-regulated by honokiol in all of the tested cell lines. The down-regulation of c-FLIP by honokiol could be prevented by the proteasome inhibitor MG132. Moreover, honokiol increased c-FLIP ubiquitination. These results indicate that honokiol down-regulates c-FLIP by facilitating its degradation through a ubiquitin/proteasome-mediated mechanism. Enforced expression of ectopic c-FLIP abolished the ability of honokiol to enhance TRAIL-induced apoptosis. Several honokiol derivatives, which exhibited more potent effects on down-regulation of c-FLIP than honokiol, showed better efficacy than honokiol in inhibiting the growth and enhancing TRAIL-induced apoptosis as well. Collectively, we conclude that c-FLIP down-regulation is a key event for honokiol to modulate the death receptor-induced apoptosis.

Topics: Apoptosis; Biphenyl Compounds; Carcinoma, Non-Small-Cell Lung; CASP8 and FADD-Like Apoptosis Regulating Protein; Cell Line, Tumor; Cell Proliferation; Cell Survival; Down-Regulation; Humans; JNK Mitogen-Activated Protein Kinases; Lignans; Lung Neoplasms; Proteasome Endopeptidase Complex; Protein Processing, Post-Translational; Receptors, Death Domain; TNF-Related Apoptosis-Inducing Ligand; Ubiquitin

Honokiol is a major bioactive compound extracted from Magnolia. The present study was designed to determine whether liposomal honokiol has the antitumor activity against human lung cancer as well as potentiates the antitumor activity of cisplatin in A549 lung cancer xenograft model, if so, to examine the possible mechanism in the phenomenon.. human A549 lung cancer-bearing nude mice were treated with liposomal honokiol, liposomal honokiol plus DDP or with control groups. Apoptotic cells and vessels were evaluated by fluorescent in situ TUNEL assay and by immunohistochemistry with an antibody reactive to CD31 respectively.. The present study showed that liposomal honokiol alone resulted in effective suppression of the tumor growth, and that the combined treatment with honokiol plus DDP had the enhanced inhibition of the tumor growth and resulted in a significant increase in life span. The more apparent apoptotic cells in the tumors treated with honokiol plus DDP was found in fluorescent in situ TUNEL assay, compared with the treatment with control groups. In addition, the combination of honokiol and DDP apparently reduced the number of vessels by immunolabeling of CD31 in the tissue sections, compared with control groups.. In summary, our data suggest that honokiol alone had the antitumor activity against human lung cancer in A549 lung cancer xenograft model, and that the combination of honokiol with DDP can enhance the antitumor activity, and that the enhanced antitumor efficacy in vivo may in part result from the increased induction of the apoptosis and the enhanced inhibition of angiogenesis in the combined treatment. The present findings may be of importance to the further exploration of the potential application of the honokiol alone or the combined approach in the treatment of lung carcinoma.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cisplatin; Humans; In Situ Nick-End Labeling; Lignans; Liposomes; Lung Neoplasms; Mice; Neoplasm Transplantation; Neovascularization, Pathologic; Plant Extracts; Platelet Endothelial Cell Adhesion Molecule-1

The effect of honokiol, an active component of Magnolia officinalis, on glutamate release from isolated nerve terminals (synaptosomes) was examined. Honokiol potently inhibited 4-aminopyridine (4-AP)-evoked glutamate release in a concentration-dependent manner, and this effect resulted from a reduction of vesicular exocytosis and not from an inhibition of Ca(2+)-independent efflux via glutamate transporter. The inhibitory action of honokiol was not due to decreasing synaptosomal excitability or directly interfering with the release process at some point subsequent to Ca(2+) influx, because honokiol did not alter the 4-AP-evoked depolarization of the synaptosomal plasma membrane potential or Ca(2+) ionophore ionomycin-induced glutamate release. Rather, examination of the effect of honokiol on cytosolic [Ca(2+)] revealed that the diminution of glutamate release could be attributed to a reduction in voltage-dependent Ca(2+) influx. Consistent with this, the honokiol-mediated inhibition of 4-AP-evoked glutamate release was completely prevented in synaptosomes pretreated with a wide-spectrum blocker of N-, P-, and Q-type Ca(2+) channels, omega-conotoxin MVIIC. In addition, honokiol modulation of 4-AP-evoked glutamate release appeared to involve a protein kinase C (PKC) signaling cascade, in so far as pretreatment of synaptosomes with the PKC inhibitors Ro318220 or GF109203X all effectively occluded the inhibitory effect of honokiol. Furthermore, honokiol attenuated 4-AP-induced phosphorylation of PKC. Together, these results suggest that honokiol effects a decrease in PKC activation, which subsequently attenuates the Ca(2+) entry through voltage-dependent N- and P/Q-type Ca(2+) channels to cause a decrease in evoked glutamate exocytosis. These actions of honokiol may contribute to its neuroprotective effect in excitotoxic injury.

Topics: Animals; Biphenyl Compounds; Cerebral Cortex; Dose-Response Relationship, Drug; Glutamic Acid; Lignans; Male; Neural Inhibition; Presynaptic Terminals; Rats; Rats, Sprague-Dawley; Synaptosomes

To assess the effects of honokiol on proliferation and apoptosis of human cervical carcinoma cell line Hela in vitro.. Cultured HeLa cells were treated with different concentrations of honokiol for the varieties of period (24, 48, 72, 96 h). Cell proliferation was assessed by MTT colorimetric assay. Cell apoptosis was determined by flow cytometry (FCM), Hoechst 33258 fluorescent staining and DNA ladder respectively.. MTT assay demonstrated that the proliferation of Hela cells were suppressed significantly by honokiol in dose-and time-dependent manner. FCM analysis showed that the apoptosis rates of Hela cells treated with 10 microg/mL and 20 microg/mL honokiol for 24 h were 22.5% and 62.2%, respectively, while that of the control group cells was 8.7%. After treatment with honokiol, typically morphologic changes of apoptosis were observed by Hoechst 33258 fluorescence staining; Genomic DNA from Hela cells treated with honokiol displayed a characteristic ladder pattern on agarose gel electrophoresis.. honokiol can inhibit the proliferation and induce apoptosis of human cervical carcinoma cell line Hela.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Proliferation; Cell Survival; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Flow Cytometry; HeLa Cells; Humans; Lignans; Time Factors

An HPLC-photodiode array (PDA) detection method was established for the simultaneous determination of 12 components in Xiao-Yao-San-Jia-Wei (XYSJW): geniposide, puerarin, paeoniflorin, ferulic acid, liquiritin, hesperidin, naringin, paeonol, daidzein, glycyrrhizic acid, honokiol, and magnolol. These were separated in less than 70 min using a Waters Symmetry Shield RP 18 column with gradient elution using (A) acetonitrile, (B) water, and (C) acetic acid at a flow rate of 1 ml/min, and with a PDA detector. All calibration curves showed good linear regression (r(2)>0.9992) within the test ranges. The method was validated for specificity, accuracy, precision, and limits of detection. The proposed method enables in a single run the simultaneous identification and determination for quality control of 12 multi-structural components of XYSJW forming the basis of its therapeutic effect.

Topics: Acetophenones; Benzoates; Biphenyl Compounds; Bridged-Ring Compounds; Coumaric Acids; Drugs, Chinese Herbal; Flavanones; Glucosides; Glycyrrhizic Acid; Hesperidin; Iridoids; Isoflavones; Lignans; Medicine, Chinese Traditional; Molecular Structure; Monoterpenes; Quality Control; Reference Standards; Reproducibility of Results; Sensitivity and Specificity

To reveal the relationship between the storage time of the bark of Magnolia officinalis and the content of phenols in it, and lay a theoretical foundation for the harvest, processing, management and storage.. The contents of magnolol and honokoiol in 15 bark samples, collected from the main producing areas in China, were determined in the time of freshly harvest and 3 and 10 years after respectively by HPLC method.. It showed that within a certain period of time, bark storage was favorable to conversion and accumulation of phenols, that the content of magnolol tended to increase from year 0 to year 3, then followed by slight decrease with years on account of volatilization of phenols, but was still higher when the bark was stored for 10 years than that that when the bark was freshly harvested, and the content of honokoiol still tended to increase when the bark had been stored for 10 years.. The phenols in bark of M. officinalis is quite stable and the bark can be stored for 10 years or longer.

Topics: Biphenyl Compounds; Drug Storage; Drugs, Chinese Herbal; Lignans; Magnolia; Plant Bark; Time Factors

Honokiol is an active compound purified from magnolia that has been shown to induce cell differentiation, apoptosis, and anti-angiogenesis effects, as well as an enhancement in tumor growth delay in combination with chemotherapeutic agents in several mouse xenograft models. Our goal was to investigate the radiosensitization effect of honokiol on lung carcinoma. The radiosensitization effect of liposomal honokiol in Lewis lung carcinoma cells (LL/2) was analyzed using an in vitro clonogenic survival assay. For an in vivo study, Lewis lung carcinoma-bearing C57BL/6 mice were treated with either liposomal honokiol at 25 mg/kg or 5 Gy of single tumor radiation, or a combination of both over 12 days of treatment. The tumor growth delay and the survival time were evaluated. In addition, histological analysis of tumor sections was performed to examine changes by detecting the microvessel density and apoptosis in tumor tissues. In the clonogenic survival assay, LL/2 cells treated with IC(50) Lipo-HNK for 24 h showed a radiation enhancement ratio of 1.9. After 12 days of combination treatment, the tumor volume decreased 78% and produced an anti-tumor activity 1.3-fold greater than a predicted additive effect of honokiol and radiation alone. This combination treatment also caused an 8.7 day delay in tumor growth. The cell cycle distribution and histological analysis demonstrated that liposomal honokiol has an anti-tumor effect via inducing apoptosis and inhibiting angiogenesis. Liposomal honokiol can enhance tumor cell radiosensitivity in vitro and in vivo, indicating that radiotherapy combined with liposomal honokiol can lead to greater anti-tumor efficacy.

Topics: Angiogenesis Inhibitors; Animals; Apoptosis; Biphenyl Compounds; Carcinoma, Lewis Lung; Cell Cycle; Cell Line, Tumor; Combined Modality Therapy; Humans; Lignans; Liposomes; Lung Neoplasms; Magnolia; Mice; Neoplasm Transplantation; Neovascularization, Pathologic; Radiation Tolerance; Transplantation, Heterologous

The Shb adapter protein is an Src homology 2-domain containing signaling intermediate operating downstream of several tyrosine kinase receptors, including vascular endothelial growth factor receptor-2. Shb is multifunctional and apoptosis is one response that Shb regulates. Inhibition of angiogenesis can be used in cancer therapy, and one way to achieve this is by inducing endothelial cell apoptosis. The angiosarcoma cell line SVR is of endothelial origin and can be used as a tool for studying in vivo inhibition of angiogenesis, and we thus employed an Shb-knockdown strategy using an inducible lentiviral system to reduce Shb levels in SVR cells and to study their responses. Shb knockdown increases the susceptibility of SVR cells to the apoptotic agents, cisplatin and staurosporine. Simultaneously, Shb knockdown causes reduced focal adhesion kinase (FAK) activation, monitored as phosphorylation of the regulatory residues tyrosines 576/577. No detectable effects on Akt or extracellular signal-regulated kinase activity were noted. The altered FAK activity coincided with an elongated cell phenotype that was particularly noticeable in the presence of staurosporine. In order to relate the effects of Shb knockdown to in vivo tumorigenicity, cells were exposed to the angiogenesis inhibitor honokiol, and again the cells with reduced Shb content exhibited increased apoptosis. Tumor growth in vivo was strongly reduced in the Shb-knockdown cells upon honokiol treatment. It is concluded that Shb regulates apoptosis and cell shape in tumor endothelial cells via FAK, and that Shb is a potential target for inhibition of angiogenesis.

Topics: Animals; Antineoplastic Agents; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Cisplatin; Endothelial Cells; Enzyme Inhibitors; Extracellular Signal-Regulated MAP Kinases; Focal Adhesion Protein-Tyrosine Kinases; Hemangiosarcoma; In Vitro Techniques; Lignans; Mice; Mice, Nude; Mutagenesis; Neoplasm Transplantation; Neovascularization, Pathologic; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-akt; Staurosporine

In our program to synthesize a series of novel derivatives as potential analogs of honokiol for anti-tumor treatment, we have found that at least three of the derivatives of honokiol showed more potency to inhibit the proliferation of K562 leukemia cells and SPC-A1 adenocarcinoma cells. As a critical step to our further series synthesis of derivatives of honokiol, three derivatives of honokiol composed of two isomers and one compound with two formyl groups, which were hardly separated by common purification methods, needed to be rapidly separated and purified. The present work describes analytical and preparative high-speed counter-current chromatography (HSCCC) for the isolation and purification of these three C-formylation derivatives of honokiol, named 3'-formylhonokiol, 5-formylhonokiol and 3',5-diformylhonokiol, respectively. The solvent system for HSCCC separation was composed of hexane-ethyl acetate-methanol-water with the ratio of 1:0.4:1:0.4 (v/v). The one-step purification produced 157.8 mg, 121.6 mg and 21.2 mg of 3'-formylhonokiol, 5-formylhonokiol, 3',5-diformylhonokiol from crude sample of 400mg with purities of 98.6%, 99.2% and 99.6%, respectively, in an elution time of 2.5 h. The purities and structural identification were determined by HPLC, (1)H NMR, (13)C NMR and mass spectroscopy. Their anti-proliferation effects on K562, A549 and SPC-A1 cell lines were evaluated by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Countercurrent Distribution; Lignans; Magnetic Resonance Spectroscopy; Mass Spectrometry

Honokiol and magnolol are the main constituents simultaneously identified in the barks of Magnolia officinalis, which have been used in traditional Chinese medicine to treat a variety of mental disorders including depression. In the present study, we reported on the antidepressant-like effects of oral administration of the mixture of honokiol and magnolol in well-validated models of depression in rodents: forced swimming test (FST), tail suspension test (TST) and chronic mild stress (CMS) model. The mixture of honokiol and magnolol significantly decreased immobility time in the mouse FST and TST, and reversed CMS-induced reduction in sucrose consumption to prevent anhedonia in rats. However, this mixture was unable to affect ambulatory or rearing behavior in the mouse open-field test. CMS induced alterations in 5-hydroxytryptamine (5-HT) and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) levels in various brain regions of rats. An increase in serum corticosterone concentrations and a reduction in platelet adenylyl cyclase (AC) activity were simultaneously found in the CMS rats. The mixture of honokiol and magnolol at 20 and 40 mg/kg significantly attenuated CMS-induced decreases of 5-HT levels in frontal cortex, hippocampus, striatum, hypothalamus and nucleus accumbens. And it markedly increased 5-HIAA levels in frontal cortex, striatum and nucleus accumbens at 40 mg/kg and in frontal cortex at 20 mg/kg in the CMS rats. A subsequent reduction in 5-HIAA/5-HT ratio was found in hippocampus and nucleus accumbens in the CMS rats receiving this mixture. Furthermore, the mixture of honokiol and magnolol reduced elevated corticosterone concentrations in serum to normalize the hypothalamic-pituitary-adrenal (HPA) hyperactivity in the CMS rats. It also reversed CMS-induced reduction in platelet AC activity, via upregulating the cyclic adenosine monophosphate (cAMP) pathway. These results suggested that the mixture of honokiol and magnolol possessed potent antidepressant-like properties in behaviors involved in normalization of biochemical abnormalities in brain 5-HT and 5-HIAA, serum corticosterone levels and platelet AC activity in the CMS rats. Our findings could provide a basis for examining directly the interaction of the serotonergic system, the HPA axis and AC-cAMP pathway underlying the link between depression and treatment with the mixture of honokiol and magnolol.

Topics: Animals; Antidepressive Agents; Behavior, Animal; Biphenyl Compounds; Corticosterone; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Therapy, Combination; Exploratory Behavior; Hindlimb Suspension; Hydroxyindoleacetic Acid; Lignans; Magnolia; Male; Mice; Mice, Inbred ICR; Phytotherapy; Rats; Rats, Wistar; Serotonin; Stress, Physiological; Swimming

Honokiol (HNK) is an active component purified from Magnolia officinalis. HNK exhibits antitumor effects by inducing apoptosis and inhibiting the growth of many cancer cell lines, while proteins involved in antitumor effects in proteomic level are still unclear. In our study, HNK could inhibit HeLa cell proliferation and induce apoptosis in a concentration- and time-dependent manner. We utilized a quantitative proteomic technique termed SILAC (Stable isotope labeling with amino acids in cell culture)-MS (mass spectrometry) to study the differential proteomic profiling of HeLa cells treated by HNK. A total of 85 proteins were changed after HeLa cells were treated with 12 microg/ml HNK for 8 h, and 8 proteins showed up-regulation while 77 proteins down-regulated. The changed proteins were classified into 9 different categories, which covered a broad variety of cellular functions. In conclusion, HNK performs cytotoxicity to HeLa cells through co-operating of many proteins and different pathways.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Proliferation; Cytotoxins; Dose-Response Relationship, Drug; Gene Expression Regulation, Neoplastic; HeLa Cells; Humans; Lignans; Magnolia; Neoplasm Proteins; Proteome; Proteomics; Time Factors

In this study, we investigated the regulatory effects of honokiol on various inflammatory events mediated by monocytes/macrophages (U937/RAW264.7 cells) and lymphocytes (splenic lymphocytes and CTLL-2 cells) and their putative action mechanism.. In order to investigate the regulatory effects, various cell lines and primary cells (U937, RAW264.7, CTLL-2 cells, and splenic lymphocytes) were employed and various inflammatory events, such as the production of inflammatory mediators, cell adhesion, cell proliferation, and the early signaling cascade, were chosen.. Honokiol strongly inhibited various inflammatory responses, such as: (i) the upregulation of nitric oxide (NO), prostaglandin E2 and TNF-alpha production and costimulatory molecule CD80 induced by lipopolysaccharide (LPS); (ii) the functional activation of beta1-integrin (CD29) assessed by U937 cell-cell and cell-fibronectin adhesions; (iii) the enhancement of lymphocytes and CD8+CTLL-2 cell proliferation stimulated by LPS, phytohemaglutinin A (PHA), and concanavalin A or interleukin (IL)-2; and (iv) the transcriptional upregulation of inducible NO synthase, TNF-alpha, cyclooxygenase-2, IL-12, and monocyte chemoattractant protein (MCP)-1. These anti-inflammatory effects of honokiol seem to be mediated by interrupting the early activated intracellular signaling molecule phosphoinositide 3-kinase (PI3K)/Akt, but not Src, the extracellular signal-regulated kinase, and p38, according to pharmacological, biochemical, and functional analyses.. These results suggest that honokiol may act as a potent anti-inflammatory agent with multipotential activities due to an inhibitory effect on the PI3K/Akt pathway.

Topics: Anti-Inflammatory Agents; Biphenyl Compounds; Cell Line; Cell Proliferation; Dinoprostone; Fibronectins; Humans; Lignans; Lipopolysaccharides; Lymphocytes; Mitogens; Nitric Oxide; Oncogene Protein v-akt; Phosphoinositide-3 Kinase Inhibitors; Signal Transduction; Tetrazolium Salts; Thiazoles; Tumor Necrosis Factor-alpha

In this paper, honokiol nanoparticles were prepared by emulsion solvent evaporation method. The prepared honokiol nanoparticles were characterized by particle size distribution, morphology, zeta potential and crystallography. Results showed that the obtained honokiol nanoparticles at size of 33 nm might be amorphous, and could be well dispersed in water. Due to the great dispersibility in water, the obtained honokiol nanoparticles might have great potential in medical field.

Topics: Biphenyl Compounds; Chemistry, Pharmaceutical; Crystallization; Drugs, Chinese Herbal; Emulsions; Lignans; Macromolecular Substances; Materials Testing; Molecular Conformation; Nanomedicine; Nanostructures; Particle Size; Solvents

This study was undertaken to determine the efficacy of honokiol, a constituent of oriental medicinal herb Magnolia officinalis, against human prostate cancer cells in culture and in vivo.. Honokiol-mediated apoptosis was assessed by analysis of cytoplasmic histone-associated DNA fragmentation. Knockdown of Bax and Bak proteins was achieved by transient transfection using siRNA. Honokiol was administered by oral gavage to male nude mice s.c. implanted with PC-3 cells. Tumor sections from control and honokiol-treated mice were examined for apoptotic bodies (terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling assay), proliferation index (proliferating cell nuclear antigen staining), and neovascularization (CD31 staining). Levels of Bcl-2 family proteins in cell lysates and tumor supernatants were determined by immunoblotting.. Exposure of human prostate cancer cells (PC-3, LNCaP, and C4-2) to honokiol resulted in apoptotic DNA fragmentation in a concentration- and time-dependent manner irrespective of their androgen responsiveness or p53 status. Honokiol-induced apoptosis correlated with induction of Bax, Bak, and Bad and a decrease in Bcl-xL and Mcl-1 protein levels. Transient transfection of PC-3 cells with Bak- and Bax-targeted siRNAs and Bcl-xL plasmid conferred partial yet significant protection against honokiol-induced apoptosis. Oral gavage of 2 mg honokiol/mouse (thrice a week) significantly retarded growth of PC-3 xenografts without causing weight loss. Tumors from honokiol-treated mice exhibited markedly higher count of apoptotic bodies and reduced proliferation index and neovascularization compared with control tumors.. Our data suggest that honokiol, which is used in traditional oriental medicine for the treatment of various ailments, may be an attractive agent for treatment and/or prevention of human prostate cancers.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; bcl-2 Homologous Antagonist-Killer Protein; bcl-2-Associated X Protein; bcl-Associated Death Protein; bcl-X Protein; Biphenyl Compounds; Cell Line, Tumor; Humans; Immunoblotting; Lignans; Magnolia; Male; Mice; Mice, Nude; Myeloid Cell Leukemia Sequence 1 Protein; Neoplasm Proteins; Prostatic Neoplasms; Proto-Oncogene Proteins c-bcl-2

Honokiol has been receiving attention as an anticancer agent because of its anti-tumor effect. In the current study, we encapsulated honokiol with liposome and tested it on cisplatin-sensitive (A2780s) and -resistant (A2780cp) human ovarian cancer models.. The anti-tumor activity of liposomal honokiol (Lipo-HNK) was evaluated in nude mice bearing A2780s and A2780cp s.c. tumors. Mice were treated twice weekly with i.v. administration of Lipo-HNK (10 mg/kg), control liposome (10 mg/kg), 0.9% NaCl solution or weekly with intraperitoneally administered cisplatin (5 mg/kg) for 3 weeks. Tumor volume and survival time were observed. Assessment of apoptotic cells by TUNEL assay was conducted in tumor tissue. Microvessel density within tumor tissue was determined by CD34 immunohistochemistry. For in vitro study, induction of apoptosis by Lipo-HNK was examined by PI staining fluorescence microscopy, DNA fragmentation assay and flow cytometric analysis.. Administration of Lipo-HNK resulted in significant inhibition (84-88% maximum inhibition relative to controls) in the growth of A2780s and A2780cp tumor xenografts and prolonged the survival of the treated mice. These anti-tumor responses were associated with marked increases in tumor apoptosis, and reductions in intratumoral microvessel density.. The present findings suggest that Lipo-HNK may provide an effective approach to inhibit tumor growth in both cisplatin sensitive and -resistant human ovarian cancer with minimal side effects.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Cell Line, Tumor; Cisplatin; Drug Resistance, Neoplasm; Female; Humans; Immunohistochemistry; Lignans; Liposomes; Mice; Mice, Nude; Ovarian Neoplasms

A rapid, sensitive and specific liquid chromatography-electrospray ionization/tandem mass spectrometry (LC-ESI/MS/MS) method has been developed and validated for simultaneous determination of five active constituents (including magnolol, honokiol, rhein, emodin and aloe-emodin) from Da-Cheng-Qi decoction (DCQD) in rat plasma. After the addition of gliquidone as the internal standard (IS), plasma samples were prepared by one-step protein precipitation using methanol and separated by HPLC on a short reversed phase C(18) column packed with smaller particles (100 mm x 3.0 mm, 3.5 microm) using a mobile phase of methanol-0.1% formic acid aqueous solution (70:30, v/v). Analytes were determined in a triple-quadrupole mass spectrometer in the selected reaction-monitoring (SRM) mode using electrospray source with negative mode. The method was proved to be rapid, sensitive, specific, accurate and reproducible and has been successfully applied to the determination of the five compounds in rat plasma after oral administration of low dose DCQD for pharmacokinetic study.

Topics: Administration, Oral; Animals; Anthraquinones; Biphenyl Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Emodin; Female; Lignans; Male; Plant Extracts; Rats; Rats, Sprague-Dawley; Tandem Mass Spectrometry

To study the effects of xanthohumol (XN), a flavonoid found in hops (Humulus lupulus) and honokiol (HK), a lignan isolated from Magnolia officinalis, alone and in combination, on apoptotic signaling in 3T3-L1 adipocytes.. 3T3-L1 mature adipocytes were incubated with various concentrations of XN and HK alone and in combination. Viability and apoptosis were quantified using an MTS-based cell viability assay and single-stranded DNA assay, respectively. Expression of apoptosis related proteins including cleaved poly(ADP-ribose) polymerase (PARP), cytochrome c, Bcl-2, caspase-3/7, phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and Akt was analyzed by western blotting.. Combinations of XN and HK significantly decreased viability and induced apoptosis in a dose-dependent manner and more than the additive responses to XN and HK alone. Western blot analysis showed an increase in cleaved PARP and cytochrome c release and decrease in expression of Bcl-2 protein by XN plus HK, whereas XN and HK individually had no effect. Furthermore, the combination of XN and HK activated PTEN and inactivated Akt by decreasing levels of phosphorylated PTEN and phosphorylated Akt.. We demonstrated that although XN and HK showed little or no effect as individual compounds, in combination (XN plus HK) they showed enhanced activity in inducing apoptosis via the cytochrome c/caspase-3/PARP and PTEN/Akt pathways in 3T3-L1 adipocytes.

Topics: 3T3-L1 Cells; Adipocytes; Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Caspase 3; Cell Survival; Cytochromes c; Dose-Response Relationship, Drug; Drug Synergism; Flavonoids; Lignans; Mice; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerases; Propiophenones; Proto-Oncogene Proteins c-akt; PTEN Phosphohydrolase; Signal Transduction

Angiopathy is a major complication of diabetes. Abnormally high blood glucose is a crucial risk factor for endothelial cell damage. Nuclear factor-kappaB (NF-kappaB) has been demonstrated as a mediated signaling in hyperglycemia or oxidative stress-triggered apoptosis of endothelial cells. Here we explored the efficacy of honokiol, a small molecular weight natural product, on NADPH oxidase-related oxidative stress-mediated NF-kappaB-regulated signaling and apoptosis in human umbilical vein endothelial cells (HUVECs) under hyperglycemic conditions. The methods of morphological Hoechst staining and annexin V/propidium iodide staining were used to detect apoptosis. Submicromolar concentrations of honokiol suppressed the increases of NADPH oxidase activity, Rac-1 phosphorylation, p22(phox) protein expression, and reactive oxygen species production in high glucose (HG)-stimulated HUVECs. The degradation of IkappaBalpha and increase of NF-kappaB activity were inhibited by honokiol in HG-treated HUVECs. Moreover, honokiol (0.125-1 microM) also suppressed HG-induced cyclooxygenase (COX)-2 upregulation and prostaglandin E(2) production in HUVECs. Honokiol could reduce increased caspase-3 activity and the subsequent apoptosis and cell death triggered by HG. These results imply that inhibition of NADPH oxidase-related oxidative stress by honokiol suppresses the HG-induced NF-kappaB-regulated COX-2 upregulation, apoptosis, and cell death in HUVECs, which has the potential to be developed as a therapeutic agent to prevent hyperglycemia-induced endothelial damage.

Topics: Apoptosis; Biphenyl Compounds; Cyclooxygenase 2; Endothelial Cells; Endothelium, Vascular; Enzyme Activation; Glucose; Humans; Lignans; NADPH Oxidases; NF-kappa B; Oxidative Stress; Phosphorylation; rac1 GTP-Binding Protein; Reactive Oxygen Species; Signal Transduction; Umbilical Veins

To observe the anti-tumor activities of honokiol on human ovarian tumor in vitro and in vivo.. Cells were treated with honokiol, and the effects on proliferation and apoptosis were examined by MTT, DNA ladder, Hoechst staining, and flow cytometry assays. Expression of Bcl-2 members and caspase-3 were assessed. Measurements of tumor volume and microvessel densities (MVDs) were performed.. Honokiol significantly inhibited proliferation and induced apoptosis, with alteration of Bcl-2 members and caspase-3. Administration of honokiol to tumor-bearing animals decreased MVD and resulted in inhibition of tumor growth.. Honokiol could induce apoptosis and inhibit angiogenesis in vitro and in vivo, suggesting a novel and attractive therapeutic candidate for ovarian tumor treatment.

Topics: Angiogenesis Inhibitors; Animals; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Disease Models, Animal; Female; Humans; Lignans; Mice; Mice, Nude; Neovascularization, Pathologic; Ovarian Neoplasms

In this article, a novel local hydrophobic drug delivery system: nanoparticles in thermo-sensitive hydrogel, was demonstrated. First, honokiol, as a model hydrophobic drug, loaded poly(epsilon-caprolactone)-poly(ethylene glycol)-poly(epsilon-caprolactone) (PCEC) nanoparticles were prepared by emulsion solvent evaporation method, and then were incorporated into thermo-sensitive F127 hydrous matrix. The obtained injectable hydrophobic drug delivery system can act as a depot for sustained release of honokiol in situ. The lower critical solution temperature (LCST) of the composite matrix increases with increase in the mass of incorporated nanoparticles, or with decrease in the amount of residual organic solvent in the system. Honokiol release profile in vitro was studied, and the results showed that honokiol could be sustained released from the system. The described injectable drug delivery system might have great potential application for local delivery of hydrophobic drugs such as honokiol.

Topics: Animals; Biphenyl Compounds; Delayed-Action Preparations; Drug Delivery Systems; Emulsions; Excipients; Hydrogels; Hydrophobic and Hydrophilic Interactions; Injections, Subcutaneous; Lignans; Mice; Nanoparticles; Poloxamer; Polyesters; Polyethylene Glycols; Solvents; Temperature

Magnolia officinalis is a commonly used traditional Chinese medicine for treating gastrointestinal disorders. HPLC quantification analysis revealed that magnolol and honokiol were the most abundant constituents of M. officinalis extracts, with their contents in the ethanol extract being the highest, the water extract the least and the 50 % ethanol extract in between. In guinea pig isolated ileum, both magnolol and honokiol inhibited contraction to acetylcholine. The herbal extracts also produced inhibitory responses, in an order of decreasing efficacy: ethanol extract > 50 % ethanol extract > water extract. The differences in inhibitory efficacies among the three extracts were similar to the differences in their magnolol and honokiol contents. Further examination demonstrated that two mixtures containing solely magnolol and honokiol at concentrations identical to those determined in the ethanol and water extracts exhibited similar levels of anti-spasmodic effects as their respective extracts while a "blank" ethanol extract free of magnolol and honokiol failed to produce any response. These observations suggest that the magnolol and honokiol contents account for the anti-spasmodic effects of M. officinalis extracts in guinea pig isolated ileum.

Topics: Animals; Biphenyl Compounds; Guinea Pigs; Ileum; Lignans; Magnolia; Molecular Structure; Muscle Contraction; Muscle, Smooth; Plant Extracts

Honokiol is a bioactive compound extracted from the Chinese medicinal herb Magnolia officinalis. We recently demonstrated that honokiol inhibited arterial thrombosis through stimulation of prostacyclin (PGI2) generation and endothelial cell protection. The current study is designed to investigate its mechanism of stimulation of PGI2 generation and cell protection. 6-keto-PGF1alpha, the stable metabolite of PGI2, in the media of rat aortic endothelial cells was measured with radioimmunoassay kits. Indomethacin, an inhibitor of cyclooxygenase (COX) and tranylcypromine, a prostacyclin synthease inhibitor were used to ascertain the target enzyme affected by honokiol. Prostacyclin synthease protein levels in endothelial cells were determined by Western blot analysis using an anti-PGI2 synthease rabbit polyclonal antibody. Flow cytometry was used to quantify the apoptotic cells and spectrophotometry was used to test the caspase-3 activity. Honokiol (0.376-37.6 microM) increased the level of 6-keto-PGF1alpha in the media of normal endothelial cells. It counteracted the inhibitory effect of tranylcypromine on the PGI2 generation, but did not influence the effect of indomethacin; evidently, honokiol up-regulated the expression of prostacyclin synthease in the endothelial cells. These effects showed perfect concentration-dependent behavior. In addition, at lower concentration (0.376-3.76 microM), honokiol significantly decreased the percentage of apoptotic endothelial cells induced by oxidized low-density lipoprotein (ox-LDL) and significantly lowered the activity of caspase-3 stimulated by ox-LDL. A high dose of honokiol (37.6 microM), however, failed to influence either of them. In conclusion, honokiol augments PGI2 generation in normal endothelial cells; its effect on PGI2 generation attributes to up-regulation of prostacyclin synthease expression; its cell protection may be correlated with its inhibition on apoptosis of endothelial cells. These findings have partly revealed the molecular mechanism of honokiol on inhibiting arterial thrombosis.

Topics: Animals; Apoptosis; Biphenyl Compounds; Caspase 3; Cells, Cultured; Cyclooxygenase Inhibitors; Cytochrome P-450 Enzyme System; Drugs, Chinese Herbal; Endothelial Cells; Epoprostenol; Fibrinolytic Agents; Gene Expression Regulation, Enzymologic; Indomethacin; Intramolecular Oxidoreductases; Lignans; Male; Rats; Rats, Sprague-Dawley; Tranylcypromine; Up-Regulation

In this paper, a rapid separation approach has been developed using high-capacity high-speed counter-current chromatography (high-capacity HSCCC) to isolate and purify honokiol and magnolol, which are the main bioactive constituents from Houpu. The optimization of the solvent selection process, sample loading volume and flow rate is systematically studied using analytical high-capacity HSCCC. The optimized parameters obtained rapidly at analytical scale were used for a 1000 x scale-up preparative run using pilot scale high-capacity HSCCC in a MAXI-DE centrifuge. A crude sample of 43 g was successfully separated and the fractions were analysed by high-performance liquid chromatography (HPLC). This large scale preparative single step run yielded 16.9 and 19.4 g of honokiol and magnolol with purities of 98.6 and 99.9%, in only 20 min. This is the first time that high-performance counter-current chromatography has been used to purify multiple gram grade bioactive compounds in less than 1h and at such high concentrations of final products (10.8 g/l for magnolol and 7.0 g/l for honokiol).

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Countercurrent Distribution; Lignans; Pilot Projects

Capillary zone electrophoresis (CZE) method was used for analysis of magnolol and honokiol. Under the optimized condition, CZE with UV absorption detection provided that the limit of detection was at microM level. To enhance detection sensitivity of magnolol and honokiol, CZE separation system was coupled with a laser-induced fluorescence (LIF) detector for the first time. The limits of detection of magnolol and honokiol were 12 nM (3.20 ng ml(-1)) and 18 nM (4.79 ng ml(-1)), respectively, showing that the CZE-LIF system provides greater than 100-fold sensitivity improvements than does the CZE-UV system. The developed method was applied to analyze magnolol and honokiol in spiked human plasma samples, microsome incubation samples as a preliminary demonstration of its potential in pharmacokinetic studies.

Topics: Animals; Biphenyl Compounds; Electrophoresis, Capillary; Fluorescence; Humans; Lignans; Microsomes, Liver; Rats; Reference Standards; Sensitivity and Specificity; Spectrophotometry, Ultraviolet

Honokiol, a soluble nontoxic natural product derived from Magnolia spp., has been shown to induce apoptosis in malignant cells. The effect of honokiol and the combined therapy with docetaxel on prostate cancer (PCa) growth and bone metastasis was investigated in experimental models.. The in vitro proapoptotic effects of honokiol on human androgen-dependent and -independent PCa, bone marrow, bone marrow-derived endothelial, and prostate stroma cells were investigated. Honokiol-induced activation of caspases was evaluated by Western blot and FACS analysis. To confirm the cytotoxicity of honokiol, mice bone was inoculated in vivo with androgen-independent PCa, C4-2 cells and the effects of honokiol and/or docetaxel on PCa growth in bone were evaluated. Daily honokiol (100 mg/kg) and/or weekly docetaxel (5 mg/kg) were injected intraperitoneally for 6 weeks. PCa growth in mouse bone was evaluated by radiography, serum prostate-specific antigen (PSA) and tissue immunohistochemistry.. Honokiol induced apoptosis in all cell lines tested. In PCa cells honokiol induced apoptosis via the activation of caspases 3, 8, and 9 and the cleavage of poly-adenosine diphosphate ribose polymerase in a dose- and time-dependent manner. Honokiol was shown to inhibit the growth and depress serum PSA in mice harboring C4-2 xenografts in the skeleton and the combination with docetaxel showed additive effects that inhibited further growth without evidence of systemic toxicity. Immunohistochemical staining confirmed honokiol exhibited growth-inhibitory, apoptotic, and antiangiogenic effects on PCa xenografts.. The combination of honokiol and low-dose docetaxel may be used to improve patient outcome in androgen-independent prostate cancer with bone metastasis.

Topics: Animals; Antineoplastic Agents, Phytogenic; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Biphenyl Compounds; Blotting, Western; Bone Marrow Cells; Bone Neoplasms; Caspases; Cell Proliferation; Docetaxel; Drugs, Chinese Herbal; Flow Cytometry; Humans; Immunoenzyme Techniques; Lignans; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Prostate-Specific Antigen; Prostatic Neoplasms; Radiography; Taxoids; Tumor Cells, Cultured

Neuroprotective effect of honokiol (HK), orally administered, on oxidative damage in the brain of mice challenged with N-methyl-d-aspartic acid (NMDA) was examined. HK (1-100 mg/kg) was administered to Institute of Cancer Research (ICR) male mice through a gavage for 3 days consecutively, and on the third day, NMDA (150 mg/kg) was intraperitoneally (i.p.) administered. Administration of NMDA, causing a lethality of approximately 60%, resulted in a significant decrease of total glutathione (GSH) level and increase of thiobarbituric acid-reactive substances (TBARS) value in brain tissue. Meanwhile, oral administration of HK (> or = 3 mg/kg) for 3 days reduced the lethality (60%) in NMDA-treated group to 10% level, and alleviated the behavioral signs of NMDA neurotoxicity. Moreover, HK pretreatment restored the levels of total GSH and TBARS in the brain tissue to control levels (p<0.01). Additionally, GSH peroxidase activity in cytosolic portion of brain homogenate was also restored significantly (p<0.01), whereas GSH reductase activity was not. Separately, compared to vehicle-treated control, no significant changes in body and brain weight were observed in mice administered with HK. Based on these results, oral intake of HK is suggested to prevent oxidative stress in the brain of mice.

Topics: Administration, Oral; Animals; Biphenyl Compounds; Brain; Dose-Response Relationship, Drug; Injections, Intraperitoneal; Lignans; Magnolia; Male; Mice; Mice, Inbred ICR; N-Methylaspartate; Neuroprotective Agents; Oxidative Stress; Phytotherapy; Plant Bark; Plant Extracts

Honokiol (HNK), a naturally occurring biphenyl, possesses potent antineoplastic and antiangiogenic properties. We investigated the in vitro and in vivo activity of HNK against breast cancer. HNK exhibited potent anti-proliferative activity against breast cancer cell lines and enhanced the activity of other drugs used for the treatment of breast cancer. In vivo, HNK was highly effective against breast cancer in nude mice. We identified two different effects of HNK on breast cancer cells: cell cycle inhibition, observed at lower doses of HNK, and induction of apoptosis, observed at higher doses of the compound. Our data suggest that HNK is a systemically available, non-toxic inhibitor of breast cancer growth and should be examined for clinical applications.

Topics: Animals; Antineoplastic Agents, Phytogenic; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Biphenyl Compounds; Blotting, Western; Breast Neoplasms; Cell Cycle; Cell Proliferation; Doxorubicin; Drug Synergism; Female; Humans; Hydroxamic Acids; In Vitro Techniques; Lignans; Mice; Mice, Nude; Paclitaxel; Tamoxifen; Vorinostat

Previous reports have shown that honokiol induces apoptosis in numerous cancer cell lines and showed preclinical efficacies against apoptosis-resistant B-cell chronic lymphocytic leukemia and multiple myeloma cells from relapse-refractory patients. Here, we show that honokiol can induce a cell death distinct from apoptosis in HL60, MCF-7, and HEK293 cell lines. The death was characterized by a rapid loss of integrity of plasma membrane without externalization of phosphatidyl serine. The broad caspase inhibitor z-VAD-fmk failed to prevent this cell death. Consistently, caspase activation and DNA laddering were not observed. The death was paralleled by a rapid loss of mitochondrial membrane potential, which was mechanistically associated with the mitochondrial permeability transition pore regulated by cyclophilin D (CypD) based on the following evidence: (a) cyclosporin A, an inhibitor of CypD (an essential component of the mitochondrial permeability transition pore), effectively prevented honokiol-induced cell death and loss of mitochondrial membrane potential; (b) inhibition of CypD by RNA interference blocked honokiol-induced cell death; (c) CypD up-regulated by honokiol was correlated with the death rates in HL60, but not in K562 cells, which underwent apoptosis after being exposed to honokiol. We further showed that honokiol induced a CypD-regulated death in primary human acute myelogenous leukemia cells, overcame Bcl-2 and Bcl-X(L)-mediated apoptotic resistance, and was effective against HL60 cells in a pilot in vivo study. To the best of our knowledge, this is the first report to document an induction of mitochondrial permeability transition pore-associated cell death by honokiol.

Topics: Adult; Aged; Antineoplastic Agents, Phytogenic; Apoptosis Inducing Factor; Biphenyl Compounds; Cell Death; Cell Line, Tumor; Cell Nucleus; Cyclophilins; Female; HL-60 Cells; Humans; Leukemia, Myeloid, Acute; Lignans; Male; Membrane Potential, Mitochondrial; Middle Aged; Mitochondria; Mitochondrial Membrane Transport Proteins; Mitochondrial Permeability Transition Pore; Necrosis; Neoplasms; Reactive Oxygen Species

A series of new bisphenol derivatives bearing allylic moieties were synthesized as potential analogs of honokiol and/or magnolol. Certain compounds exhibited specific anti-proliferation activity against SVR cells and moderate anti-HIV-1 activity in primary human lymphocytes. Compound 5h was the most potent compound and its anti-tumor activity was evaluated in vivo.

Topics: Antineoplastic Agents; Antiviral Agents; Biphenyl Compounds; HIV-1; Lignans; Molecular Structure

To investigate the effects of the principal biologically active compounds of M. officinalis on the growth and acid generation of the main cariogenic bacteria.. Streptococcus mutans, Streptococcus sanguis, Actinomyces naeslundii, Actinomyces viscosus, Lactobacillus rhamnosus were chosen as the experimental bacteria. The active compounds (Magnolol and Honokiol) were separated from M. officinalis and then the effects of the two agents on the growth and acid generation of the bacteria were assessed. The minimal inhibitory concentration (MIC) and the minimal biofilm eradication concentration (MBEC) of the test strains were determined by MBEC-Device.. It was found that the growth of not only plantonic bacteria but also the biofilm were efficiently inhibited by Magnolol and Honokiol. The two agents could also inhibit the acid production of the test strains.. M. officinalis may be an effective anti-caries agent, and further researches will be necessary to define its usefulness in this aspect.

Topics: Acids; Bacteria; Biofilms; Biphenyl Compounds; Dental Caries; Lignans; Magnolia; Plant Extracts

Honokiol (HNK), a phenolic compound isolated and purified from magnolia, has been found to have a number of pharmacologic benefits, including anti-angiogenic and anti-inflammatory properties. HNK has long been used in traditional Asian medicine without toxic side effects. We and others have extensively studied signaling to B cells by CD40 and its Epstein Barr viral mimic, latent membrane protein 1 (LMP1), which has been implicated in exacerbation of chronic autoimmune disease. We asked whether HNK could inhibit CD40 and LMP1 inflammatory signaling mechanisms. In vivo, HNK stabilized the severity of symptomatic collagen-induced arthritis in both CD40-LMP1 transgenic mice and their congenic C57BL/6 counterparts. Ex vivo studies, including collagen-specific serum Ab and Ag recall responses, as well as CD40 or LMP1-mediated activation of splenic B cells, supported the anti-inflammatory effects of HNK. In mouse B cell lines expressing the human CD40-LMP1 chimeric receptor, CD40- and LMP1-mediated NF-kappaB and AP-1 activation were abrogated in a dose-dependent manner, with a concomitant decrease in TNF-alpha and IL-6. These promising findings suggest that the nontoxic anti-inflammatory properties of HNK could be valuable for blocking the autoimmune response.

Topics: Animals; Anti-Allergic Agents; Arthritis, Experimental; B-Lymphocytes; Biphenyl Compounds; CD40 Antigens; Cell Line; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Female; Humans; Inflammation; Lignans; Magnolia; Mice; Mice, Transgenic; NF-kappa B; Phytotherapy; Plant Preparations; TNF Receptor-Associated Factor 2; Transcription Factor AP-1

Honokiol, a component of the herb Magnolia officinalis, exhibits antioxidant, anti-inflammatory and anxiolytic properties, increases seizure threshold, and promotes neurite outgrowth. Because stroke has become the second leading cause of death in industrialized countries, an effective neuroprotectant is urgently required. In this study, we attempted to elucidate in a mouse cerebral ischaemia model whether honokiol could be a neuroprotectant. Adult male Institute of Cancer Research (ICR) mice were subjected to middle cerebral artery occlusion for 45 min. Honokiol (10 microg/kg in 0.2 ml of saline) or control vehicle was intraperitoneally administered twice, 15 min. before and 60 min. after the induction of ischaemia. Cerebral ischaemia induced by this method was associated with an increase in synaptosomal production of reactive oxygen species, with decreases in synaptosomal mitochondrial membrane potential (DeltaPsim) and synaptosomal mitochondrial metabolic function, and with reductions in Na(+), K(+)-ATPase activities of tissues isolated from selected brain regions. Administration of honokiol resulted in significant reductions in brain infarct volume and in synaptosomal production of reactive oxygen species. The decreases in synaptosomal mitochondrial membrane potential, synaptosomal mitochondrial metabolic function and tissue Na(+), K(+)-ATPase activities observed in the ischaemic brains were also attenuated by honokiol treatments. It is concluded that honokiol can protect brain against ischaemic reperfusion injury and preserve mitochondrial function from oxidative stress. Regarding therapeutic application, further studies are needed to assess the efficacy and safety of honokiol in clinical situations.

Topics: Animals; Anti-Anxiety Agents; Biphenyl Compounds; Brain Ischemia; Lignans; Male; Membrane Potential, Mitochondrial; Mice; Mice, Inbred ICR; Neuroprotective Agents; Reactive Oxygen Species; Sodium-Potassium-Exchanging ATPase; Synaptosomes

Magnolia bark extract (MBE) is an extract of the dried stem, root, or branch bark of magnolia trees that has been used historically in traditional Chinese and Japanese medicines, and more recently as a component of dietary supplements and cosmetic products. To study the genotoxic potential of MBE, a bacterial reverse mutation assay and an in vivo micronucleus test were conducted. Compositional analysis of the test substance revealed that MBE contains 94% magnolol and 1.5% honokiol. MBE exerted no mutagenic activity in various bacterial strains of Salmonella typhimurium and in Escherichia coli WP2 uvrA, either in the absence or presence of metabolic activation at all doses tested. In the micronucleus test, various doses of MBE did not affect the proportions of immature to total erythrocytes, nor did it increase the number of micronuclei in the immature erythrocytes of Swiss albino mice. The results of these studies demonstrate that MBE is not genotoxic under the conditions of the in vitro bacterial reverse mutation assay and the in vivo micronucleus test, and support the safety of MBE for dietary consumption.

Topics: Administration, Oral; Animals; Biphenyl Compounds; Bone Marrow Cells; Chromatography, High Pressure Liquid; Cyclophosphamide; Dose-Response Relationship, Drug; Escherichia coli; Female; Lignans; Magnolia; Male; Mice; Mice, Inbred Strains; Micronucleus Tests; Mutagenicity Tests; Mutation; Plant Bark; Plant Extracts; Reproducibility of Results; Salmonella typhimurium

The antinociceptive actions of honokiol and magnolol, two major bioactive constituents of the bark of Magnolia officinalis, were evaluated using tail-flick, hot-plate and formalin tests in mice. The effects of honokiol and magnolol on the formalin-induced c-Fos expression in the spinal cord dorsal horn as well as motor coordination and cognitive function were examined. Data showed that honokiol and magnolol did not produce analgesia in tail-flick, hot-plate paw-shaking and neurogenic phase of the overt nociception induced by intraplantar injection of formalin. However, honokiol and magnolol reduced the inflammatory phase of formalin-induced licking response. Consistently, honokiol and magnolol significantly decreased formalin-induced c-Fos protein expression in superficial (I-II) laminae of the L4-L5 lumbar dorsal horn. However, honokiol and magnolol did not elicit motor incoordination and memory dysfunction at doses higher than the analgesic dose. These results demonstrate that honokiol and magnolol effectively alleviate the formalin-induced inflammatory pain without motor and cognitive side effects, suggesting their therapeutic potential in the treatment of inflammatory pain.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Biphenyl Compounds; Central Nervous System Depressants; Disease Models, Animal; Drug Evaluation, Preclinical; Fixatives; Formaldehyde; Inflammation; Lignans; Lumbosacral Region; Magnolia; Memory; Mice; Pain; Posterior Horn Cells; Proto-Oncogene Proteins c-fos

The present study was undertaken to gain insights into the mechanism of cell cycle arrest caused by honokiol, a constituent of oriental herb Magnolia officinalis. The honokiol treatment decreased the viability of PC-3 and LNCaP human prostate cancer cells in a concentration- and time-dependent manner, which correlated with G0-G1 phase cell cycle arrest. The honokiol-mediated cell cycle arrest was associated with a decrease in protein levels of cyclin D1, cyclin-dependent kinase 4 (Cdk4), Cdk6, and/or cyclin E and suppression of complex formation between cyclin D1 and Cdk4 as revealed by immunoprecipitation using anti-cyclin D1 antibody followed by immunoblotting for Cdk4 protein. The honokiol-treated PC-3 and LNCaP cells exhibited a marked decrease in the levels of total and phosphorylated retinoblastoma protein (Rb), which correlated with the suppression of transcriptional activity of E2F1. Exposure of PC-3 and LNCaP cells to honokiol resulted in the induction of p21 (PC-3 and LNCaP) and p53 protein expression (LNCaP). However, small interfering RNA (siRNA)-mediated knockdown of either p21 (PC-3 and LNCaP) or p53 (LNCaP) protein failed to confer any protection against honokiol-induced cell cycle arrest. The honokiol treatment caused the generation of reactive oxygen species (ROS), and the cell cycle arrest caused by honokiol was partially but significantly attenuated in the presence of antioxidant N-acetylcysteine. In conclusion, the present study reveals that the honokiol-mediated G0-G1 phase cell cycle arrest in human prostate cancer cells is associated with the suppression of protein level/phosphorylation of Rb leading to inhibition of transcriptional activity of E2F1.

Topics: Acetylcysteine; Antineoplastic Agents, Phytogenic; Antioxidants; Biphenyl Compounds; Cell Cycle; Cell Cycle Proteins; Cell Proliferation; Cyclin-Dependent Kinase Inhibitor p21; Drugs, Chinese Herbal; E2F1 Transcription Factor; G1 Phase; Humans; Immunoprecipitation; Lignans; Luciferases; Male; Phosphorylation; Prostatic Neoplasms; Reactive Oxygen Species; Resting Phase, Cell Cycle; Retinoblastoma Protein; RNA, Small Interfering; Transcription, Genetic; Tumor Cells, Cultured; Tumor Suppressor Protein p53

Honokiol, a small molecular weight natural product, has been shown to possess potent anti-neoplastic and anti-angiogenic properties. Its molecular mechanisms and the ability of anti-gastric cancer remain unknown. It has been shown that the anti-apoptotic function of the glucose-regulated proteins (GRPs) predicts that their induction in neoplastic cells can lead to cancer progression and drug resistance. We explored the effects of honokiol on the regulation of GRPs and apoptosis in human gastric cancer cells and tumor growth.. Treatment of various human gastric cancer cells with honokiol led to the induction of GRP94 cleavage, but did not affect GRP78. Silencing of GRP94 by small interfering RNA (siRNA) could induce cell apoptosis. Treatment of cells with honokiol or chemotherapeutics agent etoposide enhanced the increase in apoptosis and GRP94 degradation. The calpain activity and calpain-II (m-calpain) protein (but not calpain-I (micro-calpain)) level could also be increased by honokiol. Honokiol-induced GRP94 down-regulation and apoptosis in gastric cancer cells could be reversed by siRNA targeting calpain-II and calpain inhibitors. Furthermore, the results of immunofluorescence staining and immunoprecipitation revealed a specific interaction of GRP94 with calpain-II in cells following honokiol treatment. We next observed that tumor GRP94 over-expression and tumor growth in BALB/c nude mice, which were inoculated with human gastric cancer cells MKN45, are markedly decreased by honokiol treatment.. These results provide the first evidence that honokiol-induced calpain-II-mediated GRP94 cleavage causes human gastric cancer cell apoptosis. We further suggest that honokiol may be a possible therapeutic agent to improve clinical outcome of gastric cancer.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Calpain; Cell Line, Tumor; Endoplasmic Reticulum Chaperone BiP; Glucose; Humans; Kinetics; Lignans; Male; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Models, Biological; Stomach Neoplasms

The antiallergic effects of magnolol and honokiol, isolated from the bark of Magnolia obovata (family Magnoliaceae), were investigated both in vitro and in vivo. Magnolol and honokiol potently inhibited passive cutaneous anaphylaxis reactions in mice induced by IgE-antigen complex as well as compound 48/80-induced scratching behaviors. These constituents exhibited not only potent inhibitory activity on the degranulation of RBL-2H3 cells induced by IgE-antigen complex, with IC(50) values of 45 and 55 muM, respectively, but also inhibited the protein expressions of IL-4 and TNF-alpha. Based on these findings, magnolol and honokiol may improve IgE-induced allergic diseases.

Topics: Animals; Anti-Allergic Agents; beta-N-Acetylhexosaminidases; Biphenyl Compounds; Cell Line, Tumor; Dose-Response Relationship, Drug; Immunoglobulin E; Inhibitory Concentration 50; Lignans; Male; Mast Cells; Mice; Mice, Inbred BALB C; Mice, Inbred ICR; Molecular Structure; p-Methoxy-N-methylphenethylamine; Passive Cutaneous Anaphylaxis; Pruritus; Rats

P-glycoprotein accounts for the most intrinsic and acquired cancer multidrug resistance. To inhibit the expression of P-glycoprotein is one of the effective ways to reverse cancer drug resistance. Honokiol, a naturally occurring compound, has been demonstrated to combat cancer through mechanisms including inhibition of angiogenesis and induction of apoptosis. Here, we show that honokiol down-regulated the expression of P-glycoprotein at mRNA and protein levels in MCF-7/ADR, a human breast MDR cancer cell line. The down-regulation of P-glycoprotein was accompanied with a partial recovery of the intracellular drug accumulation, and of the sensitivities toward adriamycin. This study reveals a novel function of honokiol as an anti-cancer agent.

Topics: Antibiotics, Antineoplastic; ATP Binding Cassette Transporter, Subfamily B, Member 1; Biphenyl Compounds; Breast Neoplasms; Cell Line, Tumor; Cell Survival; Dose-Response Relationship, Drug; Down-Regulation; Doxorubicin; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Drug Synergism; Drugs, Chinese Herbal; Flow Cytometry; Gene Expression Regulation, Neoplastic; Humans; Lignans; Magnolia; Rhodamine 123; RNA, Messenger; Time Factors

A simple, rapid, and accurate method for the separation and determination of honokiol and magnolol in Magnolia officinalis and related herbal medicines was developed by combination of flow injection (FI) and capillary zone electrophoresis (CZE). The analysis was carried out using an unmodified fused-silica capillary (50-microm I.D.; total length 7.5 cm; effective length 4.5 cm). A series of optimization steps afforded the following conditions: the sample solvent consisted of 150 mM NaOH and a running buffer composed of 10 mM sodium tetraborate/10 mM sodium dihydrogenphosphate (NaH2PO4) at pH 12 was applied for the separation of the analytes. The separation could be achieved within 5 min with a sample throughput rate of up to 28 h(-1). The repeatability (defined as the relative standard deviation, RSD) for honokiol and magnolol was 2.0% and 1.6% with peak area evaluation, 3.6% and 2.0% with peak height evaluation, and 2.0% and 1.4% with migration time evaluation, respectively. Regression equations revealed linear relationships (r = 0.9991-0.9998) between the peak area of each analyte and the concentration.

Topics: Biphenyl Compounds; Buffers; Chemistry Techniques, Analytical; Chemistry, Pharmaceutical; Electrophoresis, Capillary; Equipment Design; Herbal Medicine; Hydrogen-Ion Concentration; Lignans; Magnolia; Models, Chemical; Phosphates; Plant Extracts; Regression Analysis; Time Factors

A comparative study of preparative isolation and purification of the phenolic compounds magnolol and honokiol from the Chinese medicinal plant Magnoliae officinalis by upright counter-current chromatography (CCC) and semi-preparative HPLC is presented. The comparison reveals that with a two-phase solvent system composed of light petroleum (bp 60-90 degrees C)-ethyl acetate-tetrachloromethane-methanol-water (1:1:8:6:1, v/v), 1250 mg of honokiol and 520 mg of magnolol, with a purity of 98.7 and 99.5%, respectively, were obtained from 2.0 g of a crude sample of Magnoliae officinalis in a single CCC separation. In contrast, semi-preparative HPLC allowed isolation and purification of these two phenolic compounds with significantly lower productivity and higher solvent consumption. Structures of the purified compounds were identified by 1H and 13C NMR.

Topics: Biphenyl Compounds; Chromatography; Lignans; Magnetic Resonance Spectroscopy; Magnolia; Molecular Structure; Solvents

Honokiol, a compound extracted from Chinese medicinal herb Magnolia officinalis, has several biological effects. However, its protective effects against endothelial injury remain unclarified. In this study, we examined whether honokiol prevented oxidized low-density lipoprotein (oxLDL)-induced vascular endothelial dysfunction. Incubation of oxLDL with honokiol (2.5-20 microM) inhibited copper-induced oxidative modification as demonstrated by diene formation, thiobarbituric acid reactive substances (TBARS) assay and electrophoretic mobility assay. Expression of adhesion molecules (ICAM, VCAM and E-selectin) and endothelial NO synthase (eNOS) affected by oxLDL was investigated by flow cytometry and Western blot. We also measured the production of reactive oxygen species (ROS) using the fluorescent probe 2',7'-dichlorofluorescein acetoxymethyl ester (DCF-AM). Furthermore, several apoptotic phenomena including increased cytosolic calcium, alteration of mitochondrial membrane potential, cytochrome c release and activation of caspase-3 were also investigated. Apoptotic cell death was characterized by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) stain. The results showed that honokiol prevented the copper-induced oxidative modification of LDL. Honokiol also ameliorated the oxLDL-diminished eNOS protein expression and reduced the oxLDL-induced adhesion molecules and the adherence of THP-1 cells to HUVECs. Furthermore, honokiol attenuated the oxLDL-induced cytotoxicity, apoptotic features, ROS generation, intracellular calcium accumulation and the subsequent mitochondrial membrane potential collapse, cytochrome c release and activation of caspase-3. Our results suggest that honokiol may have clinical implications in the prevention of atherosclerotic vascular disease.

Topics: Antioxidants; Apoptosis; Biphenyl Compounds; Calcium; Caspase 3; Caspases; Cell Adhesion; Cell Line; Cytochromes c; E-Selectin; Endothelial Cells; Humans; Intercellular Adhesion Molecule-1; Lignans; Lipoproteins, LDL; Membrane Potentials; Mitochondria; Nitric Oxide Synthase Type III; Reactive Oxygen Species; Vascular Cell Adhesion Molecule-1

An optimized high-performance liquid chromatographic method coupled with tandem mass spectrometric detection (LC-MS/MS) was developed for the simultaneous determination of honokiol and magnolol in Magnolia officinalis. Honokiol and magnolol were separated from the extracts using a reversed-phase C(18) column with a mobile phase consisted of acetonitrile and water (75:25, v/v) at a flow-rate of 0.8 mL/min. Selected reaction monitoring (SRM) mode was used for all sample quantification by the precursor-ion/product ion pair m/z 265 --> m/z 224 for honokiol and m/z 265 --> m/z 247 for magnolol. Validation data showed that this method has good linearity (r(2) > 0.995) over the concentration range of 0.0025-0.5 microg/mL for honokiol and magnolol, and both intra- and inter-day variability were acceptable within 15% at the lowest concentrations for this method. This proposed method provides excellent specificity, higher sensitivity and shorter run time than conventional methods and was applied successfully to determine the contents of honokiol and magnolol in M. officinalis.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Lignans; Magnolia; Mass Spectrometry; Molecular Structure; Plant Bark; Reproducibility of Results

The aim of the present study was to investigate the neuroprotective effects of honokiol and magnolol, two major bioactive constituents of the bark of Magnolia officinalis, against neuron toxicity induced by glucose deprivation, excitatory amino acids and hydrogen peroxide (H(2)O(2)) in cultured rat cerebellar granule cells. Cell membrane damage was measured with a lactate dehydrogenase (LDH) release assay and 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-tetrazolium bromide (MTT) assay was used to assess mitochondrial activity, reflecting cell survival. Results showed that honokiol and magnolol alone did not affect mitochondrial function and cell damage, but significantly reversed glucose deprivation-induced mitochondrial dysfunction and cell damage. The glutamate receptor blocker MK-801 and antioxidant vitamin E also provided protection against this damage. Furthermore, honokiol was more potent than magnolol in protecting against glutamate-, N-methyl-D-aspartate (NMDA)- and H(2)O(2)-induced mitochondrial dysfunction. These results demonstrated that the neuroprotective effects of honokiol and magnolol may be related to their anti-oxidative actions and antagonism of excitotoxicity induced by excitatory amino acids, suggesting that both compounds may be potential therapeutic agents for neurodegenerative diseases.

Topics: Animals; Antioxidants; Biphenyl Compounds; Cells, Cultured; Cerebellum; Dizocilpine Maleate; Excitatory Amino Acid Antagonists; Glucose; Glutamic Acid; Hydrogen Peroxide; L-Lactate Dehydrogenase; Lignans; Mitochondria; N-Methylaspartate; Neurons; Neuroprotective Agents; Oxidative Stress; Rats; Rats, Sprague-Dawley; Vitamin E

Capillary electrophoresis with electrochemical detection has been employed for the determination of honokiol and magnolol in Cortex Magnoliae Officinalis (i.e. Magnolia Bark) for the first time. Effects of several important factors such as the concentration and the acidity of the running buffer, separation voltage, injection time, and detection potential were investigated to acquire the optimum conditions. The detection electrode was a 300 microm diameter carbon disc electrode at a working potential of +0.90 V (versus saturated calomel electrode (SCE)). The two analytes can be well separated within 6 min in a 40 cm length fused silica capillary at a separation voltage of 18 kV in a 50mM borate buffer (pH 9.2). The relation between peak current and analyte concentration was linear over about three orders of magnitude with the detection limits (S/N=3) of 0.38 and 0.51 microM for honokiol and magnolol, respectively. The proposed method has been successfully applied to monitor the two bioactive constituents in the real plant samples with satisfactory assay results.

Topics: Anti-Arrhythmia Agents; Biphenyl Compounds; Drugs, Chinese Herbal; Electrochemistry; Electrodes; Electrophoresis, Capillary; Hydrogen-Ion Concentration; Lignans; Magnolia; Plant Bark; Regression Analysis

A separation of honokiol 1 from the closely structurally related magnolol 2 was developed. Honokiol demonstrated weak activity against HIV-1 in human lymphocytes.

Topics: Animals; Antineoplastic Agents, Phytogenic; Antiviral Agents; Biphenyl Compounds; Cell Line, Tumor; Chlorocebus aethiops; Drug Screening Assays, Antitumor; Humans; In Vitro Techniques; Leukocytes, Mononuclear; Lignans; Vero Cells

Hydrophobic bile acid-induced apoptosis plays an important role in cholestatic liver disease, and its prevention may be of therapeutic interest. The aim of this study was to investigate the protective effect of honokiol on glycochenodeoxycholic acid-induced apoptosis in primary cultured rat hepatocytes. Glycochenodeoxycholic acid is a hydrophobic bile salt that accumulates intrahepatically during cholestasis and induces hepatocyte apoptosis at pathophysiological concentrations. Primary rat hepatocytes were pretreated with honokiol at concentrations of 40, 20 and 10 microM 5 min before glycochenodeoxycholic acid treatment. Incubation of hepatocytes with glycochenodeoxycholic acid at a concentration of 100 microM for 4 h induced apoptosis as shown by DNA fragmentation, chromatin condensation and cleavage of poly(ADP-ribose) polymerase. Pretreatment with honokiol at concentrations of 40, 20 and 10 microM significantly inhibited the generation of intracellular reactive oxygen species and reduced activation of caspases-8, -9, and -3 and cleavage of poly-(ADP-ribose) polymerase. Glycochenodeoxycholic acid treatment up-regulated phosphorylation of stress-activated protein kinase/c-jun-NH2-terminal kinase which was inhibited by honokiol treatment. Inhibition of stress-activated protein kinase/c-jun-NH2-terminal kinase phosphorylation by SP600125 protected hepatocytes from apoptosis induced by glycochenodeoxycholic acid. These data indicate that honokiol protects hepatocytes from apoptosis induced by glycochenodeoxycholic acid in vitro and this protection may be due to reduced oxidative stress and inhibition of stress-activated protein kinase/c-jun-NH2-terminal kinase phosphorylation.

Topics: Animals; Apoptosis; Biphenyl Compounds; Caspases; Cells, Cultured; Glycochenodeoxycholic Acid; Hepatocytes; JNK Mitogen-Activated Protein Kinases; Lignans; Magnolia; Oxidative Stress; Phosphorylation; Plant Bark; Poly(ADP-ribose) Polymerases; Rats

Glomerulonephritis (GN) is still the most common cause of end-stage renal disease. Accumulation of glomerular macrophages, proliferation of mesangial cells, and deposition of extracellular matrix proteins are pathobiological hallmarks of GN. Pharmacological interventions that can inhibit these insults may be beneficial in the retardation of the progression of GN. Honokiol originally isolated from Magnolia officinalis, shows antioxidative, anti-inflammatory, and antiproliferative activities in a variety of inflammation models. In this study, we first investigated the in vivo effects of honokiol on rat anti-Thy1 nephritis. Anti-Thy1 nephritis was induced in Wistar rats by injecting mouse anti-rat Thy1 antibodies intravenously. Nephritic rats were randomly assigned to receive honokiol (2.5 mg/kg, twice a day) or vehicle and were killed at various time points. Glomerular histology and immunohistopathology and urine protein excretion were studied. Western blotting was conducted for markers of proliferation. Adhesion molecules, chemokine, and extracellular matrix gene expression were evaluated by Northern blotting. Honokiol-treated nephritic rats excreted less urinary protein and had lower glomerular cellularity and sclerosis. The increased intraglomerular proliferating cell nuclear antigen and Akt phosphorylation in nephritic rats could be abolished by the treatment of honokiol. Honokiol also alleviated glomerular monocyte chemoattractant protein-1 and intracellular adhesion molecule-1, similar to type I (alpha1) collagen and fibronectin mRNA levels of nephritic rats. These results indicate that honokiol may have therapeutic potential in mesangial proliferative GN.

Topics: Animals; Apoptosis; Biphenyl Compounds; Cell Proliferation; Chemokine CCL2; Disease Models, Animal; Extracellular Matrix Proteins; Gastrointestinal Agents; Glomerulonephritis, Membranoproliferative; Intercellular Adhesion Molecule-1; Lignans; Male; Proliferating Cell Nuclear Antigen; Proteinuria; Random Allocation; Rats; Rats, Sprague-Dawley; RNA, Messenger; Thy-1 Antigens

This paper describes the development and the application of a novel carbon nanotube/poly(methyl methacrylate) (CNT/PMMA) composite electrode as a sensitive amperometric detector of CE. The composite electrode was fabricated on the basis of the in situ polymerization of a mixture of CNT and prepolymerized methylmethacrylate in the microchannel of a piece of fused-silica capillary under heat. The performance of this unique system has been demonstrated by separating and detecting honokiol and magnolol in traditional Chinese medicine, Cortex Magnoliae Officinalis. Factors influencing their separation and detection processes were examined and optimized. Honokiol and magnolol were well separated within 7 min in a 40 cm long capillary at a separation voltage of 15 kV using a 50 mM borate buffer (pH 9.2). The new CNT-based CE detector offered significantly lower operating potentials, yielded substantially enhanced S/N characteristics, and exhibited resistance to surface fouling and hence enhanced stability. It demonstrated long-term stability and reproducibility with RSDs of less than 5% for the peak current (n = 9) and should also find a wide range of applications in microchip CE, flowing injection analysis, and other microfluidic analysis systems.

Topics: Biphenyl Compounds; Catalysis; Drugs, Chinese Herbal; Electrodes; Electrophoresis, Capillary; Hydrogen-Ion Concentration; Lignans; Magnolia; Nanotubes, Carbon; Polymethyl Methacrylate; Sensitivity and Specificity

Honokiol, an active component in extracts of Magnolia officinalis, has been proposed to play a role in anti-inflammatory, antioxidant activity, anti-angiogenic and anti-tumor activity. Although honokiol has a variety of pharmacological effects on certain cell types, its effects on vascular smooth muscle cells (VSMC) are unclear. This issue was investigated in the present study, honokiol was found to inhibit cell viability and DNA synthesis in cultured VSMC. These inhibitory effects were associated with G1 cell cycle arrest. Treatment with honokiol blocks the cell cycle in the G1 phase, down-regulates the expression of cyclins and CDKs and up-regulates the expression of p21WAF1, a CDK inhibitor. While honokiol did not up-regulate p27, it caused an increase in the promoter activity of the p21WAF1 gene. Immunoblot and deletion analysis of the p21WAF1 promoter showed that honokiol induced the expression of p21WAF1 and that this expression was independent of the p53 pathway. Furthermore, the honokiol-mediated signaling pathway involved in VSMC growth inhibition was examined. Among the relevant pathways, honokiol induced a marked activation of p38 MAP kinase and JNK. The expression of dominant negative p38 MAP kinase and SB203580, a p38 MAP kinase specific inhibitor, blocked the expression of honokiol-dependent p38 MAP kinase and p21WAF1. Consistently, blockade of p38 MAPK kinase function reversed honokiol-induced VSMC proliferation and cell cycle proteins. These data demonstrate that the p38 MAP kinase pathway participates in p21WAF1 induction, subsequently leading to a decrease in the levels of cyclin D1/CDK4 and cyclin E/CDK2 complexes and honokiol-dependent VSMC growth inhibition. In conclusion, these findings concerning the molecular mechanisms of honokiol in VSMC provides a theoretical basis for clinical approaches to the use therapeutic agents in treating atherosclerosis.

Topics: Antineoplastic Agents, Phytogenic; Atherosclerosis; Base Sequence; Biphenyl Compounds; Cell Survival; Cells, Cultured; Cyclin-Dependent Kinase Inhibitor p21; Cyclins; Down-Regulation; Enzyme Activation; Enzyme Inhibitors; G1 Phase; Humans; Imidazoles; Lignans; Myocytes, Smooth Muscle; p38 Mitogen-Activated Protein Kinases; Promoter Regions, Genetic; Protein Kinases; Pyridines; Sequence Deletion

Recent reports have indicated that honokiol can induce apoptosis, suppress tumor growth, and inhibit angiogenesis. In this report, we found that honokiol potentiated the apoptosis induced by tumor necrosis factor (TNF) and chemotherapeutic agents, suppressed TNF-induced tumor cell invasion, and inhibited RANKL-induced osteoclastogenesis, all of which are known to require nuclear factor-kappaB (NF-kappaB) activation. Honokiol suppressed NF-kappaB activation induced by a variety of inflammatory stimuli, and this suppression was not cell type specific. Further studies showed that honokiol blocked TNF-induced phosphorylation, ubiquitination, and degradation of IkappaBalpha through the inhibition of activation of IkappaBalpha kinase and of Akt. This led to suppression of the phosphorylation and nuclear translocation of p65 and NF-kappaB-dependent reporter gene expression. Magnolol, a honokiol isomer, was equally active. The expression of NF-kappaB-regulated gene products involved in antiapoptosis (IAP1, IAP2, Bcl-x(L), Bcl-2, cFLIP, TRAF1, and survivin), proliferation (cyclin D1, cyclooxygenase-2, and c-myc), invasion (matrix metalloproteinase-9 and intercellular adhesion molecule-1), and angiogenesis (vascular endothelial growth factor) were also down-regulated by honokiol. Honokiol also down-regulated NF-kappaB activation in in vivo mouse dorsal skin model. Thus, overall, our results indicate that NF-kappaB and NF-kappaB-regulated gene expression inhibited by honokiol enhances apoptosis and suppresses osteoclastogenesis and invasion.

Topics: Animals; Apoptosis; Biphenyl Compounds; Carrier Proteins; Cyclin D1; Cyclooxygenase 2; Dose-Response Relationship, Drug; Drug Synergism; Genes, myc; Humans; I-kappa B Proteins; Lignans; Matrix Metalloproteinase 9; Membrane Glycoproteins; Membrane Proteins; Mice; Molecular Structure; NF-kappa B; NF-KappaB Inhibitor alpha; Osteoclasts; Osteogenesis; Phosphorylation; Promoter Regions, Genetic; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B; Synaptotagmin I; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

High-performance liquid chromatography was employed to determine the contents of the eight marker components liquiritin, naringin, hesperidin, thymol, imperatorin, honokiol, isoimperatorin, and magnolol in the traditional Chinese medicinal preparation Huoxiang-zhengqi liquid. The separation was performed on a C(18) column by stepwise gradient elution with water-methanol-acetonitrile (0.01 min, 68:30:2; 20 min, 60:38:2; 50 min, 34:64:2; 65 min, 34:64:2; 75 min, 28:70:2; 85 min, 68:30:2) as the mobile phase at a flow rate of 1 ml/min, with UV detection at 283 nm. Eight regression equations showed good linear relationships between the peak area ratio of each marker to internal standard and amounts. The recoveries of the markers listed above were 97.4, 98.5, 97.4, 98.6, 97.8, 99.2, 97.0, and 97.5%, respectively. The repeatability and reproducibility (relative standard deviation) of the method were less than 2.2 and 3.0%, respectively.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Furocoumarins; Glucosides; Hesperidin; Lignans; Reproducibility of Results; Solutions; Thymol

To optimize preparative technique for banxia-houpu effervescent tablets.. Based on the pH, disintegration time limited, taste, and rigidity of effervescent tablets, the proper proportion between citric acid and sodium bicarbonate, as well as the proper quantity of polyethylene glycol 6000 and sodium cyclamate in the effervescent tablets were determined by using orthogonal design. The content of magnolol and honokiol in effervescent tablets were measured by HPLC.. The optimal preparative technique was: cirtic acid: sodium bicarbonate = 0.65:1. The percentage of polyethylene glycol 6000 was 85%, and the percentage of sodium cyclamate was 1.0%.. The preparative technique is stable, reliable and suitable for practical use.

Topics: Biphenyl Compounds; Chemistry, Pharmaceutical; Chromatography, High Pressure Liquid; Citric Acid; Drugs, Chinese Herbal; Hydrogen-Ion Concentration; Lignans; Pinellia; Plants, Medicinal; Polyethylene Glycols; Sodium Bicarbonate; Tablets; Technology, Pharmaceutical

Investigate into transport rate and retention rate transference of principal effective constituent in Shujin Kechuang capsule, a new development Chinese patent medicine for theraphy asthma.. HPLC was applied to analyze the content of ephedrine hydrochloride and honokiol and magnolol in crude drugs and 60% ethanol extracting solution and 25% concentrated solution,50% concentrated solution, 100% concentrated solution and finished product ( Shujin Kechuang capsule).. The transport rate of ephedrine hydrochloride and honokiol and magnolol is 56. 32%, 14. 43%, 14. 56% in the finished product respectively.. should be concentrate and desiccation in the condition that decompress and low temperature.

Topics: Asthma; Biphenyl Compounds; Capsules; Chromatography, High Pressure Liquid; Drug Combinations; Drugs, Chinese Herbal; Ephedra sinica; Ephedrine; Lignans; Magnolia; Plant Structures; Plants, Medicinal; Technology, Pharmaceutical

Magnolol (MG) and honokiol (HK), two lignans showing anti-inflammatory and anti-oxidant properties and abundantly available in the medicinal plants Magnolia officinalis and M. obovata, were found to enhance HL-60 cell differentiation initiated by low doses of 1,25-dihydroxyvitamin D3 (VD3) and all-trans-retinoic acid (ATRA). Cells expressing membrane differentiation markers CD11b and CD14 were increased from 4% in non-treated control to 8-16% after being treated with 10-30 microM MG or HK. When added to 1 nM VD3, MG or HK increased markers expressing cells from approximately 30% to 50-80%. When either MG or HK was added to 20 nM ATRA, only CD11b, but not CD14, expressing cells were increased from 9% to 24-70%. Under the same conditions, adding MG or HK to VD3 or ATRA treatment further enlarged the G0/G1 cell population and increased the expression of p27(Kip1), a cyclin-dependent kinase inhibitor. Pharmacological studies using PD098059 (a MEK inhibitor), SB203580 (a p38 MAPK inhibitor) and SP600125 (a JNK inhibitor) suggested that the MEK pathway was important for VD3 and ATRA-induced differentiation and also its enhancement by MG or HK, the p38 MAPK pathway had a inhibitory effect and the JNK pathway had little influence. It is evident that MG and HK are potential differentiation enhancing agents which may allow the use of low doses of VD3 and ATRA in the treatment for acute promyelocytic leukemia.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Calcitriol; Calcium Channel Agonists; CD11b Antigen; Cell Differentiation; G1 Phase; HL-60 Cells; Humans; Lignans; Lipopolysaccharide Receptors; MAP Kinase Signaling System; Resting Phase, Cell Cycle; Tretinoin

The neolignans, magnolol 1 and honokiol 2 have been reported to inhibit the growth of several tumor cell lines in vitro and in vivo. The chemical structure of magnolol and honokiol consists of biphenyl skeleton with phenolic and allylic functionalities. Analogs of 1 and 2 containing different substitution have been studies for their effect on the growth of Hep-G2 and their structure-activity relationships were reported in this work.

Topics: Antineoplastic Agents; Biphenyl Compounds; Cell Line, Tumor; Humans; Lignans; Structure-Activity Relationship

The therapeutic goal in liver fibrosis is the reversal of fibrosis and the selective clearance of activated hepatic stellate cells (HSCs) by inducing apoptosis. Over the past several years, we have screened for natural products that mediate apoptosis in activated HSCs. Among the candidate compounds, honokiol, isolated from Magnoliae cortex, was found to induce apoptotic death in activated rat HSCs, while there was no cell viability change in hepatocytes, at concentrations of 12.5-50 microM. Apoptosis was identified by DNA fragmentation, activation of caspase-3 and -9, and the proteolytic cleavage of poly(ADP-ribose) polymerase, down-regulation of bcl-2 and the release of mitochondrial cytochrome c into the cytoplasm.

Topics: Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Caspases; Cytochromes c; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Enzyme Activation; Flow Cytometry; Hepatocytes; Lignans; Magnolia; Male; Phytotherapy; Rats; Rats, Sprague-Dawley

A simple and sensitive method has been developed for determining honokiol and magnolol in fresh Magnolia obovata (M. obovata) by micro high-performance liquid chromatography with electrochemical detection (microHPLC-ECD). Chromatography was performed using a Capcell Pak C-18 UG 120 microbore octadecylsilica (ODS) column, methanol-water-phosphoric acid (65 : 35 : 0.5, v/v/v), as a mobile phase and applied potential at +0.8 V vs. Ag/AgCl. Peak heights were found linearly related to the amounts of honokiol and magnolol injected from 0.67 pg to 2.0 ng (r>0.999). The detection limits (S/N=3) were 0.13 pg, respectively. Honokiol and magnolol of 0.27 ng were detected with relative standard deviation (RSD) of 0.73 and 1.17% (n=5), respectively. Honokiol and magnolol in Magnolia Bark of the Japanese Pharmacopoeia were extracted with 70% methanol, diluted with a mobile phase, and injected into the microHPLC-ECD for determination. Recoveries of honokiol and magnolol in Magnolia Bark exceeded 98.7% with RSD, less than 0.93% (n=5). Determination of the distributions of honokiol and magnolol in bark, phloem, wood, leaf blades, and petioles of fresh M. obovata were made using weight samples of 40-238 mg. This method is useful to determine honokiol and magnolol in M. obovata, which is a candidate for crude magnolia bark for traditional Japanese herbal medicines.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Electrochemistry; Lignans; Magnolia; Medicine, Kampo; Microfluidics; Plant Components, Aerial; Plant Extracts; Plant Leaves

B-cell chronic lymphocytic leukemia (B-CLL) remains an incurable disease that requires innovative new approaches to improve therapeutic outcome. Honokiol is a natural product known to possess potent antineoplastic and antiangiogenic properties. We examined whether honokiol can overcome apoptotic resistance in primary tumor cells derived from B-CLL patients. Honokiol induced caspase-dependent cell death in all of the B-CLL cells examined and was more toxic toward B-CLL cells than to normal mononuclear cells, suggesting greater susceptibility of the malignant cells. Honokiol-induced apoptosis was characterized by the activation of caspase-3, -8, and -9 and cleavage of poly(adenosine diphosphate-ribose) polymerase (PARP). Exposure of B-CLL cells to honokiol resulted in up-regulation of Bcl2-associated protein (Bax) and down-regulation of the expression of the key survival protein myeloid-cell leukemia sequence 1 (Mcl-1), which is associated with response to treatment in B-CLL patients. In addition, B-CLL cells pretreated with interleukin-4 (IL-4), a cytokine known to support B-CLL survival, underwent apoptosis when subsequently incubated with honokiol, indicating that honokiol could also overcome the prosurvival effects of IL-4. Furthermore, honokiol enhanced cytotoxicity induced by fludarabine, cladribine, or chlorambucil. These data indicate that honokiol is a potent inducer of apoptosis in B-CLL cells and should be examined for further clinical application either as a single agent or in combination with other anticancer agents.

Topics: Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Caspases; Chlorambucil; Cladribine; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Humans; In Vitro Techniques; Interleukin-4; Leukemia, Lymphocytic, Chronic, B-Cell; Lignans; Magnolia; Myeloid Cell Leukemia Sequence 1 Protein; Neoplasm Proteins; Phytotherapy; Proto-Oncogene Proteins c-bcl-2; Vidarabine

Propionibacterium acnes, an anaerobic pathogen, plays an important role in the pathogenesis of acne and seems to initiate the inflammatory process by producing proinflammatory cytokines. In order to demonstrate the anti-inflammatory effects and action mechanisms of magnolol and honokiol, several methods were employed. Through DPPH and SOD activity assays, we found that although both magnolol and honokiol have antioxidant activities, honokiol has relatively stronger antioxidant activities than magnolol {[for DPPH assay, % of DPPH bleaching of magnolol and honokiol (500 microM magnolol: 19.8%; 500 microM honokiol: 67.3%)]; [for SOD assay, SOD activity (200 microM magnolol: 53.4%; 200 microM honokiol: 64.3%)]}. Moreover, the production of interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-alpha) induced by P. acnes in THP-1 cells, a human monocytic cell line, was reduced by magnolol and honokiol {[for IL-8 (10 microM magnolol: 42.7% inhibition; 10 microM honokiol: 51.4% inhibition)]; [for TNF-alpha (10 microM magnolol: 20.3% inhibition; 10 microM honokiol: 39.0% inhibition)]}. Cyclooxygenase-2 (Cox-2) activity was also suppressed by them [(15 microM magnolol: 45.8% inhibition), (15 microM honokiol: 66.3% inhibition)]. Using a nuclear factor-kappaB (NF-kappaB) luciferase reporter assay system and Western analysis, we identified that magnolol and honokiol exert their anti-inflammatory effects by inhibiting the NF-kappaB element, which exists in Cox-2, IL-8, and TNF-alpha promoters [(15 microM magnolol: 44.8% inhibition), (15 microM honokiol: 42.3% inhibition)]. Of particular note is that magnolol and honokiol operate downstream of the MEKK-1 molecule. Together with their previously known antibacterial activity against P. acnes and based on these results, we suggest that magnolol and honokiol may be introduced as possible acne-mitigating agents.

Topics: Anti-Bacterial Agents; Anti-Inflammatory Agents, Non-Steroidal; Antioxidants; Biphenyl Compounds; Cytokines; Humans; Interleukin-8; Lignans; Magnolia; MAP Kinase Kinase Kinase 1; Microbial Sensitivity Tests; Monocytes; NF-kappa B; Phytotherapy; Picrates; Plant Extracts; Propionibacterium acnes; Superoxide Dismutase; Tumor Necrosis Factor-alpha

Honokiol (HNK) is an active component purified from magnolia, a plant used in traditional Chinese and Japanese medicine. Here we show that HNK significantly induces cytotoxicity in human multiple myeloma (MM) cell lines and tumor cells from patients with relapsed refractory MM. Neither coculture with bone marrow stromal cells nor cytokines (interleukin-6 and insulin-like growth factor-1) protect against HNK-induced cytotoxicity. Although activation of caspases 3, 7, 8, and 9 is triggered by HNK, the pan-caspase inhibitor z-VAD-fmk does not abrogate HNK-induced apoptosis. Importantly, release of an executioner of caspase-independent apoptosis, apoptosis-inducing factor (AIF), from mitochondria is induced by HNK treatment. HNK induces apoptosis in the SU-DHL4 cell line, which has low levels of caspase 3 and 8 associated with resistance to both conventional and novel drugs. These results suggest that HNK induces apoptosis via both caspase-dependent and -independent pathways. Furthermore, HNK enhances MM cell cytotoxicity and apoptosis induced by bortezomib. In addition to its direct cytotoxicity to MM cells, HNK also represses tube formation by endothelial cells, suggesting that HNK inhibits neovascurization in the bone marrow microenvironment. Taken together, our results provide the preclinical rationale for clinical protocols of HNK to improve patient outcome in MM.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Bone Marrow Cells; Caspases; Cell Cycle; Cell Line, Tumor; Cell Proliferation; DNA; Dose-Response Relationship, Drug; Drug Resistance, Multiple; Endothelial Cells; Humans; Insulin-Like Growth Factor I; Interleukin-6; Lignans; Multiple Myeloma; Neovascularization, Pathologic; Stromal Cells; Structure-Activity Relationship

The effects of honokiol and magnolol, two major bioactive constituents of the bark of Magnolia officinalis, on Ca(2+) and Na(+) influx induced by various stimulants were investigated in cultured rat cerebellar granule cells by single-cell fura-2 or SBFI microfluorimetry. Honokiol and magnolol blocked the glutamate- and KCl-evoked Ca(2+) influx with similar potency and efficacy, but did not affect KCl-evoked Na(+) influx. However, honokiol was more specific for blocking NMDA-induced Ca(2+) influx, whereas magnolol influenced with both NMDA- and non-NMDA activated Ca(2+) and Na(+) influx. Moreover, the anti-convulsant effects of these two compounds on NMDA-induced seizures were also evaluated. After honokiol or magnolol (1 and 5 mg/kg, i.p.) pretreatment, the seizure thresholds of NMRI mice were determined by tail-vein infusion of NMDA (10 mg/ml). Data showed that both honokiol and magnolol significantly increased the NMDA-induced seizure thresholds, and honokiol was more potent than magnolol. These results demonstrated that magnolol and honokiol have differential effects on NMDA and non-NMDA receptors, suggesting that the distinct therapeutic applications of these two compounds for neuroprotection should be considered.

Topics: Analysis of Variance; Animals; Anti-Anxiety Agents; Biphenyl Compounds; Calcium; Cells, Cultured; Cerebellum; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Interactions; Excitatory Amino Acid Agonists; Excitatory Amino Acid Antagonists; Ion Channels; Lignans; Mice; N-Methylaspartate; Neurons; Platelet Aggregation Inhibitors; Potassium Chloride; Quinoxalines; Rats; Rats, Sprague-Dawley; Seizures; Sodium

We have found that honokiol [4-allyl-2-(3-allyl-4-hydroxy-phenyl)-phenol] can promote neurite outgrowth and mobilize intracellular Ca2+ store in primary cultured rat cortical neurons. In this study, we examined the effects of honokiol on extracellular signal-regulated kinases (ERK1/2) and Akt, and their possible relationship to neurite outgrowth and Ca2+ mobilization. Honokiol-induced neurite outgrowth in the cultured rat cortical neurons was significantly reduced by PD98059, a mitogen-activated protein kinase kinase (MAPKK, MAPK/ERK kinase MEK, direct upstream of ERK1/2) inhibitor, but not by LY294002, a phosphoinositide 3-kinase (PI3K, upstream of Akt) inhibitor. Honokiol also significantly enhanced the phosphorylation of ERK1/2 in a concentration-dependent manner, whereas the effect of honokiol on Akt phosphorylation was characterized by transient enhancement in 10 min and lasting inhibition after 30 min. The phosphorylation of ERK1/2 enhanced by honokiol was inhibited by PD98059 as well as by KN93, a Ca2+/calmodulin-dependent kinase II (CaMK II) inhibitor. Moreover, the products of the phosphoinositide specific phospholipase C (PLC)-derived inositol 1,4,5-triphosphate (IP3) and 1,2-diacylglycerol (DAG) were measured after honokiol treatment. Together with our previous findings, these results suggest that the signal transduction from PLC, IP3, Ca2+, and CaMK II to ERK1/2 is involved in honokiol-induced neurite outgrowth.

Topics: Animals; Benzylamines; Biphenyl Compounds; Blotting, Western; Calcium-Calmodulin-Dependent Protein Kinases; Cells, Cultured; Cerebral Cortex; Chromones; Diglycerides; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Enzyme Activation; Female; Fetus; Flavonoids; Inositol 1,4,5-Trisphosphate; Lignans; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Models, Biological; Morpholines; Neurites; Neurons; Phosphoinositide-3 Kinase Inhibitors; Phosphorylation; Pregnancy; Protein Kinase Inhibitors; Rats; Rats, Sprague-Dawley; Sulfonamides

To study the effects of magnolol and honokiol on isolated smooth muscle of gastrointestinal tract and their relationship with Ca2+, and on the gastric emptying and the intestinal propulsive activity in mice.. Routine experimental methods using isolated gastric fundus strips of rats and isolated ileum segments of guinea pigs were adopted to measure the smooth muscle tension. The effects of magnolol 10(-3), 10(-4), 10(-5) mol/L, and honokiol 10(-4), 10(-5), 10(-6) mol/L on the contractility of gastric fundus strips of rats and ileum of guinea pigs induced by acetylcholine (Ach) and 5-hydroxytryptamine (5-HT) was assessed respectively. The method using nuclein and pigment methylene blue was adopted to measure the gastric retention rate of nuclein and the intestinal propulsive ratio of a nutritional semi-solid meal for assessing the effect of magnolol and honokiol (0.5, 2, 20 mg/kg) on gastric emptying and intestinal propulsion.. Magnolol and honokiol significantly inhibited the contractility of isolated gastric fundus strips of rats treated with Ach or 5-HT and isolated ileum guinea pigs treated with Ach or CaCl2, and both of them behaved as non-competitive muscarinic antagonists. Magnolol and honokiol inhibited the contraction induced by Ach in Ca2+-free medium and extracellular Ca2+-dependent contraction induced by Ach. Each group of magnolol and honokiol experiments significantly decreased the residual rate of nuclein in the stomach and increased the intestinal propulsive ratio in mice.. The inhibitory effect of magnolol and honokiol on contractility of the smooth muscles of isolated gastric fundus strips of rats and isolated ileum of guinea pigs is associated with a calcium-antagonistic effect. Magnolol and honokiol can improve the gastric emptying of a semi-solid meal and intestinal propulsive activity in mice.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Biphenyl Compounds; Drugs, Chinese Herbal; Female; Gastric Emptying; Gastrointestinal Agents; Gastrointestinal Motility; Guinea Pigs; Lignans; Male; Mice; Muscle, Smooth; Rats

To study the effect of honokiol on arterial thrombosis and endothelial cells.. Rabbit platelet aggregation was performed with Borns turbid method. Thrombosis was produced by the endothelial injury stimulated with electric current. Rat aortic endothelial cells (RAEC) were cultured and cell viability was assessed using the MTT assay. Nitric oxide (NO) concentrations in serum-free media of RAEC were determined using the kinetic cadmium-reduction method. The stable metabolite prostacyclin was measured in serum-free media of RAEC by radioimmunoassay.. Honokiol (37.6-376 micromol/L) decreased rabbit platelet aggregation in vitro in a concentration-dependent manner, while intravenously injection of honokiol (0.12-12 microg/kg) significantly inhibited rabbit platelet aggregation induced by collagen ex vivo. In the electrical current-stimulated carotid thrombosis model in rats, honokiol (5-50 microg/kg, iv) prolonged the thrombus occlusion time in a does-dependent manner. In vitro honokiol (0.376-37.6 micromol/L) effectively protected cultured RAEC against oxidized low density lipoprotein (ox-LDL) injury, and significantly increased 6-keto-PGF1alpha (the stable metabolite of prostacyclin) in serum-free media of RAEC. Honokiol also increased NO level in RAEC serum-free medium at a lower concentration range (0.0376-0.376 micromol/L), but honokiol 3.76 micromol/L decreased NO level.. Honokiol is a potent arterial thrombosis inhibitor. Endothelial cell protection and the stimulation of prostacyclin release may be its main anti-thrombosis mechanism. Stimulation of NO release in endothelial cells may play a role, but it is not a key factor.

Topics: 6-Ketoprostaglandin F1 alpha; Animals; Biphenyl Compounds; Carotid Artery Thrombosis; Cell Survival; Dose-Response Relationship, Drug; Endothelial Cells; Epoprostenol; Fibrinolytic Agents; Lignans; Magnolia; Male; Nitric Oxide; Plants, Medicinal; Platelet Aggregation; Rabbits; Rats; Rats, Sprague-Dawley

To study the chemical characteristic, to identify the different forms and to establish the new standard for the quality control of Cortex Magnoliae Officinalis.. HPLC method was used with acetonitrile-water (63:37) as the mobile phase at room temperature. The chromatographic column was Lichrospher 100 RP-18e (4.6 mm x 250 mm, 5 microm). The flow rate was 1 mL x min(-1), and the detection wavelength was 294 nm. The chromatograms of 45 individuals from 13 seed resources of Cortex Magnolia Officinalis were recorded. The chemical characteristics analysis and comparability' s calculation of seed resources were made.. It was proposed that the area ratio of peak 5 to 6 (characteristic I) and the area ratio of peak 5 and 6 to the total peak areas (characteristic II) are the identification characteristics for different seed resources of Cortex Magnoliae Officinalis.. This method can be used effectively to identify the high quality seed resource of Cortex Magnoliae Officinalis.

Topics: Biphenyl Compounds; China; Chromatography, High Pressure Liquid; Ecosystem; Lignans; Magnolia; Plant Bark; Plants, Medicinal; Quality Control

Honokiol, a small molecular weight lignan originally isolated from Magnolia officinalis, shows anti-angiogenic, anti-invasive and anti-proliferative activities in a variety of cancers. In this study, we investigated whether honokiol affects the transcription factor nuclear factor-kappa B (NF-kappaB) which controls a large number of genes involved in angiogenesis, metastasis and cell survival. We observed that the tumor necrosis factor-alpha (TNF-alpha)-induced NF-kappaB activation was blocked by honokiol in four different cancer cell lines as evidenced by EMSA. Honokiol did not directly affect the NF-kappaB-DNA binding. Immunoblot experiments demonstrated that honokiol inhibited the TNF-alpha-stimulated phosphorylation and degradation of the cytosolic NF-kappaB inhibitor IkappaBalpha. Furthermore, honokiol suppressed the intrinsic and TNF-alpha-stimulated upstream IkappaB kinases (IKKs) activities measured by a non-radioactive kinase assay using immunoprecipitated IKKs, suggesting a critical role of honokiol in abrogating the phosphorylation and degradation of IkappaBalpha. In a HeLa cell NF-kappaB-dependent luciferase reporter system, honokiol suppressed luciferase expression stimulated by TNF-alpha and by the transient transfection and expression of NIK (NF-kappaB-inducing kinase), wild type IKKbeta, constitutively active IKKalpha and IKKbeta, or the p65 subunit. Honokiol was also found to inhibit the nuclear translocation and phosphorylation of p65 subunit of NF-kappaB. RT-PCR results showed that honokiol suppressed NF-kappaB-regulated inflammatory and carcinogenic gene products including MMP-9, TNF-alpha, IL-8, ICAM-1 and MCP-1. In line with the observation that NF-kappaB activation may up-regulate anti-apoptotic genes, it was shown that honokiol enhanced TNF-alpha-induced apoptotic cell death. In summary, our results demonstrate that honokiol suppresses NF-kappaB activation and NF-kappaB-regulated gene expression through the inhibition of IKKs, which provides a possible mechanism for its anti-tumor actions.

Topics: Antineoplastic Agents; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Electrophoretic Mobility Shift Assay; Gene Expression; Genes, Reporter; Humans; I-kappa B Kinase; Lignans; Lipopolysaccharides; Luciferases; Lymphotoxin-alpha; NF-kappa B; NF-kappa B p50 Subunit; Phosphorylation; Reverse Transcriptase Polymerase Chain Reaction; Tetradecanoylphorbol Acetate; Transcription Factor AP-1; Transcription Factor RelA; Tumor Necrosis Factor-alpha

This study aims to investigate the possible hepato-protective effects of honokiol against liver damage and cirrhosis induced by carbon tetrachloride (CCl(4)) in the rat. Rats were treated acutely, or chronically with CCl(4) at 5 day intervals (0.06 mL/100 g body weight, administered as 50% vol/vol solution in liquid paraffin) by gavage, in combination with phenobarbitone in drinking water (0.5 g/L for 7 days prior to, and during CCl(4) treatment) to induce liver damage. Some were also co-treated with 0.1 mg/kg or 0.03 mg/kg honokiol (i.p.) or with appropriate vehicle. In vivo measurement of the liver sinusoidal area was performed using confocal microscopy following i.v. fluorescein isothiocyanate (FITC) dextran. Liver histology and function tests were performed, and liver and body weights were measured. Confocal microscopy showed that acute and chronic CCl(4) treatment significantly reduced the sinusoidal area. Honokiol (0.1 mg/kg, but not 0.03 mg/kg) partially reversed the decrease in the sinusoidal area after acute or chronic treatments with CCl(4). Acute and chronic CCl(4) treatment produced significant histological liver damage. Honokiol (0.1 mg/kg) significantly reduced the histological damage caused by chronic treatment. Chronic treatment with CCl(4) caused a significant increase in the bilirubin level that was not observed following the high dose of honokiol (0.1 mg/kg). In conclusion, this study showed that honokiol exhibits potent hepato-protective effects in rats treated with CCl(4).

Topics: Animals; Bilirubin; Biphenyl Compounds; Carbon Tetrachloride; Hepatic Veno-Occlusive Disease; Lignans; Liver; Liver Cirrhosis, Experimental; Magnolia; Male; Microscopy, Confocal; Rats; Rats, Long-Evans

Limited evidence is available as to whether Kampo medicine modifies gastrointestinal function in humans. We investigated the effect of a Kampo medicine, Hange-koboku-to (Banxia-houpo-tang, HKT), on patients with functional dyspepsia (FD) and on healthy volunteers with regard to gastric motility. The gastric emptying rate (GER) in FD patients was significantly lower than in the healthy subjects. GER in FD patients and in healthy volunteers showed a significant increase after 2 weeks of medication with HKT. Furthermore, gastrointestinal symptoms improved significantly in the FD patients after the administration of HKT. These results suggest that HKT improves delayed gastric emptying and acts as a prokinetic agent.

Topics: Adult; Biphenyl Compounds; Catechols; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Dyspepsia; Fatty Alcohols; Female; Gastric Emptying; Humans; Lignans; Male; Medicine, Kampo; Middle Aged; Phytotherapy; Statistics, Nonparametric; Ultrasonography

To investigate the pharmacokinetics of honokiol in rats.. Honokiol injection was delivered by vein injection to SD-rats. The blood samples were gathered at a series of time lags. Honokiol in rat plasma was determined with an RP-HPLC method and the data were analyzed with program 3P87.. After i.v. injection of honokiol, concentration-time curves were fitted to a 3-compartment model: with halftime of 2.8 min, 11.9 min, and 56.8 min.. Honokiol was quickly distributed in rats after i.v. and the concentration decreased rapidly. Our studies provided important referrence to the research on the pharmacodynamics and the pharmaceutics of Honokiol.

Topics: Animals; Biphenyl Compounds; Drugs, Chinese Herbal; Lignans; Rats; Rats, Sprague-Dawley

Honokiol, a biphenyl-type neolignan, which shows the remarkable neurotrophic effect in primary cultured rat cortical neurons, has been effectively synthesized in 21% yield over 14 steps starting from 5-bromosalicylic acid and p-hydroxybenzoic acid by utilizing Pd-catalyzed Suzuki-Miyaura coupling reaction as a key step. Additionally, the structure-activity relationship between neurite outgrowth-promoting activity and its O-methylated and/or its hydrogenated analogues was examined in the primary cultures of fetal rat cortical neurons, suggesting that 5-allyl and 4'-hydroxyl groups are essential for affecting the neurotrophic activity of honokiol.

Topics: Animals; Biphenyl Compounds; Cells, Cultured; Cerebral Cortex; Fetus; Lignans; Nerve Growth Factors; Neurites; Neurodegenerative Diseases; Neurons; Rats; Structure-Activity Relationship

High-speed counter-current chromatography was used to isolate and purify honokiol and magnolol from cortex Magnoliae Officinalis (Magnolia officinalis Rehd. et Wils.), a plant used in the traditional Chinese medicine. A crude sample, 150 mg, was successfully separated with a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (1:0.4:1:0.4, v/v), and the fractions were analyzed by high-performance liquid chromatography. The separation produced 80 and 45 mg of honokiol and magnolol with purities of 99.2 and 98.2%, respectively, in 2.5 h.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Countercurrent Distribution; Lignans; Magnetic Resonance Spectroscopy; Magnolia

To investigate the signal pathway of honokiol-induced apoptosis on human colorectal carcinoma RKO cells and to evaluate whether p53 and p53-related genes were involved in honokiol-treated RKO cells.. Cell cycle distribution and subdiploid peak were analyzed with a flow cytometer and DNA fragment with electrophoresis on agarose gels. Transcriptional level of Bax, Bcl-2, Bid and Bcl-xl was accessed by RT-PCR. Western blotting was used to measure p53 protein expression and other factors related to apoptosis. Proliferation inhibition of two cell lines (RKO, SW480) with high expression of p53 and one cell line with p53 negative expression (LS180) was monitored by MTT assay.. Honokiol induced RKO cell apoptosis in a dose-dependent manner. The mRNA expression level and protein level of Bid were up-regulated while that of Bcl-xl was down-regulated, but no changes in Bax and Bcl-2 were observed. Western blotting showed p53 expression had no remarkable changes in honokiol-induced RKO cell apoptosis. LS180 cells treated with honokiol exhibited apparent growth inhibition like RKO cells and Sw480 cells.. Honokiol can induce RKO cells apoptosis through activating caspase cascade by p53-indepenent pathway.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Line, Tumor; Colorectal Neoplasms; Humans; Lignans; Tumor Suppressor Protein p53

Honokiol and magnolol, two major phenolic constituents of Magnolia sp., have been known to exhibit antibacterial activities. However, until now, their antibacterial activity against Propionibacterium sp. has not been reported. To this end, the antibacterial activities of honokiol and magnolol were detected using the disk diffusion method and a two-fold serial dilution assay. Honokiol and magnolol showed strong antibacterial activities against both Propionibacterium acnes and Propionibacterium granulosum, which are acne-causing bacteria. The minimum inhibitory concentrations (MIC) of honokiol and magnolol was 3-4 microg/ml (11.3-15 microM) and 9 microg/ml (33.8 microM), respectively. In addition, the killing curve analysis showed that magnolol and honokiol killed P. acnes rapidly, with 10(5) organisms/ml eliminated within 10 min of treatment with either 45 microg (169.2 microM) of magnolol or 20 microg (75.2 microM) of honokiol per ml. The cytotoxic effect of honokiol and magnolol was determined by a colorimetric (3-(4,5-dimetyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) (MTT) assay using two animal cell lines, human normal fibroblasts and HaCaT. In this experiment, magnolol exhibited lower cytotoxic effects than honokiol at the same concentration, but they showed similar cytotoxicity when triclosan was employed as an acne-mitigating agent. In addition, they reduced secretion of interleukin-8 and tumor necrosis factor alpha (TNF-alpha) induced by P. acnes in THP-1 cells indicating the anti-inflammatory effects of them. When applied topically, neither phenolic compound induced any adverse reactions in a human skin primary irritation test. Therefore, based on these results, we suggest the possibility that magnolol and honokiol may be considered as attractive acne-mitigating candidates for topical application.

Topics: Adult; Animals; Anti-Bacterial Agents; Anti-Inflammatory Agents, Non-Steroidal; Biphenyl Compounds; Cell Line; Dose-Response Relationship, Drug; Female; Humans; Lignans; Microbial Sensitivity Tests; Propionibacterium; Skin Irritancy Tests

To investigate the inhibitory effects of Magnolol and Honokiol on the activity of streptococcal glucosyltransferases (Gtfs).. The effect of Magnolol and Honokiol that inhibits the activities of streptococcal GtfB, GtfC, GtfD and GtfS was explored with standard assays. The results showed that both samples can efficiently inhibit the activity of all Gtfs in solution (66.4-96.3%) and adsorbed on the surface of saliva-coated hydroxyapatite (sHA) beads (65.5-92.7%) at concentrations between 1.25 and 5.0 mg ml(-1). Furthermore, Magnolol had a stronger inhibition of four kinds of Gtfs than Honokiol both in solution and adsorbed on the surface of sHA beads at concentrations between 0.04 and 0.63 mg ml(-1) (P < 0.05).. Magnolol had significant effects on the activities of streptococcal Gtfs.. Magnolol as a natural herb can be developed into a new oral hygiene product to prevent plaque formation.

Topics: Adsorption; Biphenyl Compounds; Enzyme Inhibitors; Glucans; Glucosyltransferases; Lignans; Microspheres; Streptococcus

To investigate the anticancer activity of honokiol on RKO, a human colorectal carcinoma cell line in vitro and in vivo, and to evaluate its possible use in clinic.. In vitro anticancer activity of honokiol was demonstrated by its induction of apoptosis in tumor cells. We analyzed cell proliferation with MTT assay, cell cycle with flow cytosmeter, DNA fragment with electrophoresis on agarose gels. To test the mechanism of honokiol-induced apoptosis, Western blotting was used to investigate the factors involved in this process. The pharmacokinetics study of honokiol was tested by high phase liquid chromatography. In in vivo study, Balb/c nude mice were incubated with RKO cells. Honokiol was injected intraperitoneally every other day into tumor bearing Balb/c nude mice.. Our results showed that honokiol induced apoptosis of RKO cells in a time- and dose-dependent manner. At 5-10 microg/mL for 48 h, honokiol induced apoptosis through activating Caspase cascades. Pharmacokinetics study demonstrated that, honokiol could be absorbed quickly by intraperitoneal injection, and maintained in plasma for more than 10 h. In nude mice bearing RKO-incubated tumor, honokiol displayed anticancer activity by inhibiting tumor growth and prolonging the lifespan of tumor bearing mice.. With its few toxicity to normal cells and potent anticancer activity in vitro and in vivo, honokiol might be a potential chemotherapy candidate in treating human colorectal carcinoma.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Blotting, Western; Caspase 3; Caspase 9; Caspases; Cell Division; Cell Line, Tumor; Colorectal Neoplasms; DNA Fragmentation; Drugs, Chinese Herbal; Humans; Lignans; Mice; Mice, Inbred BALB C; Mice, Nude; Xenograft Model Antitumor Assays

The aim of this study was to investigate the protective effect of honokiol and magnolol on hepatocyte injury induced by either tertiary butyl hydroperoxide (tBH)- or D-galactosamine (GalN). The cellular leakage of LDH and AST, and cell death by treatment with 1.5 mM tBH for 1 h, were significantly inhibited by treatment with honokiol (40 and 20 microM) or magnolol (40 microM). Treatment with honokiol or magnolol significantly inhibited lipid peroxidation in both cells and media, the generation of intracellular reactive oxygen species (ROIs), and intracellular glutathione (GSH) depletion induced by tBH. The cellular leakage of LDH and AST, and cell death, by 24-hour treatment with 30 mM GalN were significantly inhibited by treatment with honokiol (20, 5 and 1 microM) or magnolol (20, 5 and 1 microM). Treatment with honokiol (20, 5 and 1 microM) or magnolol (20 and 5 microM) significantly inhibited the intracellular GSH depletion induced by GalN. The hepatoprotective effects of honokiol and magnolol on oxidative stress induced by tBH were probably the result of their antioxidant activity. Honokiol and magnolol also had a protective effect against GalN-induced hepatotoxicity, which was used as an alternate model to oxidative stress, acting by inhibiting intracellular GSH depletion.

Topics: Animals; Biphenyl Compounds; Cells, Cultured; Galactosamine; Hepatocytes; Lignans; Liver; Male; Protective Agents; Rats; Rats, Sprague-Dawley; tert-Butylhydroperoxide

Honokiol, a constituent extracted from Magnolia officinalis, had been shown be an antioxidant and an anti-platelet agent in biological systems with an anti-arrhythmic effect and a myocardial protective effect on ischemia-reperfusion injury. We examined the neuroprotective effect of honokiol in rats subjected to focal cerebral ischemia. Honokiol was administered intravenously either at fifteen minutes before right middle cerebral artery occlusion (pretreatment groups) or when both common carotid arteries clips were removed (treatment groups). The results showed that there was no significant hemodynamic change after intravenous infusion of honokiol at the dosages of 10(-8), 10(-7) and 10(-6) g/kg in both groups. However, honokiol significantly reduced the total volume of infarction at the doses of 10(-7) or 10(-6) g/kg in both pretreatment and treatment groups. This study suggests that honokiol is a potent neuroprotective agent in focal cerebral ischemia. This beneficial effect may be related to its antioxidant effect and anti-platelet aggregation activity.

Topics: Animals; Antioxidants; Biphenyl Compounds; Brain Ischemia; Dose-Response Relationship, Drug; Infarction, Middle Cerebral Artery; Infusions, Intravenous; Lignans; Magnolia; Male; Phytotherapy; Platelet Aggregation Inhibitors; Random Allocation; Rats; Rats, Long-Evans

Natural products comprise a major source of small molecular weight angiogenesis inhibitors. We have used the transformed endothelial cell line SVR as an effective screen of natural product extracts to isolate anti-angiogenesis and anti-tumor compounds. Aqueous extracts of Magnolia grandiflora exhibit potent activity in our SVR proliferation assays. We found that the small molecular weight compound honokiol is the active principle of magnolia extract. Honokiol exhibited potent anti-proliferative activity against SVR cells in vitro. In addition, honokiol demonstrated preferential inhibition of primary human endothelial cells compared with fibroblasts and this inhibition was antagonized by antibodies against TNF alpha-related apoptosis-inducing ligand. In vivo, honokiol was highly effective against angiosarcoma in nude mice. Our preclinical data suggests that honokiol is a systemically available and non-toxic inhibitor of angiogenesis and should be further evaluated as a potential chemotherapeutic agent.

Topics: Angiogenesis Inhibitors; Animals; Apoptosis; Biphenyl Compounds; Cell Division; Cell Line; Cell Line, Transformed; Endothelium, Vascular; Hemangiosarcoma; Humans; Lignans; Magnolia; MAP Kinase Signaling System; Mice; Mice, Nude; Phytotherapy; Plant Extracts; Transplantation, Heterologous

Magnolol and honokiol are the two major phenolic constituents of the plant medicine "Houpo" ( Magnolia obovata), which is used in the treatment of chest tightness and asthma. The aim of this study was to investigate the influence of magnolol and honokiol on smooth muscle tone in porcine trachea. Magnolol and honokiol (0.1 - 100 microM) inhibited carbachol- and high K +-induced muscle contractions in a concentration-dependent fashion, but did not affect basal muscle tension. After washout of these pretreatments, carbachol- and high K +-evoked muscle contractions were still abolished, suggesting that the inhibition was irreversible. Magnolol and honokiol also concentration-dependently decreased the Ca 2+-dependent muscle contraction induced by high K + depolarization. Ca 2+ channel antagonists attenuated carbachol-induced muscular response by approximately 30 %, but did not further potentiate the inhibitory actions of magnolol and honokiol on muscle contraction. However, the inhibitory effects of magnolol and honokiol on carbachol-evoked muscular contractile response were partially reversed after removal of Ca 2+ channel antagonist pretreatment. Alternatively, caffeine-elicited muscle contractions were not altered by magnolol, honokiol, and verapamil. In conclusion, the relaxant effects of magnolol and honokiol on porcine tracheal smooth muscle suggest an association with the blockade of Ca 2+ influx through voltage-operated Ca 2+ channels instead of Ca 2+ release from intracellular Ca 2+ stores. The magnolol- and honokiol-induced inhibitions on tracheal smooth muscle contraction may be relevant to the claimed therapeutic effects of the extract from magnolia bark and contribute to their pharmacological effects by acting as anti-asthmatic agents.

Topics: Animals; Asthma; Biphenyl Compounds; Bronchodilator Agents; Calcium; Carbachol; Dose-Response Relationship, Drug; Lignans; Magnolia; Male; Muscle Contraction; Muscle, Smooth; Phytotherapy; Potassium Chloride; Swine; Trachea; Verapamil

We examined the intracellular Ca(2+) response in primary cultured rat cortical neurons and human neuroblastoma SH-SY5Y cells by Fluo 3 fluorescence imaging analysis. In these two kinds of neuronal cells, honokiol and magnolol increased cytoplasmic free Ca(2+) with a characteristic lag phase. The cytoplasmic free Ca(2+) increase was independent of extracellular Ca(2+), but dependent on activation of phospholipase C and inositol 1,4,5-triphosphate (IP(3)) receptors. These results suggest that honokiol and magnolol increase cytoplasmic free Ca(2+) through a phospholipase C-mediated pathway, and that the release of Ca(2+) from intracellular stores mainly contributes to the increase in cytoplasmic free Ca(2+). Thus, honokiol and magnolol may be involved in a new activation mechanism closely associated with intracellular Ca(2+) mobilization.

Topics: Animals; Biphenyl Compounds; Calcium; Cell Line, Tumor; Cells, Cultured; Cerebral Cortex; Female; Humans; Lignans; Neuroblastoma; Neurons; Pregnancy; Rats; Rats, Sprague-Dawley

Reactive oxygen species produced by neutrophils contribute to the pathogenesis of focal cerebral ischemia/reperfusion injury and signal the inflammatory response. We have previously shown that honokiol, an active principle extracted from Magnolia officinalis, has a protective effect against focal cerebral ischemia/reperfusion injury in rats that paralleled a reduction in reactive oxygen species production by neutrophils. To elucidate the underlying mechanism(s) of the antioxidative effect of honokiol, peripheral neutrophils isolated from rats were activated with phorbol-12-myristate-13-acetate (PMA) or N-formyl-methionyl-leucyl-phenylalanine (fMLP) in the presence or absence of honokiol. In this study, we found that honokiol inhibited PMA- or fMLP-induced reactive oxygen species production by neutrophils by three distinct mechanisms: (1) honokiol diminished the activity of assembled-NADPH oxidase, a major reactive oxygen species producing enzyme in neutrophils by 40% without interfering with its protein kinase C (PKC)-dependent assembly; (2) two other important enzymes for reactive oxygen species generation in neutrophils, i.e., myeloperoxidase and cyclooxygenase, were also inhibited by honokiol by 20% and 70%, respectively; and (3) honokiol enhanced glutathione (GSH) peroxidase activity by 30%, an enzyme that triggers the metabolism of hydrogen peroxide (H2O2). These data suggested that honokiol, acting as a potent reactive oxygen species inhibitor/scavenger, could achieve its focal cerebral ischemia/reperfusion injury protective effect by modulating enzyme systems related to reactive oxygen species production or metabolism, including NADPH oxidase, myeloperoxidase, cyclooxygenase, and GSH peroxidase in neutrophils.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Biphenyl Compounds; Lignans; Male; Neutrophils; Rats; Rats, Long-Evans; Reactive Oxygen Species

The effects of honokiol and magnolol extracted from the Magnolia officinalis on muscular contractile responses and intracellular Ca(2+) mobilization were investigated in the non-pregnant rat uterus. Honokiol and magnolol (1-100 micromol/l) were observed to inhibit spontaneous and uterotonic agonists (carbachol, PGF(2alpha), and oxytocin)-, high K(+)-, and Ca(2+) channel activator (Bay K 8644)-induced uterine contractions in a concentration-dependent manner. The inhibition rate of honokiol on spontaneous contractions appeared to be slower than that of magnolol-induced response. The time periods that were required for honokiol and magnolol, at 100 micromol/l, to abolish 50% spontaneous contractions were approximately 6 min. Furthermore, honokiol and magnolol at 10 micromol/l also blocked the Ca(2+)-dependent oscillatory contractions. Consistently, the increases in intracellular Ca(2+) concentrations ([Ca(2+)](i)) induced by PGF(2alpha) and high K(+) were suppressed by both honokiol and magnolol at 10 micromol/l. After washout of these treatments, the rise in [Ca(2+)](i) induced by PGF(2alpha) and high K(+) was still partially abolished. In conclusion, the inhibitory effects of honokiol and magnolol on uterine contraction may be mediated by blockade of external Ca(2+) influx, leading to a decrease in [Ca(2+)](i). Honokiol and magnolol may be considered as putative Ca(2+) channel blockers and be of potential value in the treatment of gynecological dysfunctions associated with uterine muscular spasm and dysmenorrhea.

Topics: Animals; Biphenyl Compounds; Calcium; Carbachol; Central Nervous System Depressants; Dinoprost; Female; Fluorescent Dyes; Fura-2; In Vitro Techniques; Lignans; Muscarinic Agonists; Myometrium; Oxytocics; Oxytocin; Potassium Chloride; Rats; Uterine Contraction; Uterus

We have previously shown that honokiol, an active component of Magnolia officinalis, displayed protective effect against focal cerebral ischemia-reperfusion (FCI/R) injury in rats. Production of reactive oxygen species (ROS) and infiltration of neutrophils to injured tissue play deleterious roles during cerebral ischemia. To study the mechanism(s) in mediating neuroprotective effect of honokiol, FCI/R-induced neutrophil infiltration and lipid peroxidation in brain tissue, and activation of neutrophils in-vitro were examined. Intravenous administration of honokiol (0.01-1.0 microg/kg) 15 min before (pretreatment) or 60 min after (post-treatment) middle cerebral artery occlusion reduced the total infarcted volume by 20-70% in dose-dependent manner. Pretreatment or post-treatment of honokiol at concentration of 0.1 and 1.0 microg/kg significantly decreased the neutrophil infiltration in the infarcted brain. Time course of neutrophil infiltration was performed in parallel with the lipid peroxidation in infracted brain tissue during FCI/R injury. The results indicate that honokiol can protect brain tissue against lipid peroxidation and neutrophil infiltration during FCI/R injury and cerebral infarction induced by FCI/R is accompanied with a prominent neutrophil infiltration to the infarcted area during FCI/R course. In-vitro, honokiol (0.1-10 microM) significantly diminished fMLP (N-formyl-methionyl-leucyl-phenylalanine)- or PMA (phorbol-12-myristate-13-acetate)-induced neutrophil firm adhesion, a prerequisite step behind neutrophil infiltration, and ROS production in neutrophils. Intracellular calcium overloading activates calcium-stimulated enzymes and further exaggerates FCI/R injury. Honokiol (0.1-10 microM) impeded the calcium influx induced by fMLP (a receptor agonist), AlF(4)(-) (a G-protein activator) or thapsigargin (an intracellular calcium pool releaser). Therefore, we conclude that the amelioration of FCI/R injury by honokiol can be attributed to its anti-oxidative and anti-inflammatory actions through, at least in part, limiting lipid peroxidation and reducing neutrophil activation/infiltration by interfering firm adhesion, ROS production, and calcium overloading that may be primed/activated during FCI/R injury.

Topics: Animals; Biphenyl Compounds; Brain Ischemia; Calcium; Calcium Signaling; Cell Adhesion; Cerebral Infarction; Chemotaxis, Leukocyte; Dose-Response Relationship, Drug; Infarction, Middle Cerebral Artery; Lignans; Lipid Peroxidation; Male; N-Formylmethionine Leucyl-Phenylalanine; Neuroprotective Agents; Neutrophils; Rats; Rats, Long-Evans; Reactive Oxygen Species; Reperfusion Injury; Tetradecanoylphorbol Acetate

To study the HPLC-FPS of Cortex Magnoliae Officinalis, the substitute species and counterfeits from different habitats, and to obtain the sameness and differences.. HPLC-FPS was used.. There were sameness and differences between two certified Cortex Magnoliae Officinalis, which were easily distinguished from their substitute species and counterfeits.. The HPLC-FPS can provide the useful information for the quality estimation and plant source of Cortex Magnoliae Officinalis.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Lignans; Magnoliaceae; Quality Control

To lay a theoretical foundation for studies on strategies for improvement of Magnolia officinalis and select superior gemplasm resources to meet the demand for modernization, industrialization and internationalization of Chinese medicine.. Seeds of Magnolia officinalis from 13 main habitats of 7 provinces were collected and strewn in a place of Jingning County, Zhejiang. At the age of seven, 195 samples were collected from the same height of the trunk of 15 individual trees of each provenance, and assayed for effective ingredients with HPLC.. Differences in the content of phenols were significant among the seed sources and even greater among individuals within a seed source.. 3 seed sources viz. Wufeng, Hefeng and Enshi of Hubei were obviously superior to other seed sources on account of high contents of magnolol, honokoiol and total phenols. Extension and application of these 3 seed sources is an effective path leading to quality improvement of Magnolia officinalis. With great differences in the content of phenols existing among individuals within each source, there is a big gap between production of medicinal materials by merely using superior seed sources of Magnolia officinalis and the demand of stable and controllable quality for modernization and internationalization of Chinese medicine. But the great difference has laid a material foundation and brought about a great potential for genetic improvement of Magnolia officinalis. Therefore, the superior individuals within a superior seed source are an excellent material for the breeding of Magnolia officinalis.

Topics: Biphenyl Compounds; China; Drugs, Chinese Herbal; Ecosystem; Lignans; Magnolia; Phenols; Plant Bark; Plants, Medicinal; Quality Control; Seeds; Species Specificity

Honokiol, a main biphenyl neolignan of the traditional crude medicine, Magnoliae cortex, was found to show neurotrophic activity on the cultures of rat cortical neurons at concentration from 0.1 to 10 microM. In the cortical neurons cultured in serum-free medium supplemented with B27, honokiol could promote neurite outgrowth. In addition, the survival and growth of neurons were significantly enhanced by adding honokiol to the primary cultures in serum-free medium supplemented with N2. Its neurotrophic activity was comparable to 40 ng mL(-1) of bFGF at concentration of 10 microM.

Topics: Animals; Biphenyl Compounds; Cells, Cultured; Cerebral Cortex; Culture Media, Serum-Free; Lignans; Neurites; Neurons; Rats

Honokiol is a phenolic compound purified from Magnolia officinalis, which induced the apoptotic cell death in several types of human cancer cells. In the present study, the molecular mechanism of honokiol-mediated apoptotic process was examined in human squamous lung cancer CH27 cells. Here, we found that honokiol-induced apoptotic cell death was accompanied by upregulation of Bad and downregulation of Bcl-XL, while honokiol had no effect on the levels of Bcl-2, Bcl-XS, Bag-1, Bax and Bak proteins. Moreover, honokiol treatment caused the release of mitochondrial cytochrome c to cytosol and sequential activation of caspases. Proteolytic activation of caspase-3 and cleavage of PARP, an in vivo substrate for caspase-3, were observed in honokiol-treated CH27 cells. Furthermore, treatment with caspase inhibitors z-DEVD-fmk and z-VAD-fmk markedly blocked honokiol-induced apoptosis. These results demonstrated that modulation of Bcl-XL and Bad proteins, release of mitochondrial cytochrome c and activation of caspase-3, participated in honokiol-triggered apoptotic process in human squamous lung cancer CH27 cells.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; bcl-X Protein; Biphenyl Compounds; Caspases; Cytochrome c Group; Enzyme Activation; Humans; Lignans; Lung Neoplasms; Proto-Oncogene Proteins c-bcl-2; Tumor Cells, Cultured

The antimicrobial activity of honokiol and magnolol, the main constituents of Magnolia officinalis was investigated. The antimicrobial activity was assayed by the agar dilution method using brain heart infusion medium and the minimum inhibitory concentration (MIC) were determined for each compound using a twofold serial dilution assay. The results showed that honokiol and magnolol have a marked antimicrobial effect (MIC = 25 microg/mL) against Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Micrococcus luteus and Bacillus subtilis, but did not show antimicrobial activity (MIC > or = 100 microg/mL) for Shigella flexneii, Staphylococcus epidermidis, Enterobacter aerogenes, Proteus vulgaris, Escherichia coli and Pseudomonas aeruginosa. Our results indicate that honokiol and magnolol, although less potent than tetracycline, show a significant antimicrobial activity for periodontal pathogens. Hence we suggest that honokiol and magnolol might have the potential to be an adjunct in the treatment of periodontitis.

Topics: Actinobacillus; Anti-Bacterial Agents; Bacillus; Bacteria; Biphenyl Compounds; Drugs, Chinese Herbal; Humans; Lignans; Magnoliopsida; Microbial Sensitivity Tests; Micrococcus; Periodontal Diseases; Plants, Medicinal; Porphyromonas; Prevotella

Honokiol and magnolol have been identified as modulators of the GABAA receptors in vitro. Our previous study suggested a possible selectivity of honokiol and magnolol on GABAA receptor subtypes. This possibility was examined in the current study by 3H-muscimol and 3H-flunitrazepam binding assays on various rat brain membrane preparations and human recombinant GABA(A) receptor subunit combinations expressed by the Sf-9/baculovirus system. Generally, honokiol and magnolol have a similar enhancing effect on (3)H-muscimol binding to various membrane preparations in nonsaturation binding assays. Honokiol and magnolol preferentially increased (3)H-muscimol binding to hippocampus compared to cortex and cerebellum (with a maximum enhancement of 400% of control). As for subunit combinations, honokiol and magnolol have a more potent enhancing effect on alpha2 subunit containing combinations (with a maximum enhancement of 400-450% of control). This action was independent of the gamma subunit. In saturation binding assays, magnolol affected either the number of binding sites (ca. 4-fold on alpha2 containing combinations) or the binding affinity (on alpha1 containing combinations) of (3)H-muscimol binding to various GABAA receptor subunit combinations. In contrast, honokiol increased only binding sites on alpha2beta3gamma2s and alpha2beta3 combinations, but both the number of binding sites and the binding affinity on alpha1beta2gamma2S and alpha(1)beta2 combinations. These results indicate that honokiol and magnolol have some selectivity on different GABAA receptor subtypes. The property of interacting with GABAA receptors and their selectivity could be responsible for the reported in vivo effects of these two compounds.

Topics: Animals; Biphenyl Compounds; Cell Line; Cerebellum; Drugs, Chinese Herbal; Flunitrazepam; GABA Agonists; GABA Modulators; Hippocampus; Humans; Insecta; Lignans; Male; Muscimol; Nitric Oxide Synthase; Protein Subunits; Rats; Rats, Wistar; Receptors, GABA-A; Recombinant Proteins; Tritium

We investigated the inhibitory effect of Magnolia obovata Thunb. bark ethanol extracts on human fibrosarcoma HT-1080 cells invasion in a reconstituted basement membrane [Matrigel (MG)]. We found that the effective components of the bark ethanol extracts were magnolol and honokiol, two biphenyl compounds. The extracts, magnolol and honokiol, did not affect HT-1080 cells adhesion to MG, but did inhibit HT-1080 cells migration at a high concentration (100 microM). HT-1080 cells secrete matrix metalloproteinase (MMP)-9, which degrades the extracellular matrix as a part of the invasive process. Magnolol and honokiol inhibited the activity of MMP-9, which may have been responsible, in part, for the inhibition of tumor cell invasiveness.

Topics: Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Fibrosarcoma; Humans; Lignans; Magnoliaceae; Matrix Metalloproteinase Inhibitors; Neoplasm Invasiveness; Phytotherapy; Plant Bark; Tumor Cells, Cultured

We have previously reported that saiboku-to, an Oriental herbal remedy composed of a mixture of 10 different herbal extracts, possesses anti-histamine release effect on mast cells in rats. This effect may be due mainly to the extract of the bark of Magnolia obovata (M. obovata), a constituent herb of saiboku-to. In the present study, it was demonstrated that the bark extract inhibited compound 48/80 (C48/80)-induced histamine release from mast cells in a concentration-dependent manner (50 % inhibitory concentration, IC(50) = 56.98 microg/ml). Furthermore, the inhibitory activity was found in the methanol fraction, but not in water and 50 % aqueous methanol fractions derived from the bark extract. Magnolol and honokiol isolated from the methanol fraction inhibited C48/80-induced histamine release from mast cells. The potency of magnolol (IC(50) = 1.04 microg/ml) was greater than that of honokiol (IC(50) = 2.77 microg/ml). Furthermore, the actual amount of magnolol (49.76 +/- 1.14 mg) contained in the bark of M. obovata (5 g) was greater than that (8.58 +/- 0.19 mg) of honokiol. Taken together, the present results suggest that magnolol may be responsible for the biological efficacy of the bark extract of M. obovata.

Topics: Animals; Biphenyl Compounds; Drugs, Chinese Herbal; Histamine H1 Antagonists; Histamine Release; Lignans; Magnoliaceae; Mast Cells; Medicine, Kampo; p-Methoxy-N-methylphenethylamine; Peritoneal Cavity; Plant Bark; Plant Extracts; Rats

To understand the main factors influencing the bark quality of Magnolia officinalis so as to theoretically establish a basis for quality assessment, genetic improvement and layout of bark producing areas.. Eighty-two samples from the main bark producing areas(11 counties of 7 provinces such as Zhejiang, Fujian, Sichuan, Hunan, Guangxi, Jiangxi and Hubei) were collected. Totally there were 121 samples, including 39 from the trial stand located in Jingning of Zhejiang the obtained out of the seeds from the bark producing areas mentioned above. HPLC was used in the analysis of phenols contained in the bark of Magnolia officinalis.. The main factors influencing the bark quality have been made clear.. The quality is affected by provenance, leaf shape, DBH, tree height, crown size, age, bark thickness color of bark powder, oiliness, grounding nature, bark type, position of sampling, etc., of which provenance, leaf shape, powder color, bark thickness and DBH are the most influential factors. These factors should be fully considered when making quality assessment and genetic improvement of the bark of Magnolia officinalis.

Topics: Biphenyl Compounds; Lignans; Magnolia; Plant Bark; Plants, Medicinal; Quality Control

A reversed-phase high performance liquid chromatographic method for simultaneous determination of magnolol and honokiol in serum and urine of rat has been established. Two drugs were determined within 15 minutes by the method on the column with spherisorb C18, by using a mobile phase consisted of methanol-water-glacial acetic acid (70:30:1, V/V) at 1 mL/min, monitored at 294 nm and with a sensitivity of 0.005 AUFS. After 0.25, 1 and 8 hour of administration of the drugs, protein in serum and urine of Wistar rat was precipitated by methanol and magnolol and honokiol in acidified body fluid were determined after being extracted by a mixture of ethyl acetate and ether. Good linear relationship between concentration in serum and urine and peak area in the ranges of 0.05-2 mg/L for magnolol and 0.025-1 mg/L for honokiol was obtained. Good precision and reproducibility were found too. The average recoveries of the two drugs were 95.6% (RSD = 3.85%), 93.8% (RSD = 3.95%) in serum and 96.0% (RSD = 3.83%), 94.9% (RSD = 3.54%) for urine respectively. The lower limit of the method was 0.02 mg/L of magnolol and 0.04 mg/L of honokiol respectively. The results showed that this method is suitable for the determination of magnolol and honokiol in body fluids.

Topics: Animals; Biphenyl Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Lignans; Magnolia; Male; Platelet Aggregation Inhibitors; Rats; Rats, Wistar

It's found that a significant correlation between the samples of Magnoliae officinalis from the provenance in phenols content and varieties or forms. The total content of magnolol and honokiol in its bark is usually used as an index to measure the quality. Therefore, Cortex Magnoliae officinalis produced in Sichuan and Hubei being considered genuinenss and good in quality, should be actually referred to local varieties. This conclusion can also reveals the nature and scientific connotation of genuineness on Cortex Magnoliae Officinalis.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Lignans; Magnolia; Plant Leaves; Plants, Medicinal; Quality Control

Two neolignan compounds, magnolol (5,5'-diallyl-2,2'-dihydroxybiphenyl, 1) and honokiol (5,5'-diallyl-2,4'-dihydroxybiphenyl, 2), were isolated from the stem bark of Magnolia obovata and evaluated for antifungal activity against various human pathogenic fungi. Compound 1 and 2 showed significant inhibitory activities against Trichophyton mentagrophytes, Microsporium gypseum, Epidermophyton floccosum, Aspergillus niger, Cryptococcus neoformans, and Candida albicans with minimum inhibitory concentrations (MIC) in a range of 25-100 microg/ml. Therefore, compound 1 and 2 could be used as lead compounds for the development of novel antifungal agents.

Topics: Antifungal Agents; Biphenyl Compounds; Humans; Lignans; Microbial Sensitivity Tests

The crude extract of magnolia bark, an herbal drug, inhibited the secretion of catecholamines from bovine adrenal chromaffin cells stimulated by acetylcholine (ACh) in a concentration-dependent manner (200-900 microg/mL). The extract also diminished the secretion induced by high K(+), which is a stimulus directly depolarizing the plasma membranes, but its inhibition was weaker than that of ACh-evoked secretion. beta-Eudesmol, honokiol, magnolol, and bornyl acetate, but not alpha- and beta-pinenes, all of which are ingredients of magnolia bark, greatly reduced ACh-evoked secretion. beta-Eudesmol and magnolol also inhibited high K(+)-induced secretion to an extent similar to that of ACh-evoked secretion. However, honokiol and bornyl acetate inhibited the secretion induced by high K(+) much less than the secretion evoked by ACh. ACh-induced Na(+) influx and ACh- or high K(+)-induced Ca(2+) influx into the cells were diminished by beta-eudesmol or honokiol. These results indicate that magnolia bark contains some effective components inhibiting the secretion of catecholamines from bovine adrenal chromaffin cells stimulated by ACh due to the antagonism of Na(+) and Ca(2+) influxes into the cells. However, inhibition by the extract of magnolia bark seems to be attributable to honokiol and bornyl acetate. Furthermore, the results indicate that the inhibitory effect of magnolia bark may be associated with its pharmacological effect on activities of the nervous system.

Topics: Acetylcholine; Adrenal Glands; Animals; Biphenyl Compounds; Calcium; Camphanes; Catecholamines; Cattle; Chromaffin Cells; In Vitro Techniques; Lignans; Magnoliopsida; Plant Extracts; Potassium; Sesquiterpenes, Eudesmane; Sodium; Terpenes

Nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) are the major mediators produced in activated macrophages which contribute to the circulatory failure associated with septic shock. An activity-guided fractionation of an MeOH extract of stem bark of Magnolia obovata afforded two inhibitors of NO production in lipopolysaccharides (LPS)-activated macrophages by the suppression of i-NOS expression. Their structures were elucidated by spectroscopic methods to be magnolol and honokiol with IC50 values of 16.8 and 6.4 microM, respectively. They also inhibited the production of TNF-alpha in LPS-activated macrophages. Thus, these compounds may be possible candidates for the development of new drugs to treat endotoxemia accompanied by the overproduction of NO and TNF-alpha.

Topics: Animals; Biphenyl Compounds; Blotting, Western; Cell Line; Enzyme Inhibitors; Lignans; Lipopolysaccharides; Macrophage Activation; Macrophages; Magnetic Resonance Spectroscopy; Magnoliopsida; Mice; Nitric Oxide; Nitric Oxide Synthase; Tumor Necrosis Factor-alpha

Honokiol and magnolol, phenolic compounds isolated from the stem bark of Magnolia officinalis, have been demonstrated to increase choline acetyltransferase activity, inhibit acetylcholinesterase, promote potassium-induced acetylcholine release and exhibit neurotrophic function in in vitro studies. The objective of the present study was to determine the effect of these compounds on hippocampal acetylcholine release in conscious, freely-moving rats. 10(-4) M-10(-6) M of honokiol or magnolol was perfused into rat hippocampus via a dialysis probe. The results showed that at 10(-4) M concentration, honokiol and magnolol markedly increased extracellular acetylcholine release to 165.5+/-5.78% and 237.83+/-9.47% of the basal level, respectively. However, lower concentrations of either compounds failed to elicit significant acetylcholine release. This result suggests that a high dose of honokiol or magnolol may enhance in vivo hippocampal acetylcholine release.

Topics: Acetylcholine; Animals; Biphenyl Compounds; Drugs, Chinese Herbal; Hippocampus; Lignans; Male; Rats; Rats, Sprague-Dawley

Honokiol has previously been shown to be an effective anxiolytic-like agent in mice when administered for 7 days at 0.2 mg/kg/day prior to evaluation in an elevated plus-maze, while 20 mg/kg is required for efficacy as a single oral dose. The aim of this study was to find analogs of honokiol that are more effective for acute administration. Among the eight analogs evaluated, one partially reduced derivative of honokiol [3'-(2-propenyl)-5-propyl-(1,1'-biphenyl)-2,4'-diol] exhibited significant anxiolytic-like activity at 0.04 mg/kg. Following oral administration of 1 mg/kg of this analog, anxiolytic-like activity was clearly evident at 1 h, peaked at 3 h, and remained significant for longer than 4 h after treatment. Combined administration of the derivative with diazepam led to enhanced anxiolytic-like efficacy. Moreover, as with diazepam, the anxiolytic-like effect of the analog was reduced by flumazenil. In contrast, bicuculline, a GABA(A) antagonist, had no effect on the activity of the derivative. Taken together, these results suggest that this analog of honokiol acts at the benzodiazepine recognition site of the GABA(A)-benzodiazepine receptor complex.

Topics: Animals; Anti-Anxiety Agents; Anxiety; Biphenyl Compounds; Diazepam; Drug Evaluation, Preclinical; Drug Therapy, Combination; Lignans; Male; Mice

An improved elevated plus-maze test in mice revealed that seven daily treatments with two different traditional Chinese medicines, known as Kampo medicines in Japan, Hange-koboku-to (composed of extracts of 5 plants) and Saiboku-to (composed of extracts of 10 plants), produced an anxiolytic effect, and the effect was mainly due to the presence of honokiol derived from magnolia. This study was carried out to evaluate the anxiolytic potential of honokiol, Hange-koboku-to and Saiboku-to, which were prescribed with two different magnolia samples: Kara-koboku (Magnoliae officinalis) (KA) or Wa-koboku (Magnoliae obovata) (WA). The doses of test samples were adjusted to ensure a constant dose of honokiol at 0.2 mg kg(-1). Although the doses of magnolol (an isomer of honokiol), as well as those of undetermined chemicals, varied among samples, the seven daily treatments with 9 out of 10 test samples produced an anxiolytic effect almost equivalent to that produced by 0.2 mg kg(-1) honokiol. The only exception was the sample containing the lowest amount of honokiol. Magnolia-free preparations of Hange-koboku-to or Saiboku-to did not have any anxiolytic effect. These results confirm that honokiol derived from magnolia is the causal chemical of the anxiolytic effect of Hange-koboku-to and Saiboku-to.

Topics: Animals; Anti-Anxiety Agents; Behavior, Animal; Biphenyl Compounds; Dose-Response Relationship, Drug; Lignans; Male; Maze Learning; Mice; Plant Extracts

Contents of magnolol and honokiol in 76 samples of Magnolia officinalis collected from 11 counties in Zhejiang, Fujian, Sichuan, Guangxi, Hunan, Jiangxi and Hubei are analyzed by means of HPLC. A study on the effect of tree age on effective ingredients in Magnolia offcinalis shows that the influence of tree age on the content of magnolol and honokiol is correlated with the varieties under cultivation, but a small effect on the content of the phenols with an indent at the tip of leaves. The content of magnolol in other types of leaves increases rapidly with increase in age, diameter of the trunk and thickness of the bark, with not many changes found at the age of 12 or up. Increase in age may be favorable to the full expression of oily characteristic. These results provide a scientific base for the determination of optimal time for harvesting bark from artificially established Magnolia officinalis stands.

Topics: Age Factors; Biphenyl Compounds; China; Chromatography, High Pressure Liquid; Lignans; Magnolia; Plant Bark; Plant Leaves; Plants, Medicinal

Use of the elevated plus-maze experiment and activity and traction tests in mice have revealed that seven daily treatments with 0.2 mg kg(-1) and higher doses of honokiol, a neolignane derivative extracted from Magnolia bark, had an anxiolytic effect without change in motor activity or muscle tone. Diazepam, 1 mg kg(-1), had the same anxiolytic potential as 0.2 mg kg(-1) honokiol but induced muscle relaxation. The aim of this study was to determine whether honokiol had diazepam-like side-effects. Mice treated with 1-10 mg kg(-1) diazepam, but not those treated with 0.1-2 mg kg(-1) honokiol, for 12 days showed withdrawal symptoms characterized by hyperactivity and running-fit when they were challenge-administered intraperitoneal flumazenil (10 mg kg(-1)) 24 h after the last treatment with diazepam. Oral diazepam (0.5-2 mg kg(-1), 10 min before) dose-dependently prolonged hexobarbital (100 mg kg(-1), i.p.)-induced sleeping, disrupted learning and memory, and inhibited (+)-bicuculline (40 mg kg(-1), i.p.)-induced death. Honokiol (0.2-20 mg kg(-1), p.o., 3 h before) had no such effects. The prolongation by diazepam (1 mg kg(-1)) of hexobarbital-induced sleeping was not modified by honokiol (0.2-20 mg kg(-1)). These results suggest that honokiol is less likely than diazepam to induce physical dependence, central depression and amnesia at doses eliciting the anxiolytic effect. It is also considered that honokiol might have no therapeutic effect in the treatment of convulsion.

Topics: Animals; Anti-Anxiety Agents; Behavior, Animal; Bicuculline; Biphenyl Compounds; Diazepam; Hexobarbital; Learning; Lignans; Male; Memory; Mice; Mice, Inbred BALB C; Sleep; Substance-Related Disorders

This study was designed to evaluate the in vivo effect of magnolol and honokiol on the acute phase of coronary ligation in the presence of nitric oxide inhibitor (L-NAME) or cyclooxygenase inhibitor (aspirin). After Sprague-Dawley rats were anesthetized with urethane, the changes of ventricular arrhythmia induced by coronary ligation for 30 min were determined with or without pretreatment with study medications. The incidence and duration of ventricular arrhythmia were significantly reduced after intravenous pretreatment (15 min before coronary ligation) with 10(-7) g/kg magnolol or 10(-7) g/kg honokiol. However, the antiarrhythmic effect of magnolol or honokiol could be abolished with the pretreatment of 1 mg/kg L-NAME, but not with pretreatment of 100 mg/kg aspirin. The abolishment of the myocardial beneficial effect of magnolol and honokiol by L-NAME, instead of aspirin, suggests an involvement of an increased nitric oxide synthesis in the protection offered by magnolol and honokiol against arrhythmia during myocardial ischemia.

Topics: Anesthesia; Animals; Anti-Arrhythmia Agents; Anti-Inflammatory Agents, Non-Steroidal; Arrhythmias, Cardiac; Aspirin; Biphenyl Compounds; Blood Pressure; Coronary Disease; Enzyme Inhibitors; Lignans; Male; NG-Nitroarginine Methyl Ester; Nitric Oxide Synthase; Rats; Rats, Sprague-Dawley

In our previous study using an improved elevated plus-maze in mice, the oriental herbal medicine Saiboku-to prolonged the time spent in open arms, showing an anxiolytic effect, and the effect was mainly caused by honokiol derived from magnolia. This study was carried out to compare the anxiolytic potentials of honokiol and water extracts of three magnolia samples; two being Kara-koboku (Magnolia officinalis) (KA: from Zhejiang-sheng, China; honokiol 0.25% and magnolol 1.16%, and KB: from Sichuan-sheng, China; honokiol 1.72% and magnolol 1.71%), and one being Wa-koboku (Magnolia obovata) (WA: from Iwate-ken, Japan; honokiol 0.32% and magnolol 0.81%). Seven daily treatments with 0.1-1 mg/kg honokiol, but not 0.2 and 1 mg/kg magnolol, revealed an anxiolytic effect with the peak potential at 0. 2 mg/kg. The anxiolytic potentials of 40 and 80 mg/kg KA, which contained the highest amount of magnolol, were almost equivalent to those of 0.1 and 0.2 mg/kg honokiol, respectively. KB, at 11.6 mg/kg, and 62.5 mg/kg WA resulted in almost the same anxiolytic potential as that of 0.2 mg/kg honokiol. No significant change in the ambulatory activity was produced by any drug treatment. These results suggest that honokiol is the chemical responsible for the anxiolytic effect of the water extract of magnolia and that the other chemicals including magnolol in magnolia scarcely influence the effect of honokiol. It is also considered that the elevated plus-maze test is applicable for evaluation of the content of honokiol in magnolia.

Topics: Animals; Biphenyl Compounds; Lignans; Male; Maze Learning; Mice; Plant Extracts; Plants, Medicinal; Water

1. The bark of the root and stem of various Magnolia species has been used in Traditional Chinese Medicine to treat a variety of disorders including anxiety and nervous disturbances. The biphenolic compounds honokiol (H) and magnolol (M), the main components of the Chinese medicinal plant Magnolia officinalis, interact with GABA(A) receptors in rat brain in vitro. We compared the effects of H and M on [3H]muscimol (MUS) and [3H]flunitrazepam (FNM) binding using EDTA/water dialyzed rat brain membranes in a buffer containing 150 mM NaCl plus 5 mM Tris-HCl, pH 7.5 as well as [35S]t-butylbicyclophosphorothionate (TBPS) in 200 mM KBr plus 5 mM Tris-HCl, pH 7.5. H and M had similar enhancing effects on [3H]MUS as well as on [3H]FNM binding to rat brain membrane preparations, but H was 2.5 to 5.2 times more potent than M. 2. [3H]FNM binding. GABA alone almost doubled [3H]FNM binding with EC50 = 450 nM and 200 nM using forebrain and cerebellar membranes, respectively. In the presence of 5 microM H or M the EC50 values for GABA were decreased to 79 and 89 nM, respectively, using forebrain, and 39 and 78 nM, using cerebellar membranes. H and M potently enhanced the potentiating effect of 200 nM GABA on [3H]FNM binding with EC50 values of 0.61 microM and 1.6 microM using forebrain membranes, with maximal enhancements of 33 and 47%, respectively. Using cerebellar membranes, the corresponding values were 0.25 and 1.1 microM, and 22 and 34%. 3. [3H]MUS binding. H and M increased [3H]MUS binding to whole forebrain membranes about 3-fold with EC50 values of 6.0 and 15 microM. Using cerebellar membranes, H and M increased [3H]MUS binding approximately 68% with EC50 values of 2.3 and 12 microM, respectively. Scatchard analysis revealed that the enhancements of [3H]MUS binding were due primarily to increases in the number of binding sites (Bmax values) with no effect on the high affinity binding constants (Kd values). The enhancing effect of H and M were not additive. 4. [35S]TBPS binding. H and M displaced [35S]TBPS binding from sites on whole rat forebrain membranes with IC50 values of 7.8 and 6.0 microM, respectively. Using cerebellar membranes, the corresponding IC50 values were 5.3 and 4.8 microM. These inhibitory effects were reversed by the potent GABA(A) receptor blocker R5135 (10 nM), suggesting that H and M allosterically increase the affinity of GABA(A) receptors for GABA and MUS by binding to sites in GABA(A) receptor complexes. 5. Two monophenols, the anestheti

Topics: Allosteric Regulation; Androstanes; Animals; Azasteroids; Binding Sites; Biphenyl Compounds; Cell Membrane; Central Nervous System Depressants; Cerebellum; Diflunisal; Drugs, Chinese Herbal; Flunitrazepam; GABA Antagonists; gamma-Aminobutyric Acid; Kinetics; Lignans; Muscimol; Picrotoxin; Propofol; Prosencephalon; Rats; Receptors, GABA-A; Tritium

An analysis of effective ingredients in 20 to 29 year old Magnolia officinalis of three different leaf types coming from various sources shows that the content of phenols differ extremely significantly in various leaf types which are a key to determination of quality of Magnolia officinalis also varies significantly among different sources which only have a significant effect on the content of magnolol. Varieties and types cultivated in different places are responsible for the variation in content of magnolol. The conclusions obtained from this study coincide with the traditional insight into quality of Magnolia officinalis.

Topics: Biphenyl Compounds; China; Chromatography, High Pressure Liquid; Lignans; Magnolia; Plant Leaves; Plants, Medicinal; Quality Control; Species Specificity

The principal active anxiolytic components in Saiboku-to, an Oriental herbal medicine, have been isolated and identified as magnolol (5,5'-di-2-propenyl-1,1'-biphenyl-2,2'-diol) and honokiol (3',5-di-2-propenyl-1,1'-biphenyl-2,4'-diol). Evaluation by means of an elevated plus-maze test showed that honokiol was at least 5000 times more potent than Saiboku-to when mice were treated orally for 7 days.

Topics: Animals; Anti-Anxiety Agents; Anxiety; Biphenyl Compounds; Drugs, Chinese Herbal; Lignans; Male; Medicine, Kampo; Mice; Mice, Inbred Strains; Plant Extracts

Magnolol (1) and honokiol (2), main compounds from the stem bark of Magnolia obovata Thunb., were evaluated for an antimicrobial activity against periodontopathic microorganisms, Porphyromonas gingivalis, Prevotella gingivalis, Actinobacillus actinomycetemcomitans, Capnocytophaga gingivalis, and Veillonella disper, and a cytotoxicity against human gingival fibroblasts and epithelial cells. Our results indicate that magnolol and honokiol, although less potent than chlorhexidine, show a significant antimicrobial activity against these microorganisms, and a relatively low cytotoxic effect on human gingival cells. Thus, it is suggested that magnolol and honokiol may have a potential therapeutic use as a safe oral antiseptic for the prevention and the treatment of periodontal disease.

Topics: Aggregatibacter actinomycetemcomitans; Anti-Bacterial Agents; Biphenyl Compounds; Capnocytophaga; Cells, Cultured; Epithelial Cells; Fibroblasts; Gingiva; Humans; Lignans; Porphyromonas gingivalis; Prevotella; Veillonella

Honokiol, a neolignane derivative of Magnolia bark, has central depressant action and, at much lower doses, anxiolytic activity. We have investigated the characteristics of the behavioural effects of honokiol by means of an elevated plus-maze test. In the plus-maze test a single oral dose of 20 mg kg(-1) honokiol significantly prolonged the time spent in the open arms of the maze, suggesting anxiolytic effect. Moreover, when honokiol was administered daily for seven days and the plus-maze test was conducted 3 or 24 h after the last administration, significant prolongation of the time in the open arms was manifested even for doses of 0.2 mg kg(-1). The maximum effect was observed for doses of 0.5 mg kg(-1). Honokiol at any dose in both single and repeated administration schedules caused neither change in motor activity nor disruption of traction performance. Orally administered diazepam, 0.5-2 mg kg(-1), caused dose-dependent prolongation of the time spent in the open arms of the maze with a significant increase in motor activity at 1 mg kg(-1), and dose-dependent disruption of traction performance. The changes in the plus-maze performance after treatment for seven days with 0.2 mg kg(-1) honokiol and after a single treatment with 1 mg kg(-1) diazepam were almost equivalent. The effect of honokiol (0.2 mg kg(-1), treatment for seven days) was inhibited by subcutaneous flumazenil (0.3 mg kg(-1)) and (+)-bicuculline (0.1 mg kg(-1)) and by intraperitoneal CCK-4 (50 microg kg(-1)) and caffeine (30 mg kg(-1)). The anxiolytic effect of diazepam (1 mg kg(-1)) was also inhibited by flumazenil and bicuculline. However, the combined administration of diazepam with caffeine enhanced the effect, and diazepam completely reversed the effect of CCK-4. These results suggest that, in contrast with diazepam, honokiol selectively induces an anxiolytic effect with less liability of eliciting motor dysfunction and sedation or disinhibition. The combined effects of the drug also revealed that the mechanism of anxiolytic effect of honokiol is partially different from that of diazepam.

Topics: Administration, Oral; Animals; Anti-Anxiety Agents; Bicuculline; Biphenyl Compounds; Caffeine; Central Nervous System Depressants; Central Nervous System Stimulants; Diazepam; Dose-Response Relationship, Drug; Flumazenil; GABA Antagonists; GABA Modulators; Injections, Subcutaneous; Lignans; Male; Maze Learning; Mice; Mice, Inbred BALB C; Motor Activity; Tetragastrin; Therapeutic Equivalency

The exposure of human lymphoid leukemia Molt 4B cells to honokiol led to both growth inhibition and the induction of apoptosis. Morphological change showing apoptotic bodies was observed in the cells treated with honokiol. The fragmentation by honokiol of DNA to oligonucleosomal-sized fragments that are characteristics of apoptosis was observed to be concentration- and time-dependent. These findings suggest that growth inhibition by honokiol of Molt 4B cells results from the induction of apoptosis in the cells.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Biphenyl Compounds; Cell Division; Dose-Response Relationship, Drug; Humans; Leukemia, Lymphoid; Lignans; Tumor Cells, Cultured

The Rhizoma Atractylodis, Cortex Magnoliae Officinalis and Pericarpium Citri Reticulatae in Huoxiang-zhengqi Liquid were identified by TLC. The total contents of magnolol and honokiol were determined by HPLC to be no less than 1.9 mg/ml, with an average recovery of 104.4% and 103.3%, RSD 1.23% and 0.91% respectively.

Topics: Asteraceae; Biphenyl Compounds; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Citrus; Drug Combinations; Drugs, Chinese Herbal; Lignans; Magnoliopsida; Quality Control

Honokiol and magnolol, neolignans in Magnolia obovata, have been evaluated as antioxidants. Microsomal lipid peroxidation induced by Fe(III)-ADP/NADPH and mitochondrial lipid peroxidation induced by Fe(III)-ADP/NADH were inhibited by these compounds. These neolignans protected mitochondrial respiratory chain enzyme activity against NADPH-induced peroxidative stress and protected red cells against oxidative haemolysis. The anti-oxidative activity of honokiol was more potent than that of magnolol. Neolignans in M. obovata were shown to be effective in protecting biological systems and functions against oxidative stress.

Topics: Animals; Antioxidants; Biphenyl Compounds; Drugs, Chinese Herbal; Erythrocytes; Hemolysis; Lignans; Lipid Peroxidation; Male; Microsomes, Liver; Mitochondria, Liver; Rats; Rats, Wistar; Trees

Honokiol, a compound extracted from the Chinese medicinal herb Magnolia officinalis, has a strong antioxidant effect on the inhibition of lipid peroxidation in rat heart mitochondria. To investigate the protective effect of honokiol on hepatocytes from peroxidative injury, oxygen consumption and malondialdehyde formation for in vitro iron-induced lipid peroxidation were assayed, and the mitochondrial respiratory function for in vivo ischemia-reperfusion injury were evaluated in rat liver, respectively. The inhibitory effect of honokiol on oxygen consumption and malondialdehyde formation during iron-induced lipid peroxidation in liver mitochondria showed obvious dose-dependent responses with a concentration of 50% inhibition being 2.3 x 10(-7) M and 4.96 x 10(-7) M, respectively, that is, 550 times and 680 times more potent than alpha-tocopherol, respectively. When rat livers were introduced with ischemia 60 min followed by reperfusion for 60 min, and then pretreated with honokiol (10 micrograms/kg BW), the mitochondrial respiratory control ratio (the quotient of the respiration rate of State 3 to that of State 4) and ADP/O ratio from the honokiol-treated livers were significantly higher than those of non-treated livers during reperfusion. The dose-dependent protective effect of honokiol on ischemia-reperfusion injury was 10 microgram-100 micrograms/Kg body weight. We conclude that honokiol is a strong antioxidant and shed insight into clinical implications for protection of hepatocytes from ischemia-reperfusion injury.

Topics: Animals; Biphenyl Compounds; Gastrointestinal Agents; Lignans; Lipid Peroxidation; Liver; Liver Diseases; Male; Malondialdehyde; Mitochondria, Liver; Oxidation-Reduction; Phosphorylation; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Vitamin E

The effects of honokiol, a diphenyl compound extracted from a Chinese herbal medicine, on leukotriene (LT) synthesis were evaluated in rat basophilic leukemia (RBL) cells. The production of LTC4 and LTB4 stimulated by the Ca2+ ionophore A23187 was measured in RBL-1 cells by high-performance liquid chromatography. Honokiol inhibited the production of LTC4 and LTB4 stimulated by A23187 in RBL-1 cells. Honokiol did not inhibit either phospholipase A2 activity, measured by the release of 3H-arachidonic acid (AA), or LTC4 synthase and LTA4 hydrolase activities, measured with LTA4-free acid as substrate. The synthesis of LTC4 and LTB4 from AA in RBL-1 cell lysates in the presence of Ca2+ was inhibited by honokiol. These results indicate that honokiol blocks LT synthesis by inhibiting 5-lipoxygenase activity. Honokiol also inhibited immunoglobulin E-mediated production of these LTs in RBL-2H3 cells, which was measured by a specific radioimmunoassay (RIA). These results suggest that honokiol may exhibit antiallergic actions by inhibiting LT synthesis in immediate-type hyperreactivity.

Topics: Animals; Anti-Allergic Agents; Biphenyl Compounds; Leukemia, Basophilic, Acute; Leukotriene Antagonists; Leukotrienes; Lignans; Rats; Tumor Cells, Cultured

Honokiol is an active component of Magnolia officinalis. It was reported to be 1000 times more potent than alpha-tocopherol in inhibiting lipid peroxidation in rat heart mitochondria. In this study, we investigated the in vivo antiarrhythmic and antiischemic effects of honokiol in coronary ligated rats. Male Sprague-Dawley rats were anesthetized with urethane. Honokiol, at dosages of 10(-7) g/kg, 10(-8) g/kg, and 10(-9) g/kg, was administered intravenously 15 min before ligation of the coronary artery. Incidence and duration of ventricular tachycardia and ventricular fibrillation during 30 min coronary ligation were significantly reduced by 10(-7) g/kg honokiol. Ventricular arrhythmia during 10 min reperfusion after the relief of coronary ligation was also reduced. In rats subjected to 4 hours coronary ligation, 10(-7) g/kg, 10(-8) g/kg, and 10(-9) g/kg honokiol significantly reduced the infarct zone. We concluded that honokiol may protect the myocardium against ischemic injury and suppress ventricular arrhythmia during ischemia and reperfusion.

Topics: Animals; Anti-Arrhythmia Agents; Biphenyl Compounds; Drugs, Chinese Herbal; Heart; Lignans; Male; Myocardial Infarction; Rats; Rats, Sprague-Dawley

To study the possible mechanism through which honokiol and magnolol elicit their central depressant effects, we examined the influence of these two phenolic compounds on 25 mM K(+)-stimulated release of [3H]acetylcholine (ACh) from the rat's hippocampal slices. Honokiol, but not magnolol, elicited a concentration-dependent enhancement of K+-evoked ACh release. Addition of either tetrodotoxin, pilocarpine, or methoctramine had no effect on honokiol-enhanced ACh release. These results suggest that honokiol enhanced K(+)-evoked ACh release directly on hippocampal cholinergic terminals via receptors other than the M2 cholinergic subtypes.

Topics: Acetylcholine; Animals; Biphenyl Compounds; Central Nervous System Depressants; Hippocampus; In Vitro Techniques; Lignans; Rats; Rats, Sprague-Dawley

Topics: Animals; Biphenyl Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Glucuronidase; Lignans; Male; Rats; Rats, Sprague-Dawley; Spectrometry, Fluorescence

A HPLC method for the determination of magnolol and honokiol in Baoji Pill has been established. The method can be used to control the quality of Boji Pill.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Drug Combinations; Drugs, Chinese Herbal; Gastrointestinal Agents; Lignans; Quality Control

To identify the inhibitor of prednisolone metabolism contained in Saiboku-To, we conducted in-vitro experiments of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD), using rat liver homogenate and cortisol as a typical substrate. We studied the effects of ten herbal constituents on 11 beta-HSD. Five herbal extracts showed inhibitory activity with Glycyrrhiza glabra > Perillae frutescens > Zizyphus vulgaris > Magnolia officinalis > Scutellaria baicalensis. This suggests that unknown 11 beta-HSD inhibitors are contained in four herbs other than G. glabra which contains a known inhibitor, glycyrrhizin (and glycyrrhetinic acid). Seven chemical constituents which have been identified as the major urinary products of Saiboku-To in healthy and asthmatic subjects were studied; magnolol derived from M. officinalis showed the most potent inhibition of the enzyme (IC50, 1.8 x 10(-4) M). Although this activity was less than that of glycyrrhizin, the inhibition mechanism (non-competitive) was different from a known competitive mechanism. These results suggest that magnolol might contribute to the inhibitory effects of Saiboku-To on prednisolone metabolism through inhibition of 11 beta-HSD.

Topics: 11-beta-Hydroxysteroid Dehydrogenases; Animals; Antineoplastic Agents, Phytogenic; Asthma; Biphenyl Compounds; Chromatography, High Pressure Liquid; Cortisone; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Glucocorticoids; Glycyrrhetinic Acid; Glycyrrhizic Acid; Hydrocortisone; Hydroxysteroid Dehydrogenases; Immunosuppressive Agents; Lignans; Liver; Male; Medicine, Kampo; Rats; Rats, Wistar

A simple and sensitive high-performance liquid chromatographic method for the identification and determination of honokiol in rat plasma has been developed. Up to 0.1 ml of plasma containing honokiol was deproteinized with acetonitrile, which contained an internal standard (paeonol). The supernatant was injected onto a reversed-phase C18 column using acetonitrile-water (70:30, v/v, adjusted to pH 2.5-2.8 with orthophosphoric acid) as the mobile phase and ultraviolet detection at 290 nm, followed by UV spectrum identification (between 220 and 380 nm) with a photodiode-array detector. The method was applied to pharmacokinetic studies of honokiol in rat following 5 or 10 mg/kg intravenous administration. A biphasic process consisting of a rapid distribution phase followed by a slower elimination phase was observed from the plasma concentration-time curves. Compartmental analysis yielded a two-compartment model.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biphenyl Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Hydrogen-Ion Concentration; Indicators and Reagents; Injections, Intravenous; Lignans; Male; Models, Biological; Rats; Rats, Sprague-Dawley; Spectrophotometry, Ultraviolet

In isolated rat heart mitochondria lipid peroxidation was induced with ADP and ferrous sulfate (FeSO4). Oxygen consumption and malondialdehyde (MDA) production were measured to quantitate lipid peroxidation. The antioxidant effects of magnolol and honokiol purified from Magnolia officinalis were 1000 times higher than that of alpha-tocopherol. The IC50 values of magnolol and honokiol for inhibition of oxygen consumption were 8.0 x 10(-8) M and 1.0 x 10(-7) M, respectively, while that of alpha-tocopherol was 1.0 x 10(-4) M. Magnolol at 0.5 microM inhibited 71.4 +/- 9.4% of oxygen consumption and 59.3 +/- 4.6% MDA production. At the same concentration, honokiol inhibited 78.1 +/- 4.7% of oxygen consumption and 86.9 +/- 4.0% of MDA production. Of conjugated diene formation 48.4 +/- 4.6% and 53.1 +/- 3.4% were inhibited by 0.5 microM magnolol and honokiol, respectively. Also both magnolol and honokiol exhibited free radical scavenging activities as shown by the diphenyl-p-picrylhydrazyl assay, but they were less potent than alpha-tocopherol.

Topics: Animals; Antioxidants; Biphenyl Compounds; Lignans; Lipid Peroxidation; Male; Malondialdehyde; Mitochondria, Heart; Oxygen Consumption; Phytotherapy; Plants, Medicinal; Rats; Rats, Sprague-Dawley; Vitamin E

Magnolol and honokiol, biphenyl compounds, were isolated as anti-emetic principles from the methanolic extract of Magnolia obovata bark. [6]-, [8]-, and [10]-shogaols and [6]-, [8]-, and [10]-gingerols were isolated from the methanolic extract of Zingiber officinale rhizome as anti-emetic principles. Some phenyl-propanoids with allyl side-chains were found to show the same activity. They inhibited the emetic action induced by the oral administration of copper sulfate pentahydrate to leopard and ranid frogs.

Topics: Animals; Antiemetics; Anura; Biphenyl Compounds; Catechols; Fatty Alcohols; Female; Lignans; Male; Plants, Medicinal

A method was developed for determining the activity of acetyl-CoA:1-alkyl-sn-glycero-3-phosphocholine acetyltransferase (EC 2.3.1.67), a key enzyme in the biosynthesis of platelet-activating factor (PAF, 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine). The assay involves measurement of the radioactivity in the trichloroacetic acid (TCA)-precipitated complex of radioactive product and albumin after incubation of 1-alkyl-sn-glycero-3-phosphocholine and [3H]acetyl-CoA with rat spleen microsomes or membrane fractions of human polymorphonuclear leukocytes (PMNs). The radioactive product associated with the precipitate was identified as PAF using an ultrahigh-sensitivity TV camera system after extraction and separation by TLC. This TCA method was then used to screen the components of crude preparations that inhibited acetyltransferase activity. Major components from the cortex of Magnoliae (magnolol and honokiol), which have anti-inflammatory and anti-bacterial actions, inhibited the acetyltransferase activity in rat spleen microsomes (IC50, 150 and 150 microM, respectively) and membrane fractions of human PMNs (IC50, 70 and 60 microM, respectively). The inhibitory action of magnolol and honokiol was reversible, and similar to or higher than that of nordihydroguaiaretic acid. PAF production in human PMNs stimulated by the ionophore A23187 was also suppressed dose dependently by magnolol and honokiol. These activities may be relevant to the claimed therapeutic effects of the extract from Magnoliae cortex.

Topics: Acetyltransferases; Animals; Biphenyl Compounds; Humans; In Vitro Techniques; Lignans; Male; Plant Extracts; Rats; Rats, Wistar; Reproducibility of Results

A simple method using ion-pair high-performance liquid chromatography was established for the rapid and precise determination of honokiol(3',5-di-2-propenyl-1,1'-biphenyl-2,4'-diol) and magnolol(5,5'-di-2-propenyl-1,1'-biphenyl-2,2'-diol) in eighteen species of oriental pharmaceutical decoctions containing Magnolia bark. An ODS column and a mixed solvent system of water involving 10 mM tetra-n-amyl-ammonium bromide (TAA) and acetonitrile (4:6) as a mobile phase were used for the separation. Honokiol and magnolol were eluted without interference of other coexisting components within 12 min.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Lignans

A simple method using ion-pair high-performance liquid chromatography was established for the rapid and precise simultaneous determination of honokiol (3', 5-di-2-propenyl-1, 1'-biphenyl-2,4'-diol) and magnolol (5,5'-di-2-propenyl-1,1'-biphenyl-2,2'-diol) in oriental pharmaceutical decoctions containing Magnolia bark. An ODS column and a mixture of water involving 10 mM tetra-n-amylammonium bromide (TAA) and acetonitrile (4:6) as a mobile phase were used for the separation. Honokiol and magnolol were eluted without interference of other co-existing components within 12 min.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Lignans

HPLC was applied to determine and compare 17 samples of Houpo processed in different ways: sauted with ginger juice, dipped with ginger juice, boiled with ginger juice, and so on. The results provide a scientific basis for screening the best processing technology of Houpo.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Hot Temperature; Lignans; Technology, Pharmaceutical

Based on the method of assessing the growing age of trees by "annual ring in the bark", the active ingredients magnolol and honokiol in 3 species Houpo (Magnolia officinalis, M. biloba, M. rostrata) of different ages, were determined by HPLC. The results present a reference for defining the reasonable period of collection and quality evaluation of Houpo.

Topics: Biphenyl Compounds; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Lignans; Plants, Medicinal; Quality Control; Species Specificity; Time Factors

This paper adds TLC identification of binglang and other three cruds drugs to the quality control of kakxiong shunqi pills incorporated in Chinese Pharmacopoeia. The contents of magnolol and honokinol were determined by TLC scanning. The method is simple, rapid accurate, and useful in controlling the quality of kaixiong shunqi pills.

Topics: Biphenyl Compounds; Chromatography, Thin Layer; Densitometry; Drug Combinations; Drugs, Chinese Herbal; Lignans; Quality Control

Topics: Biphenyl Compounds; Lignans; Oils, Volatile; Pharmacognosy; Plants, Medicinal; Trees

Data from a survey of the drug market and investigation of the original plant of "Tu-hou-po", after careful botanical examinations, showed that the drugs were derived from 5 species of the genus Manglietia of Magnoliaceae, viz. Manglietia chingii Dandy, M. insignis (Wall.) Bl., M. duclouxii Finet et Gagnep., M. yuyuanensis Law and M. szechuanica Hu. Comparisons of the main characteristics of the plants, Tu-hou-po and Hou-po crude drugs and chemical components showed that Manglietia is taxonomically the closest to Magnolia and contained similar components (tab 1-2 and fig 1). The results of HPLC analysis demonstrated that they contained magnolol, honokiol, magnocurine and salicifoline, in different quantities. However, no magnosprengerine was detected. Besides, it was also found that the percentage of magnolol and honokiol contents were higher, while that of magnocurine was lower in Hou-po. On the contrary, the content of magnocurine was higher, while that of magnolol and honokiol were lower in Tu-hou-po. Manglietia chingii (Tu-hou-po) is being used as the Chinese traditional drug "Hou-po" in the clinic in Guangxi. Therefore, M. chingii is noteworthily exploited as a new resource of Hou-po for further research.

Topics: Biphenyl Compounds; Drugs, Chinese Herbal; Isoquinolines; Lignans; Neuromuscular Nondepolarizing Agents; Plants, Medicinal

This study reports on the connexion of the content of the efficacious constituents, magnolol and honokiol of Magnolia officinalis var. biloba with age, diameter of the trunk and various parts of the tree, thus presenting a basis for its collection, quality control and clinical applications.

Topics: Biphenyl Compounds; Lignans; Plants, Medicinal; Quality Control; Time Factors

The molecular mechanism of honokiol, extracted from the bark of Magnolia obovata, was studied using bovine adrenal chromaffin cells as a model system. Honokiol inhibits catecholamine secretion induced by carbachol and DMPP and that induced by exposure to high K+ and Ba2+ but to a lesser extent. The inhibitory effects of trifluoperazine and honokiol on carbachol-, high K(+)- and Ba2(+)- induced secretion were not additive. The results suggest that honokiol interferes with the interaction between the acetylcholine receptor and its agonists and that honokiol may also affect the steps in exocytosis after intracellular calcium has been raised, possibly at the site(s) where calmodulin acts.

Topics: Adrenal Medulla; Animals; Barium; Biphenyl Compounds; Calcium; Carbachol; Catecholamines; Cattle; Cells, Cultured; Chromaffin System; Digitonin; Dimethylphenylpiperazinium Iodide; Exocytosis; Lignans; Potassium; Receptors, Cholinergic; Trifluoperazine

Magnolol and honokiol are two position isomers isolated from the bark of Magnolia officinalis. Both inhibited the aggregation and ATP release of rabbit platelet-rich plasma induced by collagen and arachidonic acid without affecting that induced by ADP, PAF or thrombin. Aggregation of washed platelets was more markedly inhibited than that of platelet-rich plasma, while the aggregation of whole blood was least affected by both inhibitors. Thromboxane B2 formation caused by collagen, arachidonic acid or thrombin was in each case inhibited by magnolol and honokiol. The rise of intracellular calcium caused by arachidonic acid or collagen was also suppressed by both agents. Collagen-induced intracellular calcium increase in the presence of indomethacin was suppressed by magnolol. It is concluded that the antiplatelet effect of magnolol and honokiol is due to an inhibitory effect on thromboxane formation and also an inhibition of intracellular calcium mobilization.

Topics: Animals; Biphenyl Compounds; Calcium; In Vitro Techniques; Lignans; Medicine, Chinese Traditional; Platelet Aggregation Inhibitors; Platelet Function Tests; Rabbits; Thromboxane B2

Topics: Animals; Aorta, Thoracic; Biphenyl Compounds; Calcium Channel Blockers; In Vitro Techniques; Lignans; Male; Medicine, Chinese Traditional; Medicine, East Asian Traditional; Muscle Contraction; Muscle, Smooth, Vascular; Plants, Medicinal; Rats; Rats, Inbred Strains

Topics: Biphenyl Compounds; Hot Temperature; Lignans; Medicine, Chinese Traditional; Medicine, East Asian Traditional; Plants, Medicinal; Tissue Distribution

Topics: Biphenyl Compounds; China; Lignans; Plants, Medicinal

Topics: Animals; Biphenyl Compounds; Central Nervous System Depressants; Chickens; Lignans; Male; Mice; Rats; Rats, Inbred Strains

Effects of some diphenyl and monophenyl compounds on grip strength in mice and spinal reflexes in young chicks were investigated in order to study structure-activity relationships between muscle relaxant activity and neolignane compounds, magnolol and honokiol extracted from Magnolia officinalis THUNB. Diphenyl produced a long-lasting suppression in the spinal reflex and relatively weak inhibition in the grip strength. An introduction of a hydroxyl into 2-position of diphenyl, o-phenylphenol, increased the muscle relaxant activity and accelerated the onset, although the duration was still long. In the spinal reflex preparation the duration of action became short. The introduction of two hydroxyls into 2- and 2'-position of diphenyl, 2,2'-dihydroxydiphenyl, further strengthened the activity and shortened the duration of the inhibitory effect on the grip strength and the spinal reflex. When two allyls are introduced into 5,5'-position of 2,2'-dihydroxydiphenyl, it corresponds to magnolol. Magnolol, 5,5'-diallyl-2,2'-dihydroxydiphenyl, produced potent inhibitory effects of gradual onset and of long duration on the two test preparations. Position of allyls and hydroxyls in honokiol, 5,3'-diallyl-2,4'-dihydroxydiphenyl, is different from magnolol, although the pharmacological characteristics are quite similar to magnolol. These results suggest that a hydroxyl accelerates the onset and shortens the muscle relaxant activity of diphenyl and an allyl influences the activity in the opposite direction. Both radicals appear to intensify the activity.

Topics: Animals; Biphenyl Compounds; Chemical Phenomena; Chemistry; Chickens; Eugenol; Lignans; Male; Mephenesin; Mice; Muscle Contraction; Muscle Relaxants, Central; Reflex; Spinal Cord; Structure-Activity Relationship

Topics: Biphenyl Compounds; China; Lignans; Plants, Medicinal