iridoids and geniposide

Gardenia jasminoides J.Ellis is widely used to treat liver diseases in traditional Chinese medicine. Geniposide, a major active constituent of Gardenia jasminoides J.Ellis, exerts therapeutic effects against liver injury, however, it also induces hepatotoxicity.. This meta-analysis was designed to determine the mechanisms of both the hepatoprotective and hepatotoxic effects of geniposide.. The articles analysed in this meta-analysis were primarily obtained from five databases. The 10-item SYRCLE risk-of-bias tool was used to evaluate the quality of the included articles. STATA (version 15.1) was used to evaluate the total effect or toxicity sizes. In addition, three-dimensional (3D) dose/time-effect and mechanistic analyses were performed to assess the therapeutic and toxic effects of geniposide.. A total of 25 studies involving 479 animals were included. Meta-analysis revealed that geniposide not only significantly (P < 0.001) increased liver injury indices including ALT and AST levels but also improved liver function by decreasing the levels of ALT, AST and inflammatory factors in animal models of liver injury. The 3D dose/time-effect analysis revealed that geniposide administered at a dose of 20-150 mg/kg for 5-28 days effectively protected the liver without inducing toxicity. Mechanistically, geniposide exerts protective or toxic effects by regulating the TNF-α/NF-κB pathway to control oxidative stress and inflammatory responses.. Geniposide exhibits dual pharmacological activity in liver injury. It exerts potent hepatoprotective effects when administered at a dose of 20-150 mg/kg for 5-28 days.

Topics: Animals; Gardenia; Iridoids; Liver; NF-kappa B; Tumor Necrosis Factor-alpha

At present, the potential of natural products in new drug development has attracted more and more scientists' attention, and natural products have become an important source for the treatment of various diseases or important lead compounds. Geniposide, as a novel iridoid glycoside compound, is an active natural product isolated from the herb

Topics: Biological Products; Diabetes Mellitus; Gardenia; Iridoids

Geniposide is a naturally sourced active ingredient that has diverse pharmacological effects and great potential in improving or treating different kinds of diseases. In recent years, more and more studies have confirmed that geniposide can improve glucose and lipid metabolism disorder, which is an increasingly prevalent health problem causing various metabolic diseases globally. Our review aims to summarize basic information on the pharmacological effects of geniposide on glucolipid metabolism. Geniposide increases glucose utilization and insulin production, protects pancreatic islet β cells, inhibits insulin resistance and hepatic glucose production, and suppresses gluconeogenesis. While in the aspect of lipid metabolism, geniposide can promote lipolysis, inhibit lipogenesis, and regulate lipid transport. Geniposide ameliorates lipid and glucose metabolic disorders, improving the entire glycolipid metabolism network in a three-dimensional manner at the level of molecular mechanism. Growing evidence revealed that geniposide may serve as an effective drug to combat metabolic diseases for the time to come.

Topics: Gardenia; Glucose; Glycolipids; Insulins; Iridoids; Lipid Metabolism; Lipids; Metabolic Diseases

Geniposide (GE) is ubiquitous in nearly 40 species of plants, among which Gardenia jasminoides J. Ellis has the highest content, and has been used ethnopharmacologically to treat chronic inflammatory diseases. As a traditional Chinese medicine, Gardenia jasminoides J. Ellis has a long history of usage in detumescence and sedation, liver protection and cholestasis, hypotension and hemostasis. It is commonly used in the treatment of diabetes, hypertension, jaundice hepatitis, sprain and contusion. As a type of iridoid glycosides extracted from Gardenia jasminoides J. Ellis, GE has many pharmacological effects, such as anti-inflammatory, anti-angiogenesic, anti-oxidative, etc. AIM OF THE REVIEW: In this article, we reviewed the sources, traditional usage, pharmacokinetics, toxicity and therapeutic effect of GE on chronic inflammatory diseases, and discussed its potential regulatory mechanisms and clinical application.. GE is a common iridoid glycoside in medicinal plants, which has strong activity in the treatment of chronic inflammatory diseases. A large number of in vivo and in vitro experiments confirmed that GE has certain therapeutic value for a variety of chronic inflammation disease. Its mechanism of function is mainly based on its anti-inflammatory, anti-oxidant, neuroprotective properties, as well as regulation of apoptotsis. GE plays a role in the treatment of chronic inflammatory diseases by regulating cell proliferation and apoptosis, realizing the dynamic balance of pro/anti-inflammatory factors, improving the state of oxidative stress, and restoring abnormally expressed inflammation-related pathways.. According to its extensive pharmacological effects, GE is a promising drug for the treatment of chronic inflammatory diseases.

Topics: Animals; Chronic Disease; Humans; Inflammation; Iridoids; Phytotherapy; Plants

Rehmannia glutinosa, the fresh or dried root of Rehmannia glutinosa (Gaertn.) Libosch. ex Fisch. & Mey., and Gardenia, the fruit of Gardenia jasminoides Ellis from Rubiaceae, both are famous traditional Chinese medicines that have been traditionally used in China. Catalpol and geniposide, as two kinds of iridoid glycosides with high activities, are the main bioactive components in Rehmannia glutinosa and Gardenia jasminoides Ellis, respectively. Over the past few decades, catalpol and geniposide have been widely studied for their therapeutic effects. The preclinical experiments demonstrated that they possessed significant neuroprotective activities against Alzheimer's disease, Parkinson's disease, stroke, and depression, etc. In this paper, the pharmacological effects and mechanisms of catalpol and geniposide on Alzheimer's disease and Parkinson's disease from 2005 to now were systematically summarized and comprehensively analyzed. At the same time, the pharmacokinetic characteristics of the analyzed compounds were also described, hoping to provide some enlightenment for the design, research, and development of iridoid glycosides.

Topics: Alzheimer Disease; Animals; Antiparkinson Agents; Drugs, Chinese Herbal; Gardenia; Humans; Iridoid Glucosides; Iridoids; Medicine, Chinese Traditional; Parkinson Disease; Rehmannia

Geniposide, an iridoid glucoside, is a major component in the fruit of

Topics: Animals; Antioxidants; Cardiovascular Diseases; Diabetes Mellitus; Drug Evaluation, Preclinical; Humans; Iridoids; Models, Molecular

Iridoid glycosides are natural products occurring widely in many herbal plants. Geniposide (C

Topics: Animals; Humans; Iridoids; Molecular Structure; Phytotherapy; Plant Extracts; Rubiaceae

Yīn-Chén-Hāo decoction (YCHD) is a traditional Chinese medicine formula composed of capillaris (

Topics: Animals; Anthraquinones; Anti-Inflammatory Agents; Antiviral Agents; Artemisia; Ascites; Chlorogenic Acid; Clinical Trials as Topic; Coumarins; Drugs, Chinese Herbal; Emodin; Fatty Liver; Gardenia; Humans; Iridoids; Liver Diseases; Plant Extracts; Rheum

A growing body of evidence has linked two of the most common aged-related diseases: type 2 diabetes mellitus (T2DM) and Alzheimer's disease (AD). It has led to the notion that drugs developed for the treatment of T2DM may be beneficial in modifying the pathophysiology of AD. As a receptor agonist of glucagon-like peptide-1 (GLP-1R), which is a newer drug class to treat T2DM, geniposide shows clear effects in inhibiting pathological processes underlying AD, such as promoting neurite outgrowth. In the present article, we review the possible molecular mechanisms of geniposide to protect the brain from pathologic damages underlying AD: reducing amyloid plaques, inhibiting τ phosphorylation, preventing memory impairment and loss of synapses, reducing oxidative stress and the chronic inflammatory response, and promoting neurite outgrowth via the GLP-1R signaling pathway. In summary, the Chinese herb geniposide shows great promise as a novel treatment for AD.

Topics: Alzheimer Disease; Animals; Diabetes Mellitus, Type 2; Humans; Iridoids; Neuroprotective Agents

Asperuloside (ASP) and geniposide (GP) are iridoids that have shown various biological properties, such as reduction of inflammation, oxidative stress, and neuroprotection. The aim of this study was to investigate the mechanism of action of ASP and GP through the experimental model of pilocarpine-induced seizures. Mice were treated daily with saline, valproic acid (VPA), GP (5, 25, or 50 mg/kg), or ASP (20 or 40 mg/kg) for 8 days. Pilocarpine (PILO) treatment was administered after the last day of treatment, and the epileptic behavior was recorded for 1 h and analyzed by an adapted scale. Afterward, the hippocampus and blood samples were collected for western blot analyses, ELISA and comet assay, and bone marrow to the micronucleus test. We evaluated the expression of the inflammatory marker cyclooxygenase-2 (COX-2), GluN2B, a subunit of the NMDA receptor, pGluR1, an AMPA receptor, and the enzyme GAD-1 by western blot and the cytokine TNF-α by ELISA. The treatments with GP and ASP were capable to decrease the latency to the first seizure, although they did not change the latency to status epilepticus (SE). ASP demonstrated a genotoxic potential analyzed by comet assay; however, the micronuclei frequency was not increased in the bone marrow. The GP and ASP treatments were capable to reduce COX-2 and GluN2B receptor expression after PILO exposure. This study suggests that GP and ASP have a protective effect on PILO-induced seizures, decreasing GluN2B receptor and COX-2 expression.

Topics: Animals; Cyclooxygenase 2; Disease Models, Animal; Hippocampus; Iridoids; Mice; Pilocarpine; Rats; Rats, Wistar; Receptors, N-Methyl-D-Aspartate; Seizures

Geniposide is the main medicinal component of Gardenia jasminoides, and its content is approximately 3-8% depending on its origin. Geniposide is a class of cyclic enol ether terpene glucoside compounds with strong antioxidant, free radical quenching and cancer-inhibiting activities. Many studies have reported that geniposide has hepatoprotective, cholestatic, neuroprotective, blood sugar and blood lipid regulation, soft tissue damage treatment, antithrombotic, antitumor and other effects. As a traditional Chinese medicine, gardenia, whether used as gardenia alone, as the monomer geniposide or as the effective part of cyclic either terpenoids, has been reported to have anti-inflammatory effects when used in the right amounts. Recent studies have found that geniposide has important roles in pharmacological activities such as anti-inflammation activity, inhibition of the NF-κB/IκB pathway, and cell adhesion molecule production. In this study, we predicted the anti-inflammatory and antioxidant effects of geniposide in piglets through network pharmacology based on the LPS-induced inflammatory response-regulated signaling pathway. The effects of geniposide on changes in inflammatory pathways and cytokine levels in the lymphocytes of inflammation-stressed piglets were investigated using in vivo and in vitro models of piglet lipopolysaccharide-induced oxidative stress. Network pharmacology identified 23 target genes, of which the main pathways of action were lipid and atherosclerosis, fluid shear stress and atherosclerosis, and Yersinia infection. The main relevant target genes were VEGFA, ROCK2, NOS3, and CCL2. Validation experiments showed that the interventional effects of geniposide reduced the relative expression of NF-κB pathway proteins and genes, restored the expression of COX-2 genes to normal levels, and increased the relative expression of tight junction proteins and genes in IPEC-J2 cells. This indicates that the addition of geniposide can alleviate inflammation and improve the level of cellular tight junctions.

Topics: Animals; Anti-Inflammatory Agents; Gardenia; Iridoids; Lipopolysaccharides; Network Pharmacology; NF-kappa B; Swine

Cardiac fibrosis is one of the main contributors to the pathogenesis of heart failure. Geniposide (GE), a major iridoid in gardenia fruit extract, has recently been reported to improve skeletal muscle fibrosis through the modulation of inflammation response. This investigation aimed to illuminate the cardio-protective effect and the potential mechanism of GE in cardiac fibrosis.. A transverse aortic contraction (TAC) induction mice model was established and GE (0 mg/kg; 10 mg/kg; 20 mg/kg; 40 mg/kg) was administered by oral gavage daily for 4 weeks. Hemodynamic parameters, Masson's trichrome stain, and hematoxylin-eosin (HE) staining were estimated and cardiomyocyte fibrosis, interstitial collagen levels, and hypertrophic markers were analyzed using qPCR and western blot. In vitro, H9C2 cells were exposed to the Ang II (1 μM) pretreated with GE (0.1 μM, 1 μM, and 10 μM). Cardiomyocyte apoptosis was detected. Moreover, the transforming growth factor β1 (TGF-β1)/Smad2 pathway was assessed in vivo and in vitro.. GE significantly ameliorated TAC-induced cardiac hypertrophy, ventricular remodeling, myocardial fibrosis, and improved cardiac function in vivo, and it inhibited Ang II-induced cardiomyocyte apoptosis in vitro. We further observed that the inflammatory channel TGF-β1/Smad2 pathway was suppressed by GE both in vivo and in vitro.. These results indicate that GE inhibited myocardial fibrosis and improved hypertrophic cardiomyocytes with attenuated the TGF-β1/Smad2 pathway and proposed to be an important therapeutic of cardiac fibrosis reduced by TAC.

Topics: Animals; Fibrosis; Iridoids; Mice; Myocardium; Myocytes, Cardiac; Transforming Growth Factor beta1

Gardenia jasminoides Ellis is a perennial evergreen shrub of G. jasminoides of Rubiaceae. Geniposide and Crocin are important components in the fruit of G. jasminoides. In addition to being used as medicinal materials, they are also widely used in food, medicine, cosmetics, and other fields. They have high medicinal value, economic value, and ornamental value. However, at present, the utilization rate of G. jasminoides resources is low, mainly focused on germplasm cultivation, primary processing, and clinical pharmacology, and there are few studies on the quality of Gardenia fruit.. Based on transcriptome sequencing and metabolic group analysis, the morphological and structural changes of Gardenia fruit with young fruit, middle fruit, and ripe fruit were analyzed, and the formation mechanism and content changes of Geniposide and Crocin in Gardenia fruit were studied. The content of Geniposide decreased with the development of fruit, so did the expression of the main structural gene GES, G10H, and IS in its synthesis pathway, while the content of Crocin increased with the development of fruit, and the expression of the main structural gene CCD, ALDH, and UGT in its synthesis pathway also increased. The relationship between the morphological structure of G. jasminoides and the accumulation of Geniposide and Crocin was summarized.. This study not only provides a theoretical basis for the mining and utilization of Geniposide and Crocin, but also provides a theoretical basis for genetic background for the identification and cloning of bioactive substances in gardenia fruit in future. At the same time, it provides support for increasing the dual-use value of G. jasminoides and breeding excellent germplasm resources.

Topics: Fruit; Gardenia; Iridoids; Metabolome; Plant Breeding; Transcriptome

Topics: Humans; Iridoids; Sclerosis; Tomography, X-Ray Computed

As the main effector cells of chronic inflammation and hyperplasia of synovium, fibroblast-like synoviocytes (FLSs) show abnormal proliferation and insufficient apoptosis in the hypoxic microenvironment, which is due to the increase of BNIP3-mediated autophagy. This study aimed to explore the mechanism of geniposide (GE) on hypoxia-induced hyper-proliferative FLSs with a focus on autophagy and the JNK-BNIP3 pathway.. The dynamic changes of autophagy, apoptosis, and hypoxia-related proteins in adjuvant arthritis (AA) rats were detected by immunohistochemistry and Western blot. The proliferation, autophagy, apoptosis, and mitochondrial state of FLSs were detected by CCK-8, flow cytometry, immunofluorescence, and transmission electron microscopy, respectively. Western blot, qRT-PCR, and co-immunoprecipitation were used to detect the expression of the JNK-BNIP3 pathway.. The excessive accumulation of BNIP3 in the synovium of AA rats was accompanied by inhibition of apoptosis and an increase in autophagy. GE inhibited the expression of BNIP3, enhanced apoptosis, decreased autophagy, and improved chronic inflammation and hyperplasia of synovium. The amount of autophagy under different oxygen concentrations was the key to mediating the different survival rates of FLSs, and the inhibition of autophagy triggered apoptosis. GE suppressed the proliferation of FLSs and down-regulated autophagy, leading to the accumulation of ROS and the decrease of mitochondrial membrane potential, induced the increase of apoptosis, and suppressed the accumulation of BNIP3 and the hyperphosphorylation of JNK.. GE inhibited autophagy by restoring the hypoxia-induced activated JNK-BNIP3 pathway, inducing mitochondrial oxidative damage, augmented apoptosis, and decreased survival rate of FLSs.

Topics: Animals; Apoptosis; Arthritis, Experimental; Autophagy; Fibroblasts; Hyperplasia; Hypoxia; Inflammation; Iridoids; Membrane Proteins; Mitochondrial Proteins; Rats; Synoviocytes

Osteoarthritis (OA) is a chronic joint disease characterized by cartilage degeneration. Autophagy is associated with chondrocyte homeostasis and exhibits a role in protecting against OA pathogenesis. Geniposide (GEN), an iridoid glycoside extracted from Eucommia ulmoides Oliv, acts as an activator of GLP-1R, which can stimulate autophagy. The AMPK/mTOR signaling pathway participates in the mediation of autophagy, and GLP-1R may act as an upstream factor of AMPK. However, whether GEN mediates the autophagic responses by activating the GLP-1R/AMPK/mTOR signaling pathway in OA chondrocytes is still unclear. In the current study, attenuated autophagy in MIA-induced rat OA models was observed, as shown by up-regulated expression of p62 and down-regulated expression of Beclin-1 and LC3-II/I. GEN stimulated autophagy and protected OA cartilage by up-regulating GLP-1R expression. In addition, GEN could enhance AMPK phosphorylation and down-regulate mTOR expression in IL-1β-treated C28/I2 cells. Inhibition of AMPK or activation of mTOR could reverse the stimulatory effects of GEN on autophagy. Furthermore, a GLP-1R inhibitor Exendin 9-39 could eliminate the chondroprotective effects of GEN by suppressing the AMPK/mTOR signaling pathway. Conclusively, Geniposide exhibits protective effects against osteoarthritis development by stimulating autophagy via activating the GLP-1R/AMPK/mTOR signaling pathway.

Topics: AMP-Activated Protein Kinases; Animals; Autophagy; Chondrocytes; Iridoids; Osteoarthritis; Rats; Signal Transduction; TOR Serine-Threonine Kinases

In this work, a robust method for the separation of gardenia yellow and geniposide from gardenia fruit was developed based on a molecularly imprinted solid phase extraction (MISPE) procedure. First, hydrophilic molecularly imprinted microspheres (HMIMs) were prepared using gardenia yellow as the template via reversible addition fragmentation chain transfer (RAFT) precipitation polymerization. The resultant HMIMs demonstrated the multiresponsiveness to pH, temperature, and magnetism, achieving controllable uptake and release of gardenia yellow and easy recovery by external magnets. Meanwhile, the HMIMs possessed high adsorption capacity, fast binding kinetics, specific recognition, and reusability. Finally, a MISPE approach using HMIMs as adsorbent was developed for extraction of gardenia yellow and purification of geniposide after optimization of the adsorption and elution conditions. Thus, efficient separation of gardenia yellow and geniposide with relative purities of 99.77 ± 0.05% (94.04 ± 0.10% recovered) and 94.50 ± 0.62% (95.40 ± 0.86% recovered), respectively, was achieved.

Topics: Fruit; Gardenia; Iridoids; Microspheres; Plant Extracts

Systemic insulin signal transduction is influenced by the inter-tissue crosstalk, which might be the potential therapeutic strategy for T2DM. Although anti-diabetic function of geniposide has been previously reported, the underlying mechanism was not completely clear in light of the complex pathogenesis of T2DM.. The present experiment is devoted to investigate the potential effects of geniposide on systemic insulin sensitivity mediated by hepatokine-RBP4 in high fat diet (HFD)-fed mice.. The HFD-fed wild type mice were administered with geniposide (25 or 50 mg/kg/d) by intraperitoneal injection, and the normal saline and Metformin were used as negative control group and positive control group, respectively. After administration for 4 weeks, the food intake, body weight, glucose tolerance tests, insulin tolerance tests and serum biochemical indices were examined, along with insulin signaling pathway-associated proteins and hepatic histomorphological analysis. The liver, gastrocnemius and mouse primary hepatocytes were also harvested for molecular mechanism study.. After geniposide treatment for 4 weeks, the blood glucose level was reduced in HFD-fed mice. Furthermore, geniposide treatment improved insulin sensitivity both in the liver and gastrocnemius (GAS). In terms of mechanism, geniposide disturbed circulating RBP4 level including its synthesis, secretion and homeostasis. Moreover, geniposide modified fuel selection and promoted glucose uptake in skeletal muscle and reduced glycogen storage, which were closely related to impaired circulating RBP4 homeostasis, leading to ameliorative systemic insulin sensitivity.. Our current study proposes a novel regulatory mechanism of geniposide for improving glucose homeostasis through regulating circulating RBP4 level, which also provides new strategies for the prevention and treatment of T2DM.

Topics: Adipose Tissue; Animals; Diet, High-Fat; Glucose; Homeostasis; Insulin; Insulin Resistance; Iridoids; Liver; Mice; Mice, Inbred C57BL; Retinol-Binding Proteins, Plasma

As a major active iridoid glycoside from. This study explores the effect of geniposide in diabetic wound model by anti-inflammatory action.. Diabetic rodent model in Wistar rats was induced by streptozotocin combined with high-fat feed. The selected rats were divided into control group, the diabetic model group and geniposide subgroups (200, 400 and 500 mg/kg), and orally administrated once daily with saline or geniposide. Wound area and histochemical indicators were measured on day 7 after continuous administration, to assess lesion retraction, inflammatory cells and fibroblasts.. Geniposide promoted diabetic wound healing by anti-inflammation and adjusting blood glucose. Further topical studies are required to evaluate effects on antibacterial activity and skin regeneration.

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents; Blood Glucose; Cytokines; Diabetes Mellitus, Experimental; Dose-Response Relationship, Drug; Gardenia; Iridoids; Male; Rats; Rats, Wistar; Streptozocin; Wound Healing

Eucommia ulmoides Oliver has been traditionally used for treatment of various diseases, including osteoporosis, knee pain, and paralysis. The extract of Eucommia ulmoides has been reported to stimulate the bone formation and suppress the bone resorption, leading to protection against osteoporosis (OP). Geniposide (GEN) has been considered as one of the effective compounds responsible for the therapeutic efficacy of Eucommia ulmoides against OP.. To explore whether GEN protected against dexamethasone (DEX)-induced osteoporosis (OP) by activating NRF2 expression and inhibiting endoplasmic reticulum (ER) stress.. The DEX-induced rat OP models were duplicated. The pathological changes were examined by histological/immunohistochemical evaluation and micro-computed tomography (micro-CT) assessment. Apoptosis was detected by a flow cytometer. Mitochondrial Ca. GEN effectively reversed DEX-induced pathological changes of trabecular bone in rats. In addition, the DEX-increased expression of ATF4/CHOP was also ameliorated. In MC3T3-E1 cells, DEX promoted endoplasmic reticulum (ER) stress and mitochondrial apoptosis. Inhibition of ER stress abolished the induction of apoptosis by DEX. Similarly, GEN significantly ameliorated DEX-induced mitochondrial apoptosis. The possible underlying mechanism might be associated with the pharmacological effects of GEN on activating the expression of NRF2 and alleviating ER stress in DEX-treated MC3T3-E1 cells.. GEN ameliorated DEX-induced ER stress and mitochondrial apoptosis in osteoblasts.

Topics: Animals; Apoptosis; Cell Line; Dexamethasone; Endoplasmic Reticulum Stress; Iridoids; Osteoblasts; Rats; Signal Transduction; X-Ray Microtomography

This study aimed to investigate the effects of geniposide(GP) on the expression of prokineticin(PK2) and prokineticin receptor 1(PKR1) in db/db mice with diabetic nephropathy(DN), so as to explore how the PK2 signaling pathway participated in the pathological changes of DN and whether GP exerted the therapeutic effect through this signaling pathway. Male mice were randomly divided into four groups, namely db/m, db/db, db/db+GP, and db/m+GP groups, with five in each group. The mice in the db/db+GP and db/m+GP groups were gavaged with 150 mg·kg~(-1) GP for eight successive weeks. Afterwards, all the mice were sacrificed and the renal tissues were embedded. The morphological changes in glomerulus and renal tubules were observed by Masson and PAS staining. The expression levels of PK2, PKR1, and Wilm's Tumor Protein 1(WT_1) in podocytes were detected by immunohistochemistry, and the protein expression levels of PK2 and PKR1 in mouse kidney by Western blot. The morphological results showed serious glomerular and tubular fibrosis(Masson), high glomerular and tubular injury score(PAS), increased glomerular mesangial matrix, thickened basement membrane, exfoliated brush border of renal tubules, decreased WT_1 in glomerular podocytes, and massive loss of podocytes in the db/db group. After administration with GP, the glomerular and tubular fibrosis was alleviated, accompanied by improved glomerular basement membrane and renal tubule brush edge, and up-regulated WT_1. As revealed by further protein detection, in the db/db group, the expression levels of PK2 and PKR1 and p-Akt/Akt ratio declined, whereas the ratio of Bax/Bcl-2 rose. Ho-wever, PKR2 and p-ERK/ERK ratio did not change significantly. After administration with GP, the PK2 and PKR1 expression was elevated, and p-Akt/Akt ratio was increased. There was no obvious change in PKR2, Bax/Bcl-2 ratio, or p-ERK/ERK ratio. All these have demonstrated that GP improves the renal damage in DN mice, and PK2/PKR1 signaling pathway may be involved in such protection, which has provided reference for clinical treatment of DN with GP.

Topics: Animals; Diabetes Mellitus; Diabetic Nephropathies; Iridoids; Kidney; Male; Mice; Signal Transduction

Rheumatoid arthritis (RA) is an angiogenesis-dependent disease caused by the imbalance of pro- and anti-angiogenic factors. More effective strategies to block synovial angiogenesis in RA should be studied. Geniposide (GE), a natural product isolated from the fruit of Gardenia jasminoides Ellis (GJ), is reported to have anti-inflammatory, anti-angiogenic and other pharmacological effects. However, the underlying mechanism through which GE affects synovial angiogenesis in RA remains unclear.. In this research, we aimed to elucidate the effect and potential mechanisms of GE on angiogenesis in RA.. Synovial angiogenesis in patients with RA and a rat model of adjuvant arthritis (AA) was detected by hematoxylin and eosin (HE) staining, immunohistochemistry (IHC), and western blottiing. The biological functions of vascular endothelial cells (VECs) and sphingosine kinase 1 (SphK1) translocation were checked by CCK-8, EdU, Transwell, tube formation, co-immunoprecipitation assays, and laser scanning confocal microscopy. The effect of the SphK1 gene on angiogenesis was assessed by transfection of SphK1-siRNA in cells and mices. The effect of GE on VEGF-induced angiogenesis was measured by Matrigel plug assay in a mouse model of AA.. GE effectively inhibited synovial angiogenesis and alleviated the disease process. SphK1, as a new regulatory molecule, has a potentially important relationship in regulating VEGF/VEGFR2 and S1P/S1PR1 signals. SphK1 translocation was activated via the VEGFR2/PKC/ERK1/2 pathway and was closely linked to the biological function of VECs. GE significantly reduced SphK1 translocation, thereby ameliorating the abnormal biological function of VECs. Furthermore, after transfection of SphK1 siRNA in VECs and C57BL/6 mice, silencing SphK1 caused effectively attenuation of VEGF-induced VEC biological functions and angiogenesis. In vivo, the Matrigel plug experiment indicated that GE significantly inhibited pericyte coverage, basement membrane formation, vascular permeability, and fibrinogen deposition.. Our findings suggest that GE inhibited VEGF-induced VEC biological functions and angiogenesis by reducing SphK1 translocation. Generally, studies have revealed that GE down-regulated VEGFR2/PKC/ERK1/2-mediated SphK1 translocation and inhibited S1P/S1PR1 signaling activation, thereby alleviating VEGF-stimulated angiogenesis. The above evidences indicated that angiogenesis inhibition may provide a new direction for RA treatment.

Topics: Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Endothelial Cells; Humans; Iridoids; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; Neovascularization, Pathologic; Phosphotransferases (Alcohol Group Acceptor); Rats; RNA, Small Interfering; Vascular Endothelial Growth Factor A

Rheumatoid arthritis (RA) is a systemic immune disease characterized by joint inflammation and pannus. The nascent pannus contributes to synovial hyperplasia, cartilage, and tissue damage in RA. This study aims to explore the therapeutic effect and potential mechanism of Geniposide (GE) on RA angiogenesis, involving the participation of phosphate and tension homology deleted on chromosome ten (PTEN) and downstream pathways. Clinical manifestations, synovial pathomorphology, microvessel density, and the level of angiogenesis-related factors were used to evaluate the therapeutic effect of GE on adjuvant-induced arthritis (AA) rats. The proliferation, migration, and tube formation of human umbilical vein endothelial cells (HUVECs) indicate the degree of angiogenesis in vitro. Lentivirus over-expression of PTEN was employed to elucidate the potential mechanism. The results showed that GE improved the degree of arthritis and angiogenesis in AA rats. The expression of PTEN was decreased significantly in vivo and in vitro, and over-expression of PTEN improved the biological function of HUVECs to inhibit angiogenesis. GE inhibited the proliferation, migration, and tubule formation of HUVECs and plays an anti-angiogenesis role in vitro. Mechanism study showed that PTEN expression was increased and p-PI3K and p-Akt expression was decreased with GE treatment. It suggests that GE up-regulated the expression of PTEN and inhibited the activation of PI3K-Akt signal, which plays a role in inhibiting angiogenesis in RA in vivo and in vitro.

Topics: Angiogenesis Inducing Agents; Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Cell Proliferation; Human Umbilical Vein Endothelial Cells; Humans; Iridoids; Neovascularization, Pathologic; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; PTEN Phosphohydrolase; Rats; Signal Transduction

Topics: Animals; Fibrosis; Hyperuricemia; Inflammation; Iridoids; Kidney Diseases; Mice; Molecular Docking Simulation; Transforming Growth Factor beta; Uric Acid; Xanthine Oxidase

Oxidative stress and neurocyte apoptosis are crucial pathophysiological process in early brain injury (EBI) after subarachnoid hemorrhage (SAH). Geniposide (GNP) has been reported to exert neuroprotective effects by reducing oxidative injury and neurocyte apoptosis. However, the effect of GNP has not been clarified in EBI after SAH. The study was performed to evaluate the neuroprotective effects and mechanisms of GNP in EBI after SAH.. A total of 60 male Wistar rats were randomly divided into five groups. The prechiasmatic cistern SAH model was used in this study. SAH grade was evaluated using a grading system. Neurological function was evaluated using the Garcia scores. Brain edema was measured by the wet-dry method. Blood-brain barrier (BBB) permeability was measured by the extravasation of Evans Blue (EB). The neurocyte apoptosis was observed using TUNEL assay. The levels of malondialdehyde (MDA) and superoxide dismutase (SOD), as well as the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), hemeoxygenase-1 (HO-1), glutathione S-transferase (GST) and quinone oxidoreductase-1 (NQO-1) were performed. The results showed that GNP reduced brain edema, attenuated BBB permeability, inhibited neurocyte apoptosis and improved neurological function. Moreover, GNP also decreased the levels of ROS and MDA, elevated Nrf2 expression in the temporal cortex and up-regulated the expression of NQO-1, HO-1 and GST after SAH.. GNP could ameliorate oxidative stress and neurocyte apoptosis to exert neuroprotective effects by Nrf2 pathway.

Topics: Animals; Apoptosis; Brain; Brain Edema; Brain Injuries; Glutathione Transferase; Iridoids; Male; Neuroprotective Agents; NF-E2-Related Factor 2; Oxidative Stress; Rats; Rats, Sprague-Dawley; Rats, Wistar; Subarachnoid Hemorrhage

White Spot Disease (WSD), caused by white spot syndrome virus (WSSV), is an acute and highly lethal viral disease of shrimp. Currently, there are no commercially available drugs to control WSD. It is urgent and necessary to find anti-WSSV drugs. Natural compounds are an important source of antiviral drug discovery. In this study, the anti-WSSV activity of natural compound geniposide (GP) was investigated in crayfish Procambarus clarkii. Results showed that GP had a concentration-dependent inhibitory effect on WSSV replication in crayfish at 24 h, and highest inhibition was more than 98%. In addition, GP significantly inhibited the expression of WSSV immediate-early gene ie1, early gene DNApol, late gene VP28. The mortality of WSSV-infected crayfish in control groups was 100%, while it reduced by 70.0% when treated with 50 mg/kg GP. Co-incubation, pre-treatment and post-treatment experiments showed that GP could prevent and treat WSSV infection in crayfish by significantly inhibiting WSSV multiplication. Mechanistically, the syntheses of WSSV structural proteins VP19, VP24, VP26 and VP28 were significantly inhibited by GP in S2 cells. Furthermore, GP could also suppress WSSV replication by blocking the expression of antiviral immunity-related factor STAT to reduce ie1 transcription. Moreover, GP possessed anti-inflammatory and anti-oxidative activity in crayfish. Overall, GP has the potential to be developed as a preventive or therapeutic agent against WSSV infection.

Topics: Animals; Antiviral Agents; Astacoidea; Iridoids; Survival Rate; White spot syndrome virus 1

Neuropathic pain (NP) is a common disorder among individuals worldwide, but there is still no effective treatment for NP. The EGFR pathway promotes NP nociceptive sensitization and represents a potential therapeutic target. Geniposide is abundant in natural plants and has various pharmacological activities, such as analgesia and anti-inflammation properties, which can improve NP, but the specific mechanisms have not been elucidated. The present study first predicted and molecularly docked geniposide targets, suggesting that geniposide may play a role in improving NP by targeting EGFR. This study further clarified that geniposide alleviates NP and improves the inflammatory response using a chronic constriction injury (CCI) model, whereas the administration of an EGFR agonist weakens the above effects of geniposide. Analysis of transcriptome data further suggests that geniposide not only improves CCI symptoms by reducing EGFR/PI3K/AKT pathway activity but also may exert anti-inflammatory effects by inhibiting the Ca

Topics: Animals; ErbB Receptors; Iridoids; Neuralgia; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley

Gut-derived lipopolysaccharide (LPS) leaking through the dysfunctional intestinal barrier contributes to the onset of non-alcoholic steatohepatitis (NASH) by triggering inflammation in the liver. In the present study, a combination consisting of

Topics: Animals; Atractylodes; Chlorogenic Acid; Endotoxins; Iridoids; Lipopolysaccharides; Liver; Mice; Non-alcoholic Fatty Liver Disease

Hyperuricemia is a common metabolic disease and is one of the factors that could induce chronic kidney disease (CKD). Geniposide (GEN) is a typical natural iridoid glucoside compound with a series of biological activities, but the poor bioavailability of GEN limits its clinical application. In this context, the pharmacological activity of the geniposide-phospholipid complex (GEN-PLC) in ameliorating posthyperuricemia CKD was evaluated by in vitro and in vivo experiments in this study. In vitro cell experiments showed that GEN-PLC treatment markedly decreased inflammatory cytokine levels and reactive oxygen species levels compared with those of GEN in uric acid-treated HKC cells. In vivo research results confirmed that a high concentration of uric acid could cause CKD by increasing inflammatory cytokines and reactive oxygen species in hyperuricemic mice. At the same time, GEN-PLC could regulate the PI3K/AKT/NF-κB and Keap1/Nrf2/HO-1 signaling pathways to effectively inhibit the inflammatory response and oxidative stress, thereby ameliorating posthyperuricemia CKD, and the therapeutic effect was better than that of GEN. In addition, the preparation technology of GEN-PLC was optimized, and the physiochemical analysis explained the intermolecular interactions of the two components. Based on the research results, GEN-PLC could enhance the bioavailability of GEN and become a promising candidate for clinical drug development.

Topics: Animals; Inflammation; Iridoids; Kelch-Like ECH-Associated Protein 1; Mice; NF-E2-Related Factor 2; Oxidative Stress; Phosphatidylinositol 3-Kinases; Phospholipids; Reactive Oxygen Species; Renal Insufficiency, Chronic; Uric Acid

Neovascularization in rheumatoid arthritis (RA) is a key bridge between malignant proliferative synovial tissue and pannus. In view of previous studies on the efficacy of Geniposide (GE) in experimentary arthritis, the purpose of this study was to investigate the possible mechanism of GE inhibiting angiogenesis by regulating the gene of phosphate and tension homology deleted on chromosome ten (PTEN). In this study, human umbilical vein endothelial cells (HUVEC) and adjuvant arthritis (AA) rat models were performed to research in vitro and in vivo. The results showed that GE treatment significantly reduced synovitis and angiogenesis in AA rats, which may be associated with the increased expression of PTEN with GE treatment. Meanwhile, the hypermethylation of PTEN accompanied by the over-expression of DNA methyltransferases (Dnmts) was demonstrated in TNF-α-induced HUVEC and AA rats. Knockdown of Dnmt1 by Dnmt1- siRNA significantly inhibited the tube formation of HUVEC in vitro. GE significantly restricted the angiogenesis of HUVEC by inhibiting DNA methylation, which was attributed to the down-regulation of Dnmt1 rather than Dnmt3a and Dnmt3b. The anti-angiogenesis effect of GE was further verified in AA model by the inhibition of Dnmt1. These results indicate that GE exhibited anti-angiogenesis effects in experimentary arthritis by inhibiting Dnmt1-mediated PTEN gene hypermethylation, which may brings new insights for the prevention and research of RA.

Topics: Angiogenesis Inhibitors; Animals; Arthritis, Experimental; Arthritis, Rheumatoid; DNA (Cytosine-5-)-Methyltransferase 1; DNA Methylation; Human Umbilical Vein Endothelial Cells; Humans; Iridoids; Neovascularization, Pathologic; PTEN Phosphohydrolase; Rats

Bile acids (BAs) play an important role in pre-diagnosing drug-induced liver injury (DILI). However, in clinical practice, different types of liver injury are characterized by different pathogeneses and pathological manifestations. Therefore, whether BAs can be used as biomarkers across different DILIs remains unclear. In this study, an ultra-performance chromatography-mass spectrometry (MS)/MS-based technique was developed for the simultaneous quantitative analysis of 31 BAs in the serum, liver, feces, urine, and intestinal contents of rats treated with acetaminophen (APAP) and geniposide to induce liver injury. The total extraction recovery for representative analytes ranged between 80.60% and 99.23% in the serum, urine, liver, feces, and intestinal contents. The correlation coefficients for all standard curves of the different matrices were at least 0.99. Validation of the BA analytical method including selectivity, residue, lower limit of quantification, accuracy, precision, matrix effect, and stability conformed with the biospecimen quality control standards of the Chinese Pharmacopoeia (version 2020). Serum biochemical and pathohistological analyses revealed APAP- and geniposide-induced hepatocellular and cholestatic DILI, respectively, with different effects on BA profiles in the enterohepatic circulation. Metabolomics further revealed that the trends in BA changes in the serum, feces, urine, and intestinal tissues were consistent between the geniposide- and APAP-treated groups. However, in the liver, the total BAs (TBA) concentration increased by 1.70 fold in the geniposide group but decreased by 43% in the APAP group compared with the control group. Multivariate analysis revealed differentially expressed BAs, including TCA, CA, and GCA, which are potential biomarkers for DILI, in the serum, liver, and urine following treatment with geniposide. Interestingly, the differentially expressed BAs in the APAP group were similar to those in the control group. Additionally, the magnitude of changes in the TBA in the urine (3.3 fold and 15.5 fold in the APAP and geniposide groups, respectively) was higher than that in the blood (290 fold and 640 fold in the APAP and geniposide groups, respectively). However, given the BA profiles after geniposide- and APAP-induced liver injury, BAs were found to be more suitable as biomarkers for diagnosing cholestatic liver injury. Overall, the BA assay developed in this study is rapid, simple, accurate, validated, sensit

Topics: Acetaminophen; Animals; Bile Acids and Salts; Biomarkers; Chemical and Drug Induced Liver Injury, Chronic; Chromatography; Enterohepatic Circulation; Iridoids; Liver; Rats; Tandem Mass Spectrometry

Spermatogenesis dysfunction is common in clinically infertile patients. Geniposide (GP) is one of the important active ingredients extracted from Eucommia ulmoides. However, the protective effect and mechanism of GP in the treatment of spermatogenic dysfunction is not known yet.. After cyclophosphamide-induced spermatogenic dysfunction was established in male mice, we gavaged GP for 4 weeks to evaluate spermatogenic function and anti-apoptotic effects by fertility, testicular weight, sperm quality, endoplasmic reticulum stress (ER stress), comet assay and serum testosterone level.. GP can improve the damage of fertility and reproductive organs induced by cyclophosphamide and increase the number and activity of sperm. In comet assay, it was found that GP administration could alleviate sperm DNA damage induced by cyclophosphamide. In addition, GP treatment can significantly reduce ThT fluorescence intensity and improve endoplasmic reticulum stress induced by cyclophosphamide. Besides, TUNEL staining and WB showed that GP could inhibit the excessive apoptosis of cells and protect testis. (p < 0.05, p < 0.01, p < 0.001).. The protective effect of Geniposide on cyclophosphamide-induced spermatogenic dysfunction in mice is related to the inhibition of endoplasmic reticulum stress.

Topics: Animals; Apoptosis; Cyclophosphamide; Endoplasmic Reticulum Stress; Iridoids; Male; Mice; Seeds; Spermatogenesis; Testis; Testosterone

Imbalance of macrophage polarization plays a critical role in the progression of rheumatoid arthritis (RA). Geniposide (GE) has been shown to exert anti-inflammatory effects. However, the effect of GE on macrophage polarization remains unclear. Here, we investigated the regulation of GE on the imbalance of macrophage polarization in RA and how it functions. We established a mouse model of collagen-induced arthritis (CIA) and isolated bone marrow-derived macrophages (BMDMs). The results confirmed that pro-inflammatory M1 macrophages were dominant in CIA mice, but the polarization imbalance of macrophages was restored to a certain extent after GE treatment. Furthermore, the membrane targeting of sphingosine kinase 1 (SphK1) was increased in BMDMs of CIA mice, as manifested by increased membrane and cytoplasmic expression of p-SphK1 and high secretion level of sphingosine-1-phosphate (S1P). RAW264.7 cells were stimulated with lipopolysaccharide (LPS)-interferon (IFN)-γ or interleukin (IL)-4 to induce M1 or M2 phenotype, respectively, to revalidate the results obtained in BMDMs. The results again observed SphK1 membrane targeting in LPS-IFN-γ-stimulated RAW264.7 cells. Selective inhibition of SphK1 by PF543 or inhibition of the S1P receptors by FTY720 both restored the proportion of M1 and M2 macrophages in LPS-IFN-γ-stimulated RAW264.7 cells, confirming that SphK1 membrane targeting mediated a proportional imbalance in M1 and M2 macrophage polarization. In addition, GE inhibited SphK1 membrane targeting and kinase activity. Taken together, results confirmed that the inhibition of SphK1 membrane targeting by GE was responsible for restoring the polarization balance of macrophages in CIA mice.

Topics: Animals; Anti-Inflammatory Agents; Arthritis, Experimental; Arthritis, Rheumatoid; Fingolimod Hydrochloride; Interferon-gamma; Iridoids; Lipopolysaccharides; Macrophages; Mice; Phosphotransferases (Alcohol Group Acceptor); Signal Transduction

Topics: Dose-Response Relationship, Drug; Drug Design; Enzyme Inhibitors; Gout Suppressants; Humans; Hyperuricemia; Iridoids; Molecular Structure; Structure-Activity Relationship; Xanthine Oxidase

Spinal cord injury (SCI) is a traumatic condition of the central nervous system , which can cause nerve injury and affect nerve regeneration, thus leading to severe dysfunction of motor and sensory pathways, and unfortunately these effects are irreversible. Inflammatory response constitutes one of the important mechanisms of spinal cord secondary injury. Geniposide (Gen) is reported to possess anti-inflammation and neuronal repair capacities.. To investigate the effect and mechanism of Gen on motor function and inflammatory response in SCI rats.. Sprague-Dawley (SD) rats were randomly grouped, and the SCI model was established by Allen's method. The motor function of rats was evaluated by the Basso, Beattie, and Bresnahan (BBB) scale. The protective effect of Gen on the injured spinal cord tissues was evaluated by measuring the water content, myeloperoxidase (MPO) activity, and levels of tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-6. Moreover, the protein level of the inflammation-related pathway was detected by spectrometry and Western blot assays.. Gen significantly promoted the recovery of SCI rats, decreased the edema of spinal cord tissues, reduced the area of cavity, increased the number of NF-200-positive neurons, as well as increased the number of horseradish peroxidase (HRP) retrograde tracing-positive neurons and regenerated axons with myelin sheath. Additionally, compared with the control group, the neutrophil infiltration, contents of TNF-α, IL-1β, and IL-6, the activity of inhibitor of nuclear factor κB kinase subunit β (IKKβ) kinase, and protein levels of (nuclear factor κB) NF-κB p65 and phosphorylated inhibitor of NF-κB (p-I-κB) in the Gen experimental group were significantly decreased.. Gen effectively alleviated inflammatory response after SCI by inhibiting the IKKs/NF-κB signaling pathway and promoted the recovery of motor function and axon regeneration in rats.. This study can provide novel insights for the early and effective intervention of SCI and confer basic data for the treatment of spinal cord secondary injury.

Topics: Animals; Anti-Inflammatory Agents; Cytokines; Disease Models, Animal; Female; I-kappa B Kinase; Inflammation Mediators; Iridoids; Motor Activity; NF-kappa B; Rats, Sprague-Dawley; Recovery of Function; Signal Transduction; Spinal Cord; Spinal Cord Injuries; Spinal Cord Regeneration

N-Methyl-D-aspartate receptor (NMDAR)-induced antioxidation is a significant cause of neuronal injury after ischemic stroke. In a previous work, we verified the neuroprotective roles of geniposide during tMCAO in vivo. However, it remains unknown whether geniposide ameliorates injury to hippocampal neurons during Ischemic Long Term Potentiation (iLTP) induction in vitro. After induction of cells oxygen-glucose deprivation or hydrogen peroxide, the protection of geniposide evaluated by MTT assay and electrophysiological tests. In this study, we suggested neuronal cell apoptosis was attenuated by geniposide. Furthermore, field excitatory postsynaptic potentials (fEPSCs) following postischemic LTP were assessed by electrophysiological tests. Finally, we determined that medium and high doses of geniposide attenuated oxidative stress insult and improved iLTP. Importantly, these effects were abolished by cotreatment with geniposide and the GluN2A antagonist NVP. In contrast, the GluN2B inhibitor ifenprodil failed to have an effect. In conclusion, we suggest for the first time that treatment with geniposide can attenuate postischemic LTP induction in a concentration-dependent manner. We infer that GluN2A-containing NMDARs are involved in the neuroprotection induced by geniposide treatment in ischemia.

Topics: Animals; Apoptosis; Excitatory Amino Acid Antagonists; Excitatory Postsynaptic Potentials; Hippocampus; Hydrogen Peroxide; Hypoxia-Ischemia, Brain; In Vitro Techniques; Infarction, Middle Cerebral Artery; Iridoids; Long-Term Potentiation; Neurons; Oxidants; PC12 Cells; Piperidines; Quinoxalines; Rats; Receptors, N-Methyl-D-Aspartate

Geniposide has been widely found to ameliorate many metabolic diseases. The recruitment and activation of brown/beige adipocytes are effective and promising methods for counteracting obesity and related diseases. However, the effect of geniposide on thermogenesis of adipocytes and its underlying mechanism have not yet been investigated. Here, we demonstrate that geniposide (25 mg/kg) reduces body temperature and cold tolerance of mice via suppressing thermogenic genes in interscapular brown adipose tissue (iBAT) and inguinal white adipose tissue (iWAT). Consistently, geniposide (20 mg/mL) suppresses thermogenic capacity of adipocytes (brown adipocytes and 3T3L1 preadipocyte cells) in vitro. Mechanistically, geniposide reduces the level of protein kinase A (PKA) catalytic subunit and further suppresses transcription activity and protein stability of uncoupling protein 1 (UCP1), leading to reduction of thermogenic capacity in adipocytes. Moreover, pharmacological PKA activation reverses geniposide-induced UCP1 inhibition, which indicated that geniposide suppresses thermogenesis of adipocytes via regulating PKA signaling. Together, our findings suggest that geniposide is an inhibitor of fat thermogenesis, establishing a novel function characteristic of geniposide.

Topics: 3T3-L1 Cells; Adipocytes; Adipose Tissue, Brown; Adipose Tissue, White; Animals; Body Temperature; Catalytic Domain; Cold Temperature; Cyclic AMP-Dependent Protein Kinases; Iridoids; Male; Mice; Mice, Inbred C57BL; Signal Transduction; Thermogenesis; Uncoupling Protein 1

Geniposide (GEN) is a natural antioxidant and anti-inflammatory product and plays an important role in the treatment of diabetes and diabetic complications. To explore the biological functions and mechanism of GEN in diabetic retinopathy (DR), we constructed the in vitro and in vivo model of DR by using primary cultured mouse retinal Müller cells and C57BL/6 mice, respectively. We found that GEN inhibited ROS accumulation, NF-

Topics: Animals; Diabetes Mellitus, Experimental; Diabetic Retinopathy; Ependymoglial Cells; Hyperglycemia; Inflammation; Iridoids; Male; Mice; Mice, Inbred C57BL; NF-E2-Related Factor 2; Oxidative Stress; Reactive Oxygen Species

Overexposure to glucocorticoid (GC) produces various clinical complications, including osteoporosis (OP), dyslipidemia, and hypercholesterolemia. Geniposide (GEN) is a natural iridoid compound isolated from Eucommia ulmoides. Our previous study found that GEN could alleviate dexamethasone (DEX)-induced differentiation inhibition of MC3T3-E1 cells. However, whether GEN protected against Dex-induced cholesterol accumulation in osteoblasts was still unclear.. DEX was used to induce rat OP. Micro-CT data was obtained. The ALP activity and mineralization were determined by the staining assays, and the total intracellular cholesterol was determined by the ELISA kits. The protein expression was detected by western blot.. GEN ameliorated Dex-induced micro-structure damages and cell differentiation inhibition in the bone trabecula in rats. In MC3T3-E1 cells, Dex enhanced the total intracellular cholesterol, which reduced the activity of cell proliferation and differentiation. Effectively, GEN decreased DEX-induced cholesterol accumulation, enhanced cell differentiation, and upregulated the expression of the GLP-1R/ABCA1 axis. In addition, inhibition of ABAC1 expression reversed the actions of GEN. Treatment with Exendin9-39, a GLP-1R inhibitor, could abrogate the protective activity of GEN.. GEN ameliorated Dex-induced accumulation of cholesterol and inhibition of cell differentiation by mediating the GLP-1R/ABCA1 axis in MC3T3-E1 cells.

Topics: 3T3 Cells; Animals; ATP Binding Cassette Transporter 1; Cell Differentiation; Cholesterol; Dexamethasone; Disease Models, Animal; Eucommiaceae; Gene Expression Regulation; Glucagon-Like Peptide-1 Receptor; Iridoids; Mice; Osteoblasts; Osteoporosis; Rats; Signal Transduction

To study the antidepressant-like effect and action mechanism of geniposide and eleutheroside B combination treatment on the lipopolysaccharide (LPS)-induced depression mice model.. Depression mice model was established by lipopolysaccharide (LPS) injection. Totally 48 mice were randomly divided into 6 groups (8 rats per group) according to a random number table, including normal, model, fluoxetine (20 mg/kg), geniposide (100 mg/kg) + eleutheroside B (100 mg/kg), geniposide + eleutheroside B + WAY 100635 (0.03 mg/kg), geniposide + eleutheroside B+ N-methyl-D-aspartic acid receptor (NMDA, 75 mg/kg) groups, respectively. After continuous administration for 10 days, autonomic activity tests after 30 min of administration were performed on the 10th day. On the 11th day, except for the normal group, the mice in the other groups were intraperitoneally injected with LPS (1 mg/kg), and the behavioral tests were performed 4 h later. Enzyme linked immunosorbent assay was used to detect tumor necrosis factor alpha (TNF- α) and interleukin-1 β (IL-1 β) levels in mice serum. The mRNA expression of indoleamine 2,3-dioxygenase (IDO) and nuclear transcription factor (NF- κB) were detected by real-time quantitative polymerase chain reaction. Western-blot analysis was used to detect IDO and NF- κB protein expressions in hippocampus tissue.. Compared with the normal group, a single administration of LPS increased the immobility time in the forced swimming test (FST) and tail suspension test (TST, P<0.01), without affecting autonomous activity. Compared with the model group, fluoxetine and geniposide + eleutheroside B administration significantly improved the immobility time of depressed mice in the FST and TST, decreased serum IL-1 β content, inhibited the expression levels of NF- κ B gene and protein in hippocampus tissues (P<0.05 or P<0.01). Compared with the model group, geniposide + eleutheroside B treatment significantly reduced serum TNF-α content and inhibited IDO mRNA and protein expressions in hippocampus (P<0.05 or P<0.01). In addition, NMDA partly prevented the inhibition of IDO mRNA expression by geniposide + eleutheroside B; NMDA and WAY-100635 also partly prevented the reduction of IL-1 ß content induced by geniposide + eleutheroside B treatment (P<0.05 or P<0.01).. The combination of geniposide and eleutheroside B showed a certain antidepression-like effect. Its main mechanism of action may be contributed to inhibiting the activation of NF- κB, decreasing the proinflammatory cytokines such as TNF-α, IL-1 β, and inhibiting in the neuroinflammatory reaction. Additionally, it also affects tryptophan metabolism, reduces the expression of a key enzyme of tryptophan metabolism, IDO. And this antidepressant-like effect may be mediated by 5-hydroxytryptamine and glutamate systems.

Topics: Animals; Antidepressive Agents; Depression; Glucosides; Iridoids; Lipopolysaccharides; Mice; Mice, Inbred ICR; NF-kappa B; Phenylpropionates; Rats; Tumor Necrosis Factor-alpha

Metabolic disorder particularly diabetes is one of the leading causes of psychiatric or other neurodegenerative diseases. Previous clinical and pre-clinical studies indicate anti-diabetic drugs such as GLP-1 analogs or GLP-1 receptor (GLP-1R) agonists could perform the neuroprotective effects with multiple molecular mechanisms. As one of natural compound to stimulate GLP-1R, geniposide was reported could improve cognitive behaviors in diabetes associated Alzheimer's disease rat model. Stimulating of GLP-1R could act the crosstalk downstream like neurotrophic factor mediated cAMP-response element binding protein (CREB) would be activated and exert cellular events including promotion of adult neurogenesis, which is one of important treatment targets in antidepressant. Here in this study, we employed HDF in combined with corticosterone (CORT) treatment to create diabetes associated depression model. Geniposide treatment could not only correct the metabolic pattern but could also improve the cognitive dysfunctions and depressive/anxiety symptoms. In consistent with its pro-neurogenic effects, geniposide also enhanced the activity of CREB in hippocampal tissue. Moreover, blocking CREB activity with 666-15 significantly compromised the effects of geniposide in promotion of neurogenesis and behavioral protective effects. In conclusion, this study expands the application of geniposide to treat diabetes associated depression subject and identified the underlying molecular mechanism for such effects.

Topics: Animals; Cyclic AMP Response Element-Binding Protein; Depression; Diabetes Mellitus; Iridoids; Mice; Neurogenesis; Rats

The purpose of this study was to estimate the pharmacological effect of geniposide (GEN) on depression, caused by chronic unpredictable mild stress (CUMS), and explore its potential mechanism. During the 6 week CUMS procedure, the mice were treated with GEN (10, 40 mg/kg) by gavage once daily for 3 weeks. As a result, the GEN treatment remarkably improved the behavioral manifestations and suppressed the generations of inflammatory cytokines both in vivo and in vitro. The MDA level was significantly increased, while the activities of SOD, GSH-PX were decreased in CUMS-challenged mice and corticosterone-stimulated PC12 cells. GEN administration significantly inhibited those changes. Moreover, GEN treatment could downregulate the expressions of p-BTK, TLR4, MyD88, p-NF-κB proteins, and upregulate BDNF, p-TrkB generations in CUMS-induced mice. Moreover, GEN administration inhibited the protein levels of p-BTK, TLR4, MyD88, p-NF-κB in corticosterone-induced PC12 cell. In summary, the results suggested that GEN exerted a therapeutic effect on CUMS-induced depressive mice possibly through the regulation of BTK/TLR4/NF-κB and BDNF/TrkB signaling pathways.

Topics: Animals; Brain-Derived Neurotrophic Factor; Iridoids; Male; Mice; NF-kappa B; PC12 Cells; Rats; Signal Transduction; Stress, Psychological; Toll-Like Receptor 4

Traditional Chinese medicine therapy, which can serve as adjuvant therapy for cancer treatment, has no obvious side effects on the human body. Geniposide (GEN), one of the main iridoid glycosides in gardenia fruit, has been widely reported to have anti-cancer effects. In this study, we aimed to inspect whether GEN could inhibit proliferation and promote the apoptosis of human breast cancer cells (MCF-7). In order to better predict the efficacy of GEN, we have prepared the Cs/Gel composite scaffolds by 3D printing technology to mimic the MCF-7 cell growth microenvironment. The prepared Cs/Gel scaffold has good mechanical properties and biocompatibility, which can provide a more accurate platform for drug screening. The semi-inhibitory concentration (IC50) evaluated by CCK-8 assay was 16.06 mg/mL (24 h), 14.85 mg/mL (48 h), and 13.14 mg/mL (72 h). After exposed to GEN for 48 h, the cancer cell survival rate reduced from 69.15 ± 2.86% (13 mg/mL) to 20.97 ± 3.24% (16 mg/mL). Although the inhibitory effect was weaker in the 3D culture system, it also managed to inhibit cell proliferation and induce cell apoptosis. Besides, Live/Dead staining, Hematoxylin-Eosin (H&E) staining and SEM evaluation were also conducted to estimate the anti-cancer effect of GEN in 2D and 3D cultures. The results indicate that GEN has an anti-cancer effect based on a time- and dose-dependent manner.

Topics: Breast Neoplasms; Cell Proliferation; Humans; Iridoids; Printing, Three-Dimensional; Tissue Scaffolds; Tumor Microenvironment

Thioredoxin-interacting protein (Txnip) has emerged as a key regulator of insulin resistance. In this study, we investigated the roles of geniposide and Txnip in insulin resistance in differentiated 3T3-L1 adipocytes. Our results revealed that geniposide markedly enhanced glucose uptake, increased the protein levels of insulin receptor substrate (IRS)-1 and GLUT-1, and prevented the phosphorylation of IRS-1 and Akt Thr308 induced by insulin resistance in 3T3-L1 adipocytes. We also observed that geniposide accelerated protein degradation of Txnip through proteasome pathway, and knockdown of Txnip with small interfering RNA attenuated the effect of geniposide on insulin signaling molecules, implying that Txnip played a pivotal role in the regulation of insulin signaling molecules by geniposide in 3T3-L1 adipocytes. Furthermore, geniposide induced the phosphorylation of adenosine monophosphate-activated protein kinase (AMPK) in the presence of high glucose in differentiated 3T3-L1 adipocytes, while compound C, an inhibitor of AMPK, prevented the effect of geniposide on Txnip degradation and the regulation of glucose uptake and insulin signaling molecules including p-IRS-1, IRS-1, and GLUT-1 in differentiated 3T3-L1 adipocytes. Taken together, all these findings suggest that geniposide improves the insulin signaling defect possibly by AMPK-mediated Txnip degradation in 3T3-L1 adipocytes.

Topics: 3T3-L1 Cells; AMP-Activated Protein Kinases; Animals; Carrier Proteins; Enzyme Activation; Insulin Resistance; Iridoids; Mice; Proteolysis; Signal Transduction; Thioredoxins

The purpose of this study was to investigate the antidepressant mechanism of GEN (geniposide) on depression mice induced by LPS. The mice were intragastrically treated with GEN (10 mg/kg/d or 40 mg/kg/d) or ibrutinib for continuous 7 days prior to LPS injection. The anxiety- and depression-like behaviors of mice were assessed via behavioral tests (sucrose preference test (SPT), tail suspension test (TST), forced swimming test (FST), and open-field test (OFT)). Microglial BV2 cells were treated with GEN or/and ibrutinib and stimulated with LPS. The productions of pro-inflammatory cytokines IL-6 and TNF-α in hippocampus, serum, and supernatant were detected by ELISA. The correlative proteins BTK, p-BTK, JAK2, p-JAK2, STAT1, p-STAT1, BDNF, TrkB, and p-TrkB were assessed through western blot. As a result, GEN ameliorated the anxiety- and depression-like behaviors of mice in behavioral tests. GEN treatment also regulated microglia polarization towards anti-inflammatory phenotype M2 and inhibited the production of pro-inflammatory cytokines IL-6 and TNF-α. In addition, with the application of ibrutinib, the selective inhibitor of BTK, it was proclaimed that the administration of GEN restrained the activation of JAK2/STAT1 pathway via attenuating the hyperphosphorylation of BTK both in mice and BV2 cells. Furthermore, it was also found that GEN activated BDNF/TrkB neuroprotective signaling pathway through the reduction of BTK phosphorylation. From the overall results, we suggested that GEN exerted a beneficial effect on LPS-induced depression in mice possibly through the modulation of BTK/JAK2/STAT1 signaling.

Topics: Agammaglobulinaemia Tyrosine Kinase; Animals; Behavior, Animal; Brain-Derived Neurotrophic Factor; Cell Line; Cytokines; Depression; Hippocampus; Iridoids; Janus Kinase 2; Lipopolysaccharides; Male; Mice; Mice, Inbred ICR; Neurons; Plant Extracts; Signal Transduction; STAT1 Transcription Factor

Diabetic nephropathy (DN) is a common pathological feature in patients with diabetes and the leading cause of end-stage renal disease. Although several pharmacological agents have been developed, the management of DN remains challenging. Geniposide, a natural compound has been reported for anti-inflammatory and anti-diabetic effects; however, its role in DN remains poorly understood. This study investigated the protective effects of geniposide on DN and its underlying mechanisms. We used a C57BL/6 mouse model of DN in combination with a high-fat diet and streptozotocin after unilateral nephrectomy and treated with geniposide by oral gavage for 5 weeks. Geniposide effectively improves DN-induced renal structural and functional abnormalities by reducing albuminuria, podocyte loss, glomerular and tubular injury, renal inflammation and interstitial fibrosis. These changes induced by geniposide were associated with an increase of AMPK activity to enhance ULK1-mediated autophagy response and a decrease of AKT activity to block oxidative stress, inflammation and fibrosis in diabetic kidney. In addition, geniposide increased the activities of PKA and GSK3β, possibly modulating AMPK and AKT pathways, efficiently improving renal dysfunction and ameliorating the progression of DN. Conclusively, geniposide enhances ULK1-mediated autophagy and reduces oxidative stress, inflammation and fibrosis, suggesting geniposide as a promising treatment for DN.

Topics: AMP-Activated Protein Kinases; Animals; Anti-Inflammatory Agents; Autophagy; Autophagy-Related Protein-1 Homolog; Cyclic AMP-Dependent Protein Kinases; Diabetes Complications; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Diet, High-Fat; Disease Models, Animal; Fibrosis; Glycogen Synthase Kinase 3 beta; Iridoids; Mice; Mice, Inbred C57BL; Oxidative Stress

Hypertension is closely related to myocardial injury. Long-term hypertension can cause myocardial injury. Therefore, it is very important to find drugs to treat myocardial injury caused by hypertension. The aim of present study is to investigate the effects and mechanisms of geniposide on myocardial injuries in spontaneously hypertensive rats (SHR) and H9c2 cells induced by NaCl solution.. Male Wistar-Kyoto (WKY) and SHR rats were given different doses of geniposide (25 mg/kg/d or 50 mg/kg/d) or distilled water for three consecutive weeks. Meanwhile, an H9c2 cell line-injury model was established using a solution of 150 µmol/L NaCl for 8 h. The cardiac function and related indexes of rats were detected.. The results showed that geniposide decreased the levels of COI and COIII, which promoted the phosphorylation of AMPK (p-AMPK) and enhanced the energy metabolism pathway. Geniposide improved myocardial apoptosis by regulating apoptotic proteins (p38, BAX and Bcl-2). Finally, heart function was regulated, and the markers of myocardial injury were decreased. Geniposide increased the viability of H9c2 cells treated with the NaCl solution and decreased the rate of apoptosis by regulating the levels of apoptotic proteins. Geniposide could activate energy metabolism signalling pathway (AMPK/SirT1/FOXO1) and reduce H9c2 cell apoptosis.. Our results showed that the mechanisms by which geniposide improves myocardial injury in SHR may be through regulating the energy metabolism signalling pathway (AMPK/SirT1/FOXO1) and improving myocardial apoptosis by regulating apoptotic proteins.

Topics: Animals; Apoptosis; Cells, Cultured; Dose-Response Relationship, Drug; Energy Metabolism; Gardenia; Iridoids; Male; Molecular Structure; Myocytes, Cardiac; Rats; Rats, Inbred SHR; Rats, Inbred WKY; Rats, Wistar; Signal Transduction; Structure-Activity Relationship

Sclerosing cholangitis, characterized by biliary inflammation, fibrosis, and stricturing, remains one of the most challenging conditions of clinical hepatology. Geniposide (GE) has anti-inflammatory, hepatoprotective, and cholagogic effects. Whether GE provides inhibition on the development of sclerosing cholangitis is unknown. Here, we investigated the role of GE in a mouse model in which mice were fed with 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) for 4 weeks to induce sclerosing cholangitis. The results demonstrated that the increased hepatic gene expressions of pro-inflammatory (IL-6, VCAM-1, MCP-1, and F4/80) and profibrogenic markers (Col1α1, Col1α2, TGF-β, and α-SMA) in DDC feeding mice were reversed after treatment with GE. GE also suppressed expressions of CK19 and Ki67 in DDC-fed mice, suggesting that GE could ameliorate DDC-induced hepatocytes and cholangiocytes proliferation. In addition, GE significantly increased bile acids (BAs) secretion in bile, which correlated with induced expressions of hepatic FXR, BAs secretion transporters (BSEP, MRP2, MDR1, and MDR2), and reduced CYP7A1 mRNA expression. Furthermore, higher expressions of ileal FXR-FGF15 signaling and reduced ASBT were also observed after GE treatment. Taken together, these data showed that GE could modulate inflammation, fibrosis, and BAs homeostasis in DDC-fed mice, which lead to efficiently delay the progression of sclerosing cholangitis.

Topics: Animals; Cholangitis, Sclerosing; Disease Models, Animal; Iridoids; Liver; Mice; Mice, Knockout

Genipin-1-β-d-gentiobioside (GG) is a kind of compound extracted from Gardenia jasminoides Ellis. The chemical structure of GG is similar to that of geniposide and has antidiabetic effects. We aimed to investigate the efficacy of GG on diabetic nephropathy (DN) in vivo and in vitro experiments and explore its potential mechanism.. For high-fat diet/streptozotocin-induced DN mice used in our study, the general features of mice were analysed after GG treatment. Oxidative stress parameters and inflammatory factors were also measured by commercial kits. Kidney damage was assessed using hematoxylin and eosin (H&E), periodic acid-Schiff (PAS) and Masson staining, respectively. In vitro, podocyte injury was assessed by TUNEL and flow cytometric analyses. AMP-activated protein kinase/silencing information regulator related enzyme 1 (AMPK/SIRT1)/nuclear factor-κB (NF-κB) pathway-related proteins were detected by AMPK-siRNA intervention and western blotting.. Treatment of GG could increase cell survival and attenuated kidney damage. Despite the presence of inflammatory and oxidative stress, when GG retained the expression of AMPK/SIRT1, it could be observed that the downstream NLRP3 inflammatory-related proteins were inhibited.. Results showed that the protective efficacy of GG on DN works together with hypoglycemia and suppressing oxidative stress and inflammation, which at least partly involved in APMK/SIRT1/NF-κB-dependent pathway.

Topics: AMP-Activated Protein Kinases; Animals; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Gardenia; Inflammation; Iridoids; Kidney; Male; Mice, Inbred C57BL; NF-kappa B; NLR Family, Pyrin Domain-Containing 3 Protein; Oxidative Stress; Phytotherapy; Plant Extracts; Podocytes; Sirtuin 1

Cerebral ischemia causes severe neurological disorders and neuronal dysfunction. Baicalin (BC), geniposide (GP), and their combination (BC/GP) have been shown to inhibit post-ischemic inflammatory injury by inhibiting the 5-LOX/CysLTs pathway. The aims of this study were to observe the inhibitory effects of BC/GP on the activation of microglial cells induced by oxygen glucose deprivation and reoxygenation (OGD/R) and to investigate whether the 5-LOX/LTB4 pathway was involved in these effects. Molecular docking showed that BC and GP exhibited considerable binding activity with LTB4 synthase LTA4H. BV-2 microglia were transfected with a 5-LOX overexpression lentiviral vector, and then OGD/R was performed. The effects of different concentrations of BC, GP, and BC/GP (6.25 μM, 12.5 μM, and 25 μM) on cell viability and apoptosis of microglia were evaluated by MTT and flow cytometry. The expression of TNF-α, IL-1β, NF-κB, and pNF-κB also was measured by ELISA, Western blots and immunofluorescence. Western blots and qRT-PCR analysis were used to determine the levels of CD11b, CD206, and 5-LOX pathway proteins. Results showed that BC, GP, and BC/GP reduced the apoptosis caused by OGD/R in a dose-dependent manner, and cell viability was significantly increased at a concentration of 12.5 μM. OGD/R significantly increased the release of TNF-α, IL-1β, NF-κB, pNF-κB, and CD11b. These effects were suppressed by BC, GP, and BC/GP, and the OGD/R-induced transfer of NF-κB p65 from the ctytoplasm to the nucleus was inhibited in microglia. Interestingly, the LTB4 inhibitor, U75302, exhibited the same effect. Also, BC, GP, and BC/GP significantly reduced the expression of 5-LOX pathway proteins. These results demonstrated that BC/GP inhibited OGD/R-induced polarization in BV2 microglia by regulating the 5-LOX/LTB4 signaling pathways and attenuating the inflammatory response. Our results supported the theoretical basis for additional in-depth study of the function of BC/GP and the value of determining its unique target, which might provide a new therapeutic strategy for ischemic cerebrovascular disease.

Topics: Amino Acid Sequence; Animals; Apoptosis; Arachidonate 5-Lipoxygenase; Cell Hypoxia; Cell Survival; Cells, Cultured; Epoxide Hydrolases; Flavonoids; Glucose; Humans; Inflammation; Iridoids; Mice; Microglia; Molecular Docking Simulation; Oxygen; Protein Binding; Signal Transduction

Gardeniae Fructus (Zhizi in Chinese, ZZ in brief), a commonly used herbal medicine, has aroused wide concern for hepatotoxicity, but the mechanism remains to be investigated. This study was aimed at investigating the mechanism of ZZ-induced liver injury in vivo and in vitro based on metabolomics and evaluating the hepatotoxicity prediction ability of the in vitro model. SD rats were administered with extracted ZZ and HepG2 cells were treated with genipin, the major hepatotoxic metabolite of ZZ. Liver, plasma, intracellular and extracellular samples were obtained for metabolomics analysis. As a result, ZZ caused plasma biochemical and liver histopathological alterations in rats, and induced purine and amino acid metabolism disorder in the liver and pyrimidine, primary bile acids, amino acid metabolism and pantothenate and CoA biosynthesis disorder in the plasma. Pyrimidine, purine, amino acid metabolism and pantothenate and CoA biosynthesis were also found to be disturbed in the genipin-treated HepG2 cells, which exhibited similarity with the result in vivo. This study comprehensively illustrates the underlying mechanism involved in ZZ-related hepatotoxicity from the aspect of metabolome, and provides evidence that identifying hepatotoxicity can be achieved in cells, representing a non-animal alternative for systemic toxicology.

Topics: Animals; Cell Survival; Chemical and Drug Induced Liver Injury; Fruit; Gardenia; Hep G2 Cells; Humans; Iridoids; Plant Extracts; Rats; Rats, Sprague-Dawley

Age-related macular degeneration (AMD), mainly wet AMD, is the major reason for nonreversible vision loss worldwide. Choroidal neovascularization (CNV) is a characteristic pathological manifestation of wet AMD. Stress or injury to the retinal pigment epithelium (RPE) induces proangiogenic factors that drive CNV. An iridoid glycoside extracted from the fruit of gardenia, geniposide (GEN) plays an antiangiogenic role. In this study, GEN inhibited the transcription and expression of heparin-binding epidermal growth factor (HB-EGF), a proangiogenic factor, in hypoxic RPE cells and a mouse laser-induced CNV model. Inhibition of glucagon-like peptide-1 receptor (GLP-1R), a GEN receptor blocker, eliminated the protective effect of GEN. Additionally, GEN decreased the transcription and expression of HB-EGF in hypoxia-exposed RPE cells by downregulating the miR-145-5p/NF-κB axis. Therefore, our research provides a promising novel strategy for wet AMD therapy.

Topics: Animals; Cells, Cultured; Choroidal Neovascularization; Disease Models, Animal; Down-Regulation; Gene Expression Regulation; Heparin-binding EGF-like Growth Factor; Iridoids; Male; Mice; MicroRNAs; Retinal Pigment Epithelium

Depression is a severe mental disorder. Unfortunately, more than half of patients with major depression disorder cannot achieve remission after initial treatment with an antidepressant. Geniposide, a bioactive iridoid glycoside isolated from Gardenia jasminoides Ellis, can ameliorate depressive-like behaviors in mice. However, the underlying mechanism is still not very clear.. The pharmacological methods including ELISA, immunofluorescence, and Western blot were used to investigate the role of geniposide on chronic unpredictable mild stress (CUMS)-induced depression mice.. In this study, we found that geniposide could inhibit CUMS-induced depressive-like behaviors in mice. Geniposide is able to reduce the levels of ceramide and lower the activity of acid sphingomyelinase (ASM) in hippocampus; besides, ASM inhibitor (amitriptyline) can decrease the concentration of ceramide and ameliorate depressive-like behaviors of mice. Moreover, geniposide can also alleviate CUMS-induced hippocampal neuronal apoptosis and increase the phosphorylated form of PI3K, Akt, and GSK3β. Additionally, PI3K inhibitor (LY294002) can also abolish the neuroprotective effect of geniposide on hippocampal neurons in vitro.. These results indicate that geniposide exert a potential antidepressant-like effect on CUMS mice, and its effect might be associated with activated PI3K/Akt/GSK3β signaling, reduced the level of ceramide and hippocampal neuron apoptosis.

Topics: Animals; Ceramides; Depression; Depressive Disorder, Major; Glycogen Synthase Kinase 3; Hippocampus; Humans; Iridoids; Mice; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Stress, Psychological

The VEGF/SphK1/S1P pathway is closely related to angiogenesis in rheumatoid arthritis (RA), but the precise underlying mechanisms are unclear at present. Here, we explored the involvement of the VEGF/SphK1/S1P cascade in RA models and determined the effects of GE intervention. Our results showed abnormal expression of proteins related to this pathway in RA synovial tissue. Treatment with GE effectively regulated the signal axis, inhibited angiogenesis, and alleviated RA symptoms. In vitro, TNF-ɑ enhanced the VEGF/SphK1/S1P pathway in a co-culture model of fibroblast-like synoviocytes (FLS) and vascular endothelial cells (VEC). GE induced downregulation of VEGF in FLS, restored the dynamic balance of pro-/antiangiogenic factors, and suppressed SphK1/S1P signaling in VEC, resulting in lower proliferation activity, migration ability, tube formation ability, and S1P secretion ability of VEC cells. Additionally, SphK1-specific small interfering RNA (siRNA) blocked the VEGF/SphK1/S1P cascade, which can effectively alleviate the stimulatory effect of FLS on VEC and further enhanced the therapeutic effect of GE. Taken together, our results demonstrate that GE suppresses the VEGF/SphK1/S1P pathway and alleviates the stimulation of VEC by FLS, thereby preventing angiogenesis and promoting therapeutic effects against RA.

Topics: Adaptor Proteins, Signal Transducing; Arthritis, Rheumatoid; Cell Proliferation; Cells, Cultured; Endothelial Cells; Fibroblasts; Humans; Iridoids; Neovascularization, Pathologic; Signal Transduction; Sphingosine-1-Phosphate Receptors; Synovial Membrane; Vascular Endothelial Growth Factor A

Idiopathic mesenteric phlebosclerosis (IMP) is a rare disease, and its etiology and risk factors remain uncertain.. To investigate the possible influence of Chinese herbal liquid containing geniposide on IMP.. The detailed formula of herbal liquid prescriptions of all patients was studied, and the herbal ingredients were compared to identify the toxic agent as a possible etiological factor. Abdominal computed tomography (CT) and colonoscopy images were reviewed to determine the extent and severity of mesenteric phlebosclerosis and the presence of findings regarding colitis. The disease CT score was determined by the distribution of mesenteric vein calcification and colon wall thickening on CT images. The drinking index of medicinal liquor was calculated from the daily quantity and drinking years of Chinese medicinal liquor. Subsequently, Spearman's correlation analysis was conducted to evaluate the correlation between the drinking index and the CT disease score.. The mean age of the 8 enrolled patients was 75.7 years and male predominance was found (all 8 patients were men). The patients had histories of 5-40 years of oral Chinese herbal liquids containing geniposide and exhibited typical imaging characteristics (. Long-term oral intake of Chinese herbal liquid containing geniposide may play a role in the pathogenesis of IMP.

Topics: Aged; Colon; Colonoscopy; Humans; Iridoids; Male; Mesenteric Veins

The changes of fibroblast-like synoviocytes (FLSs) and vascular endothelial cells (VECs) biological functions are closely related to angiogenesis in rheumatoid arthritis (RA). Nevertheless, how the crosstalk between FLSs and VECs interferes with RA is far from being clarified. Herein, we studied the effect of the reciprocal interactions between FLSs and VECs on angiogenesis and mechanism of geniposide (GE). After administration of GE, improvement of synovial hyperplasia in adjuvant arthritis rats was accompanied by downregulation of SphK1 and p-Erk1/2. The dynamic interaction between FLSs and VECs triggers the release of S1P by activating p-Erk1/2 and SphK1, then activating RhoA-F-actin and Ras-Erk1/2 pathways. When exposed to the inflammatory microenvironment mediated by FLSs-VECs crosstalk, proliferation, migration, and permeability of VECs were enhanced, the angiogenic factors were imbalanced. Meanwhile, the proliferation and secretory ability of FLSs increased. Interestingly, depletion of S1P or blocking of the activation of SphK1 by GE and PF-543 prevented the changes. In conclusion, S1P released during FLSs-VECs crosstalk changed their biological functions by activating RhoA-F-actin and Ras-Erk1/2 pathways. GE acted on p-Erk1/2 and SphK1, inhibited the secretion of S1P, and blocked the interplay between FLSs and VECs. These results provide new insights into the mechanism of angiogenesis in RA.

Topics: Actins; Animals; Arthritis, Experimental; Cell Movement; Cell Proliferation; Cells, Cultured; Endothelial Cells; Fibroblasts; Iridoids; Lysophospholipids; Rats; Signal Transduction; Sphingosine; Synovial Membrane

Type 2 diabetes mellitus( T2 DM) is a common chronic metabolic disease characterized by persistent hyperglycemia and insulin resistance. In pancreatic β-cells,glucose-stimulated insulin secretion( GSIS) plays a pivotal role in maintaining the balance of blood glucose level. Previous studies have shown that geniposide,one of the active components of Gardenia jasminoides,could quickly regulate the absorption and metabolism of glucose,and affect glucose-stimulated insulin secretion in pancreatic β cells,but the specific mechanism needs to be further explored. Emerging evidence indicated that glycosylation of glucose transporter( GLUT) has played a key role in sensing cell microenvironmental changes and regulating glucose homeostasis in eucaryotic cells. In this study,we studied the effects of geniposide on the key molecules of GLUT2 glycosylation in pancreatic β cells. The results showed that geniposide could significantly up-regulate the mRNA and protein levels of Glc NAc T-Ⅳa glycosyltransferase( Gn T-Ⅳa) and galectin-9 but had no signi-ficant effect on the expression of clathrin,and geniposide could distinctively regulate the protein level of Gn T-Ⅳa in a short time( 1 h) under the conditions of low and medium glucose concentrations,but had no significant effect on the protein level of galectin-9. In addition,geniposide could also remarkably affect the protein level of glycosylated GLUT2 in a short-time treatment. The above results suggested that geniposide could quickly regulate the protein level of Gn T-Ⅳa,a key molecule of protein glycosylation in INS-1 rat pancreatic βcells and affect the glycosylation of GLUT2. These findings suggested that the regulation of geniposide on glucose absorption,metabolism and glucose-stimulated insulin secretion might be associated with its efficacy in regulating GLUT2 glycosylation and affecting its distribution on the cell membrane and cytoplasm in pancreatic β cells.

Topics: Animals; Diabetes Mellitus, Type 2; Glucose; Glycosylation; Insulin; Insulin-Secreting Cells; Iridoids; Rats

Geniposide (GE) possesses excellent neuroprotective effects but with poor brain targeting and short half-life. Liposome was considered to have great potential for brain diseases. Therefore, this research aimed to develop a geniposide liposome (GE-LP) as a brain delivery system for cerebral ischemia reperfusion injury (CIRI) therapy and evaluate its characterization, pharmacokinetics, brain targeting, and neuroprotective effects in vivo. Then, a reverse-phase evaporation method was applied to develop the GE-LP and optimize the formulation. Notably, the GE-LP had suitable size, which was 223.8 nm. Subsequently, the pharmacokinetic behavior of GE solution and GE-LP in mice plasma was investigated, and the brain targeting was also researched. The results showed that GE in plasma of GE-LP displayed three folds longer distribution half-life and a higher bioavailability and brain targeting compared to GE solution. In vivo neuroprotective effects was evaluated through the middle cerebral artery occlusion (MCAO) rat model, and GE-LP exhibited a stronger tendency in preventing the injury of CIRI, which can significantly improve neurological deficits. Overall, this study demonstrates GE-LP as a new formulation with ease of preparation, sustained release, and high brain targeting, which has significant development prospects on CIRI; this is expected to improve the efficacy of GE and reduce the frequency of administration.

Topics: Animals; Brain; Iridoids; Liposomes; Mice; Rats; Rats, Sprague-Dawley; Reperfusion Injury

We aimed to investigate whether geniposide, a main component extracted from Gardenia jasminoides Ellis fruit, could exert neuroprotective functions against traumatic brain injury (TBI). Enzyme-linked immunosorbent assay (ELISA) was used for detection of plasma cytokines. Real-time polymerase chain reaction (RT-PCR) was employed for measurements of mRNA levels of cytokines. Neurological outcomes were evaluated by modified neurological severity score (mNSS) and Rota-Rod. Blood-brain barrier (BBB) integrity and brain edema were assessed. Protein expression was tested by Western blot. The plasma levels of interleukin (IL)-1β, IL-6, IL-8 and IL-10 were all elevated in patients with TBI compared to those of healthy controls. TBI rats treated with geniposide showed lower mNSS and longer fall latency time than untreated TBI rats. BBB integrity was maintained and brain edema was reduced by geniposide treatment in TBI rats. Plasma levels of IL-1β, IL-6 and IL-8 were significantly repressed by geniposide treatment in TBI rats, whereas IL-10 level was upregulated. mRNA expression levels of these cytokines in the brain tissues of TBI rats exhibited the same trends of changes. By testing p38 mitogen-activated protein kinase and NF-κB p65 activities, it was observed that phosphorylated (p)-p38 and p-p65 were dramatically inhibited by geniposide. In conclusion, geniposide exerts neuroprotective functions against TBI by inhibiting p-p38 and p-p65.

Topics: Adolescent; Adult; Animals; Anti-Inflammatory Agents; Blood-Brain Barrier; Brain Edema; Brain Injuries, Traumatic; Cytokines; Female; Humans; Imidazoles; Iridoids; Male; Middle Aged; Mitogen-Activated Protein Kinases; NF-kappa B; Phosphorylation; Pyrimidines; Rats, Sprague-Dawley; RNA, Messenger; Treatment Outcome; Young Adult

Liver cancer is distinguished as an irredeemable disease. We detected the geniposide (GEN) in HepG2 and Huh7 cell lines.. HepG2 and Huh7 cells were individually induced with GEN dilutions, and then they were transfected with microRNA (miR)-224 overproduction vector (miR-224 mimic) as well as the corresponding negative control (NC). Cell viability was detected with the CCK-8. The apoptotic rate was determined by the Annexin V-FITC/PI with flow cytometer. The migration or invasion rates were separately determined by migration assay or millicell hanging cell culture. The expression of miR-224 was quantified depending on qRT-PCR. Relative proteins were individually determined via western blot.. GEN treatment induced inhibition of HepG2 and Huh7 cells proliferation, migration and invasion but promotion of apoptosis. miR-224 was down-regulated by GEN. Transfection of miR-224 mimic led to high expression of miR-224, which partly rescued cancer cells survival by prohibiting cell apoptosis. Moreover, the production of Wnt/β-catenin and AKT proteins was notably reduced by GEN but increased by overexpressed miR-224.. GEN played anti-tumor roles by targeting miR-224 via blocking the Wnt/β-catenin and AKT cascades in the HepG2 and Huh7 cells.

Topics: Apoptosis; beta Catenin; Cell Movement; Cell Proliferation; Cell Survival; Gene Expression Regulation, Neoplastic; Hep G2 Cells; Humans; Iridoids; Liver Neoplasms; MicroRNAs; Neoplasm Invasiveness; Proto-Oncogene Proteins c-akt

The purpose of the present study was to elucidate the pharmacological effects of Geniposide (GEN) on high diet fed and streptozotocin (STZ)-caused diabetic cognitive impairment. The mice were fed with high fat diet (HFD) for 4 weeks and intraperitoneally injected with 60 mg/kg STZ for three times within 72 h. The mice with glucose level over 15 mmol/l were regarded as diabetic and selected for further studies. The animals were intragastrically treated with metformin or GEN once daily for 4 weeks. Afterwards, the animals were applied for Y maze, novel object recognition (NOR) test, step-through passive avoidance test, and Morris water maze (MWM) test. The blood glucose and body weight were examined. The SH-SY5Y cells were treated with GEN in the presence or absence of ibrutinib and stimulated with high-glucose culture medium. The tumor necrosis factor-a (TNF-α) and interleukin (IL)-6 in serum, hippocampus, and supernatant were measured using ELISA method. The protein expressions of Bruton's tyrosine kinase (BTK), Toll-like receptor 4 (TLR4), myeloid differentiating factor 88 (MyD88), nuclear factor kappa-B (NF-κB), p-NF-κB, brain-derived neurotrophic factor (BDNF), cAMP-response element binding protein (CREB), p-CREB, and glucagon-like peptide-1 receptor (GLP-1R) were detected by western blot analyses. As a result, the GEN treatment notably attenuated the body weight, blood glucose, and cognitive decline. GEN also inhibited the generations of inflammatory cytokines. Furthermore, the administrations of GEN ameliorated the alterations of BTK, TLR4, MyD88, NF-κB, and BDNF in HFD + STZ-induced mice. With the application of ibrutinib, the selective inhibitor of BTK, it was also found that BTK/TLR4/NF-κB pathway was associated with the GEN treatment in high glucose-induced SH-SY5Y cells. In summary, the results suggested that GEN exerted the protective effect on STZ-induced cognitive impairment possibly through the modulation of BTK/TLR4/NF-κB signaling.

Topics: Agammaglobulinaemia Tyrosine Kinase; Animals; Blood Glucose; Cell Line, Tumor; Cognitive Dysfunction; Diabetes Mellitus; Dose-Response Relationship, Drug; Humans; Iridoids; Male; Mice; Mice, Inbred ICR; NF-kappa B; Random Allocation; Signal Transduction; Streptozocin; Toll-Like Receptor 4; Treatment Outcome

High-throughput metabolomics can clarify the underlying molecular mechanism of diseases via the qualitative and quantitative analysis of metabolites. This study used the established Yang Huang syndrome (YHS) mouse model to evaluate the efficacy of geniposide (GEN). Urine metabolic data were quantified by ultraperformance liquid chromatography-tandem mass spectrometry. The non-target screening of the massive biological information dataset was performed, and a total of 33 metabolites, including tyramine glucuronide, aurine, and L-cysteine, were identified relating to YHS. These differential metabolites directly participated in the disturbance of phase I reaction and hydrophilic transformation of bilirubin. Interestingly, they were completely reversed by GEN. While, as the auxiliary technical means, we also focused on the molecular prediction and docking results in network pharmacological and integrated analysis part. We used integrated analysis to communicate the multiple results of metabolomics and network pharmacology. This study is the first to report that GEN indirectly regulates the metabolite "tyramine glucuronide" through its direct effect on the target heme oxygenase 1 in vivo. Meanwhile, heme oxygenase-1, a prediction of network pharmacology, was the confirmed metabolic enzyme of phase I reaction in hepatocytes. Our study indicated that the combination of high-throughput metabolomics and network pharmacology is a robust combination for deciphering the pathogenesis of the traditional Chinese medicine (TCM) syndrome.

Topics: Animals; Biomarkers; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Iridoids; Metabolic Networks and Pathways; Metabolomics; Mice

The aim of this study is to investigate the protective effects as well as the underlying molecular mechanisms of geniposide in APP/PS1 transgenic mice.. APP/PS1 mice were subjected to intragastric administration of geniposide (50 mg/kg/d) for 8 weeks (including a 2-week behavior test). The novel object recognition (NOR) and the Morris water maze (MWM) tests were used for behavioral assessments. Aβ1-40 plaques in mice cortices and hippocampi are visualized with immunohistochemistical staining. ELISA was used to quantify the levels of soluble Aβ1-40 and Aβ1-42 in the hippocampus. Western blot was used to detect p-Akt/Akt, p-mTOR/mTOR and p-4E-BP1/4E-BP1 levels. The relative mRNA levels of Akt, mTOR and 4E-BP1 were quantified using real-time PCR (RT-PCR).. Geniposide alleviated cognitive impairment by improving the ability of novel object exploration, spatial memory, and reduced the level of Aβ in the brain of APP/PS1 mice. Geniposide possibly regulates mTOR-related proteins through modification of phosphorylation. Geniposide markedly lowered p-mTOR and p-Akt expressions while elevating p-4E-BP1 expression. Geniposide obviously reduced the relative mRNA levels of Akt and mTOR and increased the relative mRNA level of 4E-BP1.. Geniposide is able to alleviate cognitive impairments and cerebral damage in APP/PS1 mice, with its neuroprotective effects likely mediated via modulation of the mTOR signaling pathway.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Cerebral Cortex; Cognitive Dysfunction; Disease Models, Animal; Hippocampus; Iridoids; Male; Mice, Inbred C57BL; Mice, Transgenic; Neuroprotective Agents; Peptide Fragments; Plaque, Amyloid; Signal Transduction; TOR Serine-Threonine Kinases

The research plans to make sure how Geniposide (GEN) functions in osteoblast proliferation and differentiation. The MC3T3-E1 and ATDC5 cells were treated with the GEN, XAV-939 and/or transfected with microRNA (miR)-214 mimic or corresponding control. Cell viability was detected with the CCK-8. The CyclinD1, Runx2, Osx, Ocn, Wnt3a and β-catenin were individually quantified via western blot. The cell cycle was tested by cell cycle analysis assay. The ALP activity was tested by ALP assay. qRT-PCR was used to examine the miR-214 expression level. The cell viability and the expressions of the CyclinD1, Runx2, Osx, Ocn Wnt3a and β-catenin, as well as the ALP activity were individually and significantly promoted by the GEN. Besides, miR-214 was down-regulated by the GEN. The XAV-939 or the miR-214 mimic destroyed the promotional effect of GEN on these elements above. In conclusion, GEN induced the proliferation and differentiation of the MC3T3-E1 and ATDC5 cells by targeting the miR-214 through Wnt/β-catenin activation.

Topics: 3T3 Cells; Animals; Cell Differentiation; Cell Proliferation; Down-Regulation; Fractures, Bone; Heterocyclic Compounds, 3-Ring; Humans; Iridoids; Mice; MicroRNAs; Osteoblasts; Osteogenesis; Wnt Signaling Pathway

Medulloblastoma is a common tumor originates from central nervous system in children with metastatic potential. Geniposide is the major active ingredient separated from the fruit of Gardenia jasminoides Ellis. Herein, we tested the possible anticancer activity of geniposide on human medulloblastoma cells, as well as the potential underlying molecular mechanisms.. Firstly, followed by geniposide incubation, cell viability, proliferation, apoptosis, migration, and invasion of medulloblastoma Daoy cells, along with microRNA-373 (miR-373) expression were tested, respectively. Then, the influences of miR-373 overexpression in the reduction of medulloblastoma cell proliferation, migration, and invasion and the elevation of apoptosis, triggered by geniposide treatment, were re-investigated. Finally, the Ras/Raf/MEK/ERK pathway activity was analyzed.. Geniposide treatment inhibited medulloblastoma cell viability, proliferation, migration, and invasion, but promoted cell apoptosis. Surprisingly, miR-373 expression in medulloblastoma cells was obviously downregulated by geniposide treatment. miR-373 overexpression reversed the effects of geniposide on Daoy cells. Furthermore, geniposide hindered the Ras/Raf/MEK/ERK pathway by downregulating miR-373 expression.. Geniposide exhibited anticancer activity on human medulloblastoma cells and blocked Ras/Raf/MEK/ERK pathway by downregulating miR-373 expression.

Topics: Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Cerebellar Neoplasms; Down-Regulation; Fruit; Gardenia; Humans; Iridoids; MAP Kinase Signaling System; Medulloblastoma; MicroRNAs; Neoplasm Invasiveness; Plant Extracts; Transfection

Uncontrolled inflammatory response and subsequent cardiomyocytes loss (apoptosis and pyroptosis) are closely involved in sepsis-induced myocardial dysfunction. Our previous study has found that geniposide (GE) can protect the murine hearts against obesity-induced inflammation. However, the effect of GE on sepsis-related cardiac dysfunction is still unknown. Mice were exposed to lipopolysaccharide (LPS) to generate sepsis-induced myocardial dysfunction. And 50 mg/kg GE was used to treat mice for consecutive 7 days. Our results showed that GE treatment significantly improved survival rate and cardiac function, and suppressed myocardial inflammatory response, as well as myocardial loss in LPS-treated mice. Those effects of GE were largely abolished in NOD-like receptor protein 3 (NLRP3)-deficient mice. Further detection revealed that the inhibition of NLRP3 inflammasome activation depended on the reduction of p47phox by GE. GE treatment restored the phosphorylation and activity of AMP-activated protein kinase α (AMPKα) in the hearts of sepsis mice, and knockout of AMPKα abolished the protection of GE against reactive oxygen species (ROS) accumulation, NLRP3 inflammasome activation and cardiomyocytes loss in sepsis mice. In conclusion, our findings revealed that GE activated AMPKα to suppress myocardial ROS accumulation, thus blocking NLRP3 inflammasome-mediated cardiomyocyte apoptosis and pyroptosis and improving cardiac function in mice with sepsis.

Topics: AMP-Activated Protein Kinases; Animals; Inflammasomes; Iridoids; Mice; NLR Family, Pyrin Domain-Containing 3 Protein; Sepsis

As a typical hypervascular tumour, hepatocellular carcinoma (HCC) is predominantly grown through angiogenesis. Geniposide is a promising anti-inflammatory compound found in Gardenia jasminoides, but its effects on the progression of HCC remain untested.. The anti-HCC effects of geniposide was investigated in cellular models and orthotopic HCC mice. Transcriptional regulation of the VEGF promoter was measured by dual-luciferase reporter assay. The anti-angiogenic action of geniposide was measured by tube formation assay. Both surface plasmon resonance techniques and human phospho-kinase array analysis were utilized to validate the relationship between targets of geniposide and hepatocarcinogenesis.. Geniposide exhibited significant disruption of HCC proliferation, invasion, angiogenesis and lung metastasis in orthotopic HCC mice. Geniposide inhibited secretion of VEGF by HCC and suppressed the migration of endothelial cells and the formation of intra-tumour blood vessels, without cytotoxicity and independently of the transcription factor HIF-1α. Direct inhibition of TLR4 by geniposide led to the shutdown of the TLR4/MyD88 pathway and STAT3/Sp1-dependent VEGF production. However, LPS, an agonist of TLR4, restored STAT3/Sp1-related VEGF production in geniposide-inhibited HCC angiogenesis.. The direct inhibitory effect of geniposide on TLR4/MyD88 activation contributes to the suppression of STAT3/Sp1-dependent VEGF overexpression in HCC angiogenesis and pulmonary metastasis. This action of geniposide was not affected by stabilization of HIF-1α. Our study offers a novel anti-VEGF mechanism for the inhibition of HCC.

Topics: Animals; Carcinoma, Hepatocellular; Cell Line, Tumor; Endothelial Cells; Hypoxia-Inducible Factor 1, alpha Subunit; Iridoids; Liver Neoplasms; Mice; Myeloid Differentiation Factor 88; Neovascularization, Pathologic; Toll-Like Receptor 4; Vascular Endothelial Growth Factor A

To assess geniposide's effects in New Zealand rabbits with high-fat diet induced atherosclerosis and to explore the underpinning mechanisms.. Aorta histological changes were evaluated by intravenous ultrasound (IVUS) and H&E staining. Lipid accumulation in the aortic was quantified by Oil Red O staining. Then, RNA sequencing (RNA-seq) was carried out for detecting differentially expressed genes in rabbit high-fat diet induced atherosclerosis. The levels of the cytokines CRP, IL-1β and IL-10 were determined by ELISA. Protein levels of iNOS and Arg-1 were assessed by Western blot and immunohistochemical staining. The mRNA expression levels of NR4A1, CD14, FOS, IL1A, iNOS and Arg-1 were detected by quantitative real-time PCR (qPCR).. Geniposide markedly reduced the degree of atherosclerotic lesions in aorta tissues. RNA-seq and qPCR demonstrated that NR4A1, CD14, FOS and IL1A mRNA amounts were overtly increased in New Zealand rabbits with high-fat diet induced atherosclerosis. Moreover, geniposide reduced iNOS (M1 phenotype) mRNA and protein amounts as well as IL-1β secretion, which were enhanced in New Zealand rabbits with high-fat diet induced atherosclerosis. Besides, Arg-1 (M2 phenotype) mRNA and protein amounts were significantly increased after geniposide treatment, as well as IL-10 secretion.. These findings suggest that geniposide could inhibit the progression of and stabilize atherosclerotic plaques in rabbits by suppressing M1 macrophage polarization and promoting M2 polarization through the FOS/MAPK signaling pathway.

Topics: Animals; Atherosclerosis; Cytokines; Diet, High-Fat; Disease Progression; Iridoids; Macrophages; Male; MAP Kinase Signaling System; Plaque, Atherosclerotic; Proto-Oncogene Proteins c-fos; Rabbits; RNA, Messenger; Signal Transduction

Thioredoxin-interacting protein (Txnip), a negative regulator of thioredoxin, has become an attractive therapeutic target to alleviate metabolic diseases. Our previous data demonstrated that geniposide improved glucose-stimulated insulin secretion by accelerating Txnip degradation and prevented the early-stage apoptosis of pancreatic β cells induced by palmitate, but the underlying mechanisms are still unclear. The objective of this study is to identify the role of Txnip in geniposide preventing the apoptosis of pancreatic β cells induced by high glucose and palmitate (HG/PA). The results revealed that geniposide attenuated HG/PA-induced cell apoptosis and the expression of Bax and caspase-3, while increasing mitochondrial membrane potential and the anti-apoptotic protein levels of heme-oxygenase-1 (HO-1) and Bcl-2 in INS-1 rat pancreatic β cells. Knockdown of the Txnip gene raised the levels of anti-apoptotic proteins HO-1 and Bcl-2 and geniposide potentiated the effect of Txnip when the INS-1 cells were challenged by HG/PA. Furthermore, geniposide enhanced the adoptive unfolded protein response by increasing the phosphorylation of PERK/eIF2α and IRE1α in HG/PA-treated INS-1 cells. The results together suggest that geniposide might be useful to antagonize glucolipotoxicity and Txnip might be a pleiotropic cellular factor in pancreatic β cells.

Topics: Animals; Apoptosis; Cell Cycle Proteins; Endoplasmic Reticulum Stress; Endoribonucleases; Glucose; Insulin Secretion; Insulin-Secreting Cells; Iridoids; Protein Serine-Threonine Kinases; Rats; Thioredoxins

Topics: Animals; Calibration; Chromatography, High Pressure Liquid; Dogs; Drugs, Chinese Herbal; Glucosides; Iridoids; Limit of Detection; Male; Mass Spectrometry; Monoterpenes; Quality Control; Reproducibility of Results

Topics: AMP-Activated Protein Kinases; Animals; Gene Expression Regulation; Hep G2 Cells; Humans; Inflammation; Iridoids; Lipids; Male; Mice; Mice, Inbred C57BL; NF-E2-Related Factor 2; Non-alcoholic Fatty Liver Disease; Oxidative Stress; Palmitic Acid; Phosphatidylinositol 3-Kinases; Phosphorylation; Polyethylene Glycols; Signal Transduction; TOR Serine-Threonine Kinases

Oxidized low‑density lipoprotein (ox‑LDL)‑induced vascular endothelial damage, oxidative stress and inflammation play a vital role in the pathophysiology of atherosclerosis. Geniposide is the primary active ingredient from Gardenia jasminoides Ellis associated with anti‑oxidative properties and cardioprotective action. However, the therapeutic mechanism of geniposide in atherosclerosis remains unclear. Hence, the present study aimed to elucidate the underlying mechanisms of geniposide in oxidative stress and inflammatory response during ox‑LDL injury in human umbilical vein endothelial cells (HUVECs), focusing particularly on the microRNA (miR)‑21/PTEN pathway. The results demonstrated that geniposide pretreatment significantly increased cell viability, decreased lactate dehydrogenase release, increased miR‑21 level and decreased PTEN expression under ox‑LDL condition. Subsequently, transfection with miR‑21 mimic enhanced the protection of geniposide on ox‑LDL‑induced cytotoxicity and apoptosis (mediated by the upregulation of apoptotic rate and caspase‑3 activity), whereas miR‑21 inhibitor reversed these effects of geniposide. In addition, geniposide resulted in an anti‑oxidant effect as evidenced by the decrease in reactive oxygen species generation, malondialdehyde content and NADPH oxidase 2 expression, and the increase in superoxide dismutase, glutathione peroxidase and catalase activities in ox‑LDL‑treated HUVECs, which were exacerbated by miR‑21 mimic and reversed by miR‑21 inhibitor. Furthermore, geniposide mitigated the ox‑LDL‑induced inflammatory response, demonstrated by a downregulation of pro‑inflammatory cytokine (IL‑1β, IL‑6, and TNF‑α) levels and an upregulation of anti‑inflammatory cytokine (IL‑10) level. However, miR‑21 mimic enhanced, whereas miR‑21 inhibitor attenuated, these effects of geniposide. In conclusion, the present results indicated that geniposide protects HUVECs from ox‑LDL injury by inhibiting oxidative stress and inflammation, and that these effects are partly due to the enhancement of the miR‑21/PTEN pathway.

Topics: Antioxidants; Apoptosis; Cardiotonic Agents; Cell Survival; Cells, Cultured; Cytokines; Down-Regulation; Human Umbilical Vein Endothelial Cells; Humans; Inflammation; Iridoids; Lipoproteins, LDL; MicroRNAs; Oxidative Stress; PTEN Phosphohydrolase; Reactive Oxygen Species; Signal Transduction; Up-Regulation

The nod-like receptor protein 3 (NLRP3) inflammasome has a critical role in cerebral ischemia. Autophagy may cause microglial inflammatory response or stimulate microglial function. The evidences clarify a direct crosstalk between autophagy and NLRP3. Geniposide could protect neurons or PC-12 cells against cerebral ischemic injury. However, a detailed understanding of molecular mechanisms on geniposide is still unclear. This study aimed to evaluate whether geniposide could inhibit the expression and activation of NLRP3 inflammasome in BV-2 microglial cells following oxygen-glucose deprivation/reoxygenation (OGD/R) and assessed whether autophagy is involved in this process. We used OGD/R model in BV2 microglial cells in order to mimic the ischemic reperfusion injury. The NLRP3 shRNA and autophagy inhibitor (3-MA) were used to suppress NLRP3 inflammation activation and autophagy. The results demonstrated geniposide decreased cell death and the levels of NLRP3, ASC, cleaved- caspase-1 and IL-1β, whereas significantly increased the conversion of LC3 and Beclin-1 expression, decreased the expression of P62. Taken together, our results suggested that the effect of geniposide could be ascribed to the reduction of the level of inflammatory cytokines via inhibiting the activation and expression of NLRP3 inflammasome and increasing autophagic activity following OGD/R in BV-2 microglial cells. We provided a new understanding of geniposide in neuroprotection by activating autophagy and promoting anti-inflammation inhibiting NLRP3 inflammasome in microglial cells. It might be helpful for geniposide on effective therapeutic strategies in ischemic stroke.

Topics: Animals; Autophagy; Brain Ischemia; Cell Line; Cytokines; Glucose; Inflammasomes; Iridoids; Mice; Microglia; Neuroprotective Agents; NLR Family, Pyrin Domain-Containing 3 Protein; Oxygen; RNA, Small Interfering

Geniposide (GP), extracted from a traditional Chinese herb Gardenia jasminoides, has extensive pharmacological effects. But the effects and the potential mechanisms of GP on myocardial ischemia/reperfusion (I/R) injury are poorly understood. In present study, we investigated the effect of GP on myocardial I/R injury in vivo and hypoxia/reoxygenation (H/R) in vitro respectively, and its mechanism. The results showed that GP reduced myocardial infarct size, alleviated acute myocardial injury, improved cardiac function, regulated apoptosis-related proteins and inhibited apoptosis. In vitro experiments revealed that GP enhanced the cell viability, regulated apoptosis-related proteins and prevented cell apoptosis during H/R in H9c2 cells. GP inhibited the expression of autophagy-related proteins and autophagosome accumulation both in vivo and in vitro. The effects of GP were blocked by rapamycin (RAPA) administration. In summary, our results showed that GP protected against myocardial I/R injury and involved inhibition of autophagy, which might be through activating AKT/mTOR signaling pathways.

Topics: Animals; Apoptosis; Autophagosomes; Autophagy; Cardiotonic Agents; Cell Line; Cell Survival; Heart; Iridoids; Male; Myocardial Infarction; Myocardial Reperfusion Injury; Myocytes, Cardiac; Proto-Oncogene Proteins c-akt; Rats, Sprague-Dawley; Signal Transduction; Sirolimus; TOR Serine-Threonine Kinases

Rheumatoid arthritis (RA) is a common inflammatory autoimmune disease characterized by the formation of joint synovitis and pannus. Sphingosine 1-phosphate (S1P) is an important mediator related to angiogenesis, inflammation and autoimmunity. As Geniposide (GE) has potent immuno-modulation function, we investigated the effects on the dynamic balance of angiogenesis-related factors and Sphingosine kinase 1 (SphK1)-S1P-S1P receptor 1 (S1PR1) signal transduction in adjuvant-induced arthritis (AA) rats.. The model evaluation was performed from paw swelling degree, arthritis index and movement score. The immunohistochemistry and enzyme-linked immunosorbent assay were used to study the microvascular density (MVD) and pro/anti-angiogenic factors levels. The cell viability was examined by cell counting kit-8 assay. SphK1, S1PR1 mRNA and protein levels in fibroblast-like synoviocytes (FLSs) were detected by quantitative real-time polymerase chain reaction and Western blotting.. The results showed that GE can apparently suppressed the inflammatory pathological status. The arthritis index, paw swelling and MVD of AA rats were decreased with dose dependence (. It indicated that GE reduces the activity of SphK1 by restoring the dynamic balance between pro/anti-angiogenic factors, thereby interfering with SphK1-S1P-S1PR1 signal transduction, reducing the formation of synovial microvessels and exerting anti-angiogenesis effect of RA.

Topics: Animals; Cell Survival; Cells, Cultured; Drug Delivery Systems; Iridoids; Lysophospholipids; Male; Neovascularization, Pathologic; Phosphotransferases (Alcohol Group Acceptor); Random Allocation; Rats; Rats, Sprague-Dawley; Signal Transduction; Sphingosine; Sphingosine-1-Phosphate Receptors

To investigate the effects and the mechanism of geniposide on the neuroinflammation occured in the neurodegeneration course of a chronic cerebral hypoperfusion rat model.. Permanent bilateral common carotid arteries occlusions was performed to induce gradient cognitive deficit in rats. The sham group was used as control group. Then 18 rats that met the Screening Criteria were randomly selected 8 weeks post surgery, and were randomly divided into three groups, the 2-VO rats with saline solution group (2-VO+saline group), 2-VO rats with 50 mg/kg per day geniposide group (2-VO+G50) and 2-VO rats with 100 mg/kg per day geniposide group (2-VO+G100). All intervention groups were daily administered with geniposide or saline for 4 weeks. The sham-operated rats were administrated with saline. Then the rats were tested for Morris water maze to evaluate the memory and learning ability. Rats were sacrificed to obtain cortex and hippocampus tissues for HE staining and to detect expression level of glial fibrillary acidic protein (GFAP), inducible nitric oxide synthase (iNOS) and nuclear factor-kappa B (NF-κB), and the level of inflammatory factors tumor necrosis factor-α (TNF-α) and interleukin (IL)-6.. These findings demonstrated that geniposide significantly prevented cognition deterioration induced by chronic cerebral hypoperfusion in rats. Geniposide inhibited neuroinflammation occurred in the process of chronic cerebral ischemia probably via reducing iNOS and NF-κB expression and suppressing the release of inflammatory factor TNF-α and IL-6.

Topics: Animals; Anti-Inflammatory Agents; Brain Ischemia; Cognition Disorders; Disease Models, Animal; Hippocampus; Iridoids; Maze Learning; Random Allocation; Rats

Herein a series of Geniposide derivatives were designed, synthesized and evaluated as protein tyrosine phosphatase 1B (PTPlB) inhibitors. Most of these compounds exhibited potent in vitro PTP1B inhibitory activities, the representative 7a and 17f were found to be the most potent inhibitors against the enzyme with IC

Topics: Biological Transport; Cell Line; Chemistry Techniques, Synthetic; Enzyme Inhibitors; Glucose; Inhibitory Concentration 50; Insulin; Iridoids; Molecular Docking Simulation; Protein Tyrosine Phosphatase, Non-Receptor Type 1; Structure-Activity Relationship; Sulfonic Acids

Geniposide (GE) can effectively inhibit diabetic nephropathy (DN), but its mechanism is unclear. The objective of this study was to explore the antidiabetic nephropathy effects of GE both in high fat diet/streptozotocin-induced DN mice and in high glucose-induced podocyte model. Renal function in DN mice was evaluated by levels of serum creatinine (Scr) and blood urea nitrogen (BUN). Renal inflammation was appraised by pro-inflammatory cytokines: Tumor necrosis factor α (TNF-α), Interleukin 6 (IL-6) and IL-1β via ELISA assay. Renal histopathology analysis was conducted via hematoxylin and eosin, Masson and periodic acid-silver metheramine staining. Cellular viability was measured by Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay. Moreover, the related proteins p-NF-κB, ASC, Cleave-IL-1β, NLRP3, Cleave-Caspase-1 and GSDMD-N in AMPK/SIRT1/NF-κB pathway were assayed by Western blotting. In order to further investigate the effects of GE on podocytes, we also assessed these protein levels in AMPK/SIRT1/NF-κB pathway after siRNA-AMPK intervention by Western blotting. GE alleviated renal dysfunction as evidenced by decreased levels of Scr, BUN, TNF-α, IL-6 and IL-1β. Histological examination revealed GE effectively attenuated kidney damage, including glomerular basement membrane thickening and inflammatory cells infiltration. AMPK, p-AMPK and SIRT1 levels were obviously decreased both in DN mice and in podocyte model, but GE reversed these changes. The protein expressions in APMK/SIRT1/NF-κB pathway were significantly decreased by GE treatment. These results suggested that GE could efficiently block oxidative stress and inflammatory responses accompanied with pyroptosis, thus inhibiting the development of DN, and its mechanism might be related to APMK/SIRT1/NF-κB pathway.

Topics: AMP-Activated Protein Kinases; Animals; Apoptosis; Blood Urea Nitrogen; Creatinine; Cytokines; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Diet, High-Fat; Iridoids; Kidney; Male; Mice; Mice, Inbred C57BL; NF-kappa B; Podocytes; RNA, Small Interfering; Signal Transduction; Sirtuin 1

Rheumatoid arthritis (RA), a chronic systemic autoimmune disease, is mainly characterized by joint lesions and permanent loss of joint function. To discover the metabolic characteristics of RA and the underlying mechanisms in treatment with geniposide (GE), untargeted metabolomic analysis based on hydrophilic interaction liquid chromatography coupled to high-resolution mass spectrometry (HILIC-HRMS) was performed using the joint synovial fluid samples from adjuvant arthritis (AA) rats. Microdialysis (MD) was utilized to collect the dialysate samples precisely from the articular cavity of AA rats. Multivariate statistical analysis was then conducted to discover the metabolite changes induced by AA and to differentiate GE-related biomarkers. The mass spectrometry data are available on the Chorus website (https://chorusproject.org/pages/index.html) with the data set identifier 1680. The results showed that 20 metabolites differed significantly between AA rats and normal rats. GE treatment recovered the altered levels of the 13 metabolites mentioned above, such as palmitoylethanolamide (PEA), Cer (d18:0/22:0), and PC (18:1(11Z)/16:1(9Z)), and normalized glycerophospholipid metabolism. As evidenced by western blotting, the changes in PEA levels adjusted by GE were associated with the down-regulated expression of

Topics: Animals; Arthritis, Experimental; Iridoids; Metabolomics; Rats; Synovial Fluid

Geniposide (GE) is the main component in gardenia fruit. This study aimed to investigate the protective effects and potential mechanisms of GE on dextran sulfate sodium (DSS)-induced colitis in mice and lipopolysaccharide (LPS)-induced RAW 264.7 cells.. The in vivo acute colitis experimental model was established by administering drinking water containing 3% DSS to the mice for 7 days. GE was administered to the mice via oral gavage at 20 and 40 mg/kg for 7 days. Colon length, colon myeloperoxidase (MPO) level, serum and colon malondialdehyde (MDA) levels, and superoxide dismutase (SOD) activity were determined, and histological evaluation was performed. The levels of interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α (TNF-α) in the serum and colon were detected. The expression of proteins of the nuclear factor E2 related factor 2 (Nrf-2)/HO-1/ NF-κB pathway in the colon was detected. The in vitro model of LPS-induced RAW 264.7 cells to simulate enteritis model. Cell viability, IL-6, IL-1β, and TNF-α levels in the cell supernatant were measured. The MPO levels in RAW 264.7 cells and DSS-induced mice and MDA and SOD levels in the cell supernatant were measured. The expression of proteins of the Nrf-2/HO-1/NF-κB pathway in RAW 264.7 cells was determined.. GE treatment resulted in significant histological changes and reduced the expression of inflammatory mediators IL-6, IL-1β, and TNF-α the in serum, colon, and cell supernatant effectively. Parenteral nutrition reduced MPO content in the colon and RAW 264.7 cells. GE treatment increased SOD levels in the serum, colon, and cell supernatant. GE restored the protein expression of the Nrf-2/HO-1/ NF-κB pathway in RAW 264.7 cells and nude mice, and these changes were blocked significantly by Nrf-2 siRNA.. These findings demonstrated that GE ameliorated inflammation and oxidative stress in experimental colitis via modulation of the Nrf-2/HO-1/NF-κB pathway. Thus, GE could serve as a potential therapeutic agent for the treatment of ulcerative colitis (UC).

Topics: Animals; Colitis; Dextran Sulfate; Iridoids; Mice; Mice, Nude; NF-E2-Related Factor 2; NF-kappa B; Signal Transduction

To investigate the effects of geniposide in an iridoid found in Gardenia jasminoides var. radicans Makino (GJRM) in spontaneous hypertensive rat (SHR) and explore the possible mechanisms.. In this study, we detected the content of geniposide in GJRM by high-performance liquid chromatography (HPLC). Then, we used acute diuretic experiments to determine whether geniposide has diuretic effect. Moreover, we carried out experiments on SHR to further study the mechanism of hypertension, while real-time PCR, Western blot and immunohistochemistry were used for the experiments in vivo test. Hypotonic model was used for in vitro test.. Our data showed that the content of geniposide in the extract of GJRM is 27.54%. Meanwhile, 50 mg/kg geniposide showed the strongest effect on promoting urine volume. Further study indicated that the extract of GJRM and geniposide could significantly reduce blood pressure and promote the excretion of urine and Na. Collectively, we would suggest that geniposide may potentially be utilized as an adjunct to existing thiazide and thiazide-like diuretics to control hypertension, mainly through inhibiting the activation of the WNK signalling pathway mediated by the estrogen receptor.

Topics: Animals; Antihypertensive Agents; Blood Pressure; Cell Line; Disease Models, Animal; Diuresis; Diuretics; Estrogen Receptor alpha; Estrogen Receptor beta; Gardenia; Hypertension; Iridoids; Kidney Tubules, Proximal; Male; Plant Extracts; Protein Serine-Threonine Kinases; Rats, Inbred SHR; Rats, Inbred WKY; Signal Transduction

Topics: Apoptosis; Case-Control Studies; Cell Line, Tumor; Cell Proliferation; Drug Screening Assays, Antitumor; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Gene Silencing; Humans; Iridoids; Lymph Nodes; Lymphoma, Large B-Cell, Diffuse; MicroRNAs; Proto-Oncogene Proteins c-met; RNA, Long Noncoding; Signal Transduction

Acute kidney injury is one of the most common complications that occurs in septic shock. An effective therapeutic intervention is urgently needed. Geniposide has been reported to possess pleiotropic activities against different diseases. However, the effect of geniposide on sepsis-induced kidney injury is unexplored. Our study aims to illustrate the mitigative effects of geniposide on sepsis-induced kidney injury and its relevant mechanisms. Sepsis was induced in mice undergoing cecal ligation and puncture (CLP) surgery. Mice were intraperitoneally injected with geniposide (10, 20 and 40 mg/kg) for treatment. The results showed that geniposide ameliorated kidney injury and dysfunction in CLP-induced septic mice, accompanied by reduction of inflammatory response and oxidative stress. We also found that geniposide significantly reduced vascular permeability and cellular apoptosis of the kidney, with increase of Bcl-2 and decrease of Bax and cleaved caspase-3. Moreover, PPARγ was found to be upregulated with the increasing concentration of geniposide. The protection of geniposide against inflammation and apoptosis was recovered by inhibition of PPARγ. Collectively, these results indicate that geniposide could significantly ameliorate acute kidney injury in CLP-induced septic mice and LPS-stimulated HK-2 cells by activating PPARγ. Geniposide might be a potential drug candidate for sepsis-induced kidney injury.

Topics: Acute Kidney Injury; Animals; Apoptosis; Cells, Cultured; Inflammation; Iridoids; Kidney; Male; Mice, Inbred BALB C; Oxidative Stress; PPAR gamma; Sepsis

Ulcerative colitis is a chronic and recurrent inflammatory bowel disease mediated by immune response. Geniposide is the main active ingredient extracted from Gardenia jasminoides, which has been suggested to exert excellent efficacy on inflammatory disease. Herein, in this study, we aimed to uncover the systematic understanding of the mechanism and effects of geniposide in ameliorating inflammatory responses in colitis. In brief, the TCMSP server and GEO DataSets were used to analyze the systematic understanding of the mechanism and effects of geniposide in ameliorating inflammatory responses in colitis. Dextran Sulfate Sodium (DSS)-induced acute colitis of mice were administered with 25-100[Formula: see text]mg/kg of geniposide for 7 days by gavage. Lipopolysaccharide (LPS)-induced Bone Marrow Derived Macrophage (BMDM) cell or RAW264.7 cell models were treated with 20, 50 and 100[Formula: see text][Formula: see text]M of geniposide for 4[Formula: see text]h. Myeloperoxidase (MPO) activity and Interleukin-1[Formula: see text] (IL-1[Formula: see text] levels were measured using MPO activity kits and IL-1[Formula: see text] levels enzyme-linked immunosorbent assay (ELISA) kits, respectively. Additionally, Western blot was used to determine the relevant protein expression. As a result, Geniposide could ameliorate inflammatory responses and prevent colitis in DSS-induced acute colitis of mice by activating AMP-activated protein kinase (AMPK)/Transcription 1 (Sirt1) dependent signaling via the suppression of nod-like receptor protein 3 (NLRP3) inflammasome. Geniposide attenuated macrophage differentiation in DSS-induced acute colitis of mice. Geniposide suppressed NLRP3 inflammasome and induced AMPK/Sirt1 signaling in LPS-induced BMDM cell or RAW264.7 cell models. In mechanism studies, the inhibition of AMPK/Sirt1 attenuated the anti-inflammatory effects of geniposide in colitis. The activation of NLRP3 attenuated the anti-inflammatory effects of geniposide in colitis. Taken together, our results demonstrated that geniposide ameliorated inflammatory responses in colitis vai the suppression of NLRP3 inflammasome in macrophages by AMPK/Sirt1-dependent signaling.

Topics: AMP-Activated Protein Kinases; Animals; Cells, Cultured; Colitis; Disease Models, Animal; Inflammasomes; Inflammation; Iridoids; Macrophages; Mice; Mice, Inbred C57BL; NLR Family, Pyrin Domain-Containing 3 Protein; Phytotherapy; RAW 264.7 Cells; Signal Transduction; Sirtuin 1

Topics: Animals; Atherosclerosis; Autophagy; Down-Regulation; Gene Expression; Iridoids; Macrophages; Male; Membrane Glycoproteins; Mice; Mice, Inbred C57BL; Phytotherapy; RAW 264.7 Cells; Receptors, Immunologic; Signal Transduction; TOR Serine-Threonine Kinases

Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disorder associated with features of metabolic syndrome and oxidative stress. We examined the mechanism by which the combined extracts of

Topics: AMP-Activated Protein Kinases; Animals; Antioxidants; Diet, High-Fat; Disease Models, Animal; Endoplasmic Reticulum Stress; Eucommiaceae; Fatty Liver; Iridoid Glucosides; Iridoids; Lipid Peroxidation; Male; Oxidative Stress; Phytotherapy; Plant Extracts; Rats, Sprague-Dawley; Reactive Oxygen Species

Perinatal hypoxic-ischemia (HI) is a leading cause of acute mortality and neurologic complications in newborns. Geniposide, a natural product extracted from the herb Gardenia jasminoides, has been shown to possess neuroprotective effects in neurologic deficits. This study aims to investigate whether Geniposide has therapeutic potential to HI brain injury and the underlying mechanisms. C57/bl6 mice were subjected to HI insult on postnatal day 10. Geniposide (20 mg/kg b.w.) was administered intragastrically every day after HI insult for 7 successional days. Then mice at P18 were sacrificed and brain tissues were collected for further analysis. Geniposide treatment significantly inhibited cell apoptosis, reduced serum IgG leakage into brain tissue, attenuated astrogliosis and microgliosis, prevented loss of pericytes, loss of tight junction and adherens junction proteins. The PI3K/Akt signaling pathway, which related proteins were downregulated after HI insult, was activated by Geniposide treatment. Geniposide treatment after neonatal HI insult attenuated HI-induced cell apoptosis, IgG leakage, microgliosis, astrogliosis, pericytes loss and junction protein degradation. Geniposide could protect against HI-induced brain injury, which might be through the activation of PI3K/Akt signaling pathway.

Topics: Animals; Apoptosis; Astrocytes; Brain; Female; Hypoxia-Ischemia, Brain; Iridoids; Male; Mice; Microglia; Neuroprotective Agents; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction

Approaches promoting fibroblast-like synoviocytes (FLS) apoptosis are considered as a meaningful strategy for rheumatoid arthritis (RA) treatment. We have previously reported the anti-arthritic effect of penta-acetyl geniposide ((Ac). Rat AIA was induced by complete Freund's adjuvant, and FLS were primary-cultured from synovial tissues. AIA FLS were treated with (Ac). (Ac)

Topics: Animals; Apoptosis; Arthritis, Experimental; Caspase 3; Cell Proliferation; Fibroblasts; Iridoids; Male; Proto-Oncogene Proteins c-bcl-2; Rats; Rats, Sprague-Dawley; Synovial Membrane; Synoviocytes; Transcription Factor RelA

Geniposide, the main medicinal ingredient of Gardenia jasminoides Ellis, is known to be a resistant agent to atherosclerosis. Some reports its mechanism against atherosclerosis remains completely unclear. Herein, we have investigated the protective effect of geniposide against atherosclerosis as well as clarified the mechanisms related with inhibiting the formation of foam cells and lowering reverse lipid transport via p38/MAPK signaling pathways. Macrophage Raw264.7 was induced by lysophosphatidic acid (LPA) to form foam cell as a cell model. ApoE

Topics: Animals; Atherosclerosis; ATP Binding Cassette Transporter 1; Biological Transport; CD36 Antigens; Dose-Response Relationship, Drug; Down-Regulation; Foam Cells; Iridoids; Lipid Metabolism; Male; MAP Kinase Signaling System; Mice; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Proto-Oncogene Proteins c-akt; Scavenger Receptors, Class A; Up-Regulation

Alcoholic steatosis is one of the most prevalent forms of liver disease, and appropriate insight and application of anti-steatosis drugs must be considered. Geniposide, the major active constituent of the

Topics: Animals; Chemical and Drug Induced Liver Injury; Drug Overdose; Fatty Liver, Alcoholic; Fruit; Gardenia; Iridoids; Male; Phytotherapy; PPAR alpha; Proteome; Proteomics; Rats, Sprague-Dawley; Transcription Factors

Topics: Animals; Apoptosis; Cell Cycle; Cell Line; Cell Survival; Chromatography, High Pressure Liquid; Gardenia; Inflammation; Iridoids; Liver; Molecular Docking Simulation; Oxidative Stress; Phytochemicals; Plant Extracts; Rats; Receptors, Tumor Necrosis Factor, Type I; Reference Standards

Atherosclerosis (AS) is characterized by the significant accumulation of low-density lipoprotein (LDL)-cholesterol in macrophages that reside in the vessel wall and the resultant inflammatory response. Therefore, inhibition of LDL-induced inflammation is a promising interference for AS. Many traditional Chinese medicine prescriptions have been developed for AS treatment. Geniposide (GEN) is an iridoid glycoside mainly found in

Topics: Animals; Atherosclerosis; Foam Cells; Iridoids; Lipoproteins, LDL; Male; Mice; Mice, Inbred C57BL; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Primary Cell Culture

Topics: Animals; Chromatography, High Pressure Liquid; Depression; Disaccharides; Disease Models, Animal; Drug Synergism; Drugs, Chinese Herbal; Flavanones; Hesperidin; Intestinal Absorption; Iridoids; Male; Rats; Rats, Sprague-Dawley

Geniposide and baicalin, the main components of Huangqin-Zhizi herb pair, have been combined to increase the efficacy. To reveal the underlying compatibility mechanism of these two components, the synergistic effects of geniposide on the enhancement of solubility, apparent oil-water partition coefficient, and intestinal absorption of baicalin were investigated. The equilibrium solubility and apparent oil-water partition coefficient of baicalin in different solvents were determined through the shake-flask and high-performance liquid chromatography with diode array detection methods. The intestinal absorption of baicalin was investigated through the in situ single-pass intestinal perfusion method. When combined with different amounts of geniposide, the solubility and apparent oil-water partition coefficient of baicalin improved to 98.74-159.03μg/mL and 0.24-0.29, respectively, which were respectively 1.25-2.02-fold and 1.6-1.9-fold higher than those parameters in the baicalin-only control. The intestinal absorption study indicated that geniposide was an absorption-enhancer for baicalin and significantly increased the absorption rate constant value and the apparent absorption constant value of baicalin, especially in duodenum and jejunum when the compatibility concentrations were 1:1 and 1:2. Geniposide had synergistic effects in enhancing the solubility, apparent oil-water partition coefficient, intestinal absorption of baicalin. The study results provide scientific information elucidating the compatibility mechanism of the Huangqin-Zhizi herb pair and its primary components.

Topics: Animals; Drugs, Chinese Herbal; Duodenum; Flavonoids; Intestinal Absorption; Iridoids; Jejunum; Male; Rats; Rats, Sprague-Dawley; Solubility

The aim of this paper was to screen out relevant genes of geniposide-induced hepatotoxicity based on genomics,in order to provide a scientific basis for the non-clinical evaluation of drugs containing Gardeniae Fructus and geniposide. Fifty-five SD rats were randomly divided into normal control group,24 h group and 72 h group. The changes of appearance,behavior and weight of rats were observed after administration by gavage for 3 days. The activities of ALT and AST were detected. Molecular mechanism of geniposideinduced hepatotoxicity was investigated by Affymetrix miRNA 4. 0 and Affymetrix Rat Gene 2. 0 to examine the gene expression levels in Sprague-Dawley rat livers at 24 h and 72 h after administration of overdose-geniposide( 300 mg·kg-1 daily),and then verified by Realtime quantitative PCR. Compared with the normal control group,the activities of ALT and AST were markedly increased. In addition,experimental results indicated that 324 genes were differentially expressed,among which 259 were up-regulated and 65 down-regulated.Nine candidate genes were verified by qRT-PCR,including Bcl2,Il1 b,Tpm3,MMP2,Col1α1,Ifit1,Aldob,Nr0 b2,Cyp2 c23. And Bcl2,Col1α1,Aldob,Nr0 b2 and Cyp2 c23 were found to be correlated with geniposide-induced hepatotoxicity. This study provides an important clue for mechanism of geniposide-induced hepatotoxicity.

Topics: Animals; Biomarkers; Chemical and Drug Induced Liver Injury; Genomics; Iridoids; Liver; Rats; Rats, Sprague-Dawley

In this work, a mesoporous Fe

Topics: Adsorption; Chromatography, Ion Exchange; Computer Simulation; Ferric Compounds; Ion Exchange; Iridoids; Models, Theoretical; Molecular Docking Simulation; Molecular Dynamics Simulation; Wastewater; Water Purification

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal).\ \ This article has been retracted at the request of the Editor-in-Chief.\ \ Given the comments of Dr Elisabeth Bik regarding this article “… the Western blot bands in all 400+ papers are all very regularly spaced and have a smooth appearance in the shape of a dumbbell or tadpole, without any of the usual smudges or stains. All bands are placed on similar looking backgrounds, suggesting they were copy/pasted from other sources, or computer generated”, the journal requested the authors to provide the raw data. However, the authors were not able to fulfil this request and therefore the Editor-in-Chief decided to retract the article.

Topics: Animals; Apoptosis; Blotting, Western; Cell Proliferation; Cell Survival; Gene Knockdown Techniques; Glucose; Hypoxia-Ischemia, Brain; Iridoids; Oxygen; PC12 Cells; Phosphatidylinositol 3-Kinase; Proto-Oncogene Proteins c-akt; Rats; RNA, Long Noncoding; Signal Transduction; Up-Regulation; Wnt Signaling Pathway

Discovering novel compounds with higher activities is a key aim of natural products research. Gardenia jasminoides Ellis is a herb with anti-inflammatory properties. Iridoid glycosides (mainly geniposide) and crocetin derivatives (crocins) are the two major active constituents in this herb and are considered its active ingredients. However, which components are responsible for the anti-inflammatory properties of gardenia have remained to be investigated.. Here, we prepared total iridoid glycocides (TIG) and total crocins (TC) from G. jasminoides Ellis, determined their main chemical constituents, and performed animal studies to evaluate their anti-adjuvant arthritis activities, thus, proposing a reasonable mechenism to explain the anti-inflammatory activities of the active components in this herbal remedy.. TIG and TC were prepared by using HPD-100 macroporous resin, and characterized by UHPLC-DAD-MS and UV-Vis spectrophotometer. Then, freund's complete adjuvant-injected rats underwent drug treatments with TIG (160 mg/kg) and TC (160 mg/kg) for 14 days, and their ankle diameters were measured. Moreover, X-ray radiographs of the adjuvant injected hind paws were evaluated. Finally, histopathological examinations of the ankle joints, spleens and thymus were carried out to evaluate inflammatory reactions, and immunohistochemical measurements were conducted to evaluate TNF-α and TGF-β1 expression in the ankle joint of the rats.. The chemical composition determination of the current study showed that TIG was mainly composed of geniposide and TC was a fraction predominantly with crocin-1, crocin-2 and crocin-3. Calculation of results showed that TIG and TC contained 58.2% total iridoid glycosides and 54.7% total crocins, respectively. Our study suggested TIG and TC treatments markedly decreased paw swelling and ankle diameters of AA rats (both p < 0.05). The radiological analysis showed that administration of TIG and TC ameliorated bone destruction, and reduced the radiological bone destruction scores (TIG p < 0.05, TC p>0.05). Moreover, data from histological assessment demonstrated considerable mitigation of inflammation in the joints (both p < 0.01), spleen and thymus of AA rats treated with TIG and TC. TNF-α and TGF-β1 protein expression according to immunohistochemistry staining also supported the anti-arthritis activities of TIG and TC (TNF-α: TIG p < 0.01 and TC p < 0.05, TGF-β1: TIG p < 0.01 and TC p>0.05).. In the current study, fractionation of gardenia prior to further in vivo investigation has for the first time provided reasonable explanation for the anti-inflammatory activity of this herbal remedy. Our study showed that both TIG and TC from gardenia have anti-inflammatory properties. Overall, these experimental findings suggest that gardenia could be regarded as a potential therapeutic target for arthritis. However, as geniposide has a higher content than crocins in this herbal drug, TIG (mainly geniposide) seems to be primarily responsible for the anti-inflammatory properties of gardenia. Taken together, this maiden attempt demonstrated that TIG (mainly geniposide) is more important in evaluating the anti-inflammatory activity of G. jasminoides Ellis.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Experimental; Carotenoids; Chromatography, High Pressure Liquid; Drug Evaluation, Preclinical; Freund's Adjuvant; Gardenia; Iridoid Glycosides; Iridoids; Male; Rats, Sprague-Dawley; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha; Vitamin A

Geniposide, an iridoid glycoside extract from the gardenia fruit, is used in traditional Chinese medicine to alleviate symptoms of liver and inflammatory diseases. Geniposide activates GLP-1 receptors, known to modulate the activity of mechanistic target of rapamycin (mTOR), a key kinase regulating energy balance, proliferation, and survival in cells. mTOR activation inhibits autophagy, which is often disrupted in age-related diseases. Modulation of mTOR function to increase autophagy and inhibit apoptosis is involved in the protective effects of pharmacologic agents targeting diabetes and Alzheimer's disease (AD). We investigated whether such mechanism could mediate geniposide's neuroprotective effects in the APP/PS1 mouse model of AD. Eight-week treatment with geniposide improved cognitive scores in behavioral tests, reduced amyloid-β 1-40 plaque deposition, and reduced soluble Aβ1-40 and Aβ1-42 levels in the APP/PS1 mouse brain.This also showed increased p-Akt/Akt, p-mTOR/mTOR and decreased p-4E-BP1/4E-BP1 expression, and these patterns were partially reversed by geniposide. Evidence for enhanced autophagy, denoted by increased expression of LC3-II and Beclin1, was also seen after treatment with geniposide. Our data suggests that down regulation of mTOR signaling, leading to enhanced autophagy and lysosomal clearance of Aβ fibrils, underlies the beneficial effects of geniposide against neuropathological damage and cognitive deficits characteristic of AD.

Topics: Alzheimer Disease; Amyloid; Amyloid beta-Protein Precursor; Animals; Autophagy; Behavior, Animal; Cognitive Dysfunction; Down-Regulation; Gene Expression Regulation; Humans; Iridoids; Mice; Mice, Transgenic; Peptide Fragments; Plaque, Amyloid; Random Allocation; TOR Serine-Threonine Kinases

Topics: Animals; Biomarkers; Chemical and Drug Induced Liver Injury; Chromatography, High Pressure Liquid; Computational Biology; Disease Models, Animal; Drugs, Chinese Herbal; Iridoids; Liver; Metabolic Networks and Pathways; Metabolome; Metabolomics; Rats; Reproducibility of Results; Tandem Mass Spectrometry; Tissue Distribution

Topics: Caffeic Acids; Chromatography, High Pressure Liquid; Discriminant Analysis; Drugs, Chinese Herbal; Iridoids; Mass Spectrometry; Multivariate Analysis

Geniposide (GE) is an iridoid glycoside compound with anti-inflammatory effect. The potential of sphingosine 1-phosphate (S1P) as a plasma marker in human diseases was suggested recently in the literature, which demonstrated that, in patients with inflammatory diseases, plasma S1P was elevated. It follows that the obstructive coronary artery disease can be predicted with serum S1P. Therefore, S1P can also be potentially used as a pharmacodynamic marker to study adjuvant arthritis (AA) rats. In the current study, a UHPLC-MS/MS method combined with the microdialysis sampling technique (using FTY720 phosphate as an internal standard) was adopted and validated to measure S1P levels in the hemodialysis fluid and joint cavity dialysates of AA rats after oral administration of GE. A S1P concentration-time curve in the dialysate was established in this study. It was demonstrated that GE exerted an anti-inflammatory effect by reducing AA-induced elevated S1P levels. It is showed that changes in S1P concentrations over time can be used to monitor the pharmacodynamic effects of GE in treating AA rats in pharmacodynamic studies.

Topics: Animals; Arthritis, Experimental; Chromatography, High Pressure Liquid; Drug Stability; Iridoids; Linear Models; Lysophospholipids; Male; Microdialysis; Random Allocation; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Sensitivity and Specificity; Sphingosine; Tandem Mass Spectrometry

Studies have shown that the NRF-2 /HO-1 pathway participates in myocardial ischemic reperfusion injury (MI/R) and that Geniposide (GEN) could protect the myocardial against MI/R. This study aims to examine the protective effects of GEN on MI/R in diabetic rats and further explore the possible mechanism of action. During MI/R in rats, NRF-2 /HO-1signals changed significantly including NRF-2 and HO-1up-regulation, resulting in heart dysfuction, histological damage and increasing oxidative stress and cell apoptosis. Treatment with GEN can significantly improve the general condition and heart function in diabetic rats with decreasing the expression of cTnI, CK-MB, blood glucose, MDA, ROS, cell apoptosis and pathological damage in MI/R. In addition, GEN precondition can also significantly increase the weight of rats and the activity of SOD, CAT and GPx with up-regulating the expression of NRF-2 and HO-1 in MI/R. This study implied that Geniposide has a protective effect on myocardial ischemia reperfusion injury in diabetic rats, and its mechanism is associated with activating NRF2/HO-1 signaling pathway to suppress oxidative stress.

Topics: Animals; Blood Glucose; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diabetic Angiopathies; Iridoids; Ischemic Preconditioning, Myocardial; Male; Myocardial Ischemia; Myocardial Reperfusion Injury; Rats; Rats, Sprague-Dawley; Severity of Illness Index; Treatment Outcome

It is well-known that imbalance state of glucose metabolism triggers many metabolic diseases and glucose uptake in skeletal muscle accounts for 90% of body weight. Geniposide is one of the major natural bioactive constituents of gardenia fruit, and the regulation of geniposide on glucose metabolism in skeletal muscle has not yet been investigated. Here, on the basis of microarray analysis, we discovered that geinposide decreased pyruvate dehydrogenase kinase 4 (PDK4) expression in skeletal muscle of mice and subsequently found that geniposide inhibited the expressions of forkhead box O1 (FoxO1), PDK4, and phosphorylated pyruvate dehydrogenase in vitro and in vivo. Moreover, geniposide promoted a switch of slow-to-fast myofiber type and glucose utilization, suggesting that geniposide improved glucose homeostasis. In addition, mechanistic studies revealed that geniposide played above roles by regulating FoxO1/PDK4, which controlled fuel selection via pyruvate dehydrogenase. Meanwhile, effects of geniposide mentioned above could be reversed by FoxO1 overexpression. Together, these results establish that geniposide confers controls on fuel usage and glucose homeostasis through FoxO1/PDK4 in skeletal muscle.

Topics: Animals; Diabetes Mellitus, Type 2; Forkhead Box Protein O1; Glucose; Homeostasis; Humans; Iridoids; Male; Mice; Mice, Inbred C57BL; Muscle Fibers, Skeletal; Muscle, Skeletal; Phosphorylation; Protein Kinases; Signal Transduction; Up-Regulation

A highly selective and efficient LC-MS/MS method was developed to determine the plasma concentration of magnolol, hesperidin, neohesperidin and geniposide following oral administration of Zhi-Zi-Hou-Po decoction in normal and depressed rats. Plasma samples were pretreated by protein precipitation with methanol. Chromatographic separation was performed on an XTerra

Topics: Administration, Oral; Animals; Biphenyl Compounds; Corticosterone; Depression; Disease Models, Animal; Drugs, Chinese Herbal; Hesperidin; Iridoids; Lignans; Limit of Detection; Linear Models; Male; Rats; Rats, Sprague-Dawley; Reproducibility of Results

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal).\ \ This article has been retracted at the request of the Editor-in-Chief.\ \ Given the comments of Dr Elisabeth Bik regarding this article “… the Western blot bands in all 400+ papers are all very regularly spaced and have a smooth appearance in the shape of a dumbbell or tadpole, without any of the usual smudges or stains. All bands are placed on similar looking backgrounds, suggesting they were copy/pasted from other sources, or computer generated”, the journal requested the authors to provide the raw data. However, the authors were not able to fulfil this request and therefore the Editor-in-Chief decided to retract the article.

Topics: Animals; Apoptosis; Cell Hypoxia; Cell Line; Cytoprotection; Down-Regulation; Gene Silencing; Heart; Iridoids; Janus Kinase 1; Myocardial Infarction; Myocytes, Cardiac; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Rats; RNA, Long Noncoding; Signal Transduction; STAT3 Transcription Factor

Fluoxetine (FXT), a selective serotonin reuptake inhibitor (SSRI), is one of the most common psychiatric medications clinically prescribed; while over-produced serotonin may suppress neurite development. The role of major iridoids like geniposide (GPS) and genipin (GNP) from Gardenia jasminoides Ellis fruit (family Rubiaceae) in ameliorating the anti-neurite outgrowth effect of FXT is poorly understood. In this study, the effects of these iridoids on FXT-suppressed neurite outgrowth in Neuro2a neuroblastoma cells were investigated.. Neuro2a cells were treated with FXT and GPS. The effect of GPS-FXT co-treatment on neurite outgrowth was observed using inverted phase-contrast microscope imaging system, while neurite outgrowth markers - microtubule-associated protein-2 (MAP2) and growth-associated protein 43 (GAP43) were analyzed using RT-PCR, Western blot and immunofluorescence staining. The transcription factor-cAMP response element binding (CREB), and signaling pathways - mitogen-activated protein kinase (MAPK) and protein kinase B/mammalian target of rapamycin (AKT/mTOR) were also analyzed with the help of Western blot.. The results showed that FXT decreased the neurite outgrowth in Neuro2a cells and also downregulated gene and protein expression of MAP2 and GAP43. It also downregulated the protein expression of phosphorylated-CREB, MAPK, and AKT/mTOR signaling pathways. In contrast, GPS counteracted the effects of FXT. GPS-FXT co-treatment increased the percentage of neurite-bearing cells by 3.6-fold at 200 μM as compared to FXT treatment only.. This study has provided the possible molecular mechanism showing how FXT exerted its detrimental side-effects on the neurite differentiation, and via the same mechanism how GPS attenuated these side effects.

Topics: Cell Differentiation; Cell Line, Tumor; Cell Survival; Cyclic AMP Response Element-Binding Protein; Fluoxetine; GAP-43 Protein; Humans; Iridoids; MAP Kinase Signaling System; Microtubule-Associated Proteins; Mitogen-Activated Protein Kinases; Neural Stem Cells; Neurites; Neuroblastoma; Neurogenesis; Neuronal Outgrowth; Phosphorylation; Proto-Oncogene Proteins c-akt; Signal Transduction; TOR Serine-Threonine Kinases

Proteinuria is one of the most important clinical features of nephrotic syndrome (NS). Injury of podocyte has been proved to contribute to the occurrence of proteinuria. This study explored the effects of geniposide (GEN) on lipopolysaccharide (LPS)-caused murine kidney podocyte MPC5 apoptosis and autophagy. Viability and apoptosis of MPC5 cells were respectively detected with the help of CCK-8 assay and Guava Nexin assay. 3-Methyladenine (3-MA) was used as an autophagy inhibitor, while rapamycin as autophagy activator. Si-Beclin-1 was transfected in MPC5 cells to down-regulate the expression of Beclin-1. We found that LPS stimulation significantly caused MPC5 cell viability reduction, apoptosis and autophagy (P < .05 or P < .01). GEN treatment remarkably alleviated the LPS-caused MPC5 cell viability reduction and apoptosis, but promoted cell autophagy (P < .05). Moreover, 3-MA incubation or si-Beclin-1 transfection notably weakened the effects of GEN on LPS-caused MPC5 cell apoptosis and autophagy (P < .05), while rapamycin had opposite effects (P < .05). Furthermore, GEN activated Ras/Raf/MEK/ERK pathway in LPS-treated MPC5 cells (P < .05). In conclusion, this research verified the protective effects of GEN on podocytes damage. GEN alleviates LPS-caused apoptosis of murine kidney podocytes by activating Ras/Raf/MEK/ERK-mediated cell autophagy. Highlights: LPS causes podocyte MPC5 apoptosis and autophagy. GEN alleviates LPS-caused MPC5 cell apoptosis, but promotes cell autophagy. 3-MA or si-Beclin-1 weakens the effects of GEN on LPS-treated MPC5 cells. Rapamycin strengthens the effects of GEN on LPS-treated MPC5 cells. GEN activates Ras/Raf/MEK/ERK pathway in LPS-treated MPC5 cells.

Topics: Animals; Apoptosis; Autophagy; Beclin-1; Cell Line; Cell Survival; Cytoprotection; Gene Silencing; Iridoids; Kidney; Lipopolysaccharides; MAP Kinase Signaling System; Mice; Podocytes; raf Kinases; ras Proteins

To investigate the " drug-guide" effect of Achyranthes bidentata saponins( ABS) and geniposide( GE) in the treatment on adjuvant arthritis( AA) rats. A UHPLC-MS/MS method for the quantitative determination of GE,zingibroside R1,ginsenoside Ro and chikusetsu saponin Ⅳa in rat blood and joint dialysate was established. After single or combined administration with ABS and GE was given to AA rat model,a microdialysis sampling method for rat joint cavity and jugular vein blood vessels was established to collect microdialysis samples. Waters Acquity HSS C_(18) column was used to separate the above four components,with mobile phase as acetonitrile-0. 1% formic acid water as mobile phase for gradient elution. ESI source was adopted for mass spectra in a negative ion scanning mode. Multiple reaction monitoring( MRM) mode was applied to detect the above four components. The methodological results showed that GE,zingibroside R1,ginsenoside Ro and chikusetsu saponin Ⅳa demonstrated a good linear relationship within the concentration ranges of 2-4 000,16-4 096,14-3 584,23-5 888 μg·L-1 respectively. The precision,accuracy,stability and matrix effect of these four ingredients reached the requirements of quantitative analysis of biological samples. The pharmacokinetic results demonstrated that the combined administration of ABS and GE( 60 mg·kg~(-1)+60 mg·kg~(-1)) can increase the degree of GE in joint cavity distribution,and the AUCjoint/AUCplasmwere twice of that of single administration of GE( 60 mg·kg~(-1)),which indicated that ABS might played a vital role in GE's distribution to joint cavity. Moreover,there was no significant difference between the distribution trend of total three ABS and GE in rats. The pharmacodynamics results showed that the combined administration of ABS and GE has stronger effects on paw swelling,arthritis index and synovial pathomorphology of AA rats than single administration of GE,which suggested that ABS might improve GE's anti-inflammatory effect in AA rats. Based on the above results,ABS has a targeting effect in increasing GE's concentration in joint cavity,with a synergy in efficacy.

Topics: Achyranthes; Animals; Arthritis, Experimental; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Iridoids; Microdialysis; Rats; Reproducibility of Results; Saponins; Tandem Mass Spectrometry

The blood-brain barrier (BBB) is involved in the pathogeneses of ischemic stroke (IS). Geniposide (GEN), an iridoid glycoside isolated from Gardenia jasminoides Ellis, has been used for the treatment of IS. However, the effects of GEN on the BBB are poorly understood. In vitro disease models of the BBB could be very helpful for the elucidation of underlying mechanisms and the development of novel therapeutic strategies. Therefore, we established an in vitro BBB model composed of primary cultures of brain microvascular endothelial cells and astrocytes. We then used this in vitro model to investigate the effect of GEN on the function of the BBB. Oxygen glucose deprivation and reoxygenation (OGD/R) significantly increased permeability and cell apoptosis in this in vitro BBB model. Notably, GEN pretreatment effectively improved the BBB function by decreasing the permeability of the BBB, promoting expression of tight junction proteins (zonula occludens-1, claudin-5, and occludin) and gamma-glutamyl transpeptidase, increasing transendothelial electrical resistance, mitigating oxidative stress damage and the release of inflammatory cytokines, downregulating the expression levels of matrix metallopeptidases-9 (MMP-9) and MMP-2, and increasing the release of brain derived neurotrophic factor and glial cell derived neurotrophic factor. Therefore, GEN can ameliorate the BBB dysfunction induced by OGD/R conditions through multiple protective mechanisms. The findings suggest that GEN may be an appropriate drug for restoring the barrier function of the BBB.

Topics: Animals; Apoptosis; Blood-Brain Barrier; Cell Hypoxia; Cytoprotection; Iridoids; Nerve Growth Factors; Oxidative Stress; Oxygen; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Tight Junction Proteins

Topics: Actins; Alanine Transaminase; Animals; Aspartate Aminotransferases; Carbon Tetrachloride; Cell Line; Cell Survival; Collagen Type I; Collagen Type I, alpha 1 Chain; Hedgehog Proteins; Hepatic Stellate Cells; Iridoids; Liver; Liver Cirrhosis; Male; Mice

The optimization of electrolytes, kinds and concentrations, in mobile phase for multiple constituents analyzing using high-performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS) was usually compromised to ensure good LC separation of partial components. However, the compromised electrolytes could lead to ionization suppression of some of the analytes. To solve the compromise of electrolytes within various components, taking phenolic acids and iridoids as a case, we used electrolyte switch in contiguous running time segments of UPLC-ESI-MS/MS to ensure chromatographic separation of chlorogenic acid, neochlorogenic acid and cryptochlorogenic acid and improve the response of geniposide. Then the method was applied for pharmacokinetic study of the four components in rat after inhaling Reduning aerosol for the first time. The complete separation of the three chlorogenic acid isomers was achieved and the LLOQs of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, and geniposide were 1, 1, 3, and 0.2 ng/mL, respectively. In conclusion, we developed a sensitive and time-saving LC-MS/MS method for the quantitative analysis of chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, and geniposide in rat plasma, and this method appears to be useful for pharmacokinetic studies of Reduning aerosol. The method provided a sight to alleviate compromise of electrolytes in mobile phase for HPLC-ESI-MS in analyzing multi-components.

Topics: Administration, Inhalation; Aerosols; Animals; Chlorogenic Acid; Chromatography, Liquid; Drugs, Chinese Herbal; Iridoids; Male; Rats; Rats, Sprague-Dawley; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

Intron retention, one of the major types of alternative splicing in plants and animals, has also been reported existing in filamentous fungi's glycoside hydrolases. In this study, an intron-retained β-glucosidase gene transcript (bgl1B) from A. niger B2 strain was obtained. Compared with the normally spliced transcript bgl1A, bgl1B had an extra 51bp insertion, which was confirmed to be the sixth (the last) intron of this β-glucosidase gene. The bgl1A and bgl1B were expressed in Pichia pastoris and the purified enzymes were used to compare their catalytic properties. The results showed that the intron retention didn't impair the catalytic function. Instead, the intron-retained enzyme BGL1B had a better thermostability with a higher optimal temperature and a longer half-life under 50 °C. Also it exhibited a little higher k

Topics: Aspergillus niger; beta-Glucosidase; Cloning, Molecular; Genes, Fungal; Introns; Iridoids; Pichia

Geniposide is a natural hepatotoxic iridoid glycoside. Its hydrolysate of intestinal microbiota, genipin, is thought to be responsible for the hepatotoxicity. However, the underlying mechanism that genipin contributes to the hepatotoxicity of geniposide is not well understood. In this study, we found that genipin spontaneously converted into a reactive dialdehyde intermediate and covalently bond to the primary amine group of free amino acids in both of the phosphate buffers and geniposide-treated rats. Furthermore, genipin dialdehyde can form the covalent linkage to the primary amino group (ε) of lysine side chains of the hepatic proteins in geniposide-treated rats. Pretreatment with β-glucosidase or antibiotics significantly modulated the genipin dialdehyde formation and protein modification, revealing the essential role of microbial glycosidases. The levels of protein adduct were that mapped onto the hepatotoxicity of geniposide. In summary, this study demonstrates that the intestinal microbiota mediated covalent modification of the hepatic protein by genipin dialdehyde may play a crucial role in the liver injury of geniposide. The study is also helpful for understanding the contribution of intestinal microbiota to the metabolic activation of xenobiotics.

Topics: Aldehydes; Amino Acids; Animals; Anti-Bacterial Agents; beta-Glucosidase; Bile; Chemical and Drug Induced Liver Injury; Gastrointestinal Microbiome; Glutathione; Glycoside Hydrolases; Iridoids; Liver; Lysine; Male; Rats; Rats, Sprague-Dawley

The growing evidence shows that in the early stage of type 2 diabetes mellitus (T2DM) development, when challenged by hyperglycemia and/or insulin resistance, pancreatic islets would produce more insulin to maintain the balance of blood sugar, but at the same time, endoplasmic reticulum (ER) stress will be initiated for the reason of over-compensation, which might be a crucial caused factor of dysfunction and death of pancreatic beta cell. In this study, we showed that high glucose induced a remarkably unfolded protein response (UPR) with the phosphorylation of PERK/eIF2α and IRE1α in INS-1 cells, but geniposide prevented the role of high glucose on the phosphorylation of PERK/eIF2α and IRE1α, respectively. Although inhibition of Txnip expression by siRNA had no significant effect on geniposide-regulating UPR, PERK and IRE1α were associated with geniposide-regulating Txnip degradation and glucose-stimulated insulin secretion (GSIS) in high glucose-cultured INS-1 cells. All these data suggest that geniposide might be an important regulator of ER stress and GSIS, and a promising compound for the treatment of T2DM. SIGNIFICANCE OF THE STUDY: Mounting evidence indicates that endoplasmic reticulum (ER) stress plays an essential role to maintain the normal cellular functions and dysfunction. In this study, we revealed that geniposide might be an important regulator of ER stress and glucose-stimulated insulin secretion in pancreatic beta cells.

Topics: Animals; Cell Line; Glucose; Insulin Secretion; Insulin-Secreting Cells; Iridoids; Rats; Unfolded Protein Response

Cerebral ischemia is one of the main causes of human neurological dysfunction. Baicalin (BC) and Geniposide (GP) and their combination (BC/GP) have an ameliorative effect on cerebral ischemia. Here, we use network pharmacology to predict the targets of BC, GP and BC/GP, then explored the protective mechanisms of the drugs on cerebral ischemia injury caused by abnormal activation of microglia cells in vitro. The results indicate that 45 targets related to cerebral ischemic injury were predicted by network pharmacology, and 26 cerebral ischemia related pathways were extracted by the KEGG database. In vitro lipopolysaccharide (LPS) stimulated BV-2 cells to establish a model of inflammatory injury induced by microglia. The effects of BC, GP and BC/GP on the expression of TNF-α, IL-1β and IL-10, TGF-β and TNF-α were verified. Network pharmacology predicts the regulation of the 5-LOX/CysLTs inflammatory pathway. Finally, we found that GP and BC/GP exert anti-inflammatory and neuroprotective effects by regulating the polarization state of microglia and down-regulating 5-LOX/CysLTs, and has certain protective effects on nerve damage following cerebral ischemia.

Topics: Animals; Arachidonate 5-Lipoxygenase; Brain Ischemia; Cell Line; Cell Polarity; Cell Proliferation; Cell Survival; Dose-Response Relationship, Drug; Drug Interactions; Flavonoids; Humans; Iridoids; Mice; Microglia; Molecular Targeted Therapy; Receptors, Leukotriene

This paper discussed the process parameters optimization of partial least-square (PLS) modeling according to quality by design (QbD) concept. D-optimal design and online near-infrared (NIR) sensor were proposed to analysis the Geniposide in Gardenia jasminoides Ellis using above process parameters to achieve robustness PLS model. Four critical model parameters (CMPs) were identified to construct a D-optimal design, which included the selection of sample set, spectra pre-processing, latent variables and variable selection methods. NIR sensor dataset was obtained under a pilot scale system. The D-optimal design optimization strategy resulted in a robust PLS model with the optimal parameters, 1/2 samples for calibration sets through Baseline spectra pre-processing with SiPLS-selecting variables under 8 factors. The critical evaluation attributes (CEAs) of PLS model were recommended as follows: the RMSEC and R

Topics: Drugs, Chinese Herbal; Equipment Design; Gardenia; Iridoids; Least-Squares Analysis; Spectroscopy, Near-Infrared

Geniposide, as a type of iridoid glycoside, has antioxidative capacity. However, the mechanism underlying the effect of geniposide in cadmium (Cd)‑induced osteoblast injury remains only partly elucidated. In the present study, Cell Counting Kit‑8 (CCK‑8) was used to determine MC‑3T3‑E1 cell viability. Flow cytometry was used to determine the rate of apoptosis and levels of reactive oxygen species (ROS). Oxidative stress‑related factors were assessed using enzyme‑linked immunosorbent method (ELISA). Quantitative real‑time polymerase chain reaction (qPCR) and western blotting were used to evaluate apoptosis‑ and bone formation‑related genes and nuclear factor erythroid 2‑related factor (Nrf2) signaling. It was demonstrated that geniposide increased the viability of the Cd‑treated MC‑3T3‑E1 cells. Geniposide decreased apoptosis and ROS accumulation compared to these parameters in the Cd group. Geniposide attenuated oxidative stress‑related factors, malondialdehyde and lactate dehydrogenase and increased antioxidant key enzyme superoxidase dismutase (SOD). The expression levels of Bax, Bcl‑2 and survivin were modulated by geniposide. Additionally, the mRNA and protein expression of the receptor activator of NF‑κB ligand (RANKL) and osterix were significantly increased, while osteoprotegerin was decreased by geniposide treatment compared to the Cd groups. Geniposide also enhanced Nrf2, heme oxygenase‑1 (HO‑1) and NAD(P)H quinone dehydrogenase 1 (NQO1) expression. The present study identified a potential agent for the treatment of Cd‑induced osteoblast injury.

Topics: Animals; Antioxidants; bcl-2-Associated X Protein; Cadmium Chloride; Cell Line; Gene Expression Regulation; Heme Oxygenase-1; Iridoids; L-Lactate Dehydrogenase; Malondialdehyde; Membrane Proteins; Mice; NAD(P)H Dehydrogenase (Quinone); NF-E2-Related Factor 2; Osteoblasts; Oxidants; Oxidative Stress; Proto-Oncogene Proteins c-bcl-2; RANK Ligand; Reactive Oxygen Species; RNA, Messenger; Signal Transduction; Superoxide Dismutase; Survivin

Acetaminophen (APAP) is a widely used over-the-counter drug for antipyretic and analgesic, but an overdose will induce acute liver injury. APAP hepatotoxicity has been the most common cause of acute liver failure in western countries with high morbidity and mortality. Geniposide (GP), an iridoid glycoside extracted from the fruit of Gardenia jasminoides, has been reported to exert a profound anti-inflammatory activity on acute and chronic diseases. However, it is never demonstrated whether GP can protect hepatocytes from APAP hepatotoxicity. In this study, we investigated the protective effect and underlying mechanism of GP against AILI. The results showed that GP pretreatment reduced the levels of ALT and AST in a dose-dependent manner and alleviated hepatocyte necrosis and apoptosis in mice exposed at APAP. Moreover, it suppressed the expression of CYP 2E1 and attenuated the exhaustion of GSH and accumulation of MDA in the liver. Furthermore, GP remarkably inhibited inflammatory cells infiltration and mitigated the release of IL-1β and TNF-α, and inhibited Toll-like receptor 4 (TLR4) expression and nuclear factor kappa (NF-κB) activation. These data suggested that GP could effectively protect hepatocytes from APAP hepatotoxicity through the down-regulation of CYP 2E1 expression and the inhibition of TLR4/NF-κB signaling pathway.

Topics: Acetaminophen; Animals; Chemical and Drug Induced Liver Injury; Cytochrome P-450 CYP2E1; Down-Regulation; Glutathione; Hepatocytes; Interleukin-1beta; Iridoids; Liver; Macrophages; Male; Malondialdehyde; Mice, Inbred C57BL; Neutrophils; NF-kappa B; Protective Agents; Signal Transduction; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

Geniposide is an active ingredient with anti-apoptotic and anti-inflammatory properties. This study was to examine the effects of geniposide on a cell model of spinal cord injury (SCI). PC12 cells were administrated with geniposide before subjected to LPS. The effects of geniposide were analyzed by utilizing CCK-8 assay, apoptosis assay, ELISA, RT-qPCR and Western blot. We found that PC12 cells viability was unchanged by treating with geniposide. However, geniposide with concentrations of 200 or 300 μg/mL significantly mitigated LPS-evoked viability loss. Meanwhile, apoptosis driven by LPS was mitigated by geniposide, which accompanied with p53, Bax and cleaved caspase-3 down-regulation, and Bcl-2 up-regulation. Besides this, the expression and release of IL-1β, IL-6, IL-8 and TNF-α evoked by LPS were mitigated by geniposide. miR-145-5p was a target of geniposide. miR-145-5p expression was up-regulated by geniposide, and geniposide did not protect PC12 cells against LPS injury when miR-145-5p was silenced. Moreover, geniposide inhibited NF-κB and JNK pathways via up-regulating miR-145-5p. In short, the present work described the neuroprotective effects of geniposide by targeting miR-145-5p. Further mechanisms involved in geniposide's beneficial effects are correlated with the inhibited NF-κB and JNK pathways. Highlights Geniposide prevents LPS-induced injury in PC12 cells; Geniposide up-regulates miR-145-5p; Geniposide protects PC12 cells via up-regulation of miR-145-5p; Geniposide inhibits NF-κB and JNK pathways via up-regulation of miR-145-5p.

Topics: Animals; Cytokines; Cytoprotection; Inflammation; Iridoids; JNK Mitogen-Activated Protein Kinases; Lipopolysaccharides; MicroRNAs; NF-kappa B; PC12 Cells; Rats; Signal Transduction; Up-Regulation

A simple gradient high-performance liquid chromatography with diode array detection (HPLC-DAD) method was used to simultaneously to analyze characteristics of six indicator compounds in the traditional Chinese medicine (TCM) formulation Wen-Qing-Yin (WQY). Separate optimization was performed using a Cosmosil C18 column gradient method with 0.1% formic acid in both mobile phases of aqueous and acetonitrile (ACN), at a flow rate, detection wavelength, and sample volume of 1.8 mL/min, 268 nm, and 10 μL, respectively. The linear regression of six active compounds berberine (BER), baicalin (BAI), ferulic acid (FER), geniposide (GEN), hydorxymethoxylfurfural (HMF), and paeoniflorin (PAE) was produced at the concentration range of 10-2000 μg/mL. The method validation revealed an acceptable precision (intra- and inter-day precision < 3.39% and 4.11%, respectively) and recovery (85.60-110.45% and 86.58-110.90%), a recovery range of 86.61-109.42%, and sensitivity (limit of detection [LOD] and limit of quantification [LOQ] values were in the range of 0.03-3.13, and 0.08-9.38 μg/mL, respectively) while the calibration curves were linear with a correlation coefficient (R

Topics: Berberine; Chromatography, High Pressure Liquid; Coumaric Acids; Drugs, Chinese Herbal; Flavonoids; Furaldehyde; Glucosides; Iridoids; Medicine, Chinese Traditional; Monoterpenes

Oxidative stress and cardiomyocyte apoptosis play critical roles in the development of doxorubicin- (DOX-) induced cardiotoxicity. Our previous study found that geniposide (GE) could inhibit cardiac oxidative stress and apoptosis of cardiomyocytes but its role in DOX-induced heart injury remains unknown. Our study is aimed at investigating whether GE could protect against DOX-induced heart injury. The mice were subjected to a single intraperitoneal injection of DOX (15 mg/kg) to induce cardiomyopathy model. To explore the protective effects, GE was orally given for 10 days. The morphological examination and biochemical analysis were used to evaluate the effects of GE. H9C2 cells were used to verify the protective role of GE in vitro. GE treatment alleviated heart dysfunction and attenuated cardiac oxidative stress and cell loss induced by DOX in vivo and in vitro. GE could activate AMP-activated protein kinase

Topics: AMP-Activated Protein Kinases; Animals; Doxorubicin; Humans; Iridoids; Male; Mice; Oxidative Stress; Signal Transduction

Topics: Allosteric Regulation; Brain Ischemia; Drug Discovery; Flavonoids; Gene Expression Profiling; Gene Regulatory Networks; Humans; Iridoids; Models, Genetic; Ursodeoxycholic Acid

Geniposide, the major active constituent of Fructus Gardeniae (FG), has been widely used to treat various diseases in China.. This chronic toxicity study was conducted to investigate the safety of geniposide.. Geniposide was administered to Sprague-Dawley (SD) rats of both sexes by oral gavage at dosages of 25, 50, or 100mg/kg in a volume of 10mL/kg once daily for 26 weeks. Endpoints included clinical observations, food consumption, body weights, blood biochemistry, haematology, and histomorphological observations.. Geniposide affected serum biochemistry, urinalysis, and haematological parameters as well as relative organ weights. The treatment also caused noticeable pathological abnormalities in liver and kidney tissues. Therefore, administration of a high dose of geniposide (100mg/kg) for 26 weeks could induced obvious liver and kidney damage.

Topics: Administration, Oral; Animals; China; Female; Iridoids; Kidney; Liver; Male; Medicine, Traditional; Rats, Sprague-Dawley; Toxicity Tests, Chronic

A novel methodology is proposed to determine the error propagation of partial least-square (PLS) for parameters optimization in near-infrared (NIR) modeling. The parameters include spectral pretreatment, latent variables and variable selection. In this paper, an open source dataset (corn) and a complicated dataset (Gardenia) were used to establish PLS models under different modeling parameters. And error propagation of modeling parameters for water quantity in corn and geniposide quantity in Gardenia were presented by both type І and type II error. For example, when variable importance in the projection (VIP), interval partial least square (iPLS) and backward interval partial least square (BiPLS) variable selection algorithms were used for geniposide in Gardenia, compared with synergy interval partial least squares (SiPLS), the error weight varied from 5% to 65%, 55% and 15%. The results demonstrated how and what extent the different modeling parameters affect error propagation of PLS for parameters optimization in NIR modeling. The larger the error weight, the worse the model. Finally, our trials finished a powerful process in developing robust PLS models for corn and Gardenia under the optimal modeling parameters. Furthermore, it could provide a significant guidance for the selection of modeling parameters of other multivariate calibration models.

Topics: Gardenia; Iridoids; Least-Squares Analysis; Limit of Detection; Models, Theoretical; Multivariate Analysis; Reference Standards; Spectroscopy, Near-Infrared; Water; Zea mays

Osteoarthritis (OA) is a chronic degenerative joint disease that is principally characterized by progressive joint dysfunction and cartilage degradation. Inflammation and apoptosis play critical roles in the progression of OA. Geniposide (GPO), one of the principal components of the fruit of Gardenia jasminoides Ellis, has been reported to have anti-inflammatory and other pharmacological effects. In this study, we performed in vitro experiments on rat chondrocytes to examine the therapeutic effects of GPO on OA and investigated its effects in vivo in a rat model of OA induced by medial meniscal tear (MMT). The results suggest that GPO can inhibit the expression of INOS, COX-2, and MMP-13 in vitro, and promote the expression of collagen II in rat chondrocytes stimulated with interleukin-1β (IL-1β). In addition, we also found that GPO can inhibit the expression of pro-apoptotic proteins such as Bax, Cyto-c, and C-caspase3 and increase the expression of the anti-apoptotic protein Bcl-2. These changes may be related to GPO-induced inhibition of the IL-1β-induced activation of the PI3K/Akt/NF-κB signaling pathway. In vivo, we also found that GPO can limit the development of OA in a rat model. Taken together, the above results indicate that GPO has potential therapeutic value for treating OA.

Topics: Animals; Apoptosis; Chondrocytes; Inflammation; Interleukin-1beta; Iridoids; NF-kappa B; Osteoarthritis; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Rats; Signal Transduction

Geniposide, an active component of Gardenia, has been reported to protect against cerebral ischemia in animals. Ginsenoside Rg1, a component of Panax notoginseng, is usually administered in combination with Gardenia for the treatment of acute ischemic stroke; however, there are unknown effects of ginsenoside Rg1 that require further investigation. In the present study, the effects of geniposide and ginsensoide Rg1 combination treatment on focal cerebral ischemic stroke were investigated. For in vivo analysis, male rats were separated into three groups, including the (control), model and geniposide + ginsenoside Rg1 groups (n=8 per group). A middle cerebral artery occlusion model was established as the model group. The treatment group was treated with geniposide (30 mg/kg, tail vein injection) + ginsenoside Rg1 (6 mg/kg, tail vein injection), and the model group received saline instead. Neurobehavioral deficits, infarct volume, brain edema, and the expression of microRNA (miR)‑155‑5p and CD11b by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and immunohistochemistry, were assessed following 24 h of ischemia. For in vitro analysis, BV2 mouse microglial cells were cultured and exposed to geniposide (40 µg/ml) + ginsenoside Rg1 (8 µg/ml) during various durations of oxygen‑glucose deprivation (OGD). The expression levels of miR‑155‑5p, pri‑miR‑155 and pre‑miR‑155 were detected by RT‑qPCR. The results demonstrated that increases in brain infarct volume, edema volume, CD11b‑positive cells and miR‑155‑5p levels were alleviated following geniposide + ginsenoside administration in rats exposed to ischemia. Furthermore, geniposide + ginsenoside Rg1 treatment suppressed the miR‑155‑5p, pri‑miR‑155 and pre‑miR‑155 expression levels in OGD‑injured BV2 microglial cells. The results of the present study demonstrated that tail vein administration of geniposide in combination with ginsenoside Rg1 protected against focal cerebral ischemia in rats through inhibition of microglial miR‑155‑5p following ischemic injury, which may serve as a novel therapeutic agent for the treatment of strokes.

Topics: Animals; Brain; Brain Ischemia; CD11b Antigen; Cell Hypoxia; Cells, Cultured; Disease Models, Animal; Ginsenosides; Glucose; Infarction, Middle Cerebral Artery; Iridoids; Male; Microglia; MicroRNAs; Neuroprotective Agents; Panax notoginseng; Rats; Rats, Sprague-Dawley

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Experimental; Cell Proliferation; Cell Survival; Immunosuppressive Agents; Interferon-gamma; Interleukin-17; Interleukin-4; Iridoids; Joints; MAP Kinase Signaling System; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Rats, Sprague-Dawley; Synoviocytes; Transforming Growth Factor beta1

Both baicalin (BA) and jasminoidin (JA) are active ingredients in Chinese herb medicine Scutellaria baicalensis and Fructus gardeniae, respectively. They have been shown to exert additive neuroprotective action in ischemic stroke models. In this study we used transcriptome analysis to explore the pure therapeutic mechanisms of BA, JA and their combination (BJ) contributing to phenotype variation and reversal of pathological processes. Mice with middle cerebral artery obstruction were treated with BA, JA, their combination (BJ), or concha margaritifera (CM). Cerebral infarct volume was examined to determine the effect of these compounds on phenotype. Using the hippocampus microarray and ingenuity pathway analysis (IPA) software, we exacted the differentially expressed genes, networks, pathways, and functions in positive-phenotype groups (BA, JA and BJ) by comparing with the negative-phenotype group (CM). In the BA, JA, and BJ groups, a total of 7, 4, and 11 specific target molecules, 1, 1, and 4 networks, 51, 59, and 18 canonical pathways and 70, 53, and 64 biological functions, respectively, were identified. Pure therapeutic mechanisms of BA and JA were mainly overlapped in specific target molecules, functions and pathways, which were related to the nervous system, inflammation and immune response. The specific mechanisms of BA and JA were associated with apoptosis and cancer-related signaling and endocrine and hormone regulation, respectively. In the BJ group, novel target profiles distinct from mono-therapies were revealed, including 11 specific target molecules, 10 functions, and 10 pathways, the majority of which were related to a virus-mediated immune response. The pure additive effects between BA and JA were based on enhanced action in virus-mediated immune response. This pure mechanistic analysis may provide a clearer outline of the target profiles of multi-target compounds and combination therapies.

Topics: Animals; Apoptosis; Disease Models, Animal; Drug Synergism; Drug Therapy, Combination; Flavonoids; Gene Expression Profiling; Gene Expression Regulation; Gene Regulatory Networks; Hippocampus; Immunity, Innate; Infarction, Middle Cerebral Artery; Iridoids; Male; Mice; Neuroprotective Agents; Oligonucleotide Array Sequence Analysis; Phenotype; RNA, Messenger; Signal Transduction; Systems Biology; Transcriptome

Altered proteostasis induced by amyloid peptide aggregation and hyperphosphorylation of tau protein, is a prominent feature of Alzheimer's disease, which highlights the occurrence of endoplasmic reticulum stress and triggers the activation of the unfolded protein response (UPR), a signaling pathway that enforces adaptive programs to sustain proteostasis. In this study, we investigated the role of geniposide in the activation of UPR induced by high glucose in primary cortical neurons. We found that high glucose induced a significant activation of UPR, and geniposide enhanced the effect of high glucose on the phosphorylation of IRE1α, the most conserved UPR signaling branch. We observed that geniposide induced the expression of HRD1, an ubiquitin-ligase E3 in a time dependent manner, and amplified the expression of HRD1 induced by high glucose in primary cortical neurons. Suppression of IRE1α activity with STF-083010, an inhibitor of IRE1 phosphorylation, prevented the roles of geniposide on the expression of HRD1 and APP degradation in high glucose-treated cortical neurons. In addition, the results from RNA interfere on HRD1 revealed that HRD1 was involved in geniposide regulating APP degradation in cortical neurons. These data suggest that geniposide might be benefit to re-establish proteostasis by enhancing the UPR to decrease the load of APP in neurons challenged by high glucose.

Topics: Amyloid beta-Peptides; Amyloid beta-Protein Precursor; Animals; Disease Models, Animal; Endoplasmic Reticulum Stress; Iridoids; Neurons; Rats, Sprague-Dawley; Signal Transduction; Ubiquitin-Protein Ligases; Unfolded Protein Response

Identifying the variation of core modules and hubs seems to be critical for characterizing variable pharmacological mechanisms based on topological alteration of disease networks. We first identified a total of eight core modules by using an approach of multiple modular characteristic fusing (MMCF) from different targeted networks in ischemic mice. Interestingly, the value of module disturbance intensity (MDI) increased in drug combination group. Second, we redefined a weak allosteric module and a strong allosteric module. Then, we identified 15 pharmacological module drivers (PMDs) by leave-one-out screening with a cutoff of two folds, which were at least, in part, validated by expression and variation of topological contribution. Finally, we revealed the fusional and emergent variation of PMD in core modules contributing to multidimensional synergistic mechanism in ischemic mice and rats. Our findings provide a new set of drivers that might promote the pharmacological modular flexibility and offer a potential avenue for disease treatment.

Topics: Animals; Cholic Acid; Cyclins; Drug Synergism; Gene Expression Regulation; Gene Regulatory Networks; Hippocampus; Infarction, Middle Cerebral Artery; Interleukin 1 Receptor Antagonist Protein; Iridoids; Mice; Models, Biological; Oligonucleotide Array Sequence Analysis; Rats

Oxidative stress is one possible pathogenic event in vitiligo that induces melanocyte destruction. Geniposide exerts certain antioxidant effects on various cells by activating the phosphoinositol 3-kinase (PI3K)-Akt signalling pathway. However, researchers have not clearly determined whether geniposide protects human melanocytes from oxidative stress or identified the underlying mechanism of such protection.. To determine whether geniposide protects melanocytes from H. The antioxidant effects of geniposide on human melanocytes were examined by measuring cell viability, apoptosis rates, reactive oxygen species (ROS) production and activities of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). We examined expression of Akt, phosphorylated Akt (p-Akt), Bcl-2, Bax, and cleaved caspase 3 and cleaved caspase 9 proteins to determine the involvement of the PI3K-Akt pathway.. Pretreatment with geniposide 5, 25, 125 or 625 μmol/L increased cell viability and decreased the apoptosis rate of H. Based on these results, geniposide protects human melanocytes from H

Topics: Antioxidants; Cells, Cultured; Humans; Hydrogen Peroxide; Iridoids; Melanocytes; Oxidative Stress; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction

Geniposide (genipin-1-O-β-d-glucoside) is a major bioactive ingredient in the fruits of gardenia [Gardenia jasminoides J. Ellis (Rubiaceae)], a traditional herbal medicine in Asian countries.. This work assesses the skin anti-photoaging potential of geniposide in human dermal fibroblasts under UV-B irradiation.. The anti-photoaging property of geniposide, at varying concentrations (5, 12 and 30 μM) treated for 30 min prior to UV-B irradiation, was evaluated by analysing reactive oxygen species (ROS), promatrix metalloproteinase-2 (proMMP-2), glutathione (GSH), superoxide dismutase (SOD), nuclear factor erythroid 2-related factor 2 (Nrf2) and cellular viability.. Geniposide suppressed the ROS elevation under UV-B irradiation, which was revealed using three ROS-sensitive fluorescent dyes. The use of 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA), dihydroethidium (DHE) and dihydrorhodamine 123 (DHR-123) elicited the IC. After the skin anti-photoaging potential of geniposide may be further verified, it can be utilized as a safer resource in the manufacture of effective anti-aging cosmetics.

Topics: Cell Line; Cell Survival; Dermis; Dose-Response Relationship, Drug; Fibroblasts; Humans; Iridoids; Oxidative Stress; Reactive Oxygen Species; Skin Aging; Ultraviolet Rays

Myocarditis is a cardiomyopathy associated with inflammatory response. It has been reported that geniposide (GEN), a traditional Chinese herb extract from Gardenia jasminoides Ellis, possesses an anti-inflammatory effect and a protective effect on cardiomyocytes. The present study aimed to explore the protective role of GEN and the underlying mechanism in LPS-injured H9c2 cells. H9c2 cells were treated with LPS to induce cell injury and then we investigated the effect of GEN. miR-145 expression was inhibited by transfection with miR-145 inhibitor and its expression was measured by RT-PCR. Cell viability and apoptotic cells were measured by CCK-8 assay and flow cytometry analysis. The levels of pro-inflammatory factors (IL-6, TNF-α, and MCP-1) were assessed by western blot and RT-PCR. Western blot was performed to detect the expression of the MEK/ERK pathway-related factors. LPS exposure reduced cell viability, increased apoptotic cells, and promoted the expression of pro-inflammatory factors in H9c2 cells. However, GEN pretreatment significantly reduced LPS-induced cell injury, as increased cell viability, reduced apoptotic cells, and inhibited the expression of pro-inflammatory factors. Moreover, we found that miR-145 expression was down-regulated by LPS exposure but was up-regulated by GEN pretreatment. The protective effect of GEN on LPS-injured H9c2 cells was blocked by miR-145 inhibitor. In addition, GEN inhibited the MEK/ERK pathway through up-regulating miR-145. Our results suggested that GEN exerted a protective role in LPS-injured H9c2 cells. The GEN-associated regulation might be related to its regulation on miR-145 and the MEK/ERK signaling pathway.

Topics: Animals; Cell Line; Cell Survival; Inflammation; Iridoids; Lipopolysaccharides; MAP Kinase Signaling System; MicroRNAs; Myocytes, Cardiac; Rats; Up-Regulation

Crocin (CRO), chlorogenic acid (CGA), geniposide (GEN), and quercetin (QUE) are all natural compounds with anti-obesity properties, in particular, hypolipidemic effects, which have been widely used for the treatment of obesity-related metabolic diseases. However, it is not yet known whether these compounds interact synergistically. Here, we investigated the effects and molecular mechanisms of CRO, CGA, GEN, QUE, and a combination of all four compounds (CCGQ), on lipid accumulation in human hepatoma (HepG2 cells).. The optimal concentration of CRO, CGA, GEN, QUE to stimulate HepG2 cells proliferation was determined using MTT assay. HepG2 cells were pretreated with 10 μmol/L simvastatin, 1 μmol/L CRO, 30 μmol/L CGA, 10 μmol/L GEN, 10 μmol/L QUE, and CCGQ (a combination of 1 μmol/L CRO, 30 μmol/L CGA, 10 μmol/L GEN, and 10 μmol/L QUE) for 24 or 48 h. Oil red O staining and extracellular TC and TG levels were detected. The RT-PCR was used to observe on cholesterol metabolism-related gene expression. Immunocytochemistry and western-blot assayed the 3-hydroxy-3-methylglutaryl-coenzyme (HMGCR) protein expression in HepG2 cells.. Compared to those of control, we demonstrated that treating HepG2 cells for 48 h with CCGQ resulted in a strong synergistic effect, causing a marked decrease in lipid deposition in comparison to individual treatments, in both triglyceride and total cholesterol (CRO, 5.74- and 1.49-folds; CGA, 3.38- and 1.12-folds; GEN, 4.04- and 1.44-folds; QUE, 3.36- and 1.24-folds; simvastatin, 5.49- and 1.83-folds; and CCGQ, 7.75- and 2.20-folds), and Oil red O staining assays. In addition, CCGQ treatment increased ATP-binding cassette transporter (ABCA1), cholesterol 7α-hydroxylase (CYP7A1), and AMP-activated protein kinase 2α (AMPKα2) mRNA expression, while decreasing sterol regulatory element binding protein 2 (SREBP2), and liver X receptor alpha (LXRα) mRNA expression. Notably, CCGQ was more effective in decreasing HMGCR expression than the individual treatments.. The CCGQ combination has potential, both as a complementary therapy for hyperlipemia, and in preventing further obesity-related complications.

Topics: Carotenoids; Chlorogenic Acid; Cholesterol; Drug Synergism; Hep G2 Cells; Humans; Iridoids; Lipid Metabolism; Phytochemicals; Quercetin

Geniposide (GP), a bioactive iridoid glycoside isolated from Gardenia jasminoides Ellis, as well as an agonist of Glucagon-like peptide-1 receptor (GLP-1R), has been reported to exhibit antidepressant-like effects in several rodent models. However, the underlying mechanisms remain obscure. In this study, we mainly investigated the antidepressant-like effects of GP and explored the possible mechanisms associated with GLP-1R signaling by using the repeated restraint stress (RRS)-induced depression model of mice. We found that GP treatment significantly ameliorated depression-like behaviors induced by RRS, such as decreased sucrose preference (SP) in sucrose preference test (SPT), reduced locomotor activity in open field test (OFT) and extended immobility time in tail suspension test (TST) and forced swimming test (FST). In addition, GP suppressed the neuronal apoptosis as well as reduced pro-inflammatory cytokines levels including Interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the hippocampus of RRS-induced mice. Moreover, GP restored the expression of GLP-1R/protein kinase B (AKT) signaling-related protein. Importantly, these effects were blocked by an antagonist of GLP-1R, Exendin(9-39) (Ex(9-39)), indicating that GLP-1R signaling pathway might be involved in the neuroprotective and antidepressant-like effecacy of GP. In conclusion, GP exerted promising antidepressant-like effects in RRS mice, and the antidepressant-like action of GP might be closely relevant to GLP-1R/AKT signaling.

Topics: Animals; Antidepressive Agents; Apoptosis; Depression; Glucagon-Like Peptide-1 Receptor; Hippocampus; Iridoids; Locomotion; Male; Mice, Inbred ICR; Neurons; Proto-Oncogene Proteins c-akt; Restraint, Physical; Signal Transduction; Stress, Psychological

The current study was designed to investigate the protective role of geniposide (GE) in liver injury in diabetic C57BL/KsJ-db/db mice and to explore the underlying mechanisms. The oral glucose tolerance test was performed, and the levels of insulin, alanine aminotransferase (ALT), aspartate aminotransaminase (AST), total cholesterol (TC) and triglyceride (TG) were determined. The levels of the pro-inflammatory cytokines interleukin (IL)-1β, IL-6 and tumour necrosis factor-α were decreased by GE, metformin and fasudil in diabetic db/db mice. Western blotting analysis showed that the expression levels of Rho, ROCK1, ROCK2, p-NF-κBp65 and p-IκBα were significantly reversed by GE treatment. These findings demonstrated that GE exhibits a protective effect on diabetic hepatic inflammation.

Topics: Alanine Transaminase; Animals; Anti-Inflammatory Agents; Aspartate Aminotransferases; Cholesterol; Cytokines; Diabetes Mellitus; Insulin; Iridoids; Liver; Mice, Inbred C57BL; rho-Associated Kinases; Triglycerides

It has been suggested that the activation of the p38 mitogen activated protein kinases (MAPKs) signaling pathway plays a significant role in the progression of OA by leading to the overexpression of proinflammatory cytokines, chemokines, and signaling enzymes in human osteoarthritis chondrocytes. However, most p38 MAPK inhibitors applied for OA have been thought to be limited due to their potential long-term toxicities. Geniposide (GE), an iridoid glycoside purified from the fruit of the herb, has been widely used in traditional medicine for the treatment of a variety of chronic inflammatory diseases. In this study, we evaluated the inhibition effect of geniposide on the inflammatory progression of the surgically induced osteoarthritis and whether the protective effect of geniposide on OA is related to the inhibition of the p38 MAPK signaling pathway.

Topics: Animals; Anti-Inflammatory Agents; Chondrocytes; Inflammation; Interleukin-1beta; Iridoids; Matrix Metalloproteinases; Nitric Oxide; Osteoarthritis; p38 Mitogen-Activated Protein Kinases; Rabbits; Signal Transduction; Synovial Fluid; Tumor Necrosis Factor-alpha

Topics: Animals; Anti-Inflammatory Agents; Arthritis, Experimental; Chromatography, High Pressure Liquid; Dialysis Solutions; Dinoprostone; Disease Models, Animal; Iridoids; Knee Joint; Male; Microdialysis; Rats; Tandem Mass Spectrometry; Treatment Outcome

Topics: Antineoplastic Agents, Phytogenic; Biomarkers, Tumor; Carcinoma, Squamous Cell; Caspases; Cell Line, Tumor; Cell Proliferation; Gene Expression Regulation; Humans; Iridoids; Lacticaseibacillus casei; Mouth Neoplasms

Geniposide is the main bioactive constituent of gardenia fruit. Skeletal-muscle fibrosis is a common and irreversibly damaging process. Numerous studies have shown that geniposide could improve many chronic diseases, including metabolic syndrome and tumors. However, the effects of geniposide on skeletal-muscle fibrosis are still poorly understood. Here, we found that crude extracts of gardenia fruit pomace could significantly decrease the expression of profibrotic genes in vitro. Moreover, geniposide could also reverse profibrotic-gene expression induced by TGF-β and Smad4, a regulator of skeletal-muscle fibrosis. In addition, geniposide treatment could significantly downregulate profibrotic-gene expression and improve skeletal-muscle injuries in a mouse model of contusion. These results together suggest that geniposide has an antifibrotic effect on skeletal muscle through the suppression of the TGF-β-Smad4 signaling pathway.

Topics: Animals; Fibrosis; Fruit; Gardenia; Gene Expression; Iridoids; Male; Mice; Muscle, Skeletal; Plant Extracts; Signal Transduction; Smad4 Protein; Transforming Growth Factor beta

Topics: Acetylation; beta-Glucosidase; Biotransformation; Cells, Immobilized; Chromatography, High Pressure Liquid; Crystallography, X-Ray; Fermentation; Iridoids; Isomerism; Models, Molecular; Protein Conformation; Proton Magnetic Resonance Spectroscopy; Temperature; Trichoderma

Recent evidence demonstrates that a double dose of Jasminoidin (2·JA) is more effective than Jasminoidin (JA) in cerebral ischemia therapy, but its dosage-effect mechanisms are unclear. In this study, the software GeneGo MetaCore was used to perform pathway analysis of the differentially expressed genes obtained in microarrays of mice belonging to four groups (Sham, Vehicle, JA, and 2·JA), aiming to elucidate differences in JA and 2·JA's dose-dependent pharmacological mechanism from a system's perspective. The top 10 enriched pathways in the 2·JA condition were mainly involved in neuroprotection (70% of the pathways), apoptosis and survival (40%), and anti-inflammation (20%), while JA induced pathways were mainly involved in apoptosis and survival (60%), anti-inflammation (20%), and lipid metabolism (20%). Regarding shared pathways and processes, 3, 1, and 3 pathways overlapped between the Vehicle and JA, Vehicle and 2·JA, and JA and 2·JA conditions, respectively; for the top ten overlapped processes these numbers were 3, 0, and 4, respectively. The common pathways and processes in the 2·JA condition included differentially expressed genes significantly different from those in JA. Seven representative pathways were only activated by 2·JA, such as

Topics: Animals; Cerebrovascular Disorders; Disease Models, Animal; Dose-Response Relationship, Drug; Gene Expression Profiling; Gene Expression Regulation; Humans; Iridoids; Male; Mice; Reperfusion Injury

Topics: Anti-Bacterial Agents; Antioxidants; Bacteria; China; Chlorogenic Acid; Chromatography, High Pressure Liquid; Eucommiaceae; Flavonoids; Fungi; Gutta-Percha; Iridoid Glucosides; Iridoids; Kaempferols; Lignans; Metabolome; Phenols; Plant Bark; Plant Extracts; Plant Leaves; Plants, Medicinal; Quercetin; Rutin

A new approach for ionic liquid based enzyme-assisted extraction coupled with in-situ hydrolysis (ILEIH) of geniposide from Eucommia ulmoides Olive barks is presented, in which enzymatic hydrolysis is used in an ionic liquid aqueous medium to prepare genipin. The method relied on the use of single cellulase to the extract and hydrolyze geniposide, which is performed continuously in the same system; genipin is easy in preparation with exempting the isolation and purification of geniposide. The mechanism of ILEIH procedure was discussed in detail to illustrate the advantage of ILEIH in the extraction process. 0.5 mol/L [C6mim]Cl aqueous solution was selected as extraction solvent. The optimum conditions of 140 min treatment time, 19.81 mL/g liquid-solid ratio, 5.15 mg/mL enzyme concentration and 5.0 pH value for the ILEIH process were obtained after investigating by single factor experiments and Box-Benhnken design in terms of the genipin increment. And the first-order kinetic model based on β-glucosidase in the three reaction medium were established to study their impacts on the reaction rate. The proposed ILEIH method was validated by stability, repeatability, and recovery experiments and shows reliable data in the extraction and hydrolysis process. Therefore, this proposed approach is promising for the in-situ production of genipin and should be potentially applied to the obtaining of other active aglycons.

Topics: beta-Glucosidase; Cellulase; Chemistry Techniques, Analytical; Eucommiaceae; Hydrolysis; Ionic Liquids; Iridoids

Gastric cancer (GC) is a serious disease with high incidence rate and high mortality. Geniposide (GEN) exhibits multiple biological properties including anti-tumor function. However, effect of GEN on GC is not well studied. Hence, the effects of GEN on GC were investigated in our study. HULC expression in GC tissue and GC cell lines (MKN45, SGC-7901, MKN28, AGS) was detected by qRT-PCR. Cell viability, colony formation, migration, invasion and cell apoptosis were examined by CCK-8 assay, survival fraction assay, modified two-chamber migration assay, Millicell Hanging Cell Culture and flow cytometry analysis, respectively. The expression of matrix metalloproteinase (MMP)-2/9 and vimentin was detected by qRT-PCR and western blot, respectively. The expression of PI3K/AKT and JNK was measured by western blot. The expression of HULC was up-regulated both in GC tissue and cell lines (P < .05, P < .01 or P < .001). GEN negatively regulated the expression of HULC in MKN45 cells (P < .05 or P < .01). GEN decreased cell viability (P < .05), colony formation (P < .01), migration (P < .05) and invasion (P < .05) while HULC overexpression led to the opposite results in GEN-treated cells. The expression of phosphatidylinositol 3' -kinase (PI3K)/ protein kinase B (AKT) and c-Jun N-terminal kinase (JNK) was down-regulated by GEN (all P < .05) while reversed by HULC overexpression. HULC was up-regulated in GC. GEN inhibited MNK45 cell viability, colony formation, migration and invasion while induced cell apoptosis by down-regulation of HULC in MKN45 cells. GEN inactivated PI3K/AKT and JNK signal pathways through down-regulation of HULC.

Topics: Adult; Apoptosis; Cell Movement; Cell Proliferation; Down-Regulation; Female; Humans; Iridoids; Male; Middle Aged; RNA, Long Noncoding; Signal Transduction; Stomach Neoplasms

To investigate the effect of different initial processing methods on the quality of Gardenia and determine the best cooking time in gardenia processing through the determination of index components content. The contents of geniposide, crocetin Ⅰ and total iridoid glycosides in Gardenia were determined before storage, six months after storage and one year after storage. During storage, the contents of geniposide, crocetin Ⅰ and total iridoid glycosides in directly dried Gardenia were 1.68%, 0.45% and 6.45% respectively. The contents of geniposide, crocetin Ⅰ and total iridoid glycosides in Gardenia with different steaming time were 1.34%-0.5%, 0.28%-0.06% and 6.09%-1.59% respectively. The contents of geniposide, crocetin Ⅰ and total iridoid glycosides in Gardenia with different boiling time (adding alum)were 1.42%-0.41%, 0.35%-0.07% and 6.40%-1.65% respectively. The direct drying of Gardenia samples could not achieve the function of killing enzyme and protecting glycosides. The enzymes from degradation of the index components were basically destroyed after steaming time of 13 min or boiling (adding alum) time of 8 min, achieving the function of killing enzyme and protecting glycosides.

Topics: Carotenoids; Chemistry, Pharmaceutical; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Gardenia; Iridoid Glycosides; Iridoids; Phytochemicals; Vitamin A

Staphylococcus aureus (S.aureus), a Gram-positive organism, is a frequent cause of subclinical mastitis. Geniposide, an iridoid glucoside isolated from the fruits of Gardenia jasminoides, has been reported to exhibit antibacterial and anti-inflammatory properties. The ability of S. aureus internalizing into bovine mammary epithelial cells (bMEC) has been responsible for the establishment of the bovine mastitis. In this study, we evaluated the effect of geniposide on S. aureus internalization into bMEC and investigated the possible mechanism of action. The results revealed that geniposide (25-100 μg/ml) reduced S. aureus internalization by 17%-67% and down-regulated the mRNA expressions of TAP and BNBD5. Furthermore, geniposide inhibited S.aureus-induced NF-κB activation in a dose-dependent manner. In summary, the potential mechanism of geniposide reducing S. aureus internalization may be by inhibiting NF-κB activation.

Topics: Animals; Anti-Bacterial Agents; Anti-Inflammatory Agents; Cattle; Cells, Cultured; Endocytosis; Epithelial Cells; Gene Expression Profiling; Iridoids; NF-kappa B; Signal Transduction; Staphylococcus aureus

Geniposide (GE) is an active component isolated from the fruit of Gardenia jasminoides Ellis that has anti-inflammatory and other pharmacological effects; however, the underlying mechanism of GE action has not been elucidated in rheumatoid arthritis (RA). Previous studies have shown that GE plays a therapeutic role in RA via regulation of the integrin beta 1 (Itgβ1)-mediated Ras-Erk1/2 signalling pathway. However, the specific mechanism of GE action on Itgβ1 has not been clarified. Recent evidence indicates that microRNAs (miRNAs) are involved in the development of RA. In this study, we developed a miRNA-124a-based synoviocyte repair strategy. We demonstrated that miRNA-124a can directly inhibit the expression of the Itgβ1 gene and decrease TNF-α-stimulated cell proliferation in vitro. MH7A cells were obtained from the patient with RA and treated with GE in the presence of TNF-α (10 ng/mL). Additionally, we demonstrated that the expression of miRNA-124a can be regulated by GE. GE upregulated the expression of miRNA-124a and decreased the expression of Itgβ1 at the mRNA and protein levels. The results of the present study are the first to suggest that GE inhibits TNF-α-stimulated cell proliferation and blocks the activation of the Ras-Erk1/2 pathway via the upregulation of miRNA-124a expression. Our study elucidates the role of miRNA-124a as a protected miRNA in RA and may provide a novel strategy for the diagnosis and treatment of RA in the future.

Topics: Anti-Inflammatory Agents; Arthritis, Rheumatoid; Cell Line; Cell Proliferation; Fibroblasts; Gardenia; Gene Expression Regulation; Humans; Integrin beta1; Iridoids; MAP Kinase Signaling System; MicroRNAs; Oncogene Protein p21(ras); Synoviocytes; Tumor Necrosis Factor-alpha

Topics: Animals; Anthraquinones; Chromatography, Liquid; Coumarins; Drug Evaluation, Preclinical; Herbal Medicine; Iridoids; Male; Rats; Reproducibility of Results; Sensitivity and Specificity; Tandem Mass Spectrometry

Hepatic gluconeogenesis plays an important role in regulating fasting plasma glucose levels and is a target of anti-diabetic drugs. Several kinds of iridoid glucosides exhibit hypoglycemic effect, whereas the mechanism was not clear.. In this study, the effects of geniposide and gentiopicroside, two natural iridoid glucosides, on hepatic gluconeogenesis were investigated.. Glucose uptake assay, MTT assay, q-PCR, luciferase assay and western blot assay were performed to investigate the pharmacological effect of geniposide and gentiopicroside on human liver cell line L02. Thereby the fast blood glucose and intraperitoneal glucose tolerance were measured in high fat diet induced hyperglycemic mice after geniposide or gentiopicroside administration.. The results showed that geniposide and gentiopicroside inhibited the transcription of G6PC and PEPCK in L02 cells and in mice. Additional experimental data indicated that these two compounds were able to inhibit the transcriptional activity of FOXO1 by inducing phosphorylation of AKT at Ser473. Furthermore, we found that these two compounds alleviated high fat diet induced hyperglycemia in mice.. Geniposide and gentiopicroside might reduce blood glucose and suppress hepatic gluconeogenesis by regulating the AKT-FOXO1 pathway, and the potential use of these two iridoid glucosides as anti-diabetic agents merits further in-depth exploration.

Topics: Animals; Blood Glucose; Cells, Cultured; Diet, High-Fat; Forkhead Box Protein O1; Gluconeogenesis; Humans; Hypoglycemic Agents; Iridoid Glucosides; Iridoids; Liver; Male; Mice; Mice, Inbred C57BL; Phosphorylation; Proto-Oncogene Proteins c-akt; Signal Transduction

To establish the high performance liquid chromatography (HPLC) fingerprint for Digeda-4 decoction (DGD-4D), determine the contents of aesculetin, geniposide, picroside Ⅰ, picroside Ⅱ and ellagicacid in DGD-4D, and provide the scientific foundation for quality control of DGD-4D. The analysis was performed on Diamonsil(2) C₁₈ (4.6 mm×250 mm,5 μm) column, with methanol-0.1% phosphoric acid aqueous solution as mobile phase for gradient elution. The flow rate was 1.0 mL·min⁻¹; injection size was 10 μL; temperature was maintained at 30 °C, and the detection wavelength was set at 254 nm. The common mode of DGD-4D HPLC fingerprint was established, and the hidden information was analyzed by Chemometrics. Chromatographic peaks for DGD-4D were identified by HPLC and quantitative analysis was conducted for characteristic peaks. There were 17 common peaks in the fingerprints and the similarity of the fingerprints was over 0.9 in all 15 batches. The samples were broadly divided into four kinds by principal component analysis and clustering analysis. Four marker compounds were verified by partial least squares discriminant analysis, and No. 9, 12 and 14 peaks were identified as geniposide, picroside Ⅱ, and picroside Ⅰ respectively. The average recoveries were in the range of 95.91%-97.31%. The HPLC fingerprint method for content determination is reliable, accurate, rapid, simple, and reproducible, and can be used as one of the effective methods to control the quality of DGD-4D.

Topics: Chromatography, High Pressure Liquid; Cinnamates; Drugs, Chinese Herbal; Iridoid Glucosides; Iridoids; Methanol; Principal Component Analysis; Quality Control

Our previous study found that geniposide, an agonist of glucagon-like peptide-1 receptor (GLP-1R), protected against cardiac hypertrophy via the activation of AMP-activated protein kinase

Topics: AMP-Activated Protein Kinases; Animals; Anti-Inflammatory Agents; Cardiomegaly; Diet, High-Fat; Heart; Inflammation; Iridoids; Male; Mice; Mice, Inbred C57BL; Myocytes, Cardiac; Obesity; Signal Transduction; Sirtuin 1

Qing'e Pills is a Chinese traditional herbal product, which is often used to strengthen muscles and bones in TCM (traditional Chinese Medicine) practice. Its two main component herbs, namely,

Topics: Benzofurans; Caco-2 Cells; Drugs, Chinese Herbal; Ficusin; Furocoumarins; Glycosides; Humans; Iridoids; Temperature

Our previous work revealed that in the pancreatic β cell line, geniposide modulated ATP production and glucose-stimulated insulin secretion (GSIS) induced by the acute stimulation of high glucose concentration. However, the effects of geniposide on functional impairment and the mass of β-cells exposed to elevated levels of glucose remains unknown. In the present study, impaired GSIS and restrained proliferation were observed in the prolonged culture of insulinoma INS-1 cells with 33mM of glucose (high glucose). Our results indicate that the glucose-induced impairment of insulin release was significantly reverted by the inclusion of 1 or 10μM of geniposide. Moreover, induction of the phosphorylation of AMP-activated protein kinase (AMPK) was observed, which promoted the utilization of nutrient stores for energy production. AMPK phosphorylation was enhanced by an increased number of INS-1 cells, and the increased expression of AMPK downstream target heme oxygenase 1 (HO-1), under high glucose concentration. Furthermore, geniposide protected rat insulinoma cells from apoptosis in high-glucose concentrations. We have shown that these effects were associated with an increased apoptosis-related Bcl-2/BAX protein ratio. In conclusion, geniposide dose dependently improves β-cell function and increases the proliferation of β-cells exposed to prolonged hyperglycemia.

Topics: Animals; Apoptosis; Cell Line, Tumor; Cell Proliferation; Cytoprotection; Glucose; Humans; Insulin; Insulinoma; Iridoids; Rats

Beneficial effects of the Chinese herbal medicine Qushi Huayu Decoction (QHD) were observed with non-alcoholic fatty liver disease (NAFLD) patients and animal models. The impact of QHD or its active components (geniposide and chlorogenic acid, GC) on NAFLD liver transcriptome and gut microbiota was examined with NAFLD rats. Increased expression for genes required for glutathione production and decreased expression for genes required for lipid synthesis was observed in NAFLD livers treated with QHD and GC. GC treatment decreased serum LPS, which could be explained by reduced mucosal damage in the colon of GC-treated rats. Further, our data suggest an increased abundance of Treg-inducing bacteria that stimulated the Treg activity in GC treated colon, which in turn down-regulated inflammatory signals, improved gut barrier function and consequently reduced hepatic exposure to microbial products. Our study suggests that QHD simultaneously enhanced the hepatic anti-oxidative mechanism, decreased hepatic lipid synthesis, and promoted the regulatory T cell inducing microbiota in the gut.

Topics: Animals; Chlorogenic Acid; Colon; Disease Models, Animal; Down-Regulation; Drugs, Chinese Herbal; Gastrointestinal Microbiome; Glutathione; Humans; Intestinal Mucosa; Iridoids; Lipid Metabolism; Lipopolysaccharides; Liver; Male; Mice; Mice, Inbred C57BL; Molecular Targeted Therapy; Non-alcoholic Fatty Liver Disease; Rats; Rats, Sprague-Dawley; Signal Transduction; T-Lymphocytes, Regulatory; Transcriptome

Ulcerative colitis (UC), an idiopathic inflammatory bowel disease, not only affects millions of patients worldwide, but also increases the risk of colon cancer. Geniposide is an iridoid glycoside and has many biological activities such as anti-inflammatory and antioxidant. However, its protective efficacy and mechanism of action against UC are still unclear. In this study, we aimed to investigate the protective effects and mechanisms of geniposide on dextran sulfate sodium (DSS)-induced experimental colitis in mice. The results revealed that geniposide alleviated body weight loss, disease activity index, colon length shortening and colonic pathological damage induced by DSS. Geniposide significantly suppressed pro-inflammatory cytokines by regulating NF-κB and PPARγ pathways in vivo and in vitro. Furthermore, geniposide also significantly regulated the expressions of ZO-1 and occludin in DSS-induced experimental colitis in mice and lipopolysaccharide (LPS)-triggered inflammation in Caco-2 cells. These findings indicated that geniposide may be a new natural chemopreventive agent to combat UC.

Topics: Animals; Anti-Inflammatory Agents; Caco-2 Cells; Colitis; Colitis, Ulcerative; Colon; Cytokines; Dextran Sulfate; Disease Models, Animal; Humans; Inflammation; Inflammation Mediators; Intestinal Mucosa; Iridoids; Male; Mice; Mice, Inbred C57BL; NF-kappa B; PPAR gamma; Signal Transduction

Hollow fiber cell fishing (HFCF) based on human hepatoma cell HepG-2 or human renal tubular cell ACHN coupled with high performance liquid chromatography/ultraviolet detection (HPLC/UV) was developed and used for simultaneous study of the major antitumor active components in a formula of Yinchenhao decoction (YCHD) in vitro and in vivo, and in its constituent herbs, namely, Artemisia capillaris Thunb, Gardenia jasminoides Ellis, Radix et Rhizoma Rhei in vitro. Before application, chlorogenic acid, geniposide, p-hydroxyacetophenone, crocin and rhein were chosen as model compounds, the various validations, e.g., cell growth and cell viability on the fiber inner wall, binding between fiber active center and component, repeatability of retention time or relative peak area of the active components were investigated. We screened and identified the major antitumor active components of YCHD, verified their synergetic or antagonistic effect, correlated the major active components between in vitro and in vivo and determined major effective components. Our study will serve as a valuable reference in probing the antitumor material basis of YCHD and its three constituent herbs, as well as in identifying novel clinical aspects of traditional Chinese medicines. The results showed that HCFC-HPLC is a simple, rapid, and reliable method that simultaneously researches the interaction between multiple targets and multiple components of herbal medicine.

Topics: Acetophenones; Animals; Anthraquinones; Antineoplastic Agents; Carotenoids; Cell Culture Techniques; Cell Proliferation; Cell Survival; Chlorogenic Acid; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Hep G2 Cells; Humans; Iridoids; Male; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Sensitivity and Specificity; Tissue Distribution

Rheumatoid arthritis (RA) is a systemic, Th1 cytokine-predominant autoimmune disease result in a chronic and inflammatory disorder. Geniposide (GE), an iridoid glycoside compound that is purified from Gardenia jasminoides Ellis, has antiinflammatory and other immunoregulatory effects, but its exact mechanism of actions on RA is unknown. The aim of this study was to elucidate antiinflammation effects of GE on adjuvant arthritis (AA) rats and its possible immune tolerance mechanisms. Male Sprague-Dawley rats were administered with GE (30, 60, and 120 mg/kg) orally from day 17 to 24 after immunization. Lymphocyte proliferation was assessed by MTT. Levels of interleukin-2 (IL-2), IL-4, and transforming growth factor-β1 were tested by ELISA. The expression of β2-AR, GRK2, and β-arrestin-1 and β-arrestin-2 was detected by western blot. Geniposide was found to relieve the secondary hind paw swelling and arthritis scores, along with attenuating histopathologic changes and decreasing IL-2 and increasing IL-4, transforming growth factor-β1 in mesenteric lymph node (MLN) lymphocytes of AA rats. In addition, GE in vivo increased the expression of β2-AR and decreased the expression of GRK2, β-arrestin-1 and β-arrestin-2, and level of cyclic adenosine monophosphate of MLN lymphocytes in AA rats. From these results, we can infer that GE on immune tolerance effects, β2-AR desensitization, and β2-AR-AC-cyclic adenosine monophosphate transmembrane signal transduction of MLN lymphocytes plays crucial roles in antiinflammatory and immunoregulatory pathogeneses of RA. Copyright © 2017 John Wiley & Sons, Ltd.

Topics: Animals; Anti-Inflammatory Agents; Arthritis, Experimental; Arthritis, Rheumatoid; beta-Arrestin 1; beta-Arrestin 2; Cell Proliferation; Cyclic AMP; G-Protein-Coupled Receptor Kinase 2; Gardenia; Immune Tolerance; Interleukin-2; Interleukin-4; Iridoids; Lymph Nodes; Lymphocytes; Male; Rats; Rats, Sprague-Dawley; Signal Transduction; Transforming Growth Factor beta1

Traditional Chinese medicine (TCM) treatment can be valuable therapeutic strategies. However, the active components and action mechanisms that account for its therapeutic effects remain elusive. Based on the hypothesis that the components of a formula which exert effect would be measurable in target tissue, a target tissue metabolomics-based strategy was proposed for screening of antipyretic components in Qingkaikling injection (QKLI). First, we detected the components of QKLI which could reach its target tissue (hypothalamus) by determining the hypothalamus microdialysate and discovered that only baicalin and geniposide could be detected. Then, by conducting hypothalamus metabolomics studies, 14 metabolites were screened as the potential biomarkers that related to the antipyretic mechanisms of QKLI and were used as its pharmacodynamic surrogate indices. Subsequently, the dynamic concentration of baicalin and geniposide in hypothalamus microdialysates and biomarkers in hypothalamus were measured and correlated with each other. The results indicated that only baicalin shown a good correlation with these biomarkers. Finally, a network pharmacology approach was established to validate the antipyretic activity of baicalin and the results elucidated its antipyretic mechanisms as well. The integrated strategy proposed here provided a powerful means for identifying active components and mechanisms contributing to pharmacological effects of TCM.

Topics: Administration, Intravenous; Animals; Antipyretics; Drugs, Chinese Herbal; Flavonoids; Hypothalamus; Iridoids; Male; Medicine, Chinese Traditional; Metabolomics; Rats; Rats, Sprague-Dawley

Fructus Gardenia (FG), containing the major active constituent Geniposide, is widely used in China for medicinal purposes. Currently, clinical reports of FG toxicity have not been published, however, animal studies have shown FG or Geniposide can cause hepatotoxicity in rats. We investigated Geniposide-induced hepatic injury in male Sprague-Dawley rats after 3-day intragastric administration of 100 mg/kg or 300 mg/kg Geniposide. Changes in hepatic histomorphology, serum liver enzyme, serum and hepatic bile acid profiles, and hepatic bile acid synthesis and transportation gene expression were measured. The 300 mg/kg Geniposide caused liver injury evidenced by pathological changes and increases in serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and γ-glutamytransferase (γ-GT). While liver, but not sera, total bile acids (TBAs) were increased 75% by this dose, dominated by increases in taurine-conjugated bile acids (t-CBAs). The 300 mg/kg Geniposide also down-regulated expression of Farnesoid X receptor (FXR), small heterodimer partner (SHP) and bile salt export pump (BSEP). In conclusion, 300 mg/kg Geniposide can induce liver injury with associated changes in bile acid regulating genes, leading to an accumulation of taurine conjugates in the rat liver. Taurocholic acid (TCA), taurochenodeoxycholic acid (TCDCA) as well as tauro-α-muricholic acid (T-α-MCA) are potential markers for Geniposide-induced hepatic damage.

Topics: Alanine Transaminase; Alkaline Phosphatase; Animals; Aspartate Aminotransferases; ATP Binding Cassette Transporter, Subfamily B, Member 11; Bile Acids and Salts; Biomarkers; Chemical and Drug Induced Liver Injury; Disease Models, Animal; Early Diagnosis; gamma-Glutamyltransferase; Gene Expression Regulation; Iridoids; Male; Rats; Rats, Sprague-Dawley; Receptors, Cytoplasmic and Nuclear

Berberine, baicalin, and jasminoidin were major active ingredients of Huang-Lian-Jie-Du-Decoction (HLJDD), a famous prescription of traditional Chinese medicine (TCM), which has been used for the treatment of ischemic stroke. The aim of the present study was to classify their roles in the treatment effects of ischemic stroke. A rat model of middle cerebral artery occlusion (MCAO) was constructed to mimic ischemic stroke and treatment effects of berberine, baicalin, and jasminoidin, and HLJDD was assessed by neurologic deficit scoring, infarct volume, histopathology, immunohistochemistry, biochemistry, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blotting. In addition, the

Topics: Berberine; Brain Ischemia; Drugs, Chinese Herbal; Flavonoids; Humans; Iridoids; Medicine, Chinese Traditional; Metabolomics; Stroke

Insulin resistance (IR) is known to be an important factor, which can lead to the onset of type 2 diabetes. Autophagy is a cellular process, which sequesters senescent or damaged proteins in autophagosomes for recycling of their products. Insulin and intracellular molecules, including mammalian target of rapamycin (mTOR), are well‑known inhibitors of autophagy. In patients with type 2 diabetes, the expression levels of glucose transporter 4 (GLUT‑4) in skeletal muscles are significantly decreased, indicating decreased glucose‑processing ability. Geniposide is an iridoid compound isolated from Gardenia jasminoides Ellis. Previously, it was reported that geniposide significantly promoted glucose uptake. In the present study, a HepG2 cell model of IR was constructed to determine whether geniposide can promote autophagy to inhibit insulin resistance in HepG2 cells via P62/nuclear factor (NF)‑κB/GLUT‑4. Cell proliferation was analyzed by performing an MTT assay, and the mRNA expression levels of NF‑κB and GLUT‑4 were assessed using semi‑quantitative polymerase chain reaction and immunohistochemical staining. In addition, the protein levels of GLUT‑4, P62 and phosphorylated‑P65 were assessed by western blotting. The expression of GLUT‑4 was initially increased following geniposide treatment, decreasing in time to its lowest level at 8 h. The expression levels of NF‑κB and GLUT‑4 in the IR cells treated with and without geniposide were significantly different, compared with those in the control group. Geniposide promoted autophagy in the IR HepG2 cells and significantly improved IR in the HepG2 cells, which may be associated with the dynamic regulation of the P62/NF‑κB/GLUT‑4 pathway.

Topics: Autophagy; Glucose Transporter Type 4; Hep G2 Cells; Humans; Insulin Resistance; Iridoids; Microscopy, Confocal; Microscopy, Electron, Transmission; Microtubule-Associated Proteins; Models, Biological; NF-kappa B; Sequestosome-1 Protein; Signal Transduction; Sirolimus

Calcium-permeable ionotropic NMDAR-mediated hyperactivity is regarded as the critical factor in modulating the development of ischaemic stroke. Recently, there has been increasing interest in preventing post-stroke neuronal death by focusing on intervening in the function of subpopulations of NMDARs and their downstream signalling. Geniposide, an iridoid glycoside, has been found to have cytoprotective functions in various conditions. However, it is still unclear whether and how geniposide affects neuronal insult under experimental stroke.. We demonstrate that dose-dependent geniposide significantly decreased the infarct volume in tMCAO models.. A medium level of geniposide improved anti-apoptotic functions and inhibited BBB leakage/haemorrhage via elevating GluN2A-containing NMDAR expression in tMCAO rats. Importantly, these effects could be eliminated by co-treatment of geniposide with the GluN2A antagonist NVP but not the GluN2B inhibitor ifenprodil. Moreover, geniposide's protection was due to the enhancement of GluN2A-dependent survival signals, including pAKT, pERK and PSD-95.. The results suggest that geniposide protects neurons against post-ischaemic neurovascular injury through the activation of GluN2A/AKT/ERK pathways. As a very promising natural agent, geniposide may be a future therapeutic for stroke patients.

Topics: Animals; Cell Death; Extracellular Signal-Regulated MAP Kinases; Infarction, Middle Cerebral Artery; Iridoids; Male; MAP Kinase Signaling System; Middle Cerebral Artery; Neurons; Neuroprotective Agents; Proto-Oncogene Proteins c-akt; Rats, Sprague-Dawley; Receptors, N-Methyl-D-Aspartate

Staphylococcus aureus (S. aureus) is a major cause of pneumonia that often affects young and immunocompetent patients. Inflammation plays an important role in the development of S. aureus-induced pneumonia. Geniposide, a major iridoid glucoside component of gardenia fruit, has been reported to have anti-inflammatory and anti-oxidative effects. The purpose of this study was to investigate the protective effects of geniposide on S. aureus-induced pneumonia in mice. Lung histopathological changes were detected by hematoxylin-eosin (H&E) staining. Lung myeloperoxidase (MPO) activity, wet-to-dry (W/D) ratio, and inflammatory cytokine levels in bronchoalveolar lavage fluid (BALF) were measured. The results showed that S. aureus-induced lung histopathological changes were attenuated by geniposide. S. aureus-induced MPO activity and lung W/D ratio were inhibited by treatment of geniposide. Furthermore, the levels of TNF-α and IL-1β in the BALF were also suppressed by geniposide. In addition, geniposide significantly inhibited S. aureus-induced nuclear factor kappa B (NF-κB) activation. Taken together, these results showed that geniposide inhibited S. aureus-induced pneumonia in mice by inhibiting NF-κB signaling pathway. Geniposide might be used as a potential agent for the treatment of S. aureus-induced pneumonia.

Topics: Animals; Anti-Inflammatory Agents; Bronchoalveolar Lavage Fluid; Cytokines; I-kappa B Proteins; Interleukin-1beta; Iridoids; Lung; Male; Mice; Mice, Inbred BALB C; NF-kappa B; Peroxidase; Pneumonia; Signal Transduction; Staphylococcal Infections; Staphylococcus aureus; Tumor Necrosis Factor-alpha

HuanglianJiedu decoction (HJD) is a classic prescription for heat-clearing away and detoxifying, which is used for the clinical treatment of sepsis, due to sepsis refers to the systemic inflammatory response induced by infection in western medicine, and infection belongs to the category of poison-heat syndrome in traditional Chinese medicine.. Previous study had elucidated the effective components from HJD with high affinity to lipid A, which can generate the release of pro-inflammatory-cytokines, resulting in sepsis. Now the anti-sepsis activities of these compounds were evaluated.. Immunofluorescence, immunohistochemical staining, ELISA and MTT methods were used to evaluated these compounds.. Immunofluorescence analysis evaluated the effects of compounds on the binding of FITC-LPS to RAW264.7 cells, and showed the fluorescence intensity was significant attenuated in geniposides, palmatine, baicalin and berberine groups (64 and 128 μg/mL) compared with model group (p < 0.05), which showed these compounds inhibit the combination of LPS with receptor of cells; immunohistochemical staining and ELISA method showed the TLR4 receptor expression, IL-6 and TNF-α levels were significant decreased in the groups treated with compounds, indicating that geniposides, baicalin, palmatine and berberine can play the role of anti-sepsis by inhibiting the expression of TLR4, the releasing of IL-6 and TNF-α; MTT assay showed that palmatine and berberine had a weak effect on cell viability, while others not, indicating that the compounds have protective activity.. It could be concluded the high affinity binding between these compounds and lipid A may be an important basis for its anti-LPS activity in vitro.

Topics: Animals; Berberine; Berberine Alkaloids; Cell Line; Drugs, Chinese Herbal; Flavonoids; Interleukin-6; Iridoids; Lipid A; Lipopolysaccharides; Mice; RAW 264.7 Cells; Sepsis; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

The aim of this study was to investigate the transdermal penetration enhancement effect of wintergreen oil and its action mechanisms. The in vitro transdermal tests were carried out to study the transdermal penetration enhancement effect of wintergreen oil by using osthole and geniposide as the lipophilic and hydriphilic model drugs. Fourier-transform infrared spectroscopy was used to investigate the effect of wintergreen oil on the molecular structure of rat stratum corneum, and the scanning electron microscope was employed to observe the change of rat skin surface after treatment by the oil. The wintergreen oil at proper concentrations could effectively promote the transdermal permeation of osthole and geniposide, and exhibited better penetration-enhancing activity for the lipophilic osthole, close to the commonly used classical penetration enhancer azone. The infrared spectroscopy study and scanning electron microscope showed that wintergreen oil mainly acted on the stratum corneum lipids, reduced dense stratum corneum, and reduced the skin barrier function. Thus, the wintergreen oil could effectively facilitate the transdermal absorption of the lipophilic and hydrophilic drugs, resulting from the lowed skin barrier function.

Topics: Administration, Cutaneous; Animals; Coumarins; Iridoids; Oils, Volatile; Plant Extracts; Rats; Salicylates; Skin; Skin Absorption

Geniposide and genipin have been found in Gardenia jasminoides Ellis, a traditional Chinese medicine that exhibits multiple biological functions. However, no report showing the effects of geniposide and genipin on gastric protection in Helicobacter pylori infections in vitro and in vivo has been done. In this study, we clarified how geniposide and genipin suppress H. pylori-mediated inflammation in gastric AGS cells and C57BL/6 mice. Our results demonstrated that genipin shows a strong ability to inhibit H. pylori growth and is able to reduce vacA and cagA gene expression of H. pylori in infected AGS cells. Genipin also attenuates the abilities of adhesion and invasion of H. pylori to AGS cells. An attenuation of interleukin (IL)-8 and IFN-γ production caused by genipin was observed to inhibit cell inflammatory responses. In the in vivo experiments, geniposide and genipin both showed suppressive effects on the vacA gene expression in mice after H. pylori infection. The serum levels of IFN-γ, IL-1β, immunoglobulin A, and Immunoglobulin M were decreased by geniposide and genipin in infected mice. The inflammatory maker COX2 was downregulated in H. pylori-infected mice after exposure to geniposide and genipin. Together, geniposide and genipin effectively exert a healthy promotion to reduce H. pylori infections in vivo by interfering with the growth and virulence of H. pylori as well as attenuating the gastric inflammation caused by an H. pylori infection.. Geniposide and genipin have a healthy promotion to eradicate H. pylori infections by interfering with the growth and virulence of H. pylori and to attenuate the gastric inflammation caused by an H. pylori infection.

Topics: Animals; Drugs, Chinese Herbal; Gardenia; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation Mediators; Iridoids; Male; Mice; Mice, Inbred C57BL

The search for compounds with functional properties from natural sources has grown in recent years as people have developed healthier habits. Therefore, the aim of this study was to evaluate the extraction of bioactive compounds from various parts of unripe genipap fruit (Genipa americana L.) by using pressurized ethanol to verify which part of the fruit provides the greatest recovery of the iridoids genipin and geniposide. Two process variables were studied: temperature (50 and 80°C) and pressure (2, 12 and 20 bar). The whole fruit and the peel, mesocarp, endocarp, endocarp+seeds and seeds of the fruit were studied. The endocarp presented with the highest recovery of genipin (48.6±0.6mg/g raw material) and the extraction from the mesocarp allowed a greater recovery of geniposide (59±1mg/g raw material). The highest values of total phenolic content were obtained with mesocarp extracts. The endocarp and mesocarp extracts presented the highest antioxidant activity as measured by FRAP and DPPH. These results are promising and support the use of unripe genipap fruit as a source of iridoids and natural antioxidants.

Topics: Antioxidants; Ethanol; Food Handling; Fruit; Iridoids; Phenols; Plant Extracts; Rubiaceae; Seeds

Neurodegenerative diseases are becoming prevalent as the population ages. Geniposide could inhibit oxidative stress, reduce apoptosis, protect neuron, and has been used for therapy of the neurodegenerative diseases. The bioavailability of geniposide by nasal route is greater than that by oral administration. However, mucociliary clearance is a rate-limiting factor for nasal route administration. The objective of this study was to develop and evaluate a mucoadhesive, thermoreversible in situ nasal gel of geniposide. The poloxamers (P407, P188) and the hydroxypropyl methylcellulose were used as thermoreversible and mucoadhesive polymers, respectively. Borneol was used as a permeation enhancer. The hydrogel was prepared with the cold method and optimized by the response surface methodology-central composite design. Gelation temperature, pH, clarity, gel strength, mucoadhesive strength, in vitro and ex vivo release kinetics of formulations were evaluated. The optimized amounts of poloxamer407 (P407), poloxamer188 (P188) and hydroxypropyl methylcellulose were determined to be 19.4-20.5%, 1.1-4.0% and 0.3-0.6% respectively. The second-order polynomial equation in terms of actual factors indicated a satisfactory correlation between the independent variables and the response (R2 = 0.9760). An ANOVA of the empirical second-order polynomial model indicated the model was significant (P<0.01). P407, P188, P407×P188, P4072 and P1882 were significant model terms. The effects of P407 on gelation temperature were greater than those of other independent variables. The pH values of all the formulations were found to be within 6.3-6.5 which was in the nasal physiological pH range 4.5-6.5. The drug content, gel strength, mucoadhesive strength of the optimized formulations were 97-101%, 25-50 sec and 4000-6000 dyn/cm2 respectively. The in vitro release kinetics of cumulative release of geniposide was fitted to the zero-order model. The ex vivo cumulative release kinetics of geniposide was fitted to the Weibull model. This study concludes that the release of geniposide is controlled by gel corrosion, and that the permeation of geniposide is time-dependent. The more residence time, mucoadhesive, thermoreversible in situ nasal gel of geniposide for neurodegenerative diseases is of compliance and potential application.

Topics: Administration, Intranasal; Chromatography, High Pressure Liquid; Gels; Humans; Iridoids; Nasal Mucosa; Neurodegenerative Diseases; Temperature

Herbal extracts have been extensively used worldwide for their application on memory improvement, especially among aged and memory-deficit populations. In the present study, the memory loss induced by human Abeta protein over-expression in fruitfly Alzheimer's disease (AD) model was rescued by multiple extracts from Gardenia jasminoides. Three extracts that rich with gardenia yellow, geniposide, and gardenoside components showed distinct rescue effect on memory loss. Further investigation on adding gardenoside into a formula of Ganoderma lucidum, Panax notoginseng and Panax ginseng (GPP) also support its therapeutic effects on memory improvement. Interestingly, the application of GPP and gardenoside did not alter the accumulation of Abeta proteins but suppressed the expression of immune-related genes in the brain. These results revealed the importance and relevancy of anti-inflammation process and the underlying mechanisms on rescuing memory deficits, suggesting the potential therapeutic use of the improved GPP formulation in improving cognition in defined population in the future.

Topics: Alzheimer Disease; Animals; Antimicrobial Cationic Peptides; Brain; Cognition; Disease Models, Animal; Drosophila; Drosophila Proteins; Gardenia; Gene Expression Regulation; Immunity, Innate; Iridoids; Plant Extracts; Polymerase Chain Reaction

Renal pathology was a commonly seen complication in patients with diabetes. Geniposide (GPO) was previously demonstrated to modulate glucose metabolism in diabetes. This study was to investigate effects of GPO in streptozotocin-induced diabetic rats and its underlying mechanism. Renal function in diabetic rats was evaluated by levels of serum creatinine (Scr), blood urea nitrogen (BUN), and urinary albumin. Renal inflammation was appraised by inflammatory cells infiltration and pro-inflammatory cytokines production. Renal monocytes, T lymphocytes infiltration, and intercellular adhesion molecule-1 (ICAM-1) expression were quantitated by immunohistochemistry. Moreover, renal nuclear factor-kappa B (NF-κB) was assayed by Western blotting. Diabetic rats showed renal dysfunction as evidenced by increased levels of Scr, BUN, urinary albumin, and elevator renal index. Histological examination revealed significant glomerular basement membrane (GBM) thickening. However, GPO notably improved renal function and diabetes-induced GBM changes. Additionally, diabetic rats showed noteworthy renal inflammation,as reflected by enhancement of monocytes and T lymphocytes infiltration, increased expression of ICAM-1, tumor necrosis factor-α, interleukin-1 (IL-1), and IL-6. Interestingly, the level of monocytes infiltration positively correlated with the severity of GBM. Further study indicated diabetic rats displayed increased activation of NF-κB, indicated by increased expression of NF-κB p65, IKKα, and p-IκBα in renal tissue. However, all the changes in renal inflammation and NF-κB pathway were obviously reversed in GPO-treated diabetic rats. Our works indicate GPO ameliorates structural and functional abnormalities of kidney in diabetic rats, which is associated with its suppression of NF-κB-mediated inflammatory response.

Topics: Animals; Blotting, Western; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Dose-Response Relationship, Drug; Hypoglycemic Agents; Iridoids; Kidney; Kidney Function Tests; Male; Metformin; NF-kappa B; Rats, Sprague-Dawley; Signal Transduction; Streptozocin

Huang-Lian-Jie-Du-Tang (HLJDT) is a classical recipe for relieving fever and toxicity for thousands of years in China. Geniposide is one of the main components in HLJDT. The present study was conducted in order to investigate the differences of absorption of geniposide after oral administration of geniposide alone and HLJDT in rats. Pharmacokinetic differences of geniposide following oral administrations of pure geniposide and HLJDT were investigated in vivo. The absorption of geniposide in pure compound and HLJDT was evaluated using intestinal perfusion and Caco-2 models. The in vivo and in vitro studies showed good relevance and consistent results. The co-occurring components in HLJDT were found to promote the absorption of geniposide from the pharmacokinetic study in vivo, intestinal perfusion, and Caco-2 model. Geniposide had better absorption in the duodenum and jejunum from the intestinal perfusion model, which was mainly absorbed by passive diffusion. Verapamil influenced the transportation of geniposide, while EDTA did not, demonstrating that geniposide might be the potential substance of P-glycoprotein in intestinal perfusion and Caco-2 models. The absorption of geniposide was studied systematically to guide the design of the oral dosage of geniposide and HLJDT in clinical therapy.

Topics: Administration, Oral; Animals; Caco-2 Cells; Drugs, Chinese Herbal; Humans; Intestinal Absorption; Iridoids; Male; Rats; Rats, Sprague-Dawley

Our previous work showed that geniposide affected glucose-stimulated insulin secretion (GSIS) via regulating glucose uptake and metabolism in pancreatic β cells; however, the molecular mechanisms remain largely unknown. Substantial evidence suggests that activation of 5'-AMP-activated protein kinase (AMPK) plays a central role in GSIS. Here, we aim to determine the role of AMPK on geniposide-regulated GSIS in rat pancreatic INS-1 cells. The results demonstrated that 6-[4-(2-piperidin-1-yletoxy)-phenyl]-3-pyridin-4-yl-pyrazolo[1,5-α] pyrimidine (Compound C; an AMPK inhibitor) significantly attenuated the effects of geniposide on glucose uptake, energy metabolism, and insulin secretion in INS-1 cells. We also observed that geniposide induced phosphorylation of acetyl-CoA carboxylase (ACC), a marker of AMPK activity, in a time-dependent manner in INS-1 cells; however, in the presence of Compound C, the influence of geniposide on ACC phosphorylation was obviously inhibited. Furthermore, the knockdown of AMPK protein with AMPK siRNA treatment decreased the effects of geniposide on glucose uptake, adenosine triphosphate production, and GSIS. All these data indicate that AMPK plays an essential role in geniposide-regulated GSIS in pancreatic β cells.

Topics: AMP-Activated Protein Kinases; Animals; Glucose; Insulin; Insulin-Secreting Cells; Iridoids; Rats; Transfection

In this paper, by coupling reversed phase liquid chromatography (RPLC) and hydrophilic interaction liquid chromatography (HILIC), a two-dimensional liquid chromatography system was developed for separation and identification of the active ingredients in Gardenia jasminoides Ellis (GJE). By applying the semi-preparative C18 column as the first dimension and the core-shell column as the second dimension, a total of 896 peaks of GJE were separated. Among the 896 peaks, 16 active ingredients including geniposide, gardenoside, gardoside, etc. were identified by mass spectrometry analysis. The results indicated that the proposed two-dimensional RPLC/HILIC system was an effective method for the analysis of GJE and might hold a high potential to become a useful tool for analysis of other complex mixtures.

Topics: Chromatography, High Pressure Liquid; Chromatography, Reverse-Phase; Gardenia; Hydrophobic and Hydrophilic Interactions; Iridoids; Plant Extracts

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by memory deficits and cognitive decline. Amyloid-β (Aβ) deposition and cholinergic defect are widely thought to be the underlying mechanism of learning and memory impairment. Geniposide, which is the main active component of the traditional Chinese herbal Gardenia jasminoides Ellis, elicits neuroprotective effects by alleviating inflammation responses and oxidative damages. In this study, we investigated the protective effect of geniposide on levels of cholinergic markers, RAGE, RAGE-dependent signalling pathways and amyloid accumulation in the APPswe/PS1dE9 AD model mouse. Geniposide suppressed MAPK signaling over-activation mediated by Aβ-RAGE interaction, resulting in reduced Aβ accumulation and amelioration of cholinergic deficits in the cerebral hippocampus. Furthermore, geniposide inhibited the toxic effect of oligomeric Aβ

Topics: Alzheimer Disease; Amyloid beta-Peptides; Amyloidosis; Animals; Brain; Cognition Disorders; Disease Models, Animal; Dose-Response Relationship, Drug; Iridoids; Male; MAP Kinase Signaling System; Memory Disorders; Mice, Inbred C57BL; Mice, Transgenic; Neurons; Neuroprotective Agents; Nootropic Agents; Peptide Fragments; Receptor for Advanced Glycation End Products

Gardenia jasminoides Ellis is a traditional Chinese medicine (TCM) that containing a variety of effective active ingredients and exhibits diverse pharmacological functions, such as anti-inflammatory, antioxidant and nerve protection.. This study investigated the effect of Gardenia jasminoides extract (GJE) and Geniposide on learning and memory improvement and neuroprotection in a rat model with chronic cerebral ischemia, as well as explore the underlying mechanisms.. The crude GJE was prepared using the methods of water extraction and alcohol precipitation, and refined by macroporous adsorption resin. The chronic cerebral ischemia model was simulated by permanent occlusion of bilateral common carotid arteries in rats. GJE was taken at three doses groups (150mg/kg, 100mg/kg, 50mg/kg), Geniposide group (50mg/kg), and oral administration for 30 days. Memory function was assessed using Morris water maze test. The morphological changes of hippocampus and related parts of brain in rats by Hematoxylin and Eosin (HE) staining were observed. Moreover, the levels of Acetylcholin Esterase (AchE), Nitric Oxide Synthase (NOS), Malondialdehyde (MDA), Superoxide Dismutase (SOD) in the brain tissue were quantified.. GJE contained 27% gardenoside and 72% total iridoid glycoside. The chronic cerebral ischemia rat model has been proved successfully. The memory function of the rats assessed using Morris water maze test showed that GJE significantly shortened the escape latency of rats, but had no significant improvement on the number of times crossing the platform and the percentage of time spent in the target quadrant. HE staining showed that the apoptosis and necrosis of the cortex and hippocampus in the GJE group were significantly reduced. In addition, it was found that GJE could significantly improved the content of SOD, inhibited NOS and AchE activity in brain tissue, but did not show a significant reduction in the content of MDA. The effect of medium dosage of GJE was the best among these three dose groups and also better than Geniposide according to the results of all the detection index.. GJE had the functions of learning and memory improvement and the neuroprotection on chronic cerebral ischemia model rats. The mechanisms were found to be strongly correlated with antioxygen free radical, reduction of NO toxicity and AChE activity, and brain neuron protective effect. GJE could be able to play a better effect on improving chronic cerebral ischemia than Geniposide.

Topics: Acetylcholinesterase; Animals; Brain; Brain Ischemia; Fruit; Gardenia; Iridoids; Learning; Male; Malondialdehyde; Neuroprotection; Neuroprotective Agents; Nitric Oxide Synthase; Phytotherapy; Plant Extracts; Rats, Sprague-Dawley; Superoxide Dismutase

Geniposide (GE) is one of the major iridoid glycosides isolated from the fruit of Gardenia jasminoides Ellis that has been used to treat hepatic disorders including cholestasis. However, the underlying mechanisms for GE ameliorating the reduction in bile acids accumulation by α-naphthylisothiocyanate (ANIT) remain unclear.. The purpose of this study is to characterize the efficacy of GE in regulation of bile acids uptake, synthesis, metabolism, and transport in ANIT-induced rats.. Sprague-Dawley rats were orally administrated with vehicle, GE (25, 50, and 100mg/kg), and ursodeoxycholic acid (UDCA) (60mg/kg) once daily for seven days. On the fifth day, a single dose of ANIT (75mg/kg) was administrated via oral gavage. Blood biochemical determination, bile flow rate and liver histopathology were measured to evaluate the protective effect of GE. The mRNA expressions and protein levels of transporters and enzymes involved in bile acids homeostasis were determined by quantitative real-time polymerase chain reaction (PCR) and western blot to study the underlying mechanism of GE against ANIT-induced rats.. GE (25, 50, and 100mg/kg, po) dose-dependently prevented ANIT-induced changes in serum markers for liver injury. GE treatment reduced basolateral bile acids uptake via repression of OATP2 (P<0.05). Bile acids biosynthesis was decreased through down-regulation of CYP7A1, CYP8B1, and CYP27A1 (P<0.05). GE significantly increased canalicular bile acids secretion via BSEP (P<0.05), subsequently stimulating bile flow during cholestasis. GE also markedly enhanced mRNA level of basolateral transporter OSTβ (P<0.01). Bile acids transported to the plasma were cleared into the urine, resulting in down-regulation of plasma bile acids. However, GE did not alter the mRNA levels of CYP3A2, UGT1A1 and SULT2A1. Furthermore, the gene and protein expression analysis demonstrated activation of FXR, PXR, and SHP after GE administration.. GE attenuates ANIT-induced hepatotoxicity and cholestasis in rats, due to regulation enzymes and transporters responsible for bile acids homeostasis.

Topics: 1-Naphthylisothiocyanate; Animals; ATP Binding Cassette Transporter, Subfamily B, Member 11; ATP-Binding Cassette Transporters; Bile Acids and Salts; Cholestasis; Cytochrome P-450 Enzyme System; Down-Regulation; Glucuronosyltransferase; Homeostasis; Iridoids; Liver; Organic Anion Transporters; Pregnane X Receptor; Protective Agents; Rats, Sprague-Dawley; Receptors, Cytoplasmic and Nuclear; Receptors, Steroid; Sulfotransferases

Genipin-1-o-glucuronic acid and genipin-monosulfate are two major metabolites from geniposide and genipin. Based on diabetic rat model, we developed a simultaneous quantification method to investigate their comparative pharmacokinetics by online mircrodialysis-ultra performance liquid chromatography-mass spectrometry (MD-UPLC-MS/MS) without their standard compounds. Online microdialysis sampling could avoid unexpected contamination or degradation of the analytes during the storage and transfer steps. Combined with good sensitivity, selectivity and selectivity of UPLC-MS/MS, online MD-UPLC-MS/MS method could real-timely monitor metabolites in rat blood for quantitative analysis. Our research found that AUC

Topics: Animals; Chromatography, High Pressure Liquid; Diabetes Mellitus, Experimental; Iridoids; Linear Models; Male; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Sensitivity and Specificity; Tandem Mass Spectrometry

To analyze the role of geniposide in the protein degradation of Txnip and to determine the impact of Txnip on geniposide-regulated GSIS in pancreatic INS-1 cells.. The content of Txnip protein was measured by western blot; insulin content and glucose uptake were determined by ELISA; and knockdown of Txnip was the method of RNA interference.. Glucose induces a rapid increase in Txnip protein, and geniposide accelerates the degradation of Txnip via proteasome pathway in the presence of high glucose (25 mM) in INS-1 pancreatic β-cells. And MG132, a proteasomal inhibitor, potentiates glucose uptake, metabolism (ATP production) and glucose-stimulated insulin secretion (GSIS) in high-glucose (25 mM)-treated INS-1 cells, but geniposide significantly prevents these effects. Furthermore, the combination of geniposide and Txnip knockdown shows substantial synergistic effects to reduce glucose uptake, metabolism and GSIS in high-glucose (25 mM)-treated INS-1 cells.. Txnip protein played an essential role in glucose uptake, metabolism and GSIS, and geniposide could accelerate the degradation via proteasome pathway in high-glucose-treated pancreatic INS-1 cells.

Topics: Animals; Carrier Proteins; Cell Cycle Proteins; Gene Expression Regulation; Glucose; Insulin; Insulin Secretion; Insulin-Secreting Cells; Insulinoma; Iridoids; Pancreatic Neoplasms; Proteasome Endopeptidase Complex; Proteolysis; Rats; Tumor Cells, Cultured

Geniposide (GE), an iridoid glycoside compound purified from Gardenia jasminoides Ellis, has antiinflammatory and other pharmacological effects, but its mechanism of actions on rheumatoid arthritis (RA) have not been clarified. The purpose of this article was to investigate the pharmacological effects of GE on collagen-induced arthritis (CIA) rats and its feasible mechanisms. Collagen-induced arthritis was induced by injection of chicken type II collagen emulsion. The rats were orally administered with GE (33, 66, and 132 mg/kg) from days 14 to 30 after immunization. The histological examination showed that GE could attenuate histopathologic changes of mesenteric lymph node (MLN) in CIA rats. Geniposide inhibited the production of Interleukin 6 (IL-6) and IL-17, while promoting the production of IL-4 and transforming growth factor-beta 1 in MLN lymphocytes (MLNLs). Moreover, the proliferation capability of MLNLs was increased after the administration of GE. In addition, the treatment with GE in vivo decreased the expressions of P-Raf, P-MEK, and P-Erk1/2 in MLNLs. These results may highlight the antiinflammatory effects and possible mechanisms of GE in MLNLs of RA. Copyright © 2017 John Wiley & Sons, Ltd.

Topics: Animals; Anti-Inflammatory Agents; Arthritis, Experimental; Iridoids; Male; Rats; Rats, Wistar

Four new iridoids, 2'-O-(E)-coumaroylshanzhiside (1), 6'-O-(E)-coumaroylshanzhiside (2), 8α-butylgardenoside B (3), 6α-methoxygenipin (4), and one new phenylpropanoid glucoside, 5-(3-hydroxypropyl)-2-methoxyphenyl β-d-glucopyranoside (5), together with sixteen known compounds, were isolated from the edible flowers of wild Gardenia jasminoides J.Ellis. Their chemical structures were characterized by extensive spectroscopic techniques, including 1D- and 2D-NMR, HR-ESI-MS, and CD experiments. The absolute configurations of the new isolates' sugar moiety were assigned by HPLC analysis of the acid hydrolysates. Furthermore, the antioxidant activities of those isolates were preliminarily evaluated by DPPH scavenging experiment. And comparison of

Topics: Antioxidants; Flowers; Fruit; Gardenia; Iridoids; Molecular Structure; Plant Extracts; Spectrum Analysis

Our previous works indicated that geniposide could regulate glucose-stimulated insulin secretion (GSIS), and improved chronic high glucose-induced dysfunctions in pancreatic β cells, but the molecular mechanisms remain largely unknown. In the present study, we investigated the role of 5'-AMP-activated protein kinase (AMPK) in high glucose induced cell injury and explored the associated molecular mechanisms in rat INS-1 pancreatic β cells. Data suggested that geniposide obviously prevented the cell damage induced by high (25 mM) glucose in INS-1 cells, which increased the protein levels of cell apoptosis-associated enzymes, including heme oxygenase-1 (HO-1), and Bcl-2, but apparently attenuated the protein level of Bax, an apoptotic protein. In addition, Compound C, an AMPK inhibitor, remarkably inhibited the effects of geniposide on the protein levels of HO-1, Bcl-2, and Bax, but AICAR, an AMPK activator, potentiated the role of geniposide on the protein levels of HO-1, Bcl-2, and Bax. More importantly, geniposide directly prevented the cleavage of caspase-3 induced by high glucose, and this effect was also evidently prohibited by the pre-incubation of compound C in high glucose-treated INS-1 cells. Furthermore, using the method of RNA interfere, we further proved that treatment with AMPK siRNA attenuated the effects of geniposide on the apoptosis-associated proteins and cell viability. All these data suggest that AMPK plays a crucial role on geniposide antagonizing high glucose-induced pancreatic β cells injury.

Topics: AMP-Activated Protein Kinases; Animals; Apoptosis; bcl-2-Associated X Protein; Caspase 3; Cell Line; Cell Survival; Cytoprotection; Enzyme Activation; Glucose; Heme Oxygenase-1; Insulin-Secreting Cells; Iridoids; Phosphorylation; Protective Agents; Rats; RNA, Small Interfering

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Body Weight; Caco-2 Cells; Colitis, Ulcerative; Cytokines; Down-Regulation; Humans; Intestinal Mucosa; Iridoids; Male; Neutrophil Infiltration; Permeability; Rats, Sprague-Dawley; Sulfasalazine; Trinitrobenzenesulfonic Acid; Up-Regulation

Geniposide is a key iridoid glycoside from Gardenia jasminoides fructus widely used in traditional Chinese herbal medicine. However, detection of this small molecule represents a significant challenge mostly due to the lack of specific molecular recognition elements. In this study, we have performed in vitro selection experiments to isolate DNA aptamers that can specifically bind geniposide. Using a stringent selection procedure, we have isolated DNA aptamers that can distinguish geniposide from genipin and glucose, two structural analogs of geniposide. Two top aptamers exhibit low micromolar binding affinity towards geniposide, but show significantly reduced affinity to genipin and glucose. These aptamers have the potential to be further developed into analytical tools for the detection of geniposide.

Topics: Aptamers, Nucleotide; Gardenia; Glucose; Iridoids; Medicine, Chinese Traditional; Plant Extracts; SELEX Aptamer Technique

Influenza A viruses (IAVs) have been a great threat to human health for centuries, without effective control. Geniposide, a main iridoid glycoside compound extracted from Gardenia jasminoides Ellis fruit, possesses various biological activities including anti-inflammation and anti-virus.. Madin-Darby canine kidney (MDCK) cells were infected with pandemic A/Jiangsu/1/2009 (H1N1) influenza virus in vitro. Cytotoxicity and antiviral activity of geniposide were estimated by MTT assay. The influenza respiratory tract infection murine model was established by intranasal instillation of pandemic A/Jiangsu/1/2009 (H1N1) influenza virus. One day after infection, the mice were administered with geniposide (5, 10 or 20 mg/kg/day) or the neuraminidase inhibitor (NAI) peramivir (30 mg/kg/day). Body weight, survival time, viral titre and lung index of the mice were measured. The sandwich enzyme-linked immunosorbent assay (ELISA) was used to examine levels of inflammatory cytokines.. The data showed that geniposide had little cytotoxicity on MDCK cells and protected them from pandemic A/Jiangsu/1/2009 (H1N1) influenza virus-induced cell injury. In the infected mice, geniposide treatment significantly restored the body weights, decreased the mortality, alleviated viral titres and virus-induced lung lesions. Geniposide substantially inhibited the virus-induced alveolar wall changes, alveolar haemorrhage and neutrophil-infiltration in lung tissues. Levels of inflammatory mediators, including tumour necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin (IL)-4, IL-6 and IL-10 were also markedly altered after treatment with geniposide.. Our investigation suggested that geniposide effectively inhibited cell damage mediated by pandemic A/Jiangsu/1/2009 (H1N1) influenza virus and mitigated virus-induced acute inflammation.

Topics: Animals; Anti-Inflammatory Agents; Antiviral Agents; Cell Line; China; Cytokines; Dogs; Female; Humans; Inflammation Mediators; Influenza A Virus, H1N1 Subtype; Influenza, Human; Iridoids; Lung; Mice; Microbial Sensitivity Tests; Severity of Illness Index

To establish a fast detection method during the purifying process of the extracts from Grardeniae using macroporous resin based on near infrared spectroscopy. First, the ethanol eluent was collected from the purification process of small size sample; and near infrared (NIR) spectrum was collected. Then the content of the geniposide was determined by HPLC method, and partial least squares (PLS) method was used to establish the quantitative model to predict the content of geniposide by NIR spectrum. This model was used to supervise the changes of geniposide concentrations in ethanol eluent during medium scale process. Experimental results showed that the NIR small scale model can accurately predict the concentrations of geniposide in the production process of medium scale. However, with the proceeding of batch processes, the prediction performance of the model was decreased, so model updating method was employed to maintain the model. After twice updates, the NIR quantitative model can accurately predict the concentrations of the geniposide during medium scale process. Therefore, through model updates, the established NIR quantitative model can be applied in different scales of macroporous resin purification processes, to improve the data utilization efficiency of small scale process and save the cost of rebuilding the quantitative model of medium scale.

Topics: Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Iridoids; Porosity; Resins, Synthetic; Rubiaceae; Spectroscopy, Near-Infrared

To observe the effect of geniposide on non-alcoholic fatty liver disease (NAFLD), and discuss the mechanism of geniposide for NAFLD from the aspect of free fatty acid, forty healthy Wistar male rats were randomly divided into normal group, model group, geniposide and Xuezhikang group. The rats in normal group were fed with normal diets, and the rats in other 3 groups were given with high-fat diet for 8 weeks to induce the NAFLD models. From the week 5 to end of week 8, the rats in geniposide and Xuezhikang group were intervened with corresponding medicines. The body weight, liver wet weight, and fat weight of the rats were recorded. Visual and pathological changes in hepatic tissues were observed with HE staining. The contents of TG, FFA, FAS, AMPK, ACCase and Malonyl-CoA in hepatic tissue, contents of CHO and LDL-C in serum and activities of AST and ALT in serum were detected by using corresponding methods. The results showed that the body weight, liver wet weight, and fat weight of the rats, CHO, LDL-C, ALT and AST levels in serum, TG, FFA, FAS, ACCase and Malonyl-CoA levels in hepatic tissues of the rats in model group were significantly higher than those in normal group (P<0.01), while AMPK activity was significantly lower than that of the normal group (P<0.01), with obvious visual and pathological steatosis in hepatic tissues, and inflammatory injury occurred in model group. Compared with the model group, body weight of the rat, fat weight, levels of FFA in hepatic tissues, ALT and AST activities in serum, liver wet weight, TG, FAS, ACCase and Malonyl-CoA levels were significantly decreased in geniposide group (P<0.01), while the AMPK activity in hepatic tissues was significantly increased (P<0.05),with improvement in visual and pathological performance. Compared with the model group, liver wet weight, fat weight, TG and FFA levels in hepatic tissues, and LDL-C level in serum were significantly decreased in Xuezhikang group (P<0.05). Compared with Xuezhikang group, the body weight of rat, fat weight and FFA level in hepatic tissues were significantly lower in geniposide group (P<0.01), but with no significant difference in other aspects. These findings indicated that geniposide was highly effective in improving the pharmacological effect of NAFLD induced by high-fat diet, and the mechanism was achieved through AMPK-ACCase-Malonyl-CoA-FFA axis.

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; Drugs, Chinese Herbal; Fatty Acids, Nonesterified; Humans; Iridoids; Liver; Male; Non-alcoholic Fatty Liver Disease; Rats; Rats, Wistar; Triglycerides

To optimize the matrix formulation of Chaizhi cataplasma (CC) and investigate its release and transdermal absorption properties in vitro. The optimized matrix formulation of cataplasma containing liquid herbal extract is determined by using D-optimal mixture design, with initial bonding strength, endurance bonding strength and gel strength as the evaluating indicators. Modified Franz diffusion cells were used to study the in vitro release and transdermal absorption of geniposide in CC. The optimized matrix formulation of CC contained NP700, aluminum glycinate, tartaric acid, glycerin, PVPK90 and water (9∶0.7∶0.8∶30∶5∶30.5). Cumulative release rate of geniposide in CC was (77.02±3.73)% in 24 h. The percutaneous penetration rate of geniposide was 7.25 μg•cm⁻²•h⁻¹ and the 24 h permeated amount was (156.22±4.90) μg•cm⁻². The optimized CC prepared by the D-optimal mixture design showed a good adhesion and formability. The in vitro release of the geniposide in CC was in accordance with the first order equation, while its in vitro transdermal absorption was close to the zero order equation.

Topics: Administration, Cutaneous; Animals; Chemistry, Pharmaceutical; Drugs, Chinese Herbal; Iridoids; Mice; Mice, Inbred ICR; Skin; Skin Absorption

Chinese herbal medicinal formulation Yinzhihuang injection is widely used in clinic for jaundice and chronic liver diseases in eastern Asian countries. However, the pharmacologically active components and the underlying mechanism are unclear. In this study, 30 male ICR mice were randomly assigned into 6 groups： normal control, model control, chlorogenic acid group, geniposide group, baicalin group and wogonoside group. The liver function, liver pathological changes and bile acid metabolism-related gene expression in mice were assayed. The serum levels of ALT, AST, ALP, TBA in chlorogenic acid group (15.89±2.53), (18.32±2.56), (26.38±9.87) U•L⁻¹, (40.63±7.67) μmol•L⁻¹, respectively and geniposide group (20.54±2.36), (24.28±5.19), (35.09±5.03) U•L⁻¹, (42.86±7.11) μmol•L⁻¹, respectively were lower than those in the model control group (59.52±10.94), (128.37±17.97),(169.52±9.62) U•L⁻¹, (132.50±33.00) μmol•L⁻¹, respectively. Hematoxylin-eosin staining showed necrosis, infiltration of inflammation cell in chlorogenic acid group and geniposide group were milder than those in the model control group. Q-PCR analysis revealed the expression of bile acid metabolism-related genes was normalized after treatment with chlorogenic acid or geniposide. Chlorogenic acid and geniposide improved the liver injury and cholestasis effectively, and reversed the mRNA expression of bile acid metabolism-related genes induced by ANIT. So, chlorogenic acid and geniposide were protective for cholestasis, suggesting their pharmacodynamic effect in Yinzhihuang injection.

Topics: Alanine Transaminase; Animals; Chlorogenic Acid; Cholestasis; Cholesterol 7-alpha-Hydroxylase; Cytochrome P450 Family 8; Drugs, Chinese Herbal; Flavonoids; Humans; Iridoids; Liver; Male; Mice; Mice, Inbred ICR

To study the chemical constituents of water extracted fraction from Oldenlandia diffusa.. The compounds were isolated and purified by column chromatography on macroporous resin,silica gel,MCI gel,Sephadex LH-20,ODS medium pressure liquid chromatography and RP-semi-preparative HPLC. The structures of compounds were elucidated on the basis of physicochemical and spectral analysis.. 16 compounds were isolated from the water extract of Oldenlandia diffusa,and their structures were identified as asperuloside( 1),deacetyl asperuloside( 2),geniposide( 3),10-dehydrogeniposide( 4),daphylloside( 5),diffusoside A( 6),diffusoside B( 7),coniferin( 8),scandoside methyl ester( 9),acetyl scandoside methyl ester( 10),deacetylasperulosidic acid methyl ester( 11),gardenoside( 12),galioside( 13),galioside 10-acetate( 14),loliolide( 15) and( +)-neo-olivil( 16),respectively.. Compounds 3,8 and 14 ~ 16 are obtained from Oldenlandia diffusa for the first time.

Topics: Chromatography, High Pressure Liquid; Cyclopentane Monoterpenes; Glucosides; Iridoids; Lignans; Oldenlandia; Pyrans

Using a system pharmacology strategy, this study evaluated the unique pharmacological characteristics of three different neuroprotective compounds for the treatment of cerebral ischemia-reperfusion. A microarray including 374 brain ischemia-related genes was used to identify the differentially expressed genes among five treatment groups: baicalin, jasminoidin, ursodeoxycholic acid, sham, and vehicle, and MetaCore analysis software was applied to identify the significantly altered pathways, processes and interaction network parameters. At pathway level, 46, 25, and 31 pathways were activated in the baicalin, jasminoidin, and ursodeoxycholic acid groups, respectively. Thirteen pathways mainly related with apoptosis and development were commonly altered in the three groups. Additionally, baicalin also targeted pathways related with development, neurophysiologic process and cytoskeleton remodeling, while jasminoidin targeted pathways related with cell cycle and ursodeoxycholic acid targeted those related with apoptosis and development. At process level, three processes were commonly regulated by the three groups in the top 10 processes. Further interaction network analysis revealed that baicalin, jasminoidin, and ursodeoxycholic acid displayed unique features either on network topological parameters or network structure. Additional overlapping analysis demonstrated that compared with ursodeoxycholic acid, the pharmacological mechanism of baicalin was more similar with that of jasminoidin in treating brain ischemia. The data presented in this study may contribute toward the understanding of the common and differential pharmacological mechanisms of these three compounds.

Topics: Animals; Flavonoids; Gene Expression Profiling; Gene Regulatory Networks; Iridoids; Ischemia; Male; Metabolic Networks and Pathways; Mice; Microarray Analysis; Middle Cerebral Artery; Neuroprotective Agents; Reperfusion Injury; Ursodeoxycholic Acid

Stimulating fibroblast-like synoviocyte (FLS) apoptosis in rheumatoid arthritis (RA) is a promising strategy for clinical treatment. Previous studies have confirmed that geniposide shows a certain anti-arthritic effect in vivo. However, whether geniposide can induce RA FLS apoptosis and the underlying mechanisms has not been elucidated. Herein, adjuvant-induced arthritis (AIA) in rat was induced and FLS was isolated from synovial tissues by tissue explant cultivation method. MTT assay, Hoechst staining, and flow cytometric apoptosis assay were applied to evaluate apoptotic effect of geniposide on AIA FLS. Bcl-2, Bax, and caspase 3 messenger RNA (mRNA) levels, and extracellular-signal-regulated kinases (ERKs) and phosphorylated ERK protein levels were examined by real-time PCR and western blot, respectively. We found that geniposide dose-dependently inhibited AIA FLS proliferation in vitro. AIA FLS treated with geniposide displayed typical apoptotic morphological characteristics including nuclear shrinkage and chromatin condensation. Flow cytometric apoptosis assay indicated that geniposide significantly increased the apoptosis rate of AIA FLS. Additionally, geniposide treatment on AIA FLS decreased Bcl-2 mRNA level and increased Bax and caspase 3 mRNA levels, accompanied by reduced protein levels of phosphorylated-ERK1/2, without affecting total ERK1/2. In conclusion, geniposide effectively induces AIA FLS apoptosis through regulating the apoptosis-related gene expressions and inhibiting ERK signal pathway.

Topics: Animals; Apoptosis; Arthritis, Experimental; Arthritis, Rheumatoid; bcl-2-Associated X Protein; Caspase 3; Disease Models, Animal; Extracellular Signal-Regulated MAP Kinases; Fibroblasts; Flow Cytometry; Iridoids; Male; Phosphorylation; Proto-Oncogene Proteins c-bcl-2; Rats; Rats, Sprague-Dawley; RNA, Messenger; Synovial Membrane

1. Geniposide (genipin 1-O-glucose), one of the major bioactive constituents isolated from Fructus Gardeniae, possesses many biological activities. In this study, an efficient strategy was developed using ultra-high-performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometer (UPLC-LTQ-Orbitrap) to profile the in vitro and in vivo metabolic patterns of geniposide in rat liver microsomes (RLMs), plasma, urine, and various tissues. And post-acquisition data-mining methods including extracted ion chromatogram (EIC), multiple mass defect filters (MMDF), fragment ion searching (FISh), and isotope pattern filtering (IPF) were adopted to characterize the known and unknown metabolites. 2. A total of 33 metabolites were detected and interpreted according to accurate mass measurement, diagnostic fragment ions, relevant drug biotransformation knowledge, and bibliography data. Among them, 17 metabolites were detected in the plasma, 31 metabolites were identified in the urine, six metabolites could be found in rat heart, 12 in liver, three in spleen, six in lung, 12 in kidney, six in brain, and four in RLMs. 3. A series of corresponding reactions such as hydrolysis, hydroxylation, taurine conjugation, hydrogenation, decarboxylation, demethylation, sulfate conjugation, cysteine S-conjugation, glucosylation, and their composite reactions were all discovered. 4. The results could provide comprehensive insights and guidance for elucidation of side effect mechanism and safety monitoring as well as for rational formulation design in drug delivery system. The newly discovered geniposide metabolites could be targets for future metabolism studies on the important chemical constituents from herbal medicines.

Topics: Animals; Chromatography, High Pressure Liquid; Iridoids; Male; Mass Spectrometry; Metabolome; Rats, Sprague-Dawley; Spectrometry, Mass, Electrospray Ionization

Our previous studies have shown that geniposide plays an essential role in glucose-stimulated insulin secretion from pancreatic β cells and also regulates the metabolism of Aβ and its deposition in neurons. In this study, we reported that insulin deficiency induced significant increase of tau phosphorylation. Administration of geniposide for 4 weeks significantly decreased the phosphorylated level of tau and the acceleration of GSK-3β phosphorylation in the brain of APP/PS1 transgenic mice induced by insulin deficiency. We also observed that geniposide decreased the phosphorylation of tau protein directly and increased the phosphorylation of Akt in primary cultured cortical neurons. Furthermore, geniposide enhanced the role of insulin on the phosphorylation of Akt, GSK-3β, and tau in primary cultured cortical neurons. And these effects of geniposide in cortical neurons could be prevented by preincubation with LY294002, an inhibitor of PI3K. Taken together, our findings provide a mechanistic and perhaps a foundational link between diabetes and Alzheimer's disease and are consistent with the notion that geniposide might play an essential role on the phosphorylation of tau protein via enhancing insulin signaling and may convey a therapeutic benefit in Alzheimer's disease.

Topics: Alzheimer Disease; Amyloid beta-Protein Precursor; Animals; Disease Models, Animal; Humans; Insulin; Iridoids; Mice; Mice, Transgenic; Phosphorylation; Presenilin-1; tau Proteins

Xingnaojing microemulsion (XNJ-M) administered intranasally is used for stroke treatment. In order to decrease the XNJ-M-induced mucosal irritation, XNJ-M modified by mPEG2000-PLA (XNJ-MM) were prepared in a previous work. The present work aimed to assess the impact of mPEG2000-PLA on pharmacokinetic features and brain-targeting ability of XNJ-M. The bioavailability and brain-target effects of borneol and geniposide in XNJ-M and XNJ-MM were compared in mice after intravenous (i.v.) and intranasal (i.n.) administrations. Gas chromatography, high-performance liquid chromatography, and ultra-performance liquid chromatography/tandem mass spectrometry methods were developed for the quantification of borneol and geniposide. Blood and brain samples were collected from mice at different time points after i.v. and i.n. treatments with borneol at 8.0 mg/kg, geniposide at 4.12 mg/kg. In addition, near-infrared fluorescence dye, 1,1'-dioctadecyl-3,3,3',3'-tetramethyl indotricarbocyanine iodide was loaded into microemulsions to evaluate the brain-targeting ability of XNJ-M and XNJ-MM by near-infrared fluorescence imaging in vivo and ex vivo. For XNJ-M and XNJ-MM, the relative brain targeted coefficients (Re) were 134.59% and 198.09% (borneol), 89.70% and 188.33% (geniposide), respectively. Besides, significant near-infrared fluorescent signal was detected in the brain after i.n. administration of microemulsions, compared with that of groups for i.v. administration. These findings indicated that mPEG2000-PLA modified microemulsion improved drug entry into blood and brain compared with normal microemulsion: the introduction of mPEG2000-PLA in microemulsion resulted in brain-targeting enhancement of both fat-soluble and water-soluble drugs. These findings provide a basis for the significance of mPEG2000-PLA addition in microemulsion, defining its effects on the drugs in microemulsion.

Topics: Administration, Intranasal; Animals; Biological Availability; Brain; Camphanes; Drug Delivery Systems; Drugs, Chinese Herbal; Emulsions; Iridoids; Male; Mice; Mice, Inbred ICR; Nasal Cavity; Polyesters; Polyethylene Glycols; Stroke

1. Although emerging evidence indicates the therapeutic effects of Zhizi Bopi Decoction, the extent to which essential ingredients are absorbed and the possible synergistic actions are poorly understood. 2. In this study, MDCK cell model was used to determine the bi-directional permeability and interaction between the main components (geniposide, berberine and glycyrrhizic acid) of Zhizi Bopi Decoction. 3. The transport of the active ingredients was concentration-dependent in both directions. Moreover, the Papp (AP-BL) values of berberine and glycyrrhizic acid were significantly reduced when co-incubation with an ATP inhibitor. Additionally, uptake of berberine, glycyrrhizic acid were clearly inhibited by the inhibitors of P-glycoprotein and MRP2, indicating that P-gp and MRP2 may be involved in the transport of berberine and glycyrrhizic acid, respectively. However, it was found that geniposide may be purely passive diffusion. Furthermore, the combined incubation of geniposide with berberine and glycyrrhizic acid had a powerful sorbefacient effect than use of a single drug alone which may be regulated by tight junctions. 4. In summary, our study provides useful information for pharmacological applications of Zhizi Bopi Decoction and offers new insights into this ancient decoction for further researches, especially in drug synergism.

Topics: Animals; ATP Binding Cassette Transporter, Subfamily B, Member 1; Berberine; Dogs; Drugs, Chinese Herbal; Glycyrrhizic Acid; Iridoids; Madin Darby Canine Kidney Cells; Permeability

Tripterygium glycosides (TG) are commonly used for basic medicine in curing rheumatoid arthritis but with a high incidence of liver injury. Geniposide (GP) has broad and diverse bioactivities, but until now it is still unknown whether GP can protect against TG-induced liver injury. This study, for the first time, observed the possible protection of GP against TG-induced liver injury in mice and its mechanisms underlying. Oral administration of TG (270 mg/kg) induced significant elevation in the levels of serum alanine / aspartate transaminase (ALT/AST), hepatic malondialdehyde (MDA) and pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-α) (all P < 0.01). On the other hand, remarkably decreased biomarkers, including hepatic glutathione (GSH) level, activities of glutathione transferase (GST), glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT), and anti-inflammatory cytokine interleukin (IL)-10, were observed following TG exposure (all P < 0.01). Nevertheless, all of these phenotypes were evidently reversed by pre-administration of GP for 7 continuous days. Further analysis showed that the mRNA expression of hepatic growth factor-beta1 (TGF-β1), one of tissue repair and regeneration cytokines, was enhanced by GP. Taken together, the current research suggests that GP protects against TG-induced liver injury in mice probably involved during attenuating oxidative stress and inflammation, and promoting tissue repair and regeneration.

Topics: Administration, Oral; Alanine Transaminase; Animals; Anti-Inflammatory Agents; Antirheumatic Agents; Chemical and Drug Induced Liver Injury; Gene Expression; Glutathione; Glycosides; Iridoids; Liver; Liver Regeneration; Male; Mice, Inbred Strains; Oxidative Stress; Phytotherapy; RNA, Messenger; Transforming Growth Factor beta1; Tripterygium; Tumor Necrosis Factor-alpha

Geniposide is one of the main compounds in Gardenia jasminoides ELLIS and has many pharmacological activities, but its anti-hyperglycemic activity has not yet been fully explored. This study was designed to determine, for the first time, how geniposide from G. jasminoides regulates hepatic glucose production, and the underlying mechanisms. During in vitro study, we found the inhibitory effect of geniposide on the hepatic glucose production is partly through AMP-activated protein kinase (AMPK) activation in HepG2 cells. Geniposide significantly inhibited hepatic glucose production in a dose-dependent manner. AMPK, acetyl coenzyme A synthetase (ACC) and forkhead box class O1 (FoxO1) phosphorylation were stimulated by different concentrations of geniposide. In addition, the enzyme activities of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) were all significantly suppressed. What is important is that these effects were partly reversed by (1) inhibition of AMPK activity by compound C, a selective AMPK inhibitor, and by (2) suppression of AMPKα expression by small interfering RNA (siRNA). In summary, geniposide potentially ameliorates hyperglycemia through inhibition of hepatic gluconeogenesis by modulation of the AMPK-FoxO1 signaling pathway. Geniposide or geniposide-containing medicinal plants could represent a promising therapeutic agent to prevent type 2 diabetes on gluconeogenesis.

Topics: AMP-Activated Protein Kinases; Cell Survival; Forkhead Box Protein O1; Glucose; Hep G2 Cells; Humans; Hypoglycemic Agents; Iridoids; Liver; RNA, Small Interfering

Activation of glucagon-like peptide-1 (GLP-1) receptor exerts a range of cardioprotective effects. Geniposide is an agonist of GLP-1 receptor, but its role in cardiac hypertrophy remains completely unknown. Here, we have investigated its protective effects and clarified the underlying molecular mechanisms.. The transverse aorta was constricted in C57/B6 mice and then geniposide was given orally for 7 weeks. Morphological changes, echocardiographic parameters, histological analyses and hypertrophic markers were used to evaluate hypertrophy.. Geniposide inhibited the hypertrophic response induced by constriction of the transverse aorta or by isoprenaline. Activation of 5'-AMP-activated protein kinase-α (AMPKα) and inhibition of mammalian target of rapamycin, ERK and endoplasmic reticulum stress were observed in hypertrophic hearts that were treated with geniposide. Furthermore, Compound C (CpC) or knock-down of AMPKα restricted protection of geniposide against cell hypertrophy and activation of mammalian target of rapamycin and ERK induced by hypertrophic stimuli. CpC or shAMPKα also abolished the protection of geniposide against endoplasmic reticulum stress induced by thapsigargin or dihtiothreitol. The cardio-protective effects of geniposide were ablated in mice subjected to CpC. GLP-1receptor blockade counteracted the anti-hypertrophic response and activation of AMPKα by geniposide. Knock-down of GLP-1 receptor also offset the inhibitory effects of geniposide on cardiac hypertrophy in vivo.. Geniposide protected against cardiac hypertrophy via activation of the GLP-1 receptor/AMPKα pathway. Geniposide is a potential therapeutic drug for cardiac hypertrophy.

Topics: AMP-Activated Protein Kinases; Animals; Cardiomegaly; Cells, Cultured; Dose-Response Relationship, Drug; Enzyme Activation; Glucagon-Like Peptide 1; Iridoids; Male; Mice; Mice, Inbred C57BL; Signal Transduction; Structure-Activity Relationship

Qingkailing injection (QKLI) is a modern Chinese medicine preparation derived from a well-known classical formulation, An-Gong-Niu-Huang Wan. Although the clinical efficacy of QKLI has been well defined, its severe adverse drug reactions (ADRs) were extensively increased. Through thorough attempts to reduce ADR rates, it was realized that the effect-based rational use plays the key role in clinical practices. Hence, the pharmacokinetic-pharmacodynamic (PK-PD) model was introduced in the present study, aiming to link the pharmacokinetic profiles with the therapeutic outcomes of QKLI, and subsequently to provide valuable guidelines for the rational use of QKLI in clinical settings. The PK properties of the six dominant ingredients in QKLI were compared between the normal treated group (NTG) and the pyrexia model group (MTG). Rectal temperatures were measured in parallel with blood sampling for NTG, MTG, model control group (MCG), and normal control group (NCG). Baicalin and geniposide exhibited appropriate PK parameters, and were selected as the PK markers to map the antipyretic effect of QKLI. Then, a PK-PD model was constructed upon the bacalin and geniposide plasma concentrations vs. the rectal temperature variation values, by a two-compartment PK model with a Sigmoid Emax PD model to explain the time delay between the drug plasma concentration of PK markers and the antipyretic effect after a single dose administration of QKLI. The findings obtained would provide fundamental information to propose a more reasonable dosage regimen and improve the level of individualized drug therapy in clinical settings.

Topics: Animals; Antipyretics; Body Temperature; Disease Models, Animal; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Fever; Flavonoids; Iridoids; Male; Medicine, Chinese Traditional; Rats; Rats, Sprague-Dawley

Fructus gardenia is widely used for treatment of stroke and infectious diseases in Chinese medicine. Geniposide is the key bioactive compound related to the pharmacodynamic actions of gardenia on ischemic stroke. The molecular mechanism by which geniposide improves the ischemic brain injury was observed in the study.. Recent studies showed that geniposide had protective activities against the inflammatory response in ischemic stroke. However, the molecular mechanism of geniposide anti-inflammatory role has not yet been fully elucidated. In this study, we investigated the effect of geniposide on the expression of P2Y14 receptor and downstream signaling pathway in brain microvascular endothelial cells (BMECs).. An in vitro model of cerebral ischemia in BMECs was established by oxygen-glucose-deprivation (OGD). To further confirm the specific effect of geniposide on P2Y14 receptor and downstream signaling pathways, we set up a UDP-glucose (an agonist of the P2Y14 receptor) stimulated model. After administration of geniposide, the expression of P2Y14 receptor, phosphorylation of RAF-1, mitogen activated protein kinase kinase1/2 (MEK1/2), extracellular signal-regulated kinase 1/2 (ERK1/2), level of interleukin-8 (IL-8), interleukin-1β (IL-1β), monocyte chemotactic protein 1 (MCP-1) in BMECs were determined.. The mRNA and protein expression of P2Y14 in the rat BMECs were up-regulated in OGD-induced injury. After administration of Geniposide, the expression of P2Y14 receptor was significantly down-regulated, the phosphorylation of RAF-1, MEK1/2, ERK1/2 were suppressed. Similar data were obtained in UDP-glc stimulated model. We also observed that geniposide markedly declined the production of IL-8, IL-1β and MCP-1 in OGD-induced BMECs.. Geniposide exerted anti-inflammatory effects by interfering with the expression of P2Y14 receptor, which subsequently inhibits the downstream ERK1/2 signaling pathways and the release of the pro-inflammatory cytokines IL-8, MCP-1, IL-1β. Therefore, this study provides the evidence for gardenia's clinical application in cerebral ischemia.

Topics: Animals; Cells, Cultured; Endothelial Cells; Extracellular Signal-Regulated MAP Kinases; Gene Expression Regulation; Glucose; Inflammation; Iridoids; Oxygen; Rats; Rats, Sprague-Dawley; Receptors, Purinergic P2Y; Signal Transduction

Baicalin (BA) and jasminoidin (JA) exert an additive effect in the treatment of cerebral ischemia, but the underlying molecular mechanism is still unclear. One hundred mice with focal cerebral ischemia/re-perfusion injury were divided into 5 groups: BA, JA, combination therapy (BJ), sham and vehicle. The differentially expressed genes identified by microarray consisting of 374 cDNAs were uploaded into GeneGo MetaCore software for pathway analyses. Networks were constructed to visualize the interactions of the differentially expressed genes. Among the top ten pathways and processes, we found 5, 3, 2 overlapping pathways and 6, 4, 6 overlapping processes between the BA and JA, BA and BJ, JA and BJ groups, respectively; of which 1 pathway and 3 processes were shared by all the three groups. Six representative pathways and 3 processes were activated only in BJ, such as Gamma-secretase proteolytic targets,etc. These BJ representative targeting pathways showed both vertical (e.g. Cytoplasmic/mitochondrial transport of proapoptotic Bid Bmf and Bim) and horizontal (e.g. Endothelin-1/EDNRA signaling) convergences with those of the BA and JA groups based on the upstream and downstream relationship of cerebral ischemia network, which may help to reveal their additive mechanism in the treatment of cerebral ischemia. Network comparison identified important transcription factors that regulated some of the other BJ related genes, such as cMyb and NF-AT. Such a systemic approach based on multiple pathways and networks may provide a robust path to understand the complex pharmacological variations of combination therapies.

Topics: Animals; Brain Ischemia; Disease Models, Animal; Drug Evaluation, Preclinical; Drug Therapy, Combination; Flavonoids; Gene Expression; Iridoids; Male; Mice; Microarray Analysis; Random Allocation; Reperfusion Injury

Traditional Mongolian Medicine (TMM) exhibits useful biological activities including antifungal, antibacterial, and anti-inflammatory actions. The mechanisms of TMM in anti-inflammation were still unclear. The aim of this study was to investigate the effects of the three main monomers (geniposide, gallate, berberine hydrochloride and a mixture of them) of a traditional Mongolian medicine on cell survival and the proinflammatory cytokines signaling pathways which are activated by bacterial lipopolysaccharides (LPS).. Mouse macrophage-like cell line RAW264.7 was used as a model of inflammation to investigate the anti-inflammatory effects of three TMM momomers and their combination. RT-PCR and Western blot was used to quantify the change of mRNA and protein levels of cytokines, Toll-like receptor-4 (TLR4) and Nuclear Factor-κB (NF-κB) and its inhibitor IκB. The non-radioactive electrophoresis mobility shift assay (EMSA) was used to evaluate the binding activity of NF-κB.. The monomers and their combination exhibited a potent anti-inflammatory effect for suppressing the LPS-evoked secretion of proinflammatory cytokines IL-1β, IL-6 and TNFα. Furthermore, the monomers and their combination attenuated activation of NF-κB and expression of TLR4 at both mRNA and protein levels, the upstream player of the LPS-TLR4-cytokines/ NF-κB signaling pathway.. The Mongolia herbal compound exerts a potent anti-inflammatory effect and could potentially be developed as a useful agent for the chemo-prevention of inflammatory diseases.

Topics: Animals; Anti-Inflammatory Agents; Berberine; Cell Line; Cell Survival; Gallic Acid; Interleukin-1beta; Interleukin-6; Iridoids; Lipopolysaccharides; Macrophages; Medicine, Mongolian Traditional; Mice; NF-kappa B; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

Huanglian Jiedu Decoction (HJD), the classical recipe for relieving fever and toxicity, has been used for treating sepsis in China for sixteen years. However, the effective components of HJD have not been elucidated until now. Therefore, there is a need to elucidate the effective components of HJD against sepsis on animal models induced by endotoxin (LPS). The affinity force of the effective components of HJD with lipid A was evaluated by a biosensor.. Lipid A is regarded as the bioactive center of LPS and is always used as a drug target. In order to obtain the effective components of HJD against sepsis, seven fractions from HJD were tested by a biosensor method for assessing the affinity for lipid A. After further separation, the components were isolated from high lipid A-binding fractions and their affinities to lipid A were assessed with the aid of a biosensor. Their activities were then assayed by an in vivo experiment administered through a tail vein injection. The levels of LPS, TNF-α, and IL-6 from the blood were found and pathology experiments were performed.. Three out of the seven fractions exhibited high lipid A-binding affinities. Berberine, baicalin and geniposide were obtained from the three high lipid A-binding fractions. The animal experiments indicated that the levels of LPS, TNF-α and IL-6 in the medicated treatment groups were much lower than that of the model group ((**)P<0.01). The medicated treatment groups exhibited stronger protective activities on varying organs in the animal model.. Berberine, baicalin and geniposide could neutralize LPS by binding with lipid A and then reduce the release of IL-6 and TNF-α induced by LPS. Furthermore, berberine, baicalin and geniposide exhibited protective activities on varying organs compared to the animal model established by the LPS-induced. These results validate that the components from HJD neutralized LPS and then depressed the release of IL-6 and TNF-α induced by LPS. This gives further evidence that HJD would be a suitable treatment for sepsis and protecting vital organs.

Topics: Animals; Berberine; Biosensing Techniques; Drugs, Chinese Herbal; Female; Flavonoids; Interleukin-6; Iridoids; Kidney; Lipid A; Lipopolysaccharides; Liver; Lung; Male; Mice, Inbred BALB C; Myocardium; Sepsis; Tumor Necrosis Factor-alpha

Mitochondrial dysfunction plays a key role in the progression of Alzheimer's disease (AD). The accumulation of amyloid-beta peptide (Aβ) in the brains of AD patients is thought to be closely related to neuronal mitochondrial dysfunction and oxidative stress. Therefore, protecting mitochondria from Aβ-induced neurotoxicity is an effective strategy for AD therapeutics. In a previous study, we found that geniposide, a pharmacologically active compound purified from gardenia fruit, has protective effects on oxidative stress and mitochondrial dysfunction in AD transgenic mouse models. However, whether geniposide has a protective effect on Aβ-induced neuronal dysfunction remains unknown. In the present study, we demonstrate that geniposide protects cultured primary cortical neurons from Aβ-mediated mitochondrial dysfunction by recovering ATP generation, mitochondrial membrane potential (MMP), and cytochrome c oxidase (CcO) and caspase 3/9 activity; by reducing ROS production and cytochrome c leakage; as well as by inhibiting apoptosis. These findings suggest that geniposide may attenuate Aβ-induced neuronal injury by inhibiting mitochondrial dysfunction and oxidative stress.

Topics: Amyloid beta-Peptides; Animals; Apoptosis; Biopolymers; Cerebral Cortex; Female; Iridoids; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; Mitochondria; Neurons; Peptide Fragments; Reactive Oxygen Species

A novel, rapid and simple analytical method was developed for the quantitative determination of crocin, crocetin and geniposide in soft drink, pastry and instant noodles. The solid samples were relatively homogenized into powders and fragments. The gardenia yellow colorants were successively extracted with methanol using ultrasound-assisted extraction. The analytes were quantitatively measured in the extracts by liquid chromatography coupled with electrospray ionization tandem mass spectrometry. High correlation coefficients (r(2)>0.995) of crocin, crocetin and geniposide were obtained within their linear ranges respectively (50-1000ng/mL, 50-1000ng/mL, 15-240ng/mL) by external standard method. The limits of detection (LODs) were 0.02μg/g for crocin, 0.01μg/g for crocetin and 0.002μg/g for geniposide. And the limits of quantitation (LOQs) were in the ranges of 0.05-0.45μg/g for crocin, and in the ranges of 0.042-0.32μg/g for crocetin, and in the ranges of 0.02-0.15μg/g for geniposide in soft drink, pastry and instant noodles samples. The average recoveries of crocin, crocetin and geniposide ranged from 81.3% to 117.6% in soft drink, pastry and instant noodles. The intra- and inter-day precisions were respectively in the range of 1.3-4.8% and 1.7-11.8% in soft drink, pastry and instant noodle. The developed methods were successfully validated and applied to the soft drink, pastry, and instant noodles collected from the located market in Beijing from China. Crocin, crocetin and geniposide were detected in the collected samples. The average concentrations ranged from 0.84 to 4.20mg/g for crocin, and from 0.62 to 3.11mg/g for crocetin, and from 0.18 to 0.79mg/g for gardenia in various food samples. The method can provide evidences for government to determine gardenia yellow pigments and geniposide in food.

Topics: Beijing; Carbonated Beverages; Carotenoids; Chromatography, High Pressure Liquid; Coloring Agents; Food Analysis; Gardenia; Iridoids; Plant Extracts; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry; Vitamin A

Geniposide, an iridoid glycoside, has antidiabetic effects. The present study aimed to evaluate whether geniposide has direct effects on insulin secretion from rat pancreatic islets. The results demonstrated that geniposide potentiated insulin secretion via activating the glucagon-like-1 receptor (GLP-1R) as well as the adenylyl cyclase (AC)/cAMP signaling pathway. Inhibition of protein kinase A (PKA) suppressed the insulinotropic effect of geniposide. Geniposide also inhibited voltage-dependent potassium (Kv) channels, and this effect could be attenuated by inhibition of GLP-1R or PKA. Current-clamp recording showed that geniposide prolonged action potential duration. These results collectively imply that inhibition of Kv channels is linked to geniposide-potentiated insulin secretion by acting downstream of the GLP-1R/cAMP/PKA signaling pathway. Moreover, activation of Ca(2+) channels by geniposide was observed, indicating that the Ca(2+) channel is also an important player in the geniposide effects. Together, these findings provide new insight into the mechanism underlying geniposide-regulated insulin secretion.

Topics: Adenylyl Cyclases; Animals; Calcium Channels; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Glucagon-Like Peptide-1 Receptor; Insulin; Insulin Secretion; Insulin-Secreting Cells; Ion Channel Gating; Ion Channels; Iridoids; Isoquinolines; Male; Potassium Channels; Rats, Sprague-Dawley; Signal Transduction; Sulfonamides; Tetraethylammonium

A novel method for the selective extraction of gardenia yellow and geniposide from Gardenia Jasminoides, based on a mechanochemical method is described. Without the need of complex separation techniques, gardenia yellow compliant with the national standard could be extracted in a simple fashion. The optimal ball-milling conditions determined were as follows: 30% g/g. active carbon milling at 200 rpm in a planetary mill for 5 min. The extraction conditions of the milled mixtures were as follows: the milled mixtures were extracted with water (liquid-solid ratio 10:1) at 20 °C for 5 min with yields 85% of total geniposide, followed by extraction with 80% ethanol solution (liquid-solid ratio 5:1) and 1% g/g. Tween 20 at 75 °C for 5 min to yield 1.45% ± 0.108% g/g of gardenia yellow. The mechanism of this selective extraction was demonstrated to follow a microstructure change of activated carbon, which occurred during milling and lead to alteration of the corresponding desorption capacities. Compared with traditional extraction methods, this novel extraction technique greatly simplifies the separation process, and proves to be advantageous in terms of low organic solvent consumption, easy operation, rapid process and high efficiency.

Topics: Biomechanical Phenomena; Chemical Phenomena; Chemistry Techniques, Analytical; Gardenia; Iridoids; Plant Extracts

This study aimed to explore whether the regulatory effect of miR-21 on α-synuclein expression in neurons is a potential mechanism by which geniopside (GP) protects the central nervous system from Parkinson disease (PD).. The human neuroblastoma cell line SH-SY5Y was induced to differentiate in vitro and treated with dimethyl sulfoxide (DMSO), N-methyl-4-phenylpyridinium iodide (MPP(+)), and MPP(+) together with GP. To identify the role of miR-21 in the regulation of lysosome-associated membrane protein 2 (LAMP2A) and α-synuclein, SH-SY5Y cells pretreated with MPP(+) were transfected with miR-21 mimic and miR-21 inhibitor. To identify whether GP could reduce the level of α-synuclein through miR-21/LAMP2A, SHSY5Y cells pretreated with GP were treated with miR-21 mimic or miR-21 inhibitor; meanwhile, a luciferase reporter assay was performed to confirm the direct target of miR-21. LAMP2A was overexpressed using a pCMV6-XL5-LAMP2A vector to confirm the role of LAMP2A in the regulation of α-synuclein by miR-21. In these in vitro experiments, the RNA and/or protein expressions of miR-21, LAMP2A, and α-synuclein in SH-SY5Y cells were determined by quantitative real-time polymerase chain reaction and/or western blotting, respectively. An in vivo PD mouse model was established through intraperitoneal injection with N-methyl-4-phenyl-l,2,3,6-tetrahydropyridine (MPTP). The mice were treated with saline, MPTP, MPTP+GP, and MPTP+GP+miR-21 agomir. The numbers of TH(+) cells in the substantia nigra in different groups of mice were compared. The RNA and/or protein expressions of miR-21, LAMP2A, and α-synuclein were also determined.. The level of miR-21 in the cells or mice models was significantly higher than that in normal cells or normal mice, respectively, and GP significantly downregulated miR-21. GP also raised the protein and mRNA expressions of LAMP2A and reduced the protein level of α-synuclein in PD models. MiR-21 upregulated the expression of α-synuclein by directly targeting 3' UTR of LAMP2A. LAMP2A overexpression abolished the upregulating effect of miR-21 mimic on α-synuclein. MiR-21 mimics/agomir reversed the GP-induced downregulation of α-synuclein; miR-21 inhibitor effectively increased the downregulation of α-synuclein caused by GP.. GP exhibits neuroprotective properties by inhibiting α-synuclein expression in PD models through the miR-21/LAMP2A axis.

Topics: alpha-Synuclein; Animals; Cell Line, Tumor; Disease Models, Animal; Dopaminergic Neurons; Humans; Iridoids; Lysosomal-Associated Membrane Protein 2; Mice; MicroRNAs; Neuroprotective Agents; Parkinson Disease; Parkinsonian Disorders; Substantia Nigra; Tyrosine 3-Monooxygenase

Traditional medicinal formulation of Yin-zhi-huang (YZH) is widely used in the clinic for the treatment of jaundice and chronic liver diseases in East Asian countries. However, the pharmacologically active components of YZH and the underlying mechanism are still unknown. Geniposide (GEN) was recently identified as one of the most abundant circulating components in YZH. In this study, we investigated the protective effect of GEN against liver injuries induced by alpha-naphthylisothiocyanate (ANIT). 50[Formula: see text]mg/kg of GEN was administered to ICR mice once daily for 5 days, and challenge of ANIT 75[Formula: see text]mg/kg was performed on the 4th day. Blood and liver tissues were collected on day 6 and subjected to biochemical, histopathological and pathway analyses. The biochemical and pathological findings showed that GEN almost totally attenuated ANIT-induced cholestasis and liver injury compared with the vehicle/ANIT group. The altered gene transcription related to bile acid metabolism and transport was normalized by co-treatment with GEN. The expressions of tumor necrosis factor-[Formula: see text] and the suppressor of cytokine signaling 3 were significantly decreased in the GEN/ANIT group. Western blot revealed that GEN inhibited the activation and expression of STAT3 and NF[Formula: see text]B. These data suggest GEN inhibits ANIT-induced hepatotoxicity. The protective effect is associated with the downregulation of STAT3 and NF[Formula: see text]B signaling.

Topics: 1-Naphthylisothiocyanate; Animals; Cholestasis, Intrahepatic; Down-Regulation; Drugs, Chinese Herbal; Humans; Iridoids; Male; Mice; Mice, Inbred ICR; NF-kappa B; Signal Transduction; STAT3 Transcription Factor; Tumor Necrosis Factor-alpha

1. Pinoresinol di-O-β-d-glucopyranoside (PDG), geniposide (GE), geniposidic acid (GA), aucubin (AN) and chlorogenic acid (CA) are the representative active ingredients in Eucommiae cortex (EC), which may be estrogenic. 2. The ultra high-performance liquid chromatography/tandem mass spectrometry (UHPLC-MS/MS) method for simultaneous determination of the five ingredients showed good linearity, low limits of quantification and high extraction recoveries, as well as acceptable precision, accuracy and stability in mice plasma and tissue samples (liver, spleen, kidney and uterus). It was successfully applied to the comparative study on pharmacokinetics and tissue distribution of PDG, GE, GA, AN and CA between normal and ovariectomized (OVX) mice. 3. The results indicated that except CA, the plasma and tissue concentrations of PDG, GE, GA in OVX mice were all greater than those in normal mice. AN could only be detected in the plasma and liver homogenate of normal mice, which was poorly absorbed in OVX mice and low in other measured tissues. PDG, GE and GA seem to be better absorbed in OVX mice than in normal mice proved by the remarkable increased value of AUC0-∞ and Cmax. It is beneficial that PDG, GE, GA have better plasma absorption and tissue distribution in pathological state.

Topics: Animals; Chlorogenic Acid; Drugs, Chinese Herbal; Estrogens; Glucosides; Iridoid Glucosides; Iridoids; Lignans; Mice; Ovariectomy; Tissue Distribution

To examine the metabolism of genipin-1-β-d-gentiobioside (GG), its distribution and biotransformation in vivo and in vitro were investigated. Urine, plasma, feces, and various organs were collected after oral administration of GG to normal rats and pseudo-germ-free rats to evaluate GG metabolism in vivo. GG was incubated with intestinal flora and primary hepatocytes in vitro to investigate microbial and hepatic metabolism. Using HPLC-Q-TOF-LC/MS, 11 metabolites of GG were absolutely or tentatively identified in terms possible elemental compositions, retention times, and characteristics of fragmentation patterns corresponding to eight biotransformations: deglycosylation, hydroxylation, sulfate conjugation, glucuronidation, hydrogenation, demethylation, glycosylation, and dehydration. Fewer metabolites were detected in pseudo-germ-free rats than in conventional rats. Moreover, geniposide and genipin were generated by the deglycoslation of intestinal bacteria. Geniposidic acid was detected in rat primary-hepatocyte incubation. This study first explores the metabolism of GG in vivo and in vitro. The results can aid the elucidation of PK profiles and clinical usage of gardenia fruit.

Topics: Animals; Chromatography, High Pressure Liquid; Glycosylation; Hepatocytes; Hydroxylation; Iridoids; Male; Mass Spectrometry; Molecular Structure; Rats; Rats, Sprague-Dawley

Clinical and preclinical data suggest that diabetes is often psychological complications such as depression. Geniposide (GP), a major compound in Gardenia jasminoides Ellis with both medicinal and nutritional values, has been previously confirmed to exert anti-diabetic and anti-depressive activities. The present study attempted to observe anti-depressive mechanisms of GP in streptozotocin (STZ) evoked diabetic mice by involving brain-derived neurotrophic factor (BDNF), for the first time. Mice were given GP daily (50, and 100 mg/kg, ig) or reference drugs FHMH [fluoxetine hydrochloride (FH, 10 mg/kg, ig) combined with metformin hydrochloride (MH, 100 mg/kg, ig)] for 3 weeks. The forced swimming test (FST) was performed to observe depression-like behavior, and serum and brain tissues were used for neurochemical and fluorescent quantitative reverse transcription PCR analyses. STZ induced excessively increased blood sugar and immobility time in FST, in a manner attenuated by GP and FHMH administration. GP administration further elevated BDNF levels, and up-regulated the mRNA expression of BDNF and tropomyosin-related kinase B (TrkB) in hippocampus of diabetic mice. In addition, STZ induced the excessive level of serum corticosterone (CORT), while GP did not influence on it in diabetic mice. Taken together, these findings indicate that GP can alleviate depression-like behavior in STZ-evoked diabetic mice, and suggest its mechanisms may partially be ascribed to up-regulating BDNF expression in brain.

Topics: Animals; Antidepressive Agents; Brain-Derived Neurotrophic Factor; Depression; Diabetes Mellitus, Experimental; Gene Expression; Hippocampus; Iridoids; Male; Mice; Random Allocation

Myocardial ischemia/reperfusion injury is a major cause of morbidity and mortality associated with coronary heart disease. Many studies have demonstrated that natural products are promising chemotherapeutic drugs counteracting the loss of cardiomyocytes. Thus, the purpose of the present study was to investigate the effects of geniposide, a traditional Chinese herb extract from Gardenia jasminoides J. Ellis, on cardiomyocyte apoptosis induced by hypoxia/reoxygenation (H/R) in H9c2 cells, and their underlying mechanisms.. Cell viability and apoptosis ratio were assessed using the cell counting kit-8 assay and Annexin V/propidium iodide (PI) staining. The concentrations of lactate dehydrogenase (LDH), intracellular total superoxide dismutase (T-SOD), and malondialdehyde (MDA) were detected by microplate reader. The production of reactive oxygen species/reactive nitrogen species (ROS/RNS), the level of mitochondrial calcium, and mitochondrial membrane potential depolarization were measured by confocal laser scanning microscopy. Mitochondrial morphology was visualized using transmission electron microscopy. The expressions of Bcl-2 mRNA and Caspase-3 mRNA were measured by reverse transcription-polymerase chain reaction (RT-PCR). The protein levels of cleaved caspase-3, Bcl-2, Bax, AKT, p-AKTserine473, cytochrome-c were detected by western bloting.. Geniposide pretreatment increased cell viability, decreased LDH levels in the supernatant, and inhibited cardiomyocyte apoptosis caused by H/R. Furthermore, geniposide reversed mitochondrial dysfunction by decreasing oxidative stress products (ROS/RNS and MDA), increasing anti-oxidative enzyme (T-SOD) level, improving mitochondrial morphology, attenuating mitochondrial calcium overload and blunting depolarization of mitochondrial membrane. Moreover, geniposide pretreatment increased Bcl-2 level and decreased Bax level, thus enhancing the Bcl-2/Bax ratio. Consistent with the above result, Bcl-2 mRNA expression was upregulated and caspase-3 mRNA expression was downregulated by geniposide. In addition, geniposide decreased the protein expression of cleaved caspase-3 and cytochrome-c and increased the level p-AKTserine473. The protective effects of geniposide were partially reversed by glucagon-like pepitide-1 receptor antagonist exendin-(9-39) and the phosphatidylinositol 3 kinase (PI3K) inhibitor LY294002.. Our results suggest that geniposide pretreatment inhibits H/R-induced myocardial apoptosis by reversing mitochondrial dysfunction, an effect in part due to activation of GLP-1R and PI3K/AKT signaling pathway.

Topics: Animals; Apoptosis; Blotting, Western; Caspase 3; Cell Hypoxia; Cell Line; Cell Survival; Drugs, Chinese Herbal; Gene Expression; Glucagon-Like Peptide-1 Receptor; Iridoids; Microscopy, Confocal; Microscopy, Electron, Transmission; Mitochondria, Heart; Myocytes, Cardiac; Oxygen; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; Rats; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction

Geniposide (GE) is the main bioactive component of Gardeniae Fructus. The hepatotoxicity of geniposide limited clinical application. In order to get a new geniposide derivative that has less hepatotoxicity and still possesses the antidepressant activity, a new C-1 hydroxyl methylation derivative named methyl genipin (MG) was synthesized from geniposide. In the present study, we demonstrated that MG did not increase the liver index, alanine aminotransferase (ALT) and aspirate aminotransferase (AST). Histopathological examination suggested that no toxic damages were observed in rats treated orally with MG (0.72 mmol/kg). More importantly, a 7-day treatment with MG at 0.13, 0.26, and 0.52 mmol/kg/day could reduce the duration of immobility. It showed that the antidepressant-like effects of MG were similar to GE in the tail suspension test and the forced swim test. Furthermore, we found MG could be detected in the brain homogenate of mice treated orally with MG 0.52 mmol/kg/day for 1 day by HPLC. The area under the curve (AUC) of MG in the brain homogenate was enhanced to 21.7 times that of GE. The brain amount and distribution speed of MG were improved significantly after oral administration. This study demonstrated that MG possessed the antidepressant effects and could cross the blood-brain barrier, but had less hepatotoxicity.

Topics: Animals; Antidepressive Agents; Blood-Brain Barrier; Body Weight; Brain; Chromatography, High Pressure Liquid; Iridoids; Liver; Molecular Structure; Permeability; Rats; Tissue Distribution

Module-based network analysis of diverse pharmacological mechanisms is critical to systematically understand combination therapies and disease outcomes. We first constructed drug-target ischemic networks in baicalin, jasminoidin, ursodeoxycholic acid, and their combinations baicalin and jasminoidin as well as jasminoidin and ursodeoxycholic acid groups and identified modules using the entropy-based clustering algorithm. The modules 11, 7, 4, 8 and 3 were identified as baicalin, jasminoidin, ursodeoxycholic acid, baicalin and jasminoidin and jasminoidin and ursodeoxycholic acid-emerged responsive modules, while 12, 8, 15, 17 and 9 were identified as disappeared responsive modules based on variation of topological similarity, respectively. No overlapping differential biological processes were enriched between baicalin and jasminoidin and jasminoidin and ursodeoxycholic acid pure emerged responsive modules, but two were enriched by their co-disappeared responsive modules including nucleotide-excision repair and epithelial structure maintenance. We found an additive effect of baicalin and jasminoidin in a divergent pattern and a synergistic effect of jasminoidin and ursodeoxycholic acid in a convergent pattern on "central hit strategy" of regulating inflammation against cerebral ischemia. The proposed module-based approach may provide us a holistic view to understand multiple pharmacological mechanisms associated with differential phenotypes from the standpoint of modular pharmacology.

Topics: Algorithms; Brain; Brain Ischemia; Drug Delivery Systems; Drug Synergism; Drug Therapy, Combination; Entropy; Flavonoids; Gene Ontology; Gene Regulatory Networks; Humans; Iridoids; Neuroprotective Agents; Transcriptome; Ursodeoxycholic Acid

Fructus Gradeniae, the fruit of Gardenia jasminoides Ellis, was used alone or in combination with other herb medicines in the treatment of type 2 diabetes mellitus in China for a long time. In present investigation, the HPLC method for the determination of geniposide in rat plasma was developed and validated, and the pharmacokinetics of geniposide in type 2 diabetic rats after oral administration of Fructus Gradeniae extract or pure was studied. The results showed that the pharmacokinetic profile (especially the area under the plasma concentration-time curve, AUC) of geniposide in type 2 diabetic rats after orally administered with Fructus Gradeniae extract or pure geniposide was remarkably different from that in normal rats. The results indicated that the increased AUC of geniposide in type 2 diabetic rats did not result from the effects of other components contained in Fructus Gradeniae. It could be speculated that the increased AUC of geniposide might result from the pathological state of type 2 diabetes mellitus which resulted in the pharmacokinetic alterations of geniposide.

Topics: Administration, Oral; Animals; Chromatography, High Pressure Liquid; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Gardenia; Iridoids; Male; Plant Extracts; Rats, Sprague-Dawley; Reference Standards

Module-based methods have made much progress in deconstructing biological networks. However, it is a great challenge to quantitatively compare the topological structural variations of modules (allosteric modules [AMs]) under different situations. A total of 23, 42, and 15 coexpression modules were identified in baicalin (BA), jasminoidin (JA), and ursodeoxycholic acid (UA) in a global anti-ischemic mice network, respectively. Then, we integrated the methods of module-based consensus ratio (MCR) and modified Z

Topics: Animals; Brain Ischemia; Flavonoids; Gene Expression Profiling; Gene Regulatory Networks; Humans; Iridoids; Mice; Ursodeoxycholic Acid

Huang-Lian-Jie-Du-Decoction (HLJDD, Oren-gedoku-to in Japanese) is commonly used in traditional Chinese medicine (TCM) to treat ischemic stroke. This study investigated the efficacy of various combinations of the major components of HLJDD, berberine (A), baicalin (B), and jasminoidin (C), on the treatment of ischemic stroke modeled by middle cerebral artery occlusion (MCAO) in rats. The effects of A, B and C individually and their combinations were investigated using proton nuclear magnetic resonance (1H NMR)-based metabolomics complemented with neurologic deficit scoring, infarct volume measurement, biochemistry, histopathology and immunohistochemistry, as well as quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting. Ischemic stroke produces severe oxidative stress, which induces further damage. Our results show that the ABC combination treatment increased levels of cellular antioxidants that scavenged reactive oxygen species during ischemia-reperfusion via the nuclear erythroid 2-related factor 2 (Nrf2) signaling cascade. These protective effects were not observed with the other treatments. These results suggest that a combination of component herbs in HLJDD exhibit stronger effects than the individual herbs alone. Our integrated metabolomics approach also provides a tractable, powerful tool for understanding the science behind TCM formulations.

Topics: Animals; Berberine; Brain; Disease Models, Animal; Drug Synergism; Drugs, Chinese Herbal; Flavonoids; Free Radical Scavengers; Infarction, Middle Cerebral Artery; Iridoids; Male; Metabolomics; Neuroprotective Agents; NF-E2-Related Factor 2; Oxidative Stress; Phytotherapy; Plants, Medicinal; Proton Magnetic Resonance Spectroscopy; Rats, Sprague-Dawley; Reperfusion Injury

Alcohol-induced liver injury (ALD) shows obvious metabolic disorders, categorized by a wide range of metabolite abnormalities. High-throughput metabolomics technology appears to be an appropriate solution. In this study, a urine metabolic profile was assessed using a UPLC-Q-TOF/HDMS (liquid chromatography coupled to high resolution mass spectrometry) approach to investigate the underlying molecular mechanisms of ALD and the therapeutic effect of geniposide. The endogenous low-molecular-weight metabolites in the mouse model of ALD were observed and 48 specific biomarkers were identified. Geniposide was found to have a regulatory effect on 32 of them. Furthermore, targeted analysis of biomarkers showed clear separation between the model and geniposide treatment group. Fifteen biomarkers with high contribution to group differentiation were screened out. Also, a comprehensive analysis of a significant disturbance of multiple metabolic pathways indicated that geniposide could modify abnormal metabolism due to ethanol exposure, during which disorders relating to amino acid metabolism and the oxidative stress state could be alleviated. At the same time, accessory examinations, including plasma biochemical indicators and liver tissue pathological analysis, showed similar results. It was suggested that geniposide was effective as a hepatoprotective agent against ethanol-induced liver damage by re-balancing a wide range of metabolic disorders.

Topics: Animals; Biomarkers; Biopsy; Chemical and Drug Induced Liver Injury; Chromatography, Liquid; Cluster Analysis; Disease Models, Animal; Ethanol; Immunohistochemistry; Iridoids; Male; Mass Spectrometry; Metabolic Networks and Pathways; Metabolome; Metabolomics; Mice; Phenotype

Topics: Anthraquinones; Chromatography, High Pressure Liquid; Coumarins; Drugs, Chinese Herbal; Iridoids; Microscopy; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

A simple, specific and sensitive ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was established and validated for simultaneous determination of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid and geniposide in rat plasma using puerarin as an internal standard (IS). Plasma samples were pretreated by a one-step direct protein precipitation procedure with acetonitrile after acidified using as little as 50 μL plasma. Chromatographic separation was performed on an Acquity BEH C18 column (100 × 2.1 mm, 1.7 µm) at a flow rate of 0.2 mL/min by a gradient elution, using 0.2% acetic acid-methanol as mobile phase. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring via electrospray ionization source with negative ion mode. Calibration curves showed good linearity (r > 0.995) over wide concentration ranges. The intra- and inter-day precisions were <15%, and the accuracy was within ±8.0%. The validated method was successfully applied to a pharmacokinetic study of the four bioactive components in rats after intravenous administration of Reduning injection.

Topics: Animals; Chlorogenic Acid; Chromatography, High Pressure Liquid; Drug Stability; Iridoids; Linear Models; Liquid-Liquid Extraction; Male; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Sensitivity and Specificity; Tandem Mass Spectrometry

The receptor for advanced glycation end products (RAGE)-mediated signaling pathway is related to Aβ-induced pathogenic responses. Geniposide, a pharmacologically active component purified from gardenia fruit, could attenuate the oligomeric Aβ(1-42)-induced inflammatory response by blocking the ligation of Aβ to RAGE and suppressing the RAGE-mediated signaling in vitro. Here, we investigated whether geniposide can exert protective effects on the neuroinflammation and memory deficits in an Alzheimer's disease (AD) mouse model. The results indicate that geniposide treatment significantly suppresses RAGE-dependent signaling (activation of ERK and IκB/NF-κB), the production of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) and cerebral Aβ accumulation in vivo. Furthermore, we demonstrate that geniposide augments synaptic plasticity by attenuating the Aβ-induced reduction of long-term potentiation and increasing the miniature excitatory postsynaptic current (mEPSC) amplitude and frequency in hippocampal neurons. In addition, the intragastric administration of geniposide improves learning and memory in APP/PS1 mice. Taken together, these studies indicate that geniposide has profound multifaceted neuroprotective effects in an AD mouse model. Geniposide demonstrates its neuroprotection by inhibiting inflammation, ameliorating amyloid pathology and improving cognition. Thus, geniposide may be a potential therapeutic agent for halting and preventing AD progression.

Topics: Alzheimer Disease; Animals; Cells, Cultured; Disease Models, Animal; Iridoids; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; Neuroprotective Agents; Receptor for Advanced Glycation End Products; Receptors, Immunologic; Treatment Outcome

Islet amyloid deposition is increasingly seen as a pathogenic feature of type 2 diabetes mellitus (T2DM), with the deposits containing the unique amyloidogenic peptide islet amyloid polypeptide (IAPP, also known as amylin). The fibril precursors of IAPP contribute to its cytotoxicity on pancreatic β cells and be important in causing β-cell dysfunction in T2DM. However, the development of effective this study, inhibitors against the toxicity of IAPP has been extremely challenging. We have found that pre-incubation with geniposide dose-dependently prevented human IAPP (hIAPP)-induced cell damage in INS-1E cells, and bacitracin, an inhibitor of IDE activity, prevented significantly the protective effects of geniposide in pancreatic INS-1E cells significantly. Geniposide induced the expression of insulin-degrading enzyme (IDE), a key degrading protein of hIAPP, but had no significant effect on the aggregation of hIAPP. These findings indicate that geniposide prevents hIAPP-induced cytotoxicity in INS-1E cells involving upregulation of IDE expression.

Topics: Animals; Apoptosis; Bacitracin; Cell Line, Tumor; Diabetes Mellitus, Type 2; Humans; Insulin-Secreting Cells; Insulysin; Iridoids; Islet Amyloid Polypeptide; Protective Agents; Rats

The aim of this study was to explore the anti-inflammatory effects of Geniposide (GE), an iridoid glycoside compound extracted from Gardenia jasminoides Ellis (GJ) fruit in adjuvant-induced arthritis (AA) rats and its pharmacokinetic (PK) basis. AA was induced by injecting with Freund's complete adjuvant (FCA). Male SD rats were subjected to treatment with GE (30, 60 and 120mg/kg) from day 17 to 24 after immunization. Fibroblast-like synoviocyte (FLS) proliferation was assessed by MTT. Interleukin (IL)-1, IL-6, TNF-α and IL-10 were determined using double-sandwich enzyme-linked immunosorbent assay (ELISA). Expression of p38 mitogen-activated protein kinases (p38MAPKs) related proteins in FLS was detected by Western blotting. PK profiles were simultaneously detected by ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) in AA rat plasma after oral administration of GE on day 17 after immunization. As a result, GE promoted the recovery of arthritis and inhibited the colonic inflammation damage in AA rats by decreasing the expression level of TNF-α, IL-1 and IL-6, increasing the production of IL-10 and inhibiting the expression of phospho-p38 (p-p38) related proteins in FLS. PK parameters (AUC, Cmax and t1/2) tended to be associated with dosage-related decreasing of efficacy index.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Experimental; Cell Proliferation; Cells, Cultured; Colon; Cytokines; Disease Models, Animal; Fibroblasts; Freund's Adjuvant; Fruit; Gardenia; Humans; Iridoids; Male; p38 Mitogen-Activated Protein Kinases; Phytotherapy; Rats; Rats, Sprague-Dawley; Signal Transduction; Synovial Membrane

This paper utilized a quantitative (1)H nuclear magnetic resonance (qHNMR) method for assessing the purity of iridoids and secoiridoids. The method was fully validated, including specificity, linearity, accuracy, precision, reproducibility, and robustness. For optimization of experimental conditions, several experimental parameters were investigated, including relaxation delay (D1), scan numbers (NS) and power length (PL1). The quantification was based on the area ratios of H-3 from analytes relative to aromatic protons from 1,4-dinitrobenzene (internal standard) with methanol-d4 as solvent. Five iridoids and secoiridoids (sweroside, swertiamarin, gentiopicroside, geniposide, genipin) were analyzed. Furthermore, the results were validated by the high performance liquid chromatography coupled with ultraviolet detection (HPLC-UV) method. It can be concluded that the qHNMR method was simple, rapid, and accurate, providing a reliable and superior method for assessing the purity of iridoids and secoiridoids.

Topics: Chromatography, High Pressure Liquid; Iridoid Glucosides; Iridoids; Magnetic Resonance Spectroscopy; Molecular Structure; Pyrones; Reproducibility of Results; Sensitivity and Specificity

The natural crosslinking agent genipin has been applied widely in biomedicines and foods nowadays. Because of the special hemiacetal ring structure in its molecule, it can only be prepared by hydrolysis of geniposide according to biocatalysis. In this research, strategies including aqueous-organic biphasic catalysis and substrate fed-batch mode were adopted to improve the biocatalysis process of genipin. A 10 L ethyl acetate-aqueous biphasic system with geniposide fed-batch led to a satisfying genipin yield. With Fusarium solani ACCC 36223, 15.7 g/l genipin in the ethyl acetate phase was obtained, corresponding to space-time yields of 0.654 g l(-1) h(-1).

Topics: Batch Cell Culture Techniques; Cross-Linking Reagents; Culture Media; Fusarium; Hydrolysis; Iridoids

An increasing number of studies have demonstrated that insulin-degrading enzyme (IDE) plays an essential role in both the degradation and its activity of β-amyloid (Aβ). Therefore, the regulation of IDE expression and/or modification of IDE-dependent actions are two emerging strategies for the treatment of Alzheimer's disease (AD). We previously observed that geniposide, a novel agonist of glucagon-like peptide 1 receptor (GLP-1R), could attenuate Aβ-induced neurotoxicity by regulating the expression of IDE in primary cortical neurons. However, the signal transduction mechanisms underlying this effect were not elucidated. The present study, therefore examined and explored the cell signaling transduction and molecular mechanisms by which geniposide induces the expression of IDE in primary cortical neurons. The current study revealed that LY294002 (an inhibitor for phosphatidyl inositol 3-kinase, PI3K), PP1 (inhibitor for c-Src), GW9662 (antagonist for peroxisome proliferator-activated receptor γ, PPARγ), H89 (an inhibitor for protein kinase A, PKA) and AG1478 (an antagonist for epidermal growth factor receptor, EGFR) prohibited the up-regulation of IDE induced by geniposide in primary cortical neurons. Further, geniposide also enhanced the phosphorylation of PPARγ and accelerated the release of phosphorylated FoxO1 (forkhead box O1) from nuclear fraction to the cytosol. Moreover, geniposide directly activated the activity of IDE promoter in PC12 cells, which confirmed the presence of the GLP-1 receptor. Taken together, our findings reveal for the first time the cell signaling transduction pathway of geniposide regulating the expression of IDE in neurons.

Topics: Animals; Cell Nucleus; Cells, Cultured; Central Nervous System Agents; Cerebral Cortex; Cyclic AMP-Dependent Protein Kinases; Cytosol; ErbB Receptors; Forkhead Transcription Factors; Genes, src; Insulysin; Iridoids; Nerve Tissue Proteins; Neurons; PC12 Cells; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Phosphorylation; PPAR gamma; Proto-Oncogene Proteins c-akt; Rats

An oriental medicinal formulation, Huanglian Jiedu Decoction (HLJDD), has been well documented in few Traditional Chinese Medicine Classics 1300 years ago for treatment of heat and dampness-related diseases. Its effect is well accepted in Asian community, including China, Japan and Korea. Recent studies have postulated HLJDD as a regimen for cancer treatment, especially liver cancer, but the underlying mechanism is unknown. The aim of this study was to examine the suppressive effect of HLJDD on the growth of hepatocellular carcinoma (HCC) and its possible underlying mechanism.. Chemical composition of HLJDD was analyzed by high performance liquid chromatography. The tumor suppressive effect of HLJDD was determined on both HCC cells and xenograft model. Nascent protein synthesis was detected with Click-IT protein labeling technology; protein expression was determined by immunoblotting and imunnohistochemical analysis.. Quality analysis revealed that HLJDD of different batches is consistent in both chemical composition and bioactivities. HLJDD inhibited HCC cell proliferation at its non-toxic doses, and suppressed growth and angiogenesis in xenografted murine model. HLJDD suppressed the synthesis of nascent protein via inactivation of eEF2 without deregulating the translation initiation factors. The major components in HLJDD, geniposide, berberine and baicalin, additively act on eEF2, and contributed to the responsible activity. HLJDD-activated eEF2 kinase (eEF2K) led to eEF2 inactivation, and activation of AMPK signaling may be responsible for the eEF2K induction. Blocked AMPK activity in HLJDD-treated HCC cells attenuated eEF2K activation as well as the inhibitory effect of the formula. In nutrient deprived HCC cells with inactivated eEF2, the inhibitory effect of HLJDD in tumor cell expansion was interfered.. Our results indicate that HLJDD has potential in blocking HCC progression with involvement of eEF2 inhibition.

Topics: Animals; Antineoplastic Agents; Berberine; Carcinoma, Hepatocellular; Cell Line, Tumor; Drugs, Chinese Herbal; Elongation Factor 2 Kinase; Female; Flavonoids; Humans; Iridoids; Liver Neoplasms; Mice, Inbred BALB C; Mice, Nude; Neovascularization, Pathologic; Phytotherapy; Xenograft Model Antitumor Assays

Geniposide (GP) is one of main compounds in Gardenia jasminoides Ellis, with both medicinal and nutritional value. This study was designed to determine, for the first time, how GP from G. jasminoides protects against acute alcohol-induced liver injury, and the underlying mechanisms. Mice were orally administered alcohol (6.0 g/kg body mass) 2 h after intragastric administration of GP and bifendate, every day for 7 continuous days. Six hours after the alcohol was administered, levels of serum alanine/aspartate transaminase (ALT/AST), hepatic lipid peroxidation (LPO), glutathione (GSH), glutathione-S-transferase (GST), glutathione peroxidase (GPx), copper- and zinc-containing superoxide dismutase (CuZn-SOD), and catalase (CAT), and mRNA expression of CuZn-SOD and CAT were assayed. The results demonstrated that GP (20.0, 40.0, or 80 mg/kg) significantly reversed the excessive, alcohol-induced elevation in both serum ALT/AST and hepatic LPO levels. Moreover, hepatic GSH, GST, GPx, CuZn-SOD, and CAT levels were all decreased in the alcohol-treated mice, whereas treatment with GP reversed these decreases. Further analysis indicated that hepatic mRNA expression of CuZn-SOD and CAT in the alcohol-treated mice was significantly down-regulated, whereas GP up-regulated such decreases. Taken together, this study shows that GP protects against acute alcohol-induced liver injury via up-regulating the expression of the main antioxidant enzymes, and thus ameliorates alcohol-induced oxidative stress injury in the liver.

Topics: Animals; Biomarkers; Catalase; Dose-Response Relationship, Drug; Enzyme Induction; Glutathione; Glutathione Peroxidase; Glutathione Transferase; Iridoids; Lipid Peroxidation; Liver; Liver Diseases, Alcoholic; Male; Mice, Inbred Strains; Oxidative Stress; Oxidoreductases; Protective Agents; Random Allocation; RNA, Messenger; Superoxide Dismutase

A specific, sensitive and high throughput ultra-high performance liquid chromatography-electrospray ionization tandem mass spectrometric method (UHPLC-ESI-MS/MS) was established and validated to assay geniposide (GE), a promising anti-inflammatory drug, in adjuvant arthritis rat plasma: application to pharmacokinetic and oral bioavailability studies and plasma protein binding ability. Plasma samples were processed by de-proteinised with ice-cold methanol and separated on an ACQUITY UPLC™ HSS C18 column (100 mm × 2.1mm i.d., 1.8 μm particle size) at a gradient flow rate of 0.2 mL/min using acetonitrile-0.1% formic acid in water as mobile phase, and the total run time was 9 min. Mass detection was performed in selected reaction monitoring (SRM) mode with negative electro-spray ionization includes the addition of paeoniflorin (Pae) as an internal standard (IS). The mass transition ion-pair was followed as m/z 387.4 → 122.4 for GE and m/z 479.4 → 449.0 for IS. The calibration curves were linear over the concentration range of 2-50,000 ng/mL with lower limit of quantification of 2 ng/mL. The intra-day and inter-day precisions (RSD, %) of the assay were less than 8.4%, and the accuracy was within ± 6.4% in terms of relative error (RE). Extraction recovery, matrix effect and stability were satisfactory in adjuvant arthritis rat plasma. The UHPLC-ESI-MS/MS method was successfully applied to a pharmacokinetic study of GE after oral administration of depurated GE at 33, 66, 132 mg/kg and intravenous injection at 33, 66, 132 mg/kg in adjuvant arthritis (AA) rats. In addition, it was found that GE has rapid absorption and elimination, low absolute bioavailability, high plasma protein binding ability in AA rats after oral administration within the tested dosage range. It suggested that GE showed slow distribution into the intra- and extracellular space, and the binding rate was not proportionally dependent on plasma concentration of GE when the concentration of GE was below 5.0 μg/mL.

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents; Arthritis, Experimental; Biological Availability; Blood Proteins; Calibration; Chromatography, High Pressure Liquid; Dose-Response Relationship, Drug; High-Throughput Screening Assays; Iridoids; Limit of Detection; Protein Binding; Rats; Rats, Sprague-Dawley; Sensitivity and Specificity; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

Liver fibrosis results from increased deposition of type-I collagen within the hepatic extracellular space and constitutes a common cardinal signature in all forms of liver injury, regardless of etiology. Transforming growth factor β1 (TGF-β1) plays a crucial role in the pathogenesis of liver fibrosis. Geniposide is recognized as being useful against hyperlipidemia and fatty liver. However, its cellular mechanism and anti-fibrotic effect in TGF-β1-induced hepatocytes have not been explored. In the present study, we investigated its anti-epithelial-mesenchymal transition (EMT) mechanism by examining the effect of geniposide on TGF-β1-induced hepatocytes. The effect of geniposide on TGF-β1-induced AML12 cells was assessed using Western blotting, quantitative real-time PCR, immunofluorescence staining and DNA binding activity. We found that geniposide significantly inhibited TGF-β1-induced mRNA and protein expression of type-I collagen. Cells treated concurrently with TGF-β1 and geniposide retained high levels of localized E-cadherin expression with no increase in vimentin. Treatment with geniposide almost completely blocked the phosphorylation of Smad2/3, extracellular signal-regulated kinase (ERK) and Akt in AML12 cells. Taken together, these results suggest that geniposide may suppress TGF-β1-induced EMT in hepatic fibrosis by inhibiting the TGFβ/Smad and ERK-mitogen-activated protein kinase (MAPK) signaling pathways. Our results may help researchers better understand the pathogenesis of liver fibrosis so they can develop novel therapeutic strategies for treatment of liver diseases.

Topics: Animals; Cell Line; Epithelial-Mesenchymal Transition; Extracellular Signal-Regulated MAP Kinases; Hepatocytes; Iridoids; Liver Cirrhosis; MAP Kinase Signaling System; Mice; Smad2 Protein; Smad3 Protein; Transforming Growth Factor beta1

Hwangryunhaedok-tang (HHT) is a traditional herbal medicine that is used for the treatment of fever, inflammation, gastritis, and hypertension. In this study, we performed simultaneous determination of the five components, geniposide (1), baicalin (2), coptisine (3), palmatine (4), and berberine (5) in HHT by using a high-performance liquid chromatography-photodiode array (HPLC-PDA) analysis. We also evaluated the antioxidative activity of HHT and compounds 1-5 by measuring their effects on low-density lipoprotein (LDL) oxidation and antiproliferative abilities in vascular smooth muscle cells (VSMCs).. Five compounds were separated within 40 min by using a Gemini C18 column (temp. 35°C; two-component gradient elution; flow rate 1.0 mL/min; detector 240 and 277 nm). The activities of HHT and compounds 1-5 were tested with the radical scavengers 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt and 2,2-diphenyl-1-picrylhydrazyl, in thiobarbituric acid reactive substance assays, and in relative electrophoretic mobility assays using CuSO4-induced LDL oxidation systems. The antiproliferative effects of samples on platelet-derived growth factor (PDGF)-induced VSMC proliferation were studied by using a cell proliferation assay.. Regression analysis of the five major compounds showed good linearity (r (2) ≥ 0.9997) in different concentration ranges. The recoveries of the five compounds were in the range 86.31-110.78%, with relative standard deviations below 2.1%; those of intra- and interday precision were 0.04-3.78% and 0.04-1.69%, respectively. HHT reduced the oxidation properties of LDL induced by CuSO4 and inhibited cell proliferation in PDGF-treated VSMCs. Among the five components, compound 2 could effectively suppress LDL oxidation and PDGF-induced VSMC proliferation.. The established HPLC-PDA method will help to improve quality control of HHT. The results demonstrate that HHT has antiatherosclerotic activity and that it functions by modulating LDL oxidation and VSMC proliferation. The effects of HHT may be attributed, at least I part, to compound 2.

Topics: Animals; Antioxidants; Atherosclerosis; Berberine; Berberine Alkaloids; Biphenyl Compounds; Cell Proliferation; Chromatography, High Pressure Liquid; Coptis; Drugs, Chinese Herbal; Flavonoids; Gardenia; Iridoids; Lipoproteins, LDL; Medicine, Korean Traditional; Muscle, Smooth, Vascular; Phellodendron; Phytotherapy; Picrates; Platelet-Derived Growth Factor; Rats; Regression Analysis; Scutellaria

Our previous studies have shown that geniposide plays an essential role in glucose-stimulated insulin secretion from pancreatic β cells and also antagonizesAβ1-42-induced cytotoxicity examined using a primary cortical neuron assay. However, the mechanism by which geniposide appears to regulate insulin signaling in the brain is presently not well understood. In this study, we administered streptozotocin (STZ) to induce insulin-deficiency in an AD transgenic mouse model, and investigated the effects of geniposide on the β-amyloidogenic processing of amyloid precursor protein (APP) using in vitro and in vivo models. Our results indicate that treatment with STZ (90 mg/kg, i.p., once daily for two consecutive days) induced significant reduction in peripheral and brain insulin levels in both wild-type and APP/PS1 transgenic mice. Administration of geniposide for 4 weeks significantly decreased the concentrations of cerebral β-amyloid peptides (Aβ1-40 and Aβ1-42) in STZ-treated AD mice. Further experiments showed that geniposide up-regulated the protein levels of β-site APP cleaving enzyme (BACE1) and insulin-degrading enzyme (IDE), and decreased the protein levels of ADAM10 when examined using a primary cultured cortical neuron assay and in STZ-induced AD mice. Meanwhile, geniposide also directly enhanced the effects of insulin by reducing Aβ1-42 levels in primary cultured cortical neurons. Taken together, our findings provide a mechanistic link between diabetes and AD, and is consistent with the notion that geniposide might play an important role on APP processing via enhancing insulin signaling and may convey a therapeutic benefit in AD.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Amyloid beta-Protein Precursor; Animals; Brain; Cells, Cultured; Diabetes Mellitus, Experimental; Disease Models, Animal; Humans; Insulin; Iridoids; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; Peptide Fragments; Presenilin-1

Geniposide as the major active component of Gardenia jasminoides Ellis has neuroprotective activity. This study elucidated the potential antidepressant-like effect of geniposide and its related mechanisms using a depression rat model induced by 3 consecutive weeks of chronic unpredictable mild stress (CUMS). Sucrose preference test, open field test (OFT) and forced swimming test (FST) were applied to evaluate the antidepressant effect of geniposide. Adrenocorticotropic hormone (ACTH) and corticosterone (CORT) serum levels, adrenal gland index and hypothalamic corticotrophin-releasing hormone (CRH) mRNA expression were measured to assess the activity of hypothalamus-pituitary-adrenal (HPA) axis. Hypothalamic glucocorticoid receptor α (GRα) mRNA expression and GRα protein expression in hypothalamic paraventricular nucleus (PVN) were also determined by real-time PCR and immunohistochemistry, respectively. We found that geniposide (25, 50, 100mg/kg) treatment reversed the CUMS-induced behavioral abnormalities, as suggested by increased sucrose intake, improved crossing and rearing behavior in OFT, shortened immobility and prolonged swimming time in FST. Additionally, geniposide treatment normalized the CUMS-induced hyperactivity of HPA axis, as evidenced by reduced CORT serum level, adrenal gland index and hypothalamic CRH mRNA expression, with no significant effect on ACTH serum level. Moreover, geniposide treatment upregulated the hypothalamic GRα mRNA level and GRα protein expression in PVN, suggesting geniposide could recover the impaired GRα negative feedback on CRH expression and HPA axis. These aforementioned therapeutic effects of geniposide were essentially similar to fluoxetine. Our results indicated that geniposide possessed potent antidepressant-like properties that may be mediated by its effects on the HPA axis.

Topics: Animals; Antidepressive Agents; Chronic Disease; Corticosterone; Corticotropin-Releasing Hormone; Depressive Disorder; Dietary Sucrose; Disease Models, Animal; Dose-Response Relationship, Drug; Feeding Behavior; Hypothalamo-Hypophyseal System; Iridoids; Male; Motor Activity; Pituitary-Adrenal System; Rats, Sprague-Dawley; Receptors, Glucocorticoid; RNA, Messenger; Stress, Psychological; Uncertainty

T-cell factor 7-like 2 (TCF7L2) is an important transcription factor of Wnt/β-catenin signaling, which has critical roles in β-cell survival and regeneration. In preliminary screening assay, we found geniposide, a naturally occurring compound, was able to increase TCF7L2 mRNA level in Min6 cells. Here we aimed to investigate the role of geniposide in β-cell and underlying mechanism involved. Geniposide was found to promote β-cell survival by increasing β-cell proliferation and decreasing β-cell apoptosis in cultured mouse islets after challenge with diabetic stimuli. Geniposide protected β-cell through activating Wnt signaling, enhanced expressions of TCF7L2 and GLP-1R, activated AKT, inhibited GSK3β activity, and promoted β-catenin nuclear translocation. The protective effect of geniposide was remarkably suppressed by siRNAs against β-catenin, or by ICG001 (β-catenin/TCF-mediated transcription inhibitor). Moreover, geniposide promoted β-cell regeneration in vivo to normalize blood glucose in high-fat diet and db/db mice. Increased β-cell proliferation was observed in pancreatic sections of geniposide-treated diabetic mice. Most importantly, geniposide triggered small islet-like cell clusters formation as a result of β-cell neogenesis from ductal epithelium, which was well correlated with the increase in TCF7L2 expression. In exocrine cells isolated from mouse pancreas, geniposide could induce duct cell differentiation through upregulating TCF7L2 expression and activating JAK2/STAT3 pathway. Taken together, we identified a novel role of geniposide in promoting β-cell survival and regeneration by mechanisms involving the activation of β-catenin/TCF7L2 signaling. Our finding highlights the potential value of geniposide as a possible treatment for type 2 diabetes.

Topics: Animals; beta Catenin; Cell Differentiation; Cell Proliferation; Cell Survival; Cells, Cultured; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Iridoids; Islets of Langerhans; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; Regeneration; RNA, Messenger; Signal Transduction; Transcription Factor 7-Like 2 Protein

To investigate the effect of different pH on rectum permeability of chlorogenic acid and geniposide.. Four kinds of Reduning suppositories of different pH were separated and put into the rectum to study the suppositories in vitro and the content of chlorogenic acid and geniposide samples was determined by HPLC to calculate the permeation in 24 hours.. With increase of pH within 2.5-7.4, the steady state flux of chlorogenic acid was increased, but the steady state flux of geniposidesamples was steady.. Adjusted the pH can increase the rectum permeability of active ingredients in Reduning auppositories.

Topics: Animals; Chlorogenic Acid; Drugs, Chinese Herbal; Hydrogen-Ion Concentration; Iridoids; Male; Permeability; Rats; Rats, Sprague-Dawley; Rectum; Suppositories

Geniposide (GP), an iridoid glucoside extracted from Gardenia jasminoides Ellis fruits, has been used as a herbal medicine to treat liver and gall bladder disorders for many years. However the mechanism of anti-inflammatory is largely unknown. In this study, GP significantly attenuated inflammation in acute liver injury (ALI) mice model and in lipopolysaccharide (LPS)-induced THP-1 cells. It was demonstrated that GP obviously decreased the expression of Methyl-CpG binding protein 2 (MeCP2) in vivo and in vitro. Knockdown of MeCP2 with siRNA suppressed the expressions of IL-6 and TNF-α, while over-expression of MeCP2 had a proinflammatory effect on the expression of IL-6 and TNF-α in LPS-induced THP-1 cells. Mechanistically, it was indicated that GP had anti-inflammatory effects at least in part, through suppressing MeCP2. Interestingly, GP could attenuate expressions of Sonic hedgehog (Shh) and GLIS family zinc finger 1 (GLIS1) but increase Ptched1 (PTCH1) expression. Similar findings were also demonstrated at the protein level by siRNA MeCP2. Furthermore, over-expression of MeCP2 obviously increased Shh and GLIS1 expressions but reduced PTCH1 expression. Taken together, GP may serve as an effective modulator of MeCP2-hedgehog pathway (Hh)-axis during the pathogenesis of inflammation. Our findings shed light on the potential therapeutic feature of GP in recovering inflammatory diseases.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Carbon Tetrachloride Poisoning; Cell Line; Chemical and Drug Induced Liver Injury; Gene Expression; Gene Knockdown Techniques; Hedgehog Proteins; Inflammation; Interleukin-6; Iridoids; Liver; Methyl-CpG-Binding Protein 2; Mice; Mice, Inbred C57BL; RNA, Small Interfering; Tumor Necrosis Factor-alpha

Parkinson's disease (PD) is a chronic neurodegenerative disease, and there is no cure for it at present. We tested the drug Geniposide, an active component of Gardenia jasminoides Ellis which is used in traditional Chinese medicine. Geniposide has shown neuroprotective and growth-factor like effects in several in vivo and in vitro studies. In the present study, Geniposide had been tested in an acute PD mouse model induced by four 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) intraperitoneal injections. Geniposide treatment (100mg/kg ip.) for 8 days after MPTP treatment (30mg/kg ip.) improved the locomotor and exploratory activity of mice (open field), and improved bradykinesia and movement balance of mice (rotarod, swim test). Geniposide treatment also restored tyrosine hydroxylase (TH) positive dopaminergic neuron numbers in the substantia nigra pars compacta. Drug treatment also increased levels of growth factor signaling molecule Bax and reduced the apoptosis signaling molecule Bcl-2. Caspase 3 activation was also reduced in the substantia nigra. We conclude that Geniposide exerted its neuroprotective effect by enhancing growth factor signaling and the reduction of apoptosis. Geniposide is an ingredient in Chinese traditional medicine with few known side effects and shows potential as a drug treatment for Parkinson's disease.

Topics: 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; Animals; Apoptosis; bcl-2-Associated X Protein; Caspase 3; Cell Proliferation; Disease Models, Animal; Iridoids; Male; Mice; Mice, Inbred C57BL; Motor Activity; Neurons; Neuroprotective Agents; Parkinson Disease; Pars Compacta; Proto-Oncogene Proteins c-bcl-2; Psychomotor Performance; Swimming; Tyrosine 3-Monooxygenase

To observe the protective effect of active fractions of Huanglian Jiedu Decoction (HJD) on primary cortical neuron injury after oxygen-glucose deprivation (OGD)/reperfusion (R) injury. Methods Using macroporous resin method, HJDFE30, HJDFE50, HJDFE75, and HJDFE95 with 30%, 50%, 75%, and 95% alcohol were respectively prepared. Then the content of active components in different HJD fractions was determined with reverse phase high-performance liquid chromatography (RP-HPLC). The OGD/R injury model was induced by sodium dithionite on primary cortical neurons in neonate rats. MTT assay was used to observe the effect of four fractions (HJDFE30, HJDFE50, HJDFE75, and HJDFE95) and seven index components of HJD on the neuron viability.. RP-HPLC showed active component(s) contained in HJDFE30 was geniposide; baicalin, palmatine, berberine, and wogonside contained in HJDFE50; baicalin, berberine, baicalein, and wogonin contained in HJDFE75. The neuron viability was decreased after OGD for 20 min and reperfusion for 1 h, (P <0. 01), and significantly increased after administered with HJD, HJDFE30, HJDFE50, and HJDFE75 (P <0. 05, P <0. 01). Geniposide, baicalin, baicalein, palmatine, wogonside, and wogonin could increase the cortical neuron viability (P <0. 05, P <0. 01).. HJDFE30, HJDFE50, and HJDFE75, as active fractions of HJD, had protective effect on primary cortical neuron injury after OGD/R. Furthermore, geniposide, baicalin, and baicalein were main active components of HJD.

Topics: Animals; Berberine; Berberine Alkaloids; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucose; Iridoids; Models, Animal; Neurons; Oxygen; Rats; Reperfusion Injury

To establish a method of quantitative analysis of multi-components, by single marker(QAMS)for simultaneously determining six ingredients in Gardenia jasminoides fruits.. A multi-wavelength segmentation detection method was used. A methodological mode was found to analysis six ingredients in Gardenia jasminoides fruits by quantitative analysis of QAMS. Taken geniposide as reference to create RCF with gardenia acid, chlorogenic acid, crocin I, crocin II and crocin III.. The good reproducibility and acceptable durability of method was validated between two HPLC systems and three columns. 20 batches of Gardenia jaminoides fruits was analysis, and the results showed good linear correlation compared to external standard method (r > 0. 999).. QAMS can be used as quality evaluation method of multi-component Gardenia jaminoides fruits.

Topics: Carotenoids; Chlorogenic Acid; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Fruit; Gardenia; Iridoids; Phytochemicals; Reproducibility of Results

We have previously demonstrated that geniposide attenuates the production of Aβ1-42 both in vitro and in vivo via enhancing leptin receptor signaling. But the role played by geniposide in the phosphorylation of tau and its underlying molecular mechanisms remain unclear. In this study, we investigated the effect of geniposide on the phosphorylation of tau and the role of leptin signaling in this process. Our data suggested that, accompanied by the up-regulation of leptin receptor expression, geniposide significantly decreased the phosphorylation of tau in rat primary cultured cortical neurons and in APP/PS1 transgenic mice, and this geniposide-induced decrease of tau phosphorylation could be prevented by leptin antagonist (LA). Furthermore, LA also prevented the phosphorylation of Akt at Ser-473 site and GSK-3β at Ser-9 site induced by geniposide. All these results indicate that geniposide may regulate tau phosphorylation through leptin signaling, and geniposide may be a promising therapeutic compound for the treatment of Alzheimer's disease in the future.

Topics: Alzheimer Disease; Animals; Gene Expression Regulation; Glycogen Synthase Kinase 3 beta; Iridoids; Leptin; Mice; Mice, Inbred C57BL; Mice, Transgenic; Neurons; Phosphorylation; Rats; Receptors, Leptin; Signal Transduction; tau Proteins

In order to characterize the pharmacokinetics, bioavailability and tissue distribution of geniposide following intravenous and peroral administration to rats, a reliable gradient HPLC-based method has been developed and validated. After p.o. administration of geniposide, the peak concentration of geniposide in plasma occurred at 1 h and plasma geniposide was eliminated nearly completely within 12 h. The AUC(0→∞) values of geniposide were 6.99 ± 1.27 h · µg/ml and 6.76 ± 1.23 h · µg/ml after i.v. administration of 10 mg/kg and p.o. administration of 100 mg/kg of geniposide, respectively. The absolute oral bioavailability (%F) of geniposide was calculated as 9.67%. After p.o. administration of geniposide, the AUC(0→4h) values in tissues were in the order of kidney > spleen > liver > heart > lung > brain. This study improved the understanding of the pharmacokinetics, bioavailability and tissue distribution of geniposide in rats and may provide a meaningful basis for clinical application of such a bioactive compound of herbal medicines.

Topics: Administration, Intravenous; Administration, Oral; Animals; Biological Availability; Brain; Drugs, Chinese Herbal; Iridoids; Kidney; Liver; Lung; Male; Myocardium; Rats, Sprague-Dawley; Spleen; Tissue Distribution

Gardenia fruits contain valuable natural food colorants including crocins (gardenia yellow) and geniposide. In this study, a process for the enrichment of crocins and geniposide simultaneously from gardenia fruits was developed using macroporous resin and RP chromatography. The performance of eight different types of macroporous resins was evaluated. Static absorption/desorption experiments revealed that LX60 possessed optimal separating capacity. Further dynamic absorption/desorption experiments on LX60 columns were conducted to obtain the optimal parameters. After one run treatment with LX60, the content of crocin-1 in gardenia yellow reached 29.6%, while geniposide in another fraction reached 83.4%. An extract of crocins was obtained from gardenia yellow in a second-stage separation using RP medium-pressure LC, with its color value to be 756 and the content of crocin-1 reaching 60.8%. The separation process was highly efficient, low cost, and compact, which may be informative for purifications of other natural products from complex plant extracts.

Topics: Carotenoids; Chromatography, High Pressure Liquid; Chromatography, Reverse-Phase; Fruit; Gardenia; Iridoids; Plant Extracts; Porosity; Resins, Synthetic

Gardeniae fructus is one of the medicinal herbs that have been used in Far Eastern countries, such as Korea, China, and Japan. Gardeniae fructus is the dried ripe fruit of Gardenia jasminoides Ellis (Rubiaceae) and has been used as a yellow dye. It is widely used as traditional herbal medicine for reducing fever, cholagogue, diuretic and antiphlogistic effects. We established an analytical method that was useful to evaluate the quality control, and standardize quantification monitoring of 68 samples of Gardeniae fructus collected from Korea and China. While numerous previous studies have focused on the simultaneous analysis of geniposide, which constitutes the higher proportion of Gardeniae fructus, and crocin, which determines its color, no simultaneous analysis of gardenoside and geniposide, the major components of Gardeniae Fructus, has been performed. However, previously reported methods are not considered accurate enough because only geniposide or gardenoside was chosen to be the marker component for the quality control of Gardeniae fructus. Thus, we developed the method using simultaneous determination of four components including geniposide, gardenoside, geniposic acid and chlorogenic acid. Against this backdrop, this study aims to propose a new calculation for gardenoside and geniposide concentrations by analyzing their concentrations in Gardeniae fructus.

Topics: Biomarkers; China; Chlorogenic Acid; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Fruit; Gardenia; Iridoid Glucosides; Iridoids; Republic of Korea

The aim of this study was to compare the oral bioavailability and pharmacokinetic data between pure compound of the major active component, single herbal extract and complex herbal formulation by determining bioavailability of geniposide in each group following intravenous and oral administrations.. A conscious and freely moving rat model was used in this experiment to avoid the stress caused by restraint or anaesthesia. The pure compound of geniposide, Gardenia fruits (Chinese name: Zhi-Zi), and extracts of a Gardenia herbal formulation (Chinese name: Zhi-Zi-Chi-Tang) were administered at doses of 200 mg/kg, 4.69 g/kg and 10.82 g/kg for oral administration and fed by gavages to rats, respectively. The earlier doses are equivalent to geniposide administration dose of 200 mg/kg.. The results show that after oral administration of geniposide, Gardenia fruits and Gardenia herbal formulation, the bioavailability were 4.23%, 32.32% and 27.17%, respectively. The results of oral bioavailability of geniposide also suggest that Gardenia fruits extract, single herb, is a more efficient way for geniposide, pure compound, absorption than traditional herbal formulation administration and direct pure compound administration.. The conclusion reveals that herbal ingredient-ingredient or herb-herb interaction may affect the oral absorption of geniposide-related herbal formulation.

Topics: Administration, Oral; Animals; Biological Availability; Drugs, Chinese Herbal; Fruit; Gardenia; Iridoids; Male; Plant Extracts; Rats, Sprague-Dawley

Intracerebral-ventricular (ICV) injection of streptozotocin (STZ) induces an insulin-resistant brain state that may underlie the neural pathogenesis of sporadic Alzheimer disease (AD). Our previous work showed that prior ICV treatment of glucagon-like peptide-1 (GLP-1) could prevent STZ-induced learning memory impairment and tau hyperphosphorylation in the rat brain. The Chinese herbal medicine geniposide is known to relieve symptoms of type 2 diabetes. Because geniposide is thought to act as a GLP-1 receptor agonist, we investigated the potential therapeutic effect of geniposide on STZ-induced AD model in rats. Our result showed that a single injection of geniposide (50 μM, 10 μL) to the lateral ventricle prevented STZ-induced spatial learning deficit by about 40% and reduced tau phosphorylation by about 30% with Morris water maze test and quantitative immunohistochemical analysis, respectively. It has been known that tau protein can be phosphorylated by glycogen synthase kinase-3 (GSK3) and STZ can increase the activity of GSK3β. Our result with Western blot analysis showed that central administration of geniposide resulted in an elevated expression of GSK3β(pS-9) but suppressed GSK3β(pY-216) indicating that geniposide reduced STZ-induced GSK3β hyperactivity. In addition, ultrastructure analysis showed that geniposide averted STZ-induced neural pathology, including paired helical filament (PHF)-like structures, accumulation of vesicles in synaptic terminal, abnormalities of endoplasmic reticulum (ER) and early stage of apoptosis. In summary, our study suggests that the water soluble and orally active monomer of Chinese herbal medicine geniposide may serve as a novel therapeutic agent for the treatment of sporadic AD.

Topics: Alzheimer Disease; Animals; Antibiotics, Antineoplastic; Apoptosis; Disease Models, Animal; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Hippocampus; Iridoids; Male; Maze Learning; Memory Disorders; Microscopy, Electron, Transmission; Phosphorylation; Rats; Rats, Sprague-Dawley; Signal Transduction; Streptozocin; tau Proteins; Vacuoles

Cerebral ischemia is considered to be a highly complex disease resulting from the complicated interplay of multiple pathways. Disappointedly, most of the previous studies were limited to a single gene or a single pathway. The extent to which all involved pathways are translated into fusing mechanisms of a combination therapy is of fundamental importance.. We report an integrative strategy to reveal the additive mechanism that a combination (BJ) of compound baicalin (BA) and jasminoidin (JA) fights against cerebral ischemia based on variation of pathways and functional communities.. We identified six pathways of BJ group that shared diverse additive index from 0.09 to 1, which assembled broad cross talks from seven pathways of BA and 16 pathways of JA both at horizontal and vertical levels. Besides a total of 60 overlapping functions as a robust integration background among the three groups based on significantly differential subnetworks, additive mechanism with strong confidence by networks altered functions.. These results provide strong evidence that the additive mechanism is more complex than previously appreciated, and an integrative analysis of pathways may suggest an important paradigm for revealing pharmacological mechanisms underlying drug combinations.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Disease Models, Animal; Drug Therapy, Combination; Drugs, Chinese Herbal; Flavonoids; Infarction, Middle Cerebral Artery; Iridoids; Male; Mice; Mice, Inbred Strains; Microarray Analysis; Molecular Sequence Data; Principal Component Analysis; Reperfusion Injury; Signal Transduction

A novel method was set up with the aim to obtain a simultaneous cross comparative evaluation of different Gardenia Jasminoides Ellis fruits by the HPTLC fingerprint approach. The main components among the iridoid, hydroxycinnamic derivative and crocin classes were identified by TLC-MS ancillary techniques. The iridoids geniposide, gardenoside and genepin-1-β-d-gentiobioside were also quantitated by densitometric scanning at 240nm. LiChrospher HPTLC Silica gel 60 RP-18 W F254, 20cm×10cm plates with acetonitrile: formic acid 0.1% (40:60 v/v) as the mobile phase was used. The method was validated giving rise to a dependable and high throughput procedure well suited to routine applications. Iridoids were quantified in the range of 240-1140ng with RSD of repeatability and intermediate precision between 0.9-2.5% and accuracy with bias 1.6-2.6%. The method was tested on six commercial Gardenia Jasminoides fruit samples.

Topics: Chromatography, Thin Layer; Fruit; Gardenia; Iridoids; Mass Spectrometry

Geniposide is a bioactive substance derived from gardenia, which has been used in traditional Chinese preparation, such as "Xing-Nao-Jing" (XNJ) for stroke treatment. Stroke and the ingredients of herbal preparation affect the pharmacokinetics of geniposide. A comparative pharmacokinetic study of geniposide in stroke and sham-operated rats after administration of XNJ and geniposide was proceeded to evaluate the effect of stroke on pharmacokinetics of geniposide, while the influence of other components in XNJ was determined by using gardenia extract and geniposide-borneol compounds in rats with stroke to compare.. Stroke was induced by middle cerebral artery occlusion followed by reperfusion 2h later. Plasma concentration of geniposide was determined by HPLC. Various pharmacokinetic parameters were estimated from the plasma concentration versus time data using non-compartmental methods.. The maximum plasma concentration (Cmax) and area under the curve (AUC0-t) in stroke after administration of XNJ were 5.97±3.82 μg/mL, and 570.06±274.32 μg·min/mL, respectively, which were 5 times compared with sham-operated rats or the stroke-afflicted rats given geniposide. In stroke, the Cmax and AUC(0-t) of geniposide-borneol group and gardenia extraction group were close to XNJ group and geniposide group, respectively. The geniposide-borneol group had a higher value.. Stroke improved the absorption of geniposide in XNJ. Borneol may be the key ingredient in XNJ improving the absorption of geniposide.

Topics: Animals; Area Under Curve; Camphanes; Chromatography, High Pressure Liquid; Disease Models, Animal; Drugs, Chinese Herbal; Gardenia; Iridoids; Male; Models, Biological; Rats; Rats, Sprague-Dawley; Stroke

To clarify the protective roles of compatibility of geniposide and ginsenoside (Rg1) in regulating ischemia injured microglia homeostasis by comparing the difference in regulatory roles of geniposide, Rg1, or ginsenoside + Rg1 in balancing secretion of oxygen glucose deprivation induced microglia inflammatory cytokines.. The mimic ischemia injured microglia model was induced by oxygen-glucose deprivation (OGD). Then geniposide, Rg1, or ginsenoside + Rg1 (Tongluo Jiunao Injection, TJI) was respectively added. The NO content was determined by Griess Reagent. The cyto activity was detected using cell count kit. Contents of TNF-alpha and TGF-beta and their expression levels were detected by ELISA and Western blot.. Geniposide + Rg1 could significantly inhibit the release of NO, elevate the TGF-beta level, and decrease the content of TNF-alpha without influencing the cell survival. The two active ingredients played different therapeutic roles. The compatible use was obviously superior to use any one of the two active ingredients alone.. Geniposide, Rg1, or Ginsenoside + Rg1 had regulating roles in balancing ischemia injured microglia homeostasis. Its mechanisms might be related to up-regulating the TGF-beta expression and down-regulating TNF-alpha expression.

Topics: Animals; Ginsenosides; Hypoxia; Iridoids; Mice; Microglia; Nitric Oxide; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Geniposide (GE), an iridoid glycoside compound, is the major active ingredient of Gardenia jasminoides Ellis (GJ) fruit which has anti-inflammatory and other important therapeutic activities. The aim of this study was to investigate the effects of GE on adjuvant arthritis (AA) rats and its possible mechanisms. AA was induced by injecting with Freund's complete adjuvant (FCA). Male SD rats were subjected to treatment with GE at 30, 60 and 120mg/kg from days 18 to 24 after immunization. Lymphocyte proliferation was assessed by MTT. Interleukin (IL)-6, IL-17, IL-4 and transforming growth factor-beta 1 (TGF-β1) were determined by ELISA. c-Jun N-terminal kinase (JNK) and phospho-JNK (p-JNK) were detected by Western blot. GE (60, 120mg/kg) significantly relieved the secondary hind paw swelling and arthritis index, along with decreased Th17-cells cytokines and increased Treg-cell cytokines in mesenteric lymph node lymphocytes (MLNL) and peripheral blood lymphocytes (PBL) of AA rats. In addition, GE decreased the expression of p-JNK in MLNL and PBL of AA rats. In vivo study, it was also observed that GE attenuated histopathologic changes of MLN in AA rats. Collectively, GE might exert its anti-inflammatory and immunoregulatory effects through inducing Th17 cell immune tolerance and enhancing Treg cell-mediated activities by down-regulating the expression of p-JNK. The mechanisms of GE on JNK signaling in MLNL and PBL may play critical roles in the pathogenesis of rheumatoid arthritis.

Topics: Animals; Anti-Inflammatory Agents; Arthritis, Experimental; Arthritis, Rheumatoid; Cell Proliferation; Cytokines; Foot Joints; Iridoids; JNK Mitogen-Activated Protein Kinases; Lymph Nodes; Lymphocytes; Male; Rats, Sprague-Dawley

In this study we examined the inhibition of hepatic dyslipidemia by Eucommia ulmoides extract (EUE). Using a screening assay for BAX inhibition we determined that EUE regulates BAX-induced cell death. Among various cell death stimuli tested EUE regulated palmitate-induced cell death, which involves lysosomal BAX translocation. EUE rescued palmitate-induced inhibition of lysosomal V-ATPase, α-galactosidase, α-mannosidase, and acid phosphatase, and this effect was reversed by bafilomycin, a lysosomal V-ATPase inhibitor. The active components of EUE, aucubin and geniposide, showed similar inhibition of palmitate-induced cell death to that of EUE through enhancement of lysosome activity. Consistent with these in vitro findings, EUE inhibited the dyslipidemic condition in a high-fat diet animal model by regulating the lysosomal localization of BAX. This study demonstrates that EUE regulates lipotoxicity through a novel mechanism of enhanced lysosomal activity leading to the regulation of lysosomal BAX activation and cell death. Our findings further indicate that geniposide and aucubin, active components of EUE, may be therapeutic candidates for non-alcoholic fatty liver disease.

Topics: Animals; bcl-2-Associated X Protein; Cathepsin B; Cell Death; Diet, High-Fat; Enzyme Inhibitors; Eucommiaceae; Female; Humans; Iridoid Glucosides; Iridoids; Liver; Lysosomes; Non-alcoholic Fatty Liver Disease; Palmitates; Rats; Rats, Sprague-Dawley

An extracellular β-glucosidase from Aspergillus niger Au0847 was purified to homogeneity by precipitation with ammonium sulfate, anion exchange, and gel filtration. The purified protein was composed of two subunits with molecular masses of 110 and 120 kDa. Au0847 β-glucosidase exhibited relatively high thermostability and pH stability, and its highest activity was obtained at 65°C and pH 4.6, respectively. As a potential metalloprotein, its enzymatic activity was potently stimulated by manganese ion and DTT. The β-glucosidase displayed avid affinity and high catalytic efficiency for geniposide. Au0847 β-glucosidase has potential value as an industrial enzyme for the hydrolysis of geniposide to genipin.

Topics: Aspergillus niger; Biotransformation; Enzyme Stability; Fungal Proteins; Glucosidases; Iridoids; Kinetics

Atherosclerosis is a systemic inflammatory disease characterized by the accumulation of dendritic cells (DCs) and other types of immune cells in atherosclerotic plaque. In this study, baicalin and geniposide were isolated from Scutellaria baicalensis Georgi and Gardenia jasminoids Ellis, which are the plants used in traditional Chinese medicine to treat a variety of inflammatory diseases. We then investigated whether baicalin and geniposide could induce regression of atherosclerotic lesions in ApoE-/- mice fed a high cholesterol diet and used as a model of atherosclerosis. Following model induction, these mice were treated with baicalin (100mg/kg), geniposide (100mg/kg), and then a mixture containing baicalin (100mg/kg) and geniposide (100mg/kg) administered daily by gavage for a period of 12weeks. The combined administration of baicalin and geniposide significantly reduced atherosclerotic lesions, and modulated the phenotype of dendritic cells in bone marrow and atherosclerotic plaque. Geniposide lowered both plasma lipid levels and DC numbers, while baicalin administered either alone or in combination with geniposide did not decrease plasma lipids. Our results suggest that baicalin and geniposide may have immune-regulatory effects and prevent the formation of atherosclerotic lesions by decreasing the DC numbers, and inhibit DC maturation in bone marrow and infiltration into lesions.

Topics: Administration, Oral; Animals; Aorta, Abdominal; Aorta, Thoracic; Apolipoproteins E; Atherosclerosis; Dendritic Cells; Diet, High-Fat; Flavonoids; Iridoids; Lipids; Male; Mice, Inbred C57BL; Mice, Knockout

The herbal medicine Tong Luo Jiu Nao (TLJN) contains geniposide (GP) and ginsenoside Rg1 at a molar ratio of 10:1. Rg1 is the major component of another herbal medicine, panax notoginseng saponin (PNS). TLJN has been shown to strengthen brain function in humans, and in animals it improves learning and memory. We have previously shown that TLJN reduces amyloidogenic processing in Alzheimer's disease (AD) mouse models. Together this suggests TLJN may be a potential treatment for patients with dementia. Because chronic damage of the central nervous system by formaldehyde (FA) has been presented as a risk factor for age-associated cognitive dysfunction, in the present study we investigated the protective effect of both TLJN and GP in neuron-like cells exposed to FA. FA-exposed murine N2a neuroblastoma cells were incubated with TLJN, its main ingredient GP, as well as PNS, to measure cell viability and morphology, the rate of apoptosis and expression of genes encoding Akt, FOXO3, Bcl2 and p53. The CCK-8 assay, cytoskeletal staining and flow cytometry were used to test cell viability, morphology and apoptosis, respectively. Fluorescent quantitative real-time PCR (qRT-PCR) was used to monitor changes in gene expression, and HPLC to determine the rate of FA clearance. Treatment of N2a cells with 0.09 mmol L(-1) FA for 24 h significantly reduced cell viability, changed cell morphology and promoted apoptosis. Both TLJN and GP conferred neuroprotection to FA-treated N2a cells, whereas PNS, which had to be used at lower concentrations because of its toxicity, did not. Our data demonstrate that TLJN can rescue neuronal damage caused by FA and that its main ingredient, GP, has a major role in this efficacy. This presents purified GP as a drug or lead compound for the treatment of AD.

Topics: Animals; Apoptosis; Cell Line, Tumor; Cell Survival; Central Nervous System; Chromatography, High Pressure Liquid; Cognition Disorders; Cytoskeleton; Dementia; Formaldehyde; Iridoids; Learning; Medicine, Chinese Traditional; Memory; Mice; Neuroblastoma; Plant Preparations; Real-Time Polymerase Chain Reaction

Geniposide has various pharmacological effects; however, low oral bioavailability limits its clinical utility. This study explores the pharmacokinetics, tissue distribution and relative bioavailability of geniposide-solid lipid nanoparticles (SLNs) following oral administration. The geniposide solution and geniposide-SLNs were orally administered to the rats, respectively. The Cmax value of geniposide in the geniposide-SLNs was significantly higher than that obtained with geniposide solution. Compared with the geniposide solution, the t1/2 and MRT were prolonged; the CL and V1/F were increased with geniposide-SLNs. The AUC0-∞values of geniposide-SLNs were 50 times greater than geniposide solution. The ratios of AUC0-8 h in the liver, spleen, heart, kidney, brain and lung of the geniposide-SLNs to geniposide solution were 25.93, 4.28, 27.91, 10.15, 5.16 and 16.22, respectively. Prepared geniposide-SLNs are very helpful for increasing the bioavailability of geniposide. These data suggest that SLNs are a promising delivery system to enhance the oral bioavailability of geniposide.

Topics: Administration, Oral; Animals; Biological Availability; Drug Carriers; Iridoids; Lipids; Nanoparticles; Organ Specificity; Rats; Rats, Sprague-Dawley

Huanglian-Zhizi couplet medicine comes from classical prescription Huang-Lian-Jie-Du-Tang (HLJDT), which has been proven by previous researches to be an effective compound for cerebral ischemia. This paper explores the integrated pharmacokinetics of gardenia acid and geniposide-time-antioxidant efficacy after the oral administration of Huanglian-Zhizi couplet medicine from HLJDT in rats with middle cerebral artery occlusion (MCAO). To investigate the differences in pharmacokinetics and antioxidant effect of Huanglian-Zhizi and HLJDT in MCAO rats, which have been scarcely reported, an oral dose, 24 crud drug g/kg, of Huanglian-Zhizi and 40 crud drug/kg of HLJDT were administered in two groups of normal rats and two groups of Sprague-Dawley (SD) MCAO rats, respectively. At different time points, concentrations of gardenia acid and geniposide were determined by high performance liquid chromatography (HPLC), and levels of superoxide dismutase (SOD) were calculated by ELIASA. Pharmacokinetic parameters including AUC, MRT, t1/2, T max , C max were estimated by statistical moment analysis using a data analysis system (DAS) 2.0. An AUC based on weighting approach was used for integrating gardenia acid and geniposide. Finally, the concentration-time efficacy profiles were obtained. The integrated pharmacokinetics profiles of index components could reveal the pharmacokinetics behavior of Huanglian-Zhizi and HLJDT, corresponding to the antioxidant efficacy.

Topics: Administration, Oral; Animals; Antioxidants; Chromatography, High Pressure Liquid; Disease Models, Animal; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Enzyme-Linked Immunosorbent Assay; Infarction, Middle Cerebral Artery; Iridoids; Phytotherapy; Rats, Sprague-Dawley; Superoxide Dismutase; Time Factors

Inflammatory responses are important to host immune reactions, but uncontrolled inflammatory mediators may aid in the pathogenesis of other inflammatory diseases. Geniposide, an iridoid glycoside found in the herb gardenia, is believed to have broad-spectrum anti-inflammatory effects in murine models but its mechanism of action is unclear. We investigated the action of this compound in murine macrophages stimulated by lipopolysaccharide (LPS), as the stimulation of macrophages by LPS is known to induce inflammatory reactions. We determined the effect of geniposide on LPS-induced production of the inflammatory mediators, nitric oxide (NO) and prostaglandin E2 (PGE2), the mRNA and protein expression of the NO and PGE2 synthases, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), respectively, and the mRNA and protein expression of the inflammatory cytokine, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Furthermore, nuclear factor (NF)-κB, mitogen-activated protein kinase (MAPK) and activator protein (AP)-1 activity were assayed. To understand the action of geniposide on the NF-κB and MAPK pathways, we studied the effect of NF-κB and MAPK inhibitors on the LPS-induced production of NO, PGE2 and TNF-α. Our findings clearly showed that geniposide mainly exerts its anti-inflammatory effects by inhibiting the LPS-induced NF-κB, MAPK and AP-1 signaling pathways in macrophages, which subsequently reduces overexpression of the inducible enzymes iNOS and COX-2 and suppresses the expression and release of the inflammatory factors, TNF-α, IL-6, NO and PGE2. Thus, geniposide shows promise as a therapeutic agent in inflammatory diseases.

Topics: Animals; Anti-Inflammatory Agents; Butadienes; Cell Line; Cyclooxygenase 2; Dinoprostone; Extracellular Signal-Regulated MAP Kinases; Gardenia; Imidazoles; Inflammation Mediators; Interleukin-6; Iridoids; Lipopolysaccharides; Macrophages, Peritoneal; Male; Mice; Mice, Inbred Strains; NF-kappa B; Nitric Oxide; Nitric Oxide Synthase Type II; Nitriles; Pyridines; Signal Transduction; Sulfones; Transcription Factor AP-1; Tumor Necrosis Factor-alpha

We recently discovered that the activation of the spinal glucagon-like peptide-1 receptors (GLP-1Rs) by the peptidic agonist exenatide produced antinociception in chronic pain. We suggested that the spinal GLP-1Rs are a potential target molecule for the management of chronic pain. This study evaluated the antinociceptive activities of geniposide, a presumed small molecule GLP-1R agonist. Geniposide produced concentration-dependent, complete protection against hydrogen peroxide-induced oxidative damage in PC12 and HEK293 cells expressing rat and human GLP-1Rs, but not in HEK293T cells that do not express GLP-1Rs. The orthosteric GLP-1R antagonist exendin(9-39) right-shifted the concentration-response curve of geniposide without changing the maximal protection, with identical pA2 values in both cell lines. Subcutaneous and oral geniposide dose-dependently blocked the formalin-induced tonic response but not the acute flinching response. Subcutaneous and oral geniposide had maximum inhibition of 72% and 68%, and ED50s of 13.1 and 52.7 mg/kg, respectively. Seven days of multidaily subcutaneous geniposide and exenatide injections did not induce antinociceptive tolerance. Intrathecal geniposide induced dose-dependent antinociception, which was completely prevented by spinal exendin(9-39), siRNA/GLP-1R and cyclic AMP/PKA pathway inhibitors. The geniposide iridoid analogs geniposidic acid, genipin methyl ether, 1,10-anhydrogenipin, loganin and catalpol effectively inhibited hydrogen peroxide-induced oxidative damage and formalin pain in an exendin(9-39)-reversible manner. Our results suggest that geniposide and its iridoid analogs produce antinociception during persistent pain by activating the spinal GLP-1Rs and that the iridoids represented by geniposide are orthosteric agonists of GLP-1Rs that function similarly in humans and rats and presumably act at the same binding site as exendin(9-39).

Topics: Analgesics; Animals; Central Nervous System Agents; Exenatide; Formaldehyde; Glucagon-Like Peptide-1 Receptor; HEK293 Cells; Heterocyclic Compounds, 3-Ring; Humans; Iridoid Glucosides; Iridoids; Male; Mice; Nociception; PC12 Cells; Peptide Fragments; Peptides; Rats; Rats, Wistar; Receptors, Glucagon; Spinal Cord; Venoms

Geniposide is a medicine isolated from Gardenia jasminoides Ellis, which is a traditional Chinese herb that is widely used in Asia for the treatment of inflammation, brain diseases, and hepatic disorders. Mastitis is a highly prevalent and important infectious disease. In this study, we used a lipopolysaccharide (LPS)-induced mouse mastitis model and LPS-stimulated primary mouse mammary epithelial cells (mMECs) to explore the anti-inflammatory effect and the mechanism of action of geniposide. Using intraductal injection of LPS as a mouse model of mastitis, we found that geniposide significantly reduced the infiltration of inflammatory cells and downregulated the production of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6). To further investigate the anti-inflammatory mechanism, we used LPS-stimulated mMECs as an in vitro mastitis model. The results of enzyme-linked immunosorbent assay (ELISA) and quantitative real-time polymerase chain reaction (qRT-PCR) showed that geniposide inhibited the expression of TNF-α, IL-1β, and IL-6 in a dose-dependent manner. Western blot analysis demonstrated that geniposide could suppress the phosphorylation of inhibitory kappa B (IκBα), nuclear factor-κB (NF-κB), p38, extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK). Geniposide also inhibited the expression of toll-like receptor 4 (TLR4) in the LPS-stimulated mMECs. In conclusion, geniposide exerted its anti-inflammatory effect by regulating TLR4 expression, which affected the downstream NF-κB and mitogen-activated protein kinase (MAPK) signaling pathways. Thus, geniposide may be a potential drug for mastitis therapy.

Topics: Animals; Anti-Inflammatory Agents; Cells, Cultured; Female; Iridoids; Lipopolysaccharides; Male; Mastitis; Mice; Mice, Inbred BALB C; Pregnancy; Signal Transduction; Toll-Like Receptor 4

Pharmacochemistry and integrated pharmacokinetics of six alkaloids (groenlandicine, berberine, palmatine, epiberberine, jatrorrhizine, and columbamine) after oral administration of Huang-Lian-Jie-Du-Tang (HLJDT) decoction were investigated in this paper. The method of plasma pharmacochemistry was applied to predict the potential bioactive components in HLJDT decoction. Based on the accurate molecular weight, 10 components including 2 flavonoids (baicalin and wogonoside), 1 iridoid glycoside (geniposide), and 7 alkaloids (above-mentioned 6 alkaloids and coptisine) were structurally identified. Then, integrated pharmacokinetics of the alkaloids in Sprague-Dawley rats after oral administration of HLJDT decoction was investigated by HPLC method. The results showed that the pharmacokinetic behaviors of the alkaloids were different although their chemical structures were similar. This study developed a method to predict the potential bioactive components in HLJDT decoction and research the pharmacokinetic behaviors of the potential bioactive components.

Topics: Administration, Oral; Alkaloids; Animals; Berberine; Berberine Alkaloids; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavanones; Flavonoids; Glucosides; Iridoids; Molecular Structure; Rats; Rats, Sprague-Dawley

The neuroinflammation induced by amyloid-β (Aβ) is one of the key events in Alzheimer's disease (AD) progress in which microglia are the main cells involved. Receptor for advanced glycation end products (RAGE) mediates and enhances Aβ-induced microglial activation and leads to induction of proinflammatory mediators, such as tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). Geniposide, a pharmacologically active component purified from gardenia fruit, exhibits a broad spectrum anti-inflammatory effect as well as neurotrophic and neuroprotective properties. However, the effects of geniposide on Aβ-mediated microglial pathways have not been fully discovered. Here, we demonstrate that geniposide treatment significantly blocks Aβ-induced RAGE-dependent signaling (activation of ERK and NF-κB) along with the production of TNF-α and IL-1β in cultured BV2 microglia cells. Notably, based on the data from coimmunoprecipitation assay, we infer that geniposide exerts protective effects on Aβ-induced inflammatroy response through blocking Aβ binding to RAGE and suppressing the RAGE-mediated signaling pathway. Taken together, these findings indicate that geniposide is a potent suppressor of neuroflammation through inhibiting RAGE-dependent signaling pathway. Thus, geniposide may be a potential therapeutic agent for the treatment of neuroinflammation that is involved in neurological diseases such as AD.

Topics: Amyloid beta-Peptides; Animals; Cell Count; Cell Line; Dose-Response Relationship, Drug; Drug Interactions; Gene Expression Regulation; Green Fluorescent Proteins; Humans; Immunoprecipitation; Interleukin-1beta; Iridoids; Mice; Microglia; Neuroblastoma; Peptide Fragments; Receptor for Advanced Glycation End Products; Receptors, Immunologic; RNA, Messenger; Signal Transduction; Transfection; Tumor Necrosis Factor-alpha

Type 2 diabetes mellitus is characterized by lack of, or relative deficiency in, insulin productions and insensitivity of target tissues to insulin. Improvement of β-cell functions is a potential strategy for the clinical management of this disease. We reported before that geniposide improved glucose-stimulated insulin secretion with the activation of glucagon-like peptide 1 receptor (GLP-1R) in INS-1 pancreatic β cells, but the cell signaling mechanism of geniposide regulating glucose-stimulated insulin secretion (GSIS) in β cells is so far poorly understood.. Effect of LY294002, a specific inhibitor of PI3K, on GSIS in the presence or absence of geniposide in INS-1 cells. In addition, the differential protein expression of geniposide treated INS-1 cells was examined by Western blot.. After pretreatment with 10 µM LY294002 for 1 hour, the insulin secretion induced by geniposide was partly abolished in INS-1 cells. After treatment with geniposide, the phosphorylation of PDK1 and Akt473 increased gradually to the maximum at 60 minutes or 120 minutes respectively. Furthermore, geniposide also inhibited the phosphorylation of downstream target GSK3β, and this effect was counteracted by preincubation with LY294002. And the expression of GLUT2 was increased after treatment with different doses geniposide.. Geniposide increases insulin secretion in pancreatic β cells in a PI3K dependent mechanism potentially through increased GLUT2 protein levels.

Topics: 3-Phosphoinositide-Dependent Protein Kinases; Animals; Cell Line; Glucagon-Like Peptide-1 Receptor; Glucose; Glucose Transporter Type 2; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Hypoglycemic Agents; Insulin; Insulin Secretion; Insulin-Secreting Cells; Iridoids; Phosphatidylinositol 3-Kinase; Phosphoinositide-3 Kinase Inhibitors; Phosphorylation; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-akt; Rats; Receptors, Glucagon; Signal Transduction; Time Factors; Up-Regulation

Geniposide (GE), also called Jasminoidin, is the major active ingredient of Gardenia jasminoides Ellis (GJ) fruit, which has long been used in traditional Chinese medicine (TCM). Growing evidences suggested that GE has a great potentiality for treating rheumatoid arthritis (RA). However, GE is rapidly metabolized, and we know little about its availability or metabolites in tissues. To elucidate the distribution of GE and its metabolites in tissues, three groups of adjuvant arthritis (AA) rats were given GE (33, 66 and 120 mg/kg) from days 18 to 24, and the biotransformation of GE in plasma, liver, spleen, synovium, urine and mesenteric lymph node (MLN) of rats was investigated by a novel approach named Information-Dependent Acquisition (IDA)-Mediated LC-MS/MS method. As a result, GE and its four major metabolites were detected as follows: GE, G1, G2 in plasma; GE, G2 in MLNs; only GE in liver and synovium; GE, G2, G3 and G4 in spleen; and GE, G1, G2 and G4 in urine. In total four metabolites (G1-G4) involved in the in vivo metabolism processes were identified. The results of this work have demonstrated the IDA-Mediated LC-MS/MS could screen rapidly and reliably the characterization of metabolites from iridoid compounds.

Topics: Animals; Arthritis, Experimental; Iridoids; Liver; Lymph Nodes; Male; Random Allocation; Rats, Sprague-Dawley; Spleen; Synovial Membrane

Cortex Eucommia has been used as a kidney-tonifying herbal medicine with a long history of compatibility with Fructus Psoraleae. Geniposide (GP) and geniposidic acid (GPA) are the two main chemical components in Cortex Eucommia. In the present study, the effects of Fructus Psoraleae extract (FPE) on intestinal absorption kinetics of GP and GPA in rat were investigated. Twenty four male Sprague-Dawley rats were randomly assigned into four groups which were treated with GP, GPA, GP mixed with FPE and GPA mixed with FPE, respectively, by in situ intestinal perfusion for 3 h. The samples of intestinal perfusion solutions were collected every 30 min, and analyzed by ultra high performance liquid chromatography (UPLC). The curves of time and residual quantities of GP and GPA (lnx) in the intestinal perfusion solution and the cumulative absorption rate were obtained. The results showed that FPE exhibited different effects on the intestinal absorption of GP and GPA in rat: it increased the intestinal absorption of GP (p<0.05), while demonstrated no significant effect on the absorption of GPA.

Topics: Animals; Chromatography, High Pressure Liquid; Intestinal Absorption; Iridoid Glucosides; Iridoids; Kinetics; Male; Plant Extracts; Psoralea; Rats; Rats, Sprague-Dawley

An HPLC method for the determination of geniposide concentration in mouse plasma was developed and the pharmacokinetics after intranasal administration of Xingnaojing microemulsion (XNJ-M) and mPEG2000-PLA modified Xingnaojing microemulsion (XNJ-MM) were investigated. Eighty mice were treated by XNJ-M and XNJ-MM nasally. The plasma samples were collected at different times and the drug in samples was detected by HPLC. The pharmacokinetic parameters were calculated by the software of Kinetica. The pharmacokinetic parameters of geniposide of XNJ-M were C(max) (4.36 +/- 2.69) mg x L(-1), t(max) 1 min, MRT (29.73 +/- 4.54) min, AUC (53.63 +/- 14.03) mg x L(-1) x min. The pharmacokinetic parameters of geniposide of XNJ-MM were C(max) (9.75 +/- 4.14) mg x L(-1), t(max) 1 min, MRT(22.34 +/- 2.90) min, AUC (131.87 +/- 40.13) mg x L(-1) x min. Geniposide can be absorbed into blood in a higher degree after intranasal administration with XNJ-MM compared to XNJ-M, which maybe caused by its less irritating and more absorption.

Topics: Animals; Drugs, Chinese Herbal; Emulsions; Iridoids; Lactic Acid; Male; Mice; Polyesters; Polyethylene Glycols; Polymers

Baicalin and geniposide, which are respectively isolated from Scutellariae radix and Gardenia jasminoides, have been known to exhibit a number of pharmacological effects, including anti-inflammatory and anti-oxidant. Here, we primarily aimed to observe the protective effects of these two Chinese herbs on inhibiting the development of atherosclerosis in apolipoprotein E knockout mice via lipids regulation and immunoregulation. After the ApoE-/- mice with high-cholesterol diet had received 12-weeks׳ oral administration of either baicalin or geniposide (100 mg/kg), atherosclerotic plaque areas in aorta were measured and exhibited a prominent decrease in the treated mice. We then assayed serum lipids levels, serum Treg-cell-associated cytokines (TGF-β1 and IL-10) and the frequency of splenic Treg cells. We found that geniposide notably decreased serum TC and LDL-c. Both baicalin and geniposide treated mice showed much more splenic Treg cells and the correlated cytokines (TGF-β1 and IL-10). Foxp3, as the marker of Treg cell, was detected in atherosclerotic lesions, and we found that Foxp3 expression at both mRNA and protein levels was up-regulated in addition to increased Foxp3 positive Treg cells detected by immunohistochemistry in baicalin or geniposide treated mice. In conclusion, baicalin and geniposide up-regulated the expression of foxp3, promoted the number and function of Treg cells and ameliorated the atherosclerotic lesions progression partly through lipids regulation and immunoregulation.

Topics: Animals; Aorta; Apolipoproteins E; Atherosclerosis; Flavonoids; Forkhead Transcription Factors; Interleukin-10; Iridoids; Lipids; Male; Mice, Inbred C57BL; Mice, Knockout; Myocardium; Phytotherapy; Spleen; T-Lymphocytes, Regulatory; Transforming Growth Factor beta1

Geniposide is widely used in the treatment of cerebral ischemic stroke and cerebrovascular diseases for its anti-thrombotic and anti-inflammatory effects. Recent studies demonstrated that geniposide could be absorbed promptly and thoroughly by intranasal administration in mice and basically transported into the brain. Here, we explored its transport mechanism and the effect of borneol and muscone on its transport by human nasal epithelial cell (HNEC) monolayer. The cytotoxicity of geniposide, borneol, muscone and their combinations on HNECs was evaluated by the MTT assay. Transcellular transport of geniposide and the influence of borneol and muscone were studied using the HNEC monolayer. Immunostaining and transepithelial electrical resistance were measured to assess the integrity of the monolayer. The membrane fluidity of HNEC was evaluated by fluorescence recovery after photobleaching. Geniposide showed relatively poor absorption in the HNEC monolayer and it was not a P-gp substrate. Geniposide transport in both directions significantly increased when co-administrated with increasing concentrations of borneol and muscone. The enhancing effect of borneol and muscone on geniposide transport across the HNEC may be attributed to the significant enhancement on cell membrane fluidity, disassembly effect on tight junction integrity and the process was reversible. These results indicated that intranasal administration has good potential to treat cerebrovascular diseases.

Topics: Actins; Biological Transport; Camphanes; Cell Membrane Permeability; Cell Survival; Cells, Cultured; Chromatography, High Pressure Liquid; Cycloparaffins; Epithelial Cells; Humans; Iridoids; Microscopy, Fluorescence; Nasal Mucosa

Oxidative stress and mitochondrial dysfunction appear early and contribute to the disease progression in Alzheimer's disease (AD), which can be detected extensively in AD patients brains as well as in transgenic AD mice brains. Thus, treatments that result in attenuation of oxidative stress and mitochondrial dysfunction may hold potential for AD treatment. Geniposide, a pharmacologically active component purified from gardenia fruit, exhibits anti-oxidative, antiinflammatory and other important therapeutic properties. However, whether geniposide has any protective effect on oxidative stress and mitochondrial dysfunction in AD transgenic mouse model has not yet been reported. Here, we demonstrate that intragastric administration of geniposide significantly reduces oxidative stress and mitochondrial dysfunction in addition to improving learning and memory in APP/PS1 mice. Geniposide exerts protective effects on mitochondrial dysfunction in APP/PS1 mice through suppressing the mitochondrial oxidative damage and increasing the mitochondrial membrane potential and activity of cytochrome c oxidase. These studies indicate that geniposide may attenuate memory deficits through the suppression of mitochondrial oxidative stress. Thus, geniposide may be a potential therapeutic reagent for halting and preventing AD progress.

Topics: Alzheimer Disease; Amyloid beta-Protein Precursor; Animals; Brain; Disease Models, Animal; Electron Transport Complex IV; Humans; Iridoids; Male; Malondialdehyde; Maze Learning; Membrane Potential, Mitochondrial; Memory Disorders; Mice, Inbred C57BL; Mice, Transgenic; Mitochondria; Nootropic Agents; Oxidative Stress; Presenilin-1; Random Allocation; Reactive Oxygen Species

To study the effects of borneol and muscone on membrane fluidity, membrane potential, Na+, K(+)-ATPase activity and calcium ions of blood brain barrier (BBB) model cells (MDCK and MDCK-MDR1) for exploring the mechanism of aromatics.. MDCK and MDCK-MDR1 cells were incubated and the experiment was performed as following. Cells were incubated with aromatic herbs for 3 h. The membrane fluidity were detected by FRAP. The changes of membrane potentials and the concentration of calcium ions were detected by flow cytometer.. Borneol (55.6, 111.2 microg/mL) and muscone (8.34, 16.68 microg/mL) significantly enhanced the cell membrane fluidity. Borneol (27.8, 55.6, 111.2 microg/mL) and muscone (4.17, 8.34, 16.68 microg/mL) made the MDCK and MDCK-MDR1 membrane potentials less negative or depolarized. Borneol increased the concentration of intracellular free calcium in MDCK while decreased the concentration of intracellular free calcium in MDCK-MDR1 cells. Muscone increased the concentration of calcium in MDCK and MDCK-MDR1 cells. Na+, K(+)-ATPase activity was significantly increased in borneol and muscone group.. The regulating effect of membrane fluidity, membrane potential, Na+, K(+)-ATPase activity and calcium ions in MDCK and MDCK-MDR1 cells might be one of the mechanisms of borneol and muscone's BBB opening function.

Topics: Animals; Biological Transport; Blood-Brain Barrier; Calcium; Camphanes; Cell Membrane; Cell Membrane Permeability; Cycloparaffins; Dogs; Dose-Response Relationship, Drug; Drug Interactions; Iridoids; Madin Darby Canine Kidney Cells; Membrane Potentials; Models, Biological; Sodium-Potassium-Exchanging ATPase

Geniposide, Geniposide, the main active component in extracts of Gardenia jasminoides ELLIS., is one of the main components of Huanglian-Jiedu-Tang (HJT). This study aimed to validate an indirect competitive enzyme-linked immunosorbent assay (icELISA) based on monoclonal antibodies (mAb) against geniposide (anti-geniposide mAb), which was developed by our lab, and apply the assay to study the pharmacokinetics of geniposide in HJT in mice. Blood samples were drawn from mice at predetermined time points after oral administration of HJT in three dosages. A linear correlation was obtained for geniposide concentrations in the range from 1.17 to 37.50 µg/mL. The intra-day and inter-day precision values of the icELISA method were well within the recommended range (≤10%). The recovery rates ranged from 99.74 to 102.40%. Stability studies showed that geniposide sample solutions were intact for 12 h. The Tmax and mean residence time (MRT) of geniposide of the three groups were consistent with previous data. The results suggest that a reliable and effective method was established and could be applied to the study of the pharmacokinetics of geniposide in HJT.

Topics: Administration, Oral; Animals; Antibodies, Monoclonal; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Enzyme-Linked Immunosorbent Assay; Iridoids; Male; Mice, Inbred BALB C; Reproducibility of Results

Topics: Animals; Brain; Brain Ischemia; Drug Synergism; Gene Regulatory Networks; Iridoids; Linear Models; Male; Mice; Models, Genetic; Oligonucleotide Array Sequence Analysis; Reperfusion Injury; Signal Transduction; Transcriptome; Ursodeoxycholic Acid

This study investigated the effects of intestinal microbiota on the metabolism of geniposide by using a rat model treated with a mixture of antibiotics. The plasma concentration of geniposide was determined after oral administration in control and antibiotics-treated rats by using liquid chromatography-tandem mass spectrometry. The maximum plasma concentrations (Cmax) of geniposide in control and antibiotics-treated rats were 0.91 ± 0.26 and 1.01 ± 0.04 μg/mL, respectively, and the area under the curve (AUC) values were 7.34 ± 3.32 and 11.9 ± 2.1 μg·h/mL (p < 0.05), respectively. The levels of geniposide in rat feces were 0.64 and 15.6 mg, respectively, in the control and antibiotics-treated groups. Thus, the systemic exposure of geniposide was greater in the antibiotics-treated rats. This may be due to the antibiotic-induced suppression of the metabolic activities of the intestinal microbiota. These results suggest that the gut microbiota may have an impact on the bioavailability of geniposide.

Topics: Administration, Oral; Animals; Biological Availability; Intestines; Iridoids; Male; Microbiota; Rats, Sprague-Dawley

Evaluating the safety of traditional medicinal herbs and their major active constituents is critical for their widespread usage. Geniposide, a major active constituent with a defined structure from the traditional medicinal herb Gardenia jasminoides ELLIS fruit, exhibits remarkable anti-inflammatory, antiapoptotic, and antifibrotic properties and has been used in a variety of medical fields, mainly for the treatment of liver diseases. However, geniposide-induced hepatotoxicity and methods for the early detection of hepatotoxicity have yet to be reported. In this study, geniposide-induced hepatotoxicity was investigated. In addition, candidate biomarkers for the earlier detection of geniposide-induced hepatotoxicity were identified using a label-free quantitative proteomics approach on a geniposide overdose-induced liver injury in a rat model. Using an accurate intensity-based, absolute quantification (iBAQ)-based, one-step discovery and verification approach, a candidate biomarker panel was easily obtained from individual samples in response to different conditions. To determine the biomarkers' early detection abilities, five candidate biomarkers were selected and tested using enzyme-linked immunosorbent assays (ELISAs). Two biomarkers, glycine N-methyltransferase (GNMT) and glycogen phosphorylase (PYGL), were found to indicate hepatic injuries significantly earlier than the current gold standard liver biomarker. This study provides a first insight into geniposide-induced hepatotoxicity in a rat model and describes a method for the earlier detection of this hepatotoxicity, facilitating the efficient monitoring of drug-induced hepatotoxicity.

Topics: Animals; Biomarkers; Chemical and Drug Induced Liver Injury; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Glycine N-Methyltransferase; Glycogen Phosphorylase; Iridoids; Liver; Male; Proteome; Proteomics; Rats, Sprague-Dawley; Reproducibility of Results; Sensitivity and Specificity

Geniposide, a major iridoid glycoside found in gardenia fruit, is widely used in Asian countries for its anti-inflammatory, anti-tumor and anti-apoptotic activities. Although the anti-inflammatory effect of geniposide has been widely reported, its anti-apoptotic role in mastitis remains unclear. In the present study, we investigated whether geniposide exerts anti-apoptotic activity in lipopolysaccharide (LPS)-induced mouse mammary glands.. We established a LPS-induced mouse mastitis model and LPS-stimulated primary mouse mammary epithelial cells (mMECs) model to investigate the anti-apoptotic effect of geniposide and the underlying mechanism of action. In the in vivo studies, apoptosis in mammary glands was detected by TUNEL. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to analyze the expression of Bax, Bcl-2, Caspase-3 and p53. In the in vitro study, the apoptosis in mammary epithelial cells was measured by Live-Dead staining. Western blot and qRT-PCR analysis were used to analyze the expression of Bax, Bcl-2, Caspase-3, p53 and TLR4.. Geniposide alleviated mammary gland apoptosis, down-regulated Bax expression, inhibited Caspase-3 cleavage and p53 phosphorylation and up-regulated Bcl-2 expression in vivo. In vitro, geniposide decreased the ratio of dead cells in a dose-dependent manner. Geniposide inhibited Bax expression and Caspase-3 cleavage, and up-regulated the expression of Bcl-2. Moreover, geniposide down-regulated the expression of TLR4 and repressed the phosphorylation of p53.. These results demonstrate that the anti-apoptotic property of geniposide is due to its modulation of TLR4 and apoptosis-related factors (p53, Bax, Bcl-2 and Caspase-3) in LPS-induced mouse mastitis.

Topics: Animals; Anti-Inflammatory Agents; Apoptosis; Cells, Cultured; Female; Gardenia; Iridoids; Lipopolysaccharides; Mammary Glands, Animal; Mastitis; Mice; Mice, Inbred BALB C; Toll-Like Receptor 4

Our former studies have suggested that TongLuoJiuNao (TLJN) is clinically efficacious in the treatment of dementia and improving learning and memory in AD models. When Aβ aggregated with oligomer, it is known to be able to induce cellular toxicity as well as cognitive impairment. We tested the possibility that TLJN affects the formation of Aβ oligomers. In our experiment, TLJN improved cell viability, inhibited LDH release, and promoted the outgrowth of neurites of neurons treated with Aβ. Geniposide, the main component of TLJN, could increase the cell viability of SY5Y-APP695sw cells. The cytotoxicity of pretreated Aβ with geniposide was decreased in a dose-dependent manner. SDS-PAGE and Western blotting showed that geniposide and TLJN stimulated Aβ oligomer assembly. Compared with the control, more and longer fibrils of Aβ in the presence of geniposide were observed under electron microscope though the fibrils became less sensitive to thioflavin T staining. In sum, geniposide is able to protect neurons from Aβ-induced damage by remodeling Aβ.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Apoptosis; Cell Line; Cell Survival; Drugs, Chinese Herbal; Hippocampus; Humans; Iridoids; Neurons; Neuroprotective Agents; Rats

A sensitive and efficient liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of gentiopicroside, geniposide, baicalin, and swertiamarin in rat plasma. To avoid the stress caused by restraint or anesthesia, a freely moving rat model was used to investigate the pharmacokinetics of herbal medicine after the administration of a traditional Chinese herbal prescription of Long-Dan-Xie-Gan-Tang (10 g/kg, p.o.). Analytes were separated by a C18 column with a gradient system of methanol-water containing 1 mM ammonium acetate with 0.1% formic acid. The linear ranges were 10-500 ng/mL for gentiopicroside, geniposide, and baicalin, and 5-250 ng/mL for swertiamarin in biological samples. The intra- and inter-day precision (relative standard deviation) ranged from 0.9% to 11.4% and 0.3% to 14.4%, respectively. The accuracy (relative error) was from -6.3% to 10.1% at all quality control levels. The analytical system provided adequate matrix effect and recovery with good precision and accuracy. The pharmacokinetic data demonstrated that the area under concentration-time curve (AUC) values of gentiopicroside, geniposide, baicalin, and swertiamarin were 1417 ± 83.8, 302 ± 25.8, 753 ± 86.2, and 2.5 ± 0.1 min µg/mL. The pharmacokinetic profiles provide constructive information for the dosage regimen of herbal medicine and also contribute to elucidate the absorption mechanism in herbal applications and pharmacological experiments.

Topics: Administration, Oral; Animals; Area Under Curve; Chromatography, Gel; Drugs, Chinese Herbal; Flavonoids; Iridoid Glucosides; Iridoids; Limit of Detection; Male; Pyrones; Rats, Sprague-Dawley; Tandem Mass Spectrometry

It has been verified that borneol could promote the accumulation of other drugs in the whole brain. In this study, a microdialysis sampling system coupled with UPLC-MS was developed to evaluate the delivery of geniposide to four brain regions (cortex, hippocampus, hypothalamus and striatum) of conscious rats in the absence/presence of borneol: rats were administrated with geniposide alone (300mg/kg, iv) or administrated with both geniposide and borneol (0.2g/kg, ig). The dialysate collected from specific brain area was analyzed by a UPLC-MS system: separated on a BEH C18 column (50mm×2.1mm id, 1.7μm) within 1.5min, and detected in positive ion electrospray mode. The calibration curve was in good linearity over the concentration range of 0.009-90μg/mL. The inter- and intra-day accuracies were within ±10%, and the precisions were within 9.13%. The established method was applied to study the brain pharmacokinetics of geniposide and the results demonstrated that borneol markedly facilitated the delivery of geniposide to hippocampus and hypothalamus, but slightly hampered its delivery in cortex.

Topics: Animals; Brain; Camphanes; Cerebral Cortex; Chromatography, High Pressure Liquid; Consciousness; Corpus Striatum; Drug Interactions; Hippocampus; Hypothalamus; Iridoids; Male; Microdialysis; Random Allocation; Rats; Rats, Sprague-Dawley; Spectrometry, Mass, Electrospray Ionization

To investigate the therapeutic effects and acting mechanism of a combination of Chinese herb active components, i.e., a combination of baicalin, jasminoidin and cholic acid (CBJC) on Alzheimer's disease (AD).. Male rats were intracerebroventricularly injected with ibotenic acid (IBO), and CBJC was orally administered. Therapeutic effect was evaluated with the Morris water maze test, FDG-PET examination, and histological examination, and the acting mechanism was studied with DNA microarrays and western blotting.. CBJC treatment significantly attenuated IBO-induced abnormalities in cognition, brain functional images, and brain histological morphology. Additionally, the expression levels of 19 genes in the forebrain were significantly influenced by CBJC; approximately 60% of these genes were related to neuroprotection and neurogenesis, whereas others were related to anti-oxidation, protein degradation, cholesterol metabolism, stress response, angiogenesis, and apoptosis. Expression of these genes was increased, except for the gene related to apoptosis. Changes in expression for 5 of these genes were confirmed by western blotting.. CBJC can ameliorate the IBO-induced dementia in rats and may be significant in the treatment of AD. The therapeutic mechanism may be related to CBJC's modulation of a number of processes, mainly through promotion of neuroprotection and neurogenesis, with additional promotion of anti-oxidation, protein degradation, etc.

Topics: Alzheimer Disease; Animals; Cholic Acid; Dementia; Disease Models, Animal; Drug Combinations; Drugs, Chinese Herbal; Flavonoids; Humans; Ibotenic Acid; Iridoids; Male; Neuroprotective Agents; Oligonucleotide Array Sequence Analysis; Rats

Until now the overlapping and diverse pharmacological protective mechanisms of different compounds in the treatment of cerebral ischemia, both on the signaling pathway and network levels have not been revealed. In order to find differential pathway networks from gene expression profiles of hippocampus of ischemic mice treated with baicalin (BA), ursodeoxycholic acid (UA) and jasminoidin (JA), a microarray comprising 16,463 genes, FDA Arraytrack software and Ingenuity Pathway Analysis, was employed. A total of 5, 8, 11, 9 networks and 6, 7, 40, 16 pathways were found in vehicle (vs sham), BA, UA and JA (vs vehicle), respectively. Only 4 and 7 overlapping pathways were shared between BA and UA, UA and JA, accounting for 9.3% and 14.3% of the total number of all pathways, respectively. BA, UA and JA all acted on Ca(2+)-dependent signaling cascades in diverse links. BA intervened in arachidonic acid metabolism. UA affected eicosanoid, cyclin-dependent kinase 5, nuclear factor-kB, and T-helper 1 cell cytokine production. It was found that JA might decrease oxidative damage via nuclear factor erythroid 2-related factor 2-mediated antioxidant response. Compared to vehicle, no overlapping pathways were found among three groups. However, the total of 60 (71.4%) overlapping functions could be approximately divided into diseases and disorders, molecular and cellular functions, physiological system development and function as categories with ratio of 1:1:1. Analysis of network functions and known pathways may be two complementary paradigms for revealing potential pharmacological mechanisms based on the same phenotype variation.

Topics: Animals; Brain Ischemia; Flavonoids; Gene Regulatory Networks; Iridoids; Male; Mice; Random Allocation; Signal Transduction; Transcriptome; Ursodeoxycholic Acid

Composed of Ginsenoside Rg1 and Geniposide, the herbal medicine TongLuoJiuNao (TLJN) injection liquid has anti-inflammatory properties and can improve learning and memory in mice. Recently, TLJN has been used to treat the patients with cerebral ischemic stroke and vascular dementia, which significantly increase the risk of developing Alzheimer's disease (AD) in the early human beings. Although beneficial effects of TLJN have been reported in the vascular-associated brain disorders, the roles of TLJN in AD brains are still not clear. In this study, we chronically administered TLJN in amyloid precursor protein (APP) Swedish mutant transgenic mice (APP23) from 6 months old of age, which is at the onset of Aβ plaques, to 12 months old. We found that TLJN significantly decreased Aβ production and deposition in the brain of APP23 mice. Furthermore, we observed that TLJN down-regulated the levels and activity of β-secretase 1 (BACE1) protein as well as the expression levels of γ-secretase complex components: PS1, nicastrin and anterior pharynx-defective 1 (APH1) but not presenilin enhancer 2 (PEN2). The results suggest an inhibitory effect of TLJN on amyloidogenic APP processing by down-regulating the cleavage enzymes BACE1 and γ-secretase.

Topics: Alzheimer Disease; Amyloid; Amyloid beta-Protein Precursor; Amyloid Precursor Protein Secretases; Animals; Aspartic Acid Endopeptidases; Central Nervous System Agents; Drug Combinations; Drugs, Chinese Herbal; Endopeptidases; Ginsenosides; Humans; Iridoids; Male; Membrane Glycoproteins; Membrane Proteins; Mice; Mice, Inbred C57BL; Mice, Transgenic; Mutation; Presenilin-1; Stroke

To study the effects of Geniposide on SNP(sodium nitroprusside)-induced apoptosis of chondrocyte in vitro and cell cycle.. The chondrocyte of three-week-old SD rats were separated and cultivated. The second generation of chondrocyte cells were involved in experiment. Chondrocyte proliferation was measured by assay; flow cytometer were adopted to observe cell cycle and apoptosis rate; NO examination adopted nitrate reductase method.. Geniposide could significantly decrease the percentage of SNP-induced chondrocytes in G0/G1 phase and increased percentage in S phase and G2/M phase. The apoptosis of chondrocyte and the concentration of NO in the culture supernatants was reduced significantly (r=0.917, P<0.01).. Geniposide could impact SNP-induced apoptosis of chondrocyte by reducing the concentration of NO in the culture supernatants, promoting proliferation of chondrocytes, which is a probable and important mechanism of Geniposide preventing osteoarthritis.

Topics: Animals; Apoptosis; Cell Cycle; Chondrocytes; Female; Iridoids; Male; Nitroprusside; Osteoarthritis; Rats; Rats, Sprague-Dawley

Although cognitive dysfunction in diabetic patients has been explored extensively, diabetic complications of the central nervous system have not been studied. We have reported previously that geniposide has neurotrophic and neuroprotective activities with the activation of glucagons-like peptide 1 receptor, and regulates glucose-stimulated insulin secretion in vitro. But the role of geniposide on diabetic complications, especially on the neurodegenerative diseases, remains to be investigated. In this study, we investigated the effect of geniposide on the level of Aβ1-42 in the hippocampi of streptozotocin-induced diabetic rats and explored its possible mechanism. The results demonstrated that, accompanied with the improvement of insulin and blood glucose, treatment with geniposide decreased the Aβ1-42 level and improved the expression of insulin-degrading enzyme, which is the key degrading enzyme of Aβ peptide. The results of present study will help to understand the biochemical mechanisms of neuronal dysfunction and death in diabetes and to develop an efficient therapeutic strategy on Alzheimer's disease.

Topics: Amyloid beta-Peptides; Animals; Blood Glucose; Blotting, Western; Cell Line, Tumor; Cholesterol; Diabetes Mellitus, Experimental; Dose-Response Relationship, Drug; Gardenia; Hippocampus; Humans; Insulin; Insulysin; Iridoids; Male; Peptide Fragments; Phytotherapy; Rats; Rats, Sprague-Dawley; Triglycerides

Formaldehyde can induce misfolding and aggregation of Tau protein and β amyloid protein, which are characteristic pathological features of Alzheimer's disease (AD). An increase in endogenous formaldehyde concentration in the brain is closely related to dementia in aging people. Therefore, the discovery of effective drugs to counteract the adverse impact of formaldehyde on neuronal cells is beneficial for the development of appropriate treatments for age-associated cognitive decline.. In this study, we assessed the neuroprotective properties of TongLuoJiuNao (TLJN), a traditional Chinese medicine preparation, against formaldehyde stress in human neuroblastoma cells (SH-SY5Y cell line). The effect of TLJN and its main ingredients (geniposide and ginsenoside Rg1) on cell viability, apoptosis, intracellular antioxidant activity and the expression of apoptotic-related genes in the presence of formaldehyde were monitored.. Cell counting studies showed that in the presence of TLJN, the viability of formaldehyde-treated SH-SY5Y cells significantly recovered. Laser scanning confocal microscopy revealed that the morphology of formaldehyde-injured cells was rescued by TLJN and geniposide, an effective ingredient of TLJN. Moreover, the inhibitory effect of geniposide on formaldehyde-induced apoptosis was dose-dependent. The activity of intracellular antioxidants (superoxide dismutase and glutathione peroxidase) increased, as did mRNA and protein levels of the antiapoptotic gene Bcl-2 after the addition of geniposide. In contrast, the expression of the apoptotic-related gene - P53, apoptotic executer - caspase 3 and apoptotic initiator - caspase 9 were downregulated after geniposide treatment.. Our results indicate that geniposide can protect SH-SY5Y cells against formaldehyde stress through modulating the expression of Bcl-2, P53, caspase 3 and caspase 9, and by increasing the activity of intracellular superoxide dismutase and glutathione peroxidase.

Topics: Apoptosis; Caspase 3; Caspase 9; Cell Line, Tumor; Drugs, Chinese Herbal; Formaldehyde; Humans; Iridoids; Neuroblastoma; Neurons; Neuroprotective Agents; Proto-Oncogene Proteins c-bcl-2

Our group recently reported the strong anti-inflammatory effects of geniposide (Gen), a bioactive iridoid glucoside derived from gardenia jasminoides, in a mouse acute lung injury model. Herein, we hypothesized that Gen might also have potential therapeutic benefits in treatment of asthma, which was tested in a mouse model of ovalbumin (Ova)-induced allergic airway inflammation. Ova-sensitized and -challenged BALB/c mice, as compared with control animals, displayed airway hyperresponsiveness (AHR), bronchoalveolar lavage eosinophilia, mucus hypersecretion, and increased T help 2 (Th2)-associated cytokine and chemokine amounts, as well as serum Ova-specific immunoglobulin E (IgE) level. Being compared with the Ova-induced hallmarks of asthma, intraperitoneal Gen treatment prevented eosinophilic pulmonary infiltration, attenuated the increases in interleukin (IL)-4, IL-5, and IL-13, and reduced eotaxin and vascular cell adhesion molecule 1 (VCAM-1) expression. Also, Gen significantly ameliorated the Ova-driven airway hyperresponsiveness, mucus hypersecretion, and allergen-specific IgE level, which are the cardinal pathophysiological symptoms in allergic airway diseases. In addition, the efficacy of Gen was comparable to that of dexamethasone (Dex), a currently available anti-asthmatic drug. Collectively, our findings reveal that the development of immunoregulatory strategies based on Gen may be considered as an effective adjuvant therapy for allergic asthma.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cell Count; Cytokines; Disease Models, Animal; Female; Immunoglobulin E; Iridoids; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Xingnaojing (XNJ) is an effective clinical drug used to treat acute stroke. Compared with injection administration, its nasal administration has better brain targeting. Therefore, through nasal administration, XNJ microemulsion could help solve the drug load of compound components of different polarities contained in large-dose and high-concentration traditional Chinese medicines, and reduce irritation to nasal mucosa In this study, the modified volume correction method and the improved rat in situ nasal perfusion model were adopted to compare the nasal absorption of geniposide contained in different XNJ preparations. The results showed that the constant absorption rate of geniposide (GE) in XNJ-D was (2.95 +/- 0.25) x 10(-3) min(-1), whereas the constant absorption rate of GE in XNJ-M was (2.16 +/- 0.21) x 10(-3) min(-1). This indicated that the rat nasal absorption of GE in different XNJ preparations complied with the first-order process and could be considered as passive absorption. GE in XNJ-D was absorbed faster than that in XNJ-M, which provided basis for the development of nasal preparations of XNJ.

Topics: Absorption; Administration, Intranasal; Animals; Drugs, Chinese Herbal; Emulsions; Iridoids; Male; Nasal Mucosa; Rats; Rats, Sprague-Dawley

A simple and rapid ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) method for quantitative analysis of geniposide (GE) in rat plasma was developed, validated and applied to determine the level of GE in rat plasma after oral administration of GE in adjuvant-induced arthritis (AA) and normal rats. The investigation showed that there were significant differences in the groups between the normal rat and AA rat in pharmacokinetics parameters, such as the area under the time versus drug concentration curve (AUC(0-∞)) (3.77 ± 0.68 versus 2.27 ± 0.42, p < 0.05), the apparent volume of distribution (V) (140.41 ± 2.07 versus 136.51 ± 1.03, p < 0.05), the mean residence time (MRT) (3.98 ± 0.90 versus 3.80 ± 0.50, p < 0.05) and the clearance from the total body (CL) (16.10 ± 2.87 versus 26.44 ± 4.94, p < 0.05). The results indicated that AA could alter the pharmacokinetics of the drug and these experimental findings could be useful for the further study of the clinical applications of GE.

Topics: Administration, Oral; Animals; Area Under Curve; Arthritis, Experimental; Chromatography, Liquid; Iridoids; Male; Rats; Rats, Wistar; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

Geniposide is an important constituent of Gardenia jasminoides Ellis, a famous Chinese medicinal plant, and has displayed bright prospects in prevention and therapy of hepatic injury (HI). Unfortunately, the working mechanisms of this compound are difficult to determine and thus remain unknown. To determine the mechanisms that underlie this compound, we conducted a systematic analysis of the therapeutic effects of geniposide using biochemistry, metabolomics and proteomics. Geniposide significantly intensified the therapeutic efficacy as indicated by our modern biochemical analysis. Metabolomics results indicate 9 ions in the positive mode as differentiating metabolites which were associated with perturbations in primary bile acid biosynthesis, butanoate metabolism, citrate cycle (TCA cycle), alanine, aspartate and glutamate metabolism. Of note, geniposide has potential pharmacological effect through regulating multiple perturbed pathways to normal state. In an attempt to address the benefits of geniposide based on the proteomics approaches, the protein-interacting networks were constructed to aid identifying the drug targets of geniposide. Six identified differential proteins appear to be involved in antioxidation and signal transduction, energy production, immunity, metabolism, chaperoning. These proteins were closely related in the protein-protein interaction network and the modulation of multiple vital physiological pathways. These data will help to understand the molecular therapeutic mechanisms of geniposide on hepatic damage rats. We also conclude that metabolomics and proteomics are powerful and versatile tools for both biomarker discovery and exploring the complex relationships between biological pathways and drug response, highlighting insights into drug discovery.

Topics: Animals; Biomarkers; Cluster Analysis; Iridoids; Liver; Male; Metabolic Networks and Pathways; Metabolomics; Plants, Medicinal; Protein Interaction Mapping; Protein Interaction Maps; Proteomics; Rats; Signal Transduction

The objective of this study was (1) to characterize geniposide transport through MDCK and MDCK-MDR1 cell lines to confirm its transport mechanism and (2) to evaluate the effect of borneol and muscone as enhancers of geniposide transport in the BBB models so as to explore the enhancement mechanism. Transport studies of geniposide were performed in both directions, from apical to basolateral and from basolateral to apical sides. Drug concentrations were analyzed by HPLC. Geniposide showed relatively poor absorption in MDCK and MDCK-MDR1 cells, apparent permeability coefficients ranging from 0.323×10(-6) to 0.422×10(-6) cm/s. The in vitro experiments showed that geniposide transport in both directions was not concentration dependent and saturable, indicating purely passive diffusion. The efflux ratio of geniposide was less than 2 in the two cell models, which suggested that geniposide was not P-gp substrates. Geniposide transport in both directions significantly increased when co-administrated with increasing concentrations of borneol and muscone. Actin staining results indicated that borneol and muscone increased geniposide transport in the BBB models may attribute to disassembly effect on tight junction integrity.

Topics: Animals; Biological Transport; Blood-Brain Barrier; Camphanes; Cell Survival; Cycloparaffins; Dogs; Drug Delivery Systems; Iridoids; Madin Darby Canine Kidney Cells; Models, Biological

To study the anti-inflammatory mechanism of geniposide and observe the effect of geniposide on the expression of Toll-like receptor 4 (TLR4), the activity of NF-κB, and the release of pro-inflammatory cytokines- TNF-α, IL-1, and IL-6-in the RAW264.7 macrophages treated with lipopolysaccharide (LPS).. There were three experimental groups, including the control group, LPS group and LPS combined with geniposide group in this study. RAW264.7 macrophage cells were treated with LPS to induce cellular inflammation. Cell proliferation was measured by CCK-8. The concentrations of TNF-α, IL-1, and IL-6 in cell culture media were measured by ELISA. mRNA levels of TLR4 and P65 were examined by real-time PCR. The protein levels of p-IκB, P65, p-P65 and TLR4 were detected by Western blotting.. Geniposide had no effect on cell proliferation. However, geniposide down-regulated the expression of TNF-α, IL-1, and IL-6, and also inhibited the expression of TLR4 and the activity of NF-κB.. Geniposide exerts its anti-inflammatory effect through inhibiting the activity of NF-κB in the TLR4-NF-κB pathway in macrophages.

Topics: Animals; Cell Proliferation; Cells, Cultured; Cytokines; Iridoids; Lipopolysaccharides; Mice; NF-kappa B; Signal Transduction; Toll-Like Receptor 4

Glucose-stimulated insulin secretion (GSIS) is essential to the control of metabolic fuel homeostasis. The impairment of GSIS is a key element of β-cell failure and one of causes of type 2 diabetes mellitus (T2DM). Although the KATP channel-dependent mechanism of GSIS has been broadly accepted for several decades, it does not fully describe the effects of glucose on insulin secretion. Emerging evidence has suggested that other mechanisms are involved. The present study demonstrated that geniposide enhanced GSIS in response to the stimulation of low or moderately high concentrations of glucose, and promoted glucose uptake and intracellular ATP levels in INS-1 cells. However, in the presence of a high concentration of glucose, geniposide exerted a contrary role on both GSIS and glucose uptake and metabolism. Furthermore, geniposide improved the impairment of GSIS in INS-1 cells challenged with a high concentration of glucose. Further experiments showed that geniposide modulated pyruvate carboxylase expression and the production of intermediates of glucose metabolism. The data collectively suggest that geniposide has potential to prevent or improve the impairment of insulin secretion in β-cells challenged with high concentrations of glucose, likely through pyruvate carboxylase mediated glucose metabolism in β-cells.

Topics: Animals; Cell Line; Diabetes Mellitus, Type 2; Glucose; Insulin; Insulin Secretion; Insulin-Secreting Cells; Iridoids; Rats

Recently, the pharmaceutical industry has shifted to pursuing combination therapies that comprise more than one active ingredient. Interestingly, combination therapies have been used for more than 2500 years in traditional Chinese medicine (TCM). Understanding optimal proportions and synergistic mechanisms of multi-component drugs are critical for developing novel strategies to combat complex diseases. A new multi-objective optimization algorithm based on least angle regression-partial least squares was proposed to construct the predictive model to evaluate the synergistic effect of the three components of a novel combination drug Yi-qi-jie-du formula (YJ), which came from clinical TCM prescription for the treatment of encephalopathy. Optimal proportion of the three components, ginsenosides (G), berberine (B) and jasminoidin (J) was determined via particle swarm optimum. Furthermore, the combination mechanisms were interpreted using PLS VIP and principal components analysis. The results showed that YJ had optimal proportion 3(G): 2(B): 0.5(J), and it yielded synergy in the treatment of rats impaired by middle cerebral artery occlusion induced focal cerebral ischemia. YJ with optimal proportion had good pharmacological effects on acute ischemic stroke. The mechanisms study demonstrated that the combination of G, B and J could exhibit the strongest synergistic effect. J might play an indispensable role in the formula, especially when combined with B for the acute stage of stroke. All these data in this study suggested that in the treatment of acute ischemic stroke, besides restoring blood supply and protecting easily damaged cells in the area of the ischemic penumbra as early as possible, we should pay more attention to the removal of the toxic metabolites at the same time. Mathematical system modeling may be an essential tool for the analysis of the complex pharmacological effects of multi-component drug. The powerful mathematical analysis method could greatly improve the efficiency in finding new combination drug from TCM.

Topics: Algorithms; Animals; Berberine; Brain Ischemia; Cerebrum; Drug Combinations; Drug Compounding; Drug Evaluation, Preclinical; Drug Synergism; Drugs, Chinese Herbal; Ginsenosides; Infarction, Middle Cerebral Artery; Iridoids; Least-Squares Analysis; Male; Medicine, Chinese Traditional; Membrane Potential, Mitochondrial; Mitochondria; Mitochondrial Swelling; Neuroprotective Agents; Principal Component Analysis; Rats; Rats, Sprague-Dawley; Regional Blood Flow

Eucommia ulmoides Oliver is a natural product widely used as a dietary supplement and medicinal plant. Here, we examined the potential regulatory effects of Eucommia ulmoides Oliver extracts (EUE) on hepatic dyslipidemia and its related mechanisms by in vitro and in vivo studies. EUE and its two active constituents, aucubin and geniposide, inhibited palmitate-induced endoplasmic reticulum (ER) stress, reducing hepatic lipid accumulation through secretion of apolipoprotein B and associated triglycerides and cholesterol in human HepG2 hepatocytes. To determine how EUE diminishes the ER stress response, lysosomal and proteasomal protein degradation activities were analyzed. Although proteasomal activity was not affected, lysosomal enzyme activities including V-ATPase were significantly increased by EUE as well as aucubin and geniposide in HepG2 cells. Treatment with the V-ATPase inhibitor, bafilomycin, reversed the inhibition of ER stress, secretion of apolipoprotein B, and hepatic lipid accumulation induced by EUE or its component, aucubin or geniposide. In addition, EUE was determined to regulate hepatic dyslipidemia by enhancing lysosomal activity and to regulate ER stress in rats fed a high-fat diet. Together, these results suggest that EUE and its active components enhance lysosomal activity, resulting in decreased ER stress and hepatic dyslipidemia.

Topics: Cholesterol; Endoplasmic Reticulum Stress; Eucommiaceae; Hep G2 Cells; Humans; Iridoid Glucosides; Iridoids; Liver; Lysosomes; Plant Extracts; Triglycerides

To verify the anti-inflammatory potency of iridoids, three iridoids (two natural, loganic acid: LA; geniposide: GE; and an artefact, 7(S)-n-butyl morroniside: BM) were investigated in vitro on the inhibition of superoxide generation in human neutrophils. All compounds showed inhibitory effect on fMLP-induced superoxide generation in a concentration-dependent manner with the following order: BM>LA>GE. BM exhibits potent inhibitory activity on superoxide anion induced by PMA, while LA and GE showed weak effect. When AA was used as stimulus, the generation of superoxide anion was suppressed by BM in a concentration-dependent manner. LA and GE exhibit both sides effect on superoxide generation.

Topics: Anti-Inflammatory Agents; Arachidonic Acid; Humans; Iridoids; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Superoxides

The safety of geniposide, mainly focusing on its hepatotoxicity in rats, was determined by liver enzymes in serum and histopathology ultrastructural preparation. The lethal dose, 50% (LD50) of per oral geniposide was 1431.1 mg kg(-1). The acute toxicity study indicated geniposide at dose of 574 mg kg(-1) or more could cause hepatic toxicity in rats and the hepatotoxicity often appeared at 24-48 h after the oral administration. The hepatotoxicity was associated with oxidative stress with decrease of total superoxide dismutase activity and increase of malondialdehyde concentration in rats' livers. Subchronic toxicity study showed geniposide did not cause hepatotoxicity at the doses of 24.3 and 72.9 mg kg(-1) orally for 90 days in rats. Thus, acute hepatotoxicity of geniposide at high doses was likely to be linked to oxidative stress, while geniposide at normal dose of 24.3 mg kg(-1) or less did not cause hepatotoxicity even in the repeated dosing study.

Topics: Animals; Female; Fruit; Gardenia; Iridoids; Liver; Male; Malondialdehyde; Oxidative Stress; Rats; Superoxide Dismutase

This study was performed to develop methods for the chromatographic determination of biomarkers in Hwangryunhaedok-tang (HHT) and the quantitative evaluation of commercial HHT. To develop an analytical method, an RP-amide column (2.7 μm, 4.6 × 100 mm, Halo: Supelco, Bellefonte, PA) was used with a gradient solvent system of mixed acetonitrile and 0.1 % phosphoric acid/water and an ultra performance liquid chromatography-diode array detector. The method was validated by specificity, linearity, accuracy (recovery) and precision tests (repeatability, intra and inter-day). The correlation coefficients (R (2)) of biomarkers were calculated as 0.9998-1.000 and their ranges were as follows: geniposide (62.5-1,000.0 μg/ml), berberine (31.3-500.0 mg/ml), palmatine (31.3-500.0 μg/ml), baicalin (125.0-1,500.0 μg/ml), baicalein (15.6-250.0 μg/ml) and wogonin (5.2-125.0 μg/ml), respectively. The limit of detection was 0.34-4.01 μg/ml, and the limit of quantification was 1.02-12.16 μg/ml. The intra-day and inter-day precision of six components were revealed as 0.02-2.48 % as a relative standard deviation (RSD). The repeatability value of biomarkers in three different concentrations of HHT was 0.29-2.98 % (RSD value) and recovery was 95.72-104.90 %. Among several extraction methods tested, biomarker content was higher with the 20 times extraction (20TE) and mixture of extract powder (MEP) methods than with any other method, and some differences among diverse pharmaceutical medicines were revealed. The validation data indicated that the method developed is suited to the determination of six marker compounds in HHT. The content of biomarkers by simultaneous analysis was evaluated in 20TE, MEP, USA formula and Taiwan formula.

Topics: Berberine; Drugs, Chinese Herbal; Flavanones; Flavonoids; Iridoids

Gardenia jasminoides J. Ellis (Rubiaceae) is a shrub tree species distributed all over the world. Now its pharmacological activities such as anti-atherosclerosis have been extensively studied.. To offer pharmacological proof for its further clinical application in cardiovascular diseases, the antithrombotic activities of the aqueous extract of G. jasminoides (GJ-ext) were studied in mouse and rat models.. GJ-ext was administrated orally to detect the effects on the models of carrageenan-induced tail thrombosis and arteriovenous shunt thrombosis. The effects of GJ-ext and geniposide (p.o.) on antiplatelet aggregation were examined. Geniposide and genipin were studied on venous thrombosis by oral administration.. GJ-ext (67, 133 and 266 mg/kg) and aspirin (50 mg/kg), respectively, decreased the length of tail thrombus with average thrombus inhibition rate of 21.9, 55.7, 65.8 and 57.6% at 48 h and 19.0, 54.5, 69.3 and 56.9% at 72 h after carrageenan injection and, meanwhile, improved thrombosis induced by arteriovenous shunt (silk thread) with 36.3, 45.5, 86.4 and 63.7% inhibition rate of thrombus respectively, and the ED(50) of GJ-ext was 160.8 mg/kg. Furthermore, GJ-ext (67 mg/kg) and geniposide (20 mg/kg) significantly inhibited platelet aggregation induced by thrombin/collagen with 45.1%/19.3% and 52.8%/26.2% aggregation rate. Geniposide (10-40 mg/kg) and genipin (5-20 mg/kg) inhibited venous thrombosis induced by tight ligation of the inferior vena cava, their ED(50) values were 18.4 and 8.6 mg/kg, respectively.. This study indicated that GJ-ext and geniposide demonstrated remarkable antithrombotic activities and supported their therapeutic uses for thrombotic diseases.

Topics: Administration, Oral; Animals; Arteriovenous Shunt, Surgical; Aspirin; Carrageenan; Disease Models, Animal; Dose-Response Relationship, Drug; Fibrinolytic Agents; Fruit; Gardenia; Iridoids; Male; Mice; Mice, Inbred ICR; Plant Extracts; Plants, Medicinal; Platelet Aggregation; Platelet Aggregation Inhibitors; Rats; Rats, Sprague-Dawley; Solvents; Time Factors; Venous Thrombosis; Water

Fructus gardeniae has long been used by traditional Chinese medical practitioners for its anti-inflammatory, anti-oxidant, anti-tumor and anti-hyperlipidemic characteristics. Here we describe our finding that F. gardeniae greatly reduces anti-enterovirus 71 (EV71) activity, resulting in significant decreases in EV71 virus yields, EV71 infections, and internal ribosome entry site activity. We also found that geniposide, a primary F. gardeniae component, inhibited both EV71 replication and viral IRES activity. Our data suggest the presence of a mechanism that blocks viral protein translation. According to our findings, F. gardeniae and geniposide deserve a closer look as potential chemopreventive agents against EV71 infections.

Topics: Antiviral Agents; Cell Line, Tumor; Drugs, Chinese Herbal; Enterovirus A, Human; Enterovirus Infections; Gardenia; Humans; Iridoids; Medicine, Chinese Traditional; Microbial Sensitivity Tests; Molecular Sequence Data; Protein Biosynthesis; Ribosomes; RNA, Viral; Virus Replication

To enhance the therapeutic efficacy and reduce the adverse effects of traditional Chinese medicine, practitioners often prescribe combinations of plant species and/or minerals, called formulae. Unfortunately, the working mechanisms of most of these compounds are difficult to determine and thus remain unknown. In an attempt to address the benefits of formulae based on current biomedical approaches, we analyzed the components of Yinchenhao Tang, a classical formula that has been shown to be clinically effective for treating hepatic injury syndrome. The three principal components of Yinchenhao Tang are Artemisia annua L., Gardenia jasminoids Ellis, and Rheum Palmatum L., whose major active ingredients are 6,7-dimethylesculetin (D), geniposide (G), and rhein (R), respectively. To determine the mechanisms underlying the efficacy of this formula, we conducted a systematic analysis of the therapeutic effects of the DGR compound using immunohistochemistry, biochemistry, metabolomics, and proteomics. Here, we report that the DGR combination exerts a more robust therapeutic effect than any one or two of the three individual compounds by hitting multiple targets in a rat model of hepatic injury. Thus, DGR synergistically causes intensified dynamic changes in metabolic biomarkers, regulates molecular networks through target proteins, has a synergistic/additive effect, and activates both intrinsic and extrinsic pathways.

Topics: Animals; Anthraquinones; Biomarkers; Drug Evaluation, Preclinical; Drug Synergism; Drug Therapy, Combination; Drugs, Chinese Herbal; Iridoids; Liver; Liver Diseases; Male; Medicine, Chinese Traditional; Metabolome; Metabolomics; Molecular Targeted Therapy; Proteome; Proteomics; Rats; Rats, Wistar; Umbelliferones

The purpose of this study was to investigate the effects of natural borneol (NB) on the pharmacokinetics and bioavailability of ophthalmic administered geniposide (Ge) in rabbits. In vitro permeability characteristics of Ge in excised rabbit corneas were evaluated using Franz-type cells. The effect of NB on Ge pharmacokinetic profiles in vivo was studied with microdialysis. Concentrations of Ge were determined with reversed-phase high performance liquid chromatography (HPLC) following ophthalmic administration of Ge alone or with NB (0.01%, 0.02%, and 0.04%) or 0.5% ethylendiaminetetraacetic acid (EDTA). Ocular irritation was evaluated using the Draize method and histological examination. Ge solution alone (control solution) had limited corneal permeability. The ratio of the apparent permeability coefficient (Papp) with respect to the control solution significantly increased by approximately 1.6-, 2.0-, and 2.4-fold at NB concentrations of 0.01, 0.02, and 0.04%, respectively. The Papp for Ge with 0.5% EDTA (positive control) was approximately 1.7-fold higher than that for control solution. Compared to control solution, Ge exhibited a 1.46-, 2.16-, and 2.47-fold greater AUC0-6h, and 2.0-, 3.5-, and 4.4-fold greater Cmax, with 0.01, 0.02, and 0.04% NB, respectively, while Tmax remained unchanged. In conclusion, the ocular bioavailability of Ge significantly increased in the presence of NB.

Topics: Absorption; Animals; Biological Availability; Camphanes; Eye; Iridoids; Ophthalmic Solutions; Rabbits

To investigate the chemical constituents of Fructus Gardeniae.. Normal phase silica gel, RP-18 silica gel and Sephadex LH-20 column chromatographies combined with recrystallization were used to isolate and purify the constituents. Their structures were identified by spectroscopic methods, including 1H-NMR, 13C-NMR, ESI-MS and EI-MS.. Seven compounds were isolated. Their structures were identified as geniposide (I), 6alpha-hydroxygeniposide (II), genipin-gentiobioside (III), adian-5-en-3alpha-ol (IV), (23Z) -cycloart-23-en-3beta,25-diol (V), 7alpha-hydroxy sitosterol (VI) and 5,8-epidioxystigmasta-6,22-dien-3-ol (VII) from ethanol extract of Fructus Gardeniae.. Compounds IV, V, VI and VII are obtained from this plant for the first time.

Topics: Ethanol; Fruit; Gardenia; Iridoids; Magnetic Resonance Spectroscopy; Molecular Structure; Plant Extracts

Zhi-Zi-Hou-Pu decoction (ZZHPD), a classic antidepressant formula, is composed of Gardenia jasminoides Ellis (ZZ), Fructus aurantii immaturus (ZS) and Cortex magnoliae officinalis (HP). ZZHPD has attracted a great deal of attention for its antidepressant effects. The aim of this study was to compare the pharmacokinetics of geniposide (one of the predominant active ingredients) after oral administration of different combinations of ZZHPD and to explore the influence of herb-herb interaction on the pharmacokinetics of geniposide. Twenty four rats were divided randomly into four groups and were administered one of the four extracts: ZZ, ZZ-HP, ZZ-ZS and ZZHPD (ZZ-HP-ZS) via intragastric gavage with approximately the same dose of 40.65 mg/kg geniposide (an effective human daily dose of ZZHPD). Plasma concentrations of geniposide were determined using an HPLC method. Pharmacokinetic parameters were calculated from the plasma concentration-time data. Compared with ZZ alone, the ZZ-ZS combination delayed T(max) and ZZ-HP, ZZ-ZS, ZZHPD remarkably shortened the T(1/2) of geniposide. In addition, ZZ-HP, ZZ-ZS, ZZHPD obviously increased the AUC of geniposide. The result illustrated that the oral bioavailability of geniposide was dramatically enhanced when ZZ was combined with HP or/and ZS. It can be deduced that herb-herb interaction may increase the absorption, and significantly improve the oral bioavailability of geniposide in rats.

Topics: Administration, Oral; Animals; Area Under Curve; Biological Availability; Citrus; Drug Combinations; Drug Synergism; Drugs, Chinese Herbal; Gardenia; Humans; Intestinal Absorption; Iridoids; Magnolia; Male; Random Allocation; Rats; Rats, Sprague-Dawley

Increasing evidence has demonstrated that multidrug combinations could amplify the therapeutic efficacies of each agent. Interestingly, the pharmacological effect of traditional Chinese medicine (TCM) is usually attributed to the drug-interaction property (synergism) of multiple active constituents. Pharmacokinetics is a useful means of evaluating the drug interactions of major active compounds in TCM. A simple, sensitive and reliable RP-HPLC-DAD method has been developed to simultaneously quantify 6,7-dimethylesculetin (D), geniposide (G) and rhein (R), which are the active ingredients in Yin-Chen-Hao-Tang (YCHT), performing drug-interaction pharmacokinetics studies in vivo. Plasma samples were prepared using methanolic precipitation, a filtration step, and then injection of the methanolic extract onto a Nova-Pak C₁₈ Guard-Pak™ guard column with a gradient mobile phase. Triple-wavelength diode array detection was set at λ(max) values of 343 nm for D, 241 nm for the G, and 259 nm for R. Our results successfully demonstrate that this method has excellent and satisfactory selectivity, sensitivity, linearity, precision, accuracy and recovery. In healthy rats, the estimated pharmacokinetic parameters (i.e. C(max) , AUC and Cl) of D, G and R, when administered with COC (a combination of D, G and R), were C(max) 16.05 mg/L, AUC 108.96 mg h/L and Cl 0.36 L/h for D; C(max) 9.35 mg/L, AUC 64.71 mg h/L and Cl 0.88 L/h for G; and C(max) 14.18 mg/L, AUC 57.98 mg h/L and Cl 1.77 L/h for R. Here, we report that the COC combination could significantly increase the plasma level and slow the elimination rate compared with any one or two of the three individual compounds, which may indicate a drug-drug interaction.

Topics: Animals; Anthraquinones; Chromatography, High Pressure Liquid; Chromatography, Reverse-Phase; Coumarins; Drug Synergism; Drugs, Chinese Herbal; Iridoids; Linear Models; Male; Models, Chemical; Rats; Rats, Wistar; Reproducibility of Results; Sensitivity and Specificity

To explore the role of the glucagon-like peptide 1 receptor (GLP-1R) in geniposide regulated insulin secretion in rat INS-1 insulinoma cells.. Rat INS-1 insulinoma cells were cultured. The content of insulin in the culture medium was measured with ELISA assay. GLP-1R gene in INS-1 cells was knocked down with shRNA interference. The level of GLP-1R protein in INS-1 cells was measured with Western blotting.. Geniposide (0.01-100 μmol/L) increased insulin secretion from INS-1 cells in a concentration-dependent manner. Geniposide (10 μmol/L) enhanced acute insulin secretion in response to both the low (5.5 mmol/L) and moderately high levels (11 mmol/L) of glucose. Blockade of GLP-1R with the GLP-1R antagonist exendin (9-39) (200 nmol/L) or knock-down of GLP-1R with shRNA interference in INS-1 cells decreased the effect of geniposide (10 μmol/L) on insulin secretion stimulated by glucose (5.5 mmol/L).. Geniposide increases insulin secretion through glucagon-like peptide 1 receptors in rat INS-1 insulinoma cells.

Topics: Animals; Cell Line, Tumor; Diabetes Mellitus, Type 2; Gardenia; Glucagon-Like Peptide 1; Insulin; Iridoids; Islets of Langerhans; Plant Extracts; Rats; RNA Interference; RNA, Small Interfering

Iridoids belong to a group of monoterpene compounds with cyclopentane ring and found as mostly the glycoside forms in nature. They act primarily as the defense substances and found in various medicinal plants.. Although many iridoids exhibit anti-inflammatory and anticancer activities, their molecular targets/pathways are not fully understood. Here, the antiproliferative effect of the hydrolyzed-iridoid product (H-iridoid) form through the STAT3 signaling pathways on tumor cells was investigated.. H-iridoids were obtained from five iridoid glycosides with β-glucosidase treatment. The effects of several H-iridoids on cell viability and cell proliferation in tumor cells were measured by the MTT assay. The phosphorylation levels of STAT3, its regulatory molecules, and apoptosis by H-geniposide treatment in DU145 cells were investigated by immunoblots and flow cytometry.. No single iridoid glycoside exerted any cytotoxicity in the tumor cells, whereas H-iridoids had significant cytotoxic, antiproliferative, and STAT3 inhibitory effects and revealed different potencies depending on their chemical structures. Among the H-iridoids tested, H-geniposide inhibited constitutive STAT3 activation through inhibiting upstream JAK1 and c-Src. Consistent with STAT3 inactivation, H-geniposide downregulated the expressions of Bcl-2, Bcl-xL, survivin, and cyclin D1; this correlated with the accumulation of cells in the sub-G1 phase of the cell cycle and the induction of apoptosis.. Our results indicate that the hydrolysis of the glycosidic bond from iridoid glycoside is required for exhibiting cytotoxicity in tumor cells. H-geniposide is the most potent agent and a novel blocker of STAT3 activation in DU145 cells.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Flow Cytometry; Gene Expression Regulation, Neoplastic; Humans; Hydrolysis; Iridoid Glycosides; Iridoids; Neoplasms; Signal Transduction; STAT3 Transcription Factor

Eucommia ulmoides Oliver is a perennial woody plant distributed widely in China. To characterize some major compounds in E. ulmoides bark extract, six compounds were identified via high-performance liquid chromatography qualitative analysis. E. ulmoides bark extract protects against cadmium-induced oxidative damage in rat kidneys. Two compounds of E. ulmoides bark extract, geniposide and genipin, which were identified both in serum and in kidney tissue, showed inhibitory effects on nitric oxide production. This study provides biological evidence supporting the usefulness of E. ulmoides bark against cadmium-induced toxic oxidative stress in rat kidney tissue.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cadmium Poisoning; Drug Discovery; Drugs, Chinese Herbal; Eucommiaceae; Iridoids; Kidney; Macrophage Activation; Male; Mice; Mice, Inbred Strains; Nitric Oxide; Oxidative Stress; Plant Bark; Protective Agents; Rats; Rats, Sprague-Dawley; Renal Insufficiency

Intestinal microflora (IM) is able to produce toxic and carcinogenic metabolites and induce more potent cytotoxicity against cells than non-metabolites. This study was performed to investigate the cytotoxic responses of geniposide (GS) and its metabolite and to determine the role of metabolism by IM in GS-induced cytotoxicity. Genipin (GP), a GS metabolite, increased cytotoxic effects in cells, but GS did not. Following GS incubation with IM for metabolic activation, increased cytotoxicity was detected compared to GS. Western blot analysis revealed that the activated GS inhibited Bcl-2 expression with a subsequent increase in Bax expression. Likewise, GS activation by IM stimulated caspase-3 and the production of reactive oxygen species (ROS). In addition, activated GS-induced apoptosis was confirmed by apoptosis and ROS assays; N-acetyl-l-cysteine (NAC) suppressed ROS production and apoptotic cell death. Activated GS induced sustained JNK phosphorylation. Moreover, activated GS-induced cell death was reversed by SP600125. Taken together, these findings suggest that human IM is able to metabolize GS into GP, and the related biological activities induce apoptosis through ROS/JNK signaling.

Topics: Apoptosis; bcl-2-Associated X Protein; Biotransformation; Blotting, Western; Caspase 3; Cell Survival; Feces; Female; Hep G2 Cells; Humans; In Situ Nick-End Labeling; Intestines; Iridoids; Male; MAP Kinase Signaling System; Molecular Structure; Proto-Oncogene Proteins c-bcl-2; Reactive Oxygen Species; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction

Geniposide, a main iridoid glucoside component of gardenia fruit, has been shown to possess anti-inflammatory activity. However, its potential use for acute lung injury (ALI) has not yet been studied. The aim of this study was to evaluate the anti-inflammatory properties of geniposide using a mouse ALI model. ALI was induced by intranasal injection of lipopolysaccharide (LPS). Pretreatment of mice with geniposide (20, 40, or 80 mg/kg) resulted in a marked reduction in inflammatory cells and total protein concentration in the bronchoalveolar lavage fluid (BALF) of mice. Levels of inflammatory mediators, including tumour necrosis factor- α (TNF- α), interleukin-6 (IL-6), and interleukin-10 (IL-10), were significantly altered after treatment with geniposide. Histological studies using hematoxylin and eosin (H&E) staining demonstrate that geniposide substantially inhibited LPS-induced alveolar wall changes, alveolar haemorrhage, and neutrophil infiltration in lung tissue, with evidence of reduced myeloperoxidase (MPO) activity. In addition, we investigated potential signal transduction mechanisms that could be implicated in geniposide activity. Our results suggest that geniposide may provide protective effects against LPS-induced ALI by mitigating inflammatory responses and that the compound's mechanism of action may involve blocking nuclear factor-kappaB (NF- κB) and mitogen-activated protein kinases (MAPK) signalling pathway activation.

Topics: Acute Lung Injury; Animals; Anti-Inflammatory Agents; Cytokines; Disease Models, Animal; Dose-Response Relationship, Drug; Fruit; Gardenia; Iridoid Glucosides; Iridoids; Lipopolysaccharides; Lung; Male; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinase Kinases; NF-kappa B; Peroxidase; Plants, Medicinal; Random Allocation; Signal Transduction

In present study, the performance and separation characteristics of nine macroporous resins for the enrichment and purification of gardenia yellow from Gardenia jasminoides var. radicans Makino have been evaluated. The adsorption and desorption properties of crude gardenia yellow solution on macroporous resins including HPD722, HPD100, HPD100A, HPD400, HPD400A, D101, AB-8, XAD-16, and NKA-9 have been compared. Then, HPD722 was chosen to purify gardenia yellow because of its strong adsorption and desorption abilities as well as high selectivity. Column packed with HPD722 resin was used to perform dynamic adsorption and desorption tests to optimize the separation process of gardenia yellow. The optimal conditions were as follows: The crude gardenia yellow solution with concentration of 15 mg/mL was loaded in column packed with HPD722 resin at the flow rate of 1.0 mL/min, and the adsorbate-laden column was washed with 800 mL water, 600 mL 15% ethanol water solution respectively at the speed of 2.5 mL/min, then desorbed with 200 mL 80% ethanol water solution at the speed of 3.5 mL/min. The colority of the product obtained were up to 300. The method developed in this study provides a new approach for scale-up separation and purification of gardenia yellow from G. jasminoides var. radicans Makino.

Topics: Adsorption; Carotenoids; Chromatography, Affinity; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Ethanol; Gardenia; Iridoids; Laboratory Chemicals; Plant Extracts; Spectrophotometry, Ultraviolet; Water

Possible role of metabolism by the intestinal bacteria in geniposide-induced cytotoxicity was investigated in human hepatoma HepG2 cells. Initially, toxic effects of geniposide and its metabolite genipin were compared. Genipin, a deglycosylated form of geniposide, was cytotoxic at the concentrations that geniposide was not. As metabolic activation systems for geniposide, human intestinal bacterial cultures, fecal preparation (fecalase) and intestinal microbial enzyme mix were employed in the present study. When geniposide was incubated with human intestinal bacteria, either Bifidobacterium longum HY8001 or Bacteroides fragilis, for 24 h, the cultured media caused cytotoxicity in HepG2 cells. Fecalase and intestinal enzyme mix were also effective to metabolically activate geniposide to its cytotoxic metabolite. The present results indicated that genipin, a metabolite of geniposide, might be more toxic than geniposide, and that intestinal bacteria might have a role in biotransformation of geniposide to its toxic metabolite. In addition, among three activation systems tested, intestinal microbial enzyme mix would be convenient to use in detecting toxicants requiring metabolic activation by intestinal bacteria.

Topics: Bacteroides fragilis; Bifidobacterium; Biotransformation; Cell Survival; Dose-Response Relationship, Drug; Feces; Hep G2 Cells; Humans; Intestines; Iridoids

The Direct Analysis in Real Time (DART) ionization source coupled with a quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS/MS) system has the capability to desorb analytes directly from samples from complex Chinese herbal preparations without sample cleanup or chromatographic separation.. In this work, a method based on DART/Q-TOF MS/MS has been developed for rapid determination of geniposide present in 'Re Du Ning Injections', a Chinese herbal preparation. The method has been evaluated for both qualitative and quantitative analysis of geniposide in Re Du Ning Injections.. Variables including polarity for ion detection, DART gas heater temperature, matrix effect and sample presentation speed were investigated. The quantitative method was validated with respect to linearity, sensitivity, repeatability, precision and accuracy by using both internal and external standards. A comparison of the results obtained using the DART-based method was made with those obtained using a conventional High-Performance Liquid Chromatography/Diode-Array Detector (HPLC/DAD) by analyzing geniposide in four batches of Re Du Ning Injections.. The DART/Q-TOF MS/MS-based method provides a rapid, efficient and powerful method to analyze compounds from complex Traditional Chinese Medicines with limited sample preparation thus reducing time and complexity of quality control for those materials.

Topics: Drugs, Chinese Herbal; Iridoids; Linear Models; Reproducibility of Results; Sensitivity and Specificity; Tandem Mass Spectrometry

Geniposide is derived from Gardenia jasminoides Ellis (Rubiaceae). Its anti-inflammatory, antithrombotic effects as well as its preventive effect against ischemic stroke have been reported. Radix notoginseng (Chinese name tienchi or sanqi) is the dried root of Panax notoginseng (Burk.) F.H. Chen, an herb noted for its promotion of blood circulation, blood stasis removal and pain alleviation, and has been widely utilized for the prevention and treatment of microcirculatory disturbances in China and other Asian countries for many years. Notoginsenoside R₁ is an effective and structurally representative bioactive constituent of R. notoginseng. In our preliminary study, notoginsenoside R₁ was able significantly to improve the bioavailability of geniposide in beagle dogs, but the underlying mechanisms remain unknown.. The present study aimed to investigate the intestinal kinetic absorptive characteristics of geniposide as well as the absorptive behavior influenced by the co-administration of notoginsenoside R₁ using an in vitro everted rat gut sac model.. The results showed good linear correlation between the geniposide absorption in sac contents and the incubation time from 0 to 120 min. The concentration dependence showed a non-linear correlation between the geniposide absorption and the concentrations 0.356-1.424 mg/mL, the absorption was saturated about 1.424 mg/mL. Notoginsenoside R₁ at 0.1 and 0.2mg/mL concentrations was able significantly to enhance the absorption of geniposide (1.424 mg/mL) by 1.7- and 1.4-fold. Moreover, verapamil, a well-known P-glycoprotein inhibitor, was able significantly to elevate the absorption of geniposide 2.4-fold. Notoginsenoside R₁ influenced geniposide's absorption in a way similar to that of a P-glycoprotein inhibitor.. In conclusion, notoginsenoside R₁ significantly enhances the intestinal absorption of geniposide. As for the mechanism underlying the improvement of geniposide's bioavailability, it is proposed that notoginsenoside R₁ was able to decrease the efflux transport of geniposide by P-glycoproteins.

Topics: Animals; ATP Binding Cassette Transporter, Subfamily B, Member 1; Ginsenosides; Intestinal Absorption; Intestinal Mucosa; Iridoids; Male; Rats; Rats, Wistar; Rhodamine 123; Verapamil

Fructus gardeniae (Zhizi), one of commonly-used traditional Chinese medicines, is derived from the ripe fruit of the evergreen shrub, Gardenia jasminoides Ellis, and is an ingredient of many traditional Chinese preparations, and has numerous pharmacological actions. Geniposide is the important bioactive element derived from F. gardeniae. This study established optimum conditions and method of ultrasound-assisted extraction for geniposide from F. gardeniae by exploring the different experimental parameters, such as type of solvent, ratio of solid/liquid, extraction temperature and extraction time. The data gained from this study is important to further extract and apply the geniposide and is also a significant reference to extract the bioactive compounds from plant materials by the ultrasound-assisted method.

Topics: Drugs, Chinese Herbal; Gardenia; Iridoids; Molecular Structure; Sonication

In order to elucidate the overlapping and diverse pharmacological protective mechanisms of different Chinese medicinal compounds, we investigated the alteration of gene expression and activation of signaling pathways in the mouse hippocampus after treatment of cerebral ischemia-reperfusion injury with various compounds. A microarray including 16,463 genes was used to identify differentially expressed genes among six treatment groups: baicalin (BA), jasminoidin (JA), cholic acid (CA), concha margaritiferausta (CM), sham, and vehicle. The US Food and Drug Administration (FDA) ArrayTrack system and Kyoto Encyclopedia of Genes and Genomes (KEGG) database were used to screen significantly altered genes and pathways (P < 0.05, fold change >1.5). Vehicle treatment alone resulted in alteration of 726 genes (283 upregulated, 443 downregulated) compared to the sham treatment group. BA, JA, and CA treatments, but not CM treatment, were effective in reducing infarct volume compared with vehicle treatment (P < 0.05). Compared with the CM group, a total of 167 (73 upregulated, 94 downregulated), 379 (211 upregulated, 168 downregulated), and 181 (76 upregulated, 105 downregulated) altered genes were found in the BA, JA, and CA groups, respectively. The numbers of overlapping genes between the BA and JA, BA and CA, and JA and CA groups were 28 (16 upregulated, 12 downregulated), 14 (4 upregulated, 10 downregulated), and 31 (8 upregulated, 23 downregulated), respectively. Three overlapping genes were identified among the BA, JA, and CA treatment groups: Il1rap, Gnb5, and Wdr38. Based on KEGG pathway analysis, two, seven, and four pathways were significantly activated in the BA, JA, and CA groups, respectively, when compared to the CM group. The ATP-binding cassette (ABC) transporters general pathway was activated by BA and JA treatment, and the mitogen-activated protein kinase (MAPK) signaling pathway was activated by JA and CA treatment. Alteration of IL-1 and Hspa1a expression was found by real time reverse transcription polymerase chain reaction, confirming the results of the microarray analysis. Our data demonstrated that polytypic profiles of 167-379 altered genes exist in the mouse hippocampus treated with different compounds known to be therapeutically effective in cerebral ischemia-reperfusion injury, and we were able to identify overlapping genes and pathways among these groups. Therefore, these different compounds may function through both overlapping and distin

Topics: Animals; ATP-Binding Cassette Transporters; Capillary Electrochromatography; Cerebral Infarction; Cholic Acid; Drugs, Chinese Herbal; Flavonoids; Gene Expression; Hippocampus; Iridoids; Male; Mice; Microarray Analysis; Mitogen-Activated Protein Kinases; Real-Time Polymerase Chain Reaction; Reperfusion Injury; RNA; Signal Transduction

Activated microglia producing reactive nitrogen species, inflammatory factors, reactive oxygen species (ROS) and other neurovirulent factors, can lead to the development of neurodegenerative diseases. Certain compounds can inhibit the activation of microglia. However, the mechanisms remain unclear. In the present study, we investigated the inhibitory effect of geniposide on the production of ROS and inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-stimulated N9 murine microglial cells through the p38, ERK1/2 and nuclear factor-κB (NF-κB) signaling pathways. After the N9 cells were pre-treated with the vehicle or geniposide and exposed to LPS for the time indicated, the MTT conversion test was used to assess cell viability. Suitable concentrations were chosen and adjusted according to the experiments. Extracellular nitric oxide (NO) release was measured by Griess reaction. The formation of ROS and intracellular NO was evaluated by fluorescence imaging. NOS activities were determined using commercially available kits. The morphology of the N9 cells was examined by hematoxylin and eosin staining. The expression of iNOS mRNA was examined by RT-PCR. The protein levels of iNOS, p38 mitogen-activated protein kinase (MAPK), ERK1/2 and NF-κB, inhibitory factor-κB-α (IκB-α) were determined by western blot analysis. The results showed that geniposide attenuated the activation of N9 cells and inhibited the overproduction of NO, intracellular ROS and the expression of iNOS induced by LPS in the cells. In addition, geniposide blocked the phosphorylation of p38, ERK1/2 and inhibited the drop-off of IκB induced by LPS in the cells. These data indicate that geniposide has therapeutic potential for the treatment of neurodegenerative diseases, and that it exerts its effects by inhibiting inflammation.

Topics: Animals; Cell Line; Cell Nucleus; Cell Survival; Extracellular Signal-Regulated MAP Kinases; Free Radical Scavengers; Gene Expression Regulation; Iridoids; Lipopolysaccharides; Mice; Microglia; NF-kappa B; Nitric Oxide; Nitric Oxide Synthase Type II; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Protein Transport; Reactive Oxygen Species; Signal Transduction

Jasminoidin and ursodeoxycholic acid are 2 bioactive compounds extracted from Chinese medicine that have been proven to exert a synergistic effect as a combined administration for the treatment of stroke. The aim of this study was to reveal the pharmacogenomic mechanism of this synergistic effect of jasminoidin and ursodeoxycholic acid.. One hundred and fifteen mice with brain damage, induced by focal cerebral ischemia/reperfusion, were divided into 5 groups: jasminoidin-treated, ursodeoxycholic acid-treated, combination-treated, vehicle group, and sham-operated group. Comparative analysis of stroke-related gene expression profiles and Kyoto Encyclopedia of Genes and Genomes pathways among the 3 treatment groups were performed to reveal the mechanism of this synergistic effect.. This study demonstrated that (1) treatment with jasminoidin alone caused similar changes in the pattern of gene expression as those treated with the combination; (2) jasminoidin treatment and the combination treatment had more overlapping changes in gene expression and activated pathways than the ursodeoxycholic acid treatment; (3) Hspa1a and Ppm1e were only up-regulated in the combination-treated group; (4) the nonoverlapping genes Fgf12, Rarα, Map3k4, paxillin (PXN) in the combination-treated group were markedly expressed, and P53 pathway was obviously activated in the combination-treated group.. These findings may suggest that jasminoidin is the major component of the combination, and the combination plays an important role of the synergistic effect in up-regulating expression of gene Hspa1a, genes Fgf12, Rarα, Map3k4 and down-regulating gene PXN, as well as activating P53 pathway.

Topics: Animals; Cluster Analysis; Coloring Agents; Databases, Genetic; DNA, Complementary; Drug Synergism; Gene Expression; Gene Expression Profiling; Iridoids; Mice; Microarray Analysis; Principal Component Analysis; Real-Time Polymerase Chain Reaction; Reperfusion Injury; RNA; Signal Transduction; Stroke; Ursodeoxycholic Acid

Idiopathic mesenteric phlebosclerosis (IMP) is a rare disease, characterised by thickening of the wall of the right hemicolon with calcification of mesenteric veins. However, the aetiology remains unknown.. To investigate the possible association of herbal medicines with IMP.. The clinical data of four of our own patients were collected. Furthermore, we searched for previous reports about similar patients with detailed descriptions of herbal prescriptions that they had taken. We compared herbal ingredients to identify the toxic agent as a possible aetiological factor.. Clinical data on a total of 25 patients were summarised. Mean age was 61.8 years and there was female predominance (6 men and 19 women). The used Kampo prescription, the number of cases, and the mean duration of use were as follows: kamisyoyosan in 12 cases for 12.8 years, inshin-iseihaito in 5 cases for 13.4 years, orengedokuto in 4 cases for 14.3 years, inchinkoto in 1 case for 20 years, kamikihitou in 1 case for 19 years, seijobofuto in 1 case for 10 years and gorinsan in 1 case for an unknown duration. Only one ingredient, sansisi, was common to the herbal medicines of all 25 patients. This crude drug called geniposide in English is a major constituent of the Gardenia fruits.. The long-term use of geniposide in herbal medicines appears to be associated with mesenteric phlebosclerosis.

Topics: Aged; Biopsy; Drugs, Chinese Herbal; Female; Humans; Intestinal Mucosa; Iridoids; Male; Mesenteric Vascular Occlusion; Mesenteric Veins; Middle Aged; Plants, Medicinal; Sclerosis; Time Factors; Tomography, X-Ray Computed

β-Cell apoptosis is considered to be a major cause of loss of β cells in diabetes. Geniposide could prevent oxidative stress-induced neuron apoptosis, and improved glucose stimulated insulin secretion by activating glucagon-like peptide 1 receptor (GLP-1R) in INS-1 cells. Here we have investigated whether geniposide can exert a direct effect against pancreatic β-cell lipoapoptosis. The results indicated that pretreatment pancreatic INS-1 cells with geniposide for 7h attenuated palmitate-induced β-cell apoptosis and active caspase-3 expression, but this effect was disappeared at 18 h. Long-term incubation with palmitate decreased GLP-1R expression in INS-1 cells, and exendin (9-39), an antagonist for GLP-1R, inhibited the effect of geniposide on palmitate-induced apoptosis in INS-1 cells. Moreover, geniposide also improved the impairment of GLP-1R signaling through enhancing the phosphorylation of Akt and Foxo1, and increased the expression of PDX-1 in palmitate-treated INS-1 cells. These results suggest that geniposide inhibits early stage of lipotoxicity-induced β-cell apoptosis, and GLP-1R plays a critical role in geniposide counteracting the action of lipotoxicity in INS-1 pancreatic β cells.

Topics: Animals; Apoptosis; Caspase 3; Cell Line, Tumor; Drug Antagonism; Glucagon-Like Peptide 1; Glucagon-Like Peptide-1 Receptor; Insulin-Secreting Cells; Iridoids; Palmitic Acid; Peptide Fragments; Rats; Receptors, Glucagon; Signal Transduction

Geniposide, an iridoid glycoside isolated from Gardenia, has neuroprotective activities against oxidative stress and inflammation. The present study investigated the in vivo protective effect of geniposide on ischemia/reperfusion-injured rats by middle cerebral artery occlusion (MCAO), and the inhibitory effects of geniposide and mechanisms against activation of microglial cells by oxygen-glucose deprivation (OGD) in vitro. Male SD rats were subjected to treatment with geniposide at 15, 30 and 60 mg/kg immediately after MCAO. Cerebral infarct volume and microglial cell activation were assessed following 24 h reperfusion. Cultured primary rat microglial cells were exposed to geniposide at the concentrations of 12.5, 25 and 50 μg/mL during 4 h of OGD. The effects of geniposide were evaluated in terms of (1) cell viability; (2) secretion of TNF-α, IL-1β, IL-6, IL-8 and IL-10 into culture media; (3) TLR4 mRNA expression; (4) protein expression of TLR4, p-ERK1/2, p-IκB, p-p38, nuclear and cytoplasmic fraction NF-κB p65; and (5) nuclear transfer of NF-κB p65. Geniposide reduced the infarct volume and inhibited the activation of microglial cells in ischemic penumbra in vivo. OGD increased cell viability and release of TNF-α, IL-1β, IL-6, IL-8 and IL-10, these effects were suppressed by geniposide. Geniposide also attenuated the increases in the OGD-induced TLR4 mRNA and protein levels. In addition, geniposide at 25 and 50 μg/mL downregulated the phosphorylation of ERK, IκB and p38, as well as inhibited nuclear transcriptional activity triggered via NF-κB p65 in microglial cells by OGD. In conclusion, geniposide displays a neuroprotective effect on ischemia/reperfusion-injured rats in vivo and inhibits OGD-induced activation of microglial cells by attenuating inflammatory factors and NF-κB activation in vitro.

Topics: Animals; Culture Media; Cytokines; Dose-Response Relationship, Drug; Glucose; Inflammation; Iridoids; Male; Microglia; Oxygen; Rats; Rats, Sprague-Dawley; Signal Transduction; Toll-Like Receptor 4

Geniposide, a main iridoid glucoside component of gardenia fruit, has been known to exhibit antibacterial, anti-inflammatory and other important therapeutic activities. The objective of this study was to investigate the protective effects of geniposide on inflammation in lipopolysaccharide (LPS) stimulated primary mouse macrophages in vitro and LPS induced lung injury model in vivo. The expression of pro-inflammatory cytokines was determined by enzyme-linked immunosorbent assay (ELISA). Nuclear factor-kappa B (NF-κB), inhibitory kappa B (IκBα) protein, p38, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and Toll-like receptor 4 (TLR4) were determined by Western blot. Further analysis was carried out in mTLR4 and mMD-2 co-transfected HEK293 cells. The results showed that geniposide markedly inhibited the LPS-induced TNF-α, IL-6 and IL-1β production both in vitro and in vivo. Geniposide blocked the phosphorylation of IκBα, p65, p38, ERK and JNK in LPS stimulated primary mouse macrophages. Furthermore, geniposide inhibited the expression of TLR4 in LPS stimulated primary mouse macrophages and inhibited the LPS-induced IL-8 production in HEK293-mTLR4/MD-2 cells. In vivo study, it was also observed that geniposide attenuated lung histopathologic changes in the mouse models. These results suggest that geniposide exerts an anti-inflammatory property by down-regulating the expression of TLR4 up-regulated by LPS. Geniposide is highly effective in inhibiting acute lung injury and may be a promising potential therapeutic reagent for acute lung injury treatment.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Bronchoalveolar Lavage Fluid; Cells, Cultured; Female; Gardenia; Gene Expression Regulation; Granulocyte Colony-Stimulating Factor; Interleukin-3; Iridoids; Macrophages, Peritoneal; Male; Mice; Mice, Inbred BALB C; Recombinant Fusion Proteins

Huang-Lian-Jie-Du-Tang (HLJDT), a famous traditional Chinese prescription with wide anti-inflammatory applications, is an aqueous extract of four herbal materials: Rhizoma coptidis, Radix scutellariae, Cortex phellodendri, and Fructus gardeniae. Its effects on the cyclooxygenase (COX)-2 and 5-lipoxygenase (5-LOX) pathways are thought to be responsible for its anti-inflammatory activity. However, our previous work found that the inhibitory effects of HLJDT act on the 5-LOX pathway but not on the COX pathway. The possibility that HLJDT inhibits COX-2- or 5-LOX-catalyzed eicosanoid generation by downregulating enzyme expression requires further investigation.. To observe the effects of HLJDT and its four major components (baicalin, baicalein, berberine and geniposide) on COX-2- or 5-LOX-catalyzed eicosanoid generation and to distinguish the effects of HLJDT on enzyme activity from those on enzyme expression.. The topical anti-inflammatory activities and inhibition of eicosanoid formation of HLJDT and its components were observed in an arachidonic acid (AA)-induced mouse ear edema model. Macrophage-based systems were established to observe the effects of the drugs on enzyme activity and enzyme expression of COX-2 and 5-LOX. Further experiments were carried out to confirm these effects at the mRNA and protein levels.. Topical treatment of HLJDT significantly inhibited AA-induced mouse ear edema and reduced PGE(2) and LTB(4) release in the edematous ears. Baicalein, geniposide, and berberine also ameliorated the symptoms and suppressed eicosanoid generation with varying efficacies. Cell-based assays showed that HLJDT and baicalein inhibited the PGE(2) levels by decreasing COX-2 enzyme expression without affecting COX-2 enzyme activity in RAW 246.7 murine macrophages. The other experiments on rat peritoneal macrophages indicated that HLJDT and baicalein exerted significant inhibition on LTB(4) production by decreasing 5-LOX enzyme activity. The real-time PCR and western blotting data demonstrated that HLJDT and baicalein reduced COX-2 expression at the mRNA and protein levels, whereas no inhibition on 5-LOX expression was observed.. HLJDT can suppress eicosanoid generation via both the COX and LOX pathways, which definitely contributes to its topical anti-inflammatory activity. We have confirmed that its dual inhibition on the COX and LOX pathways mainly result from the downregulation of COX-2 expression and direct inhibition of 5-LOX activity, respectively. Baicalein worked as a potent active component in most of the tests. These findings about the different inhibitory effects of HLJDT on COX-2 and 5-LOX help to better understand the mechanism of HLJDT and promote safer applications of drug.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Arachidonate 5-Lipoxygenase; Arachidonic Acid; Berberine; Cell Line; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Dinoprostone; Drugs, Chinese Herbal; Edema; Flavanones; Flavonoids; Iridoids; Leukotriene B4; Lipoxygenase Inhibitors; Male; Mice; Mice, Inbred ICR; Rats; Rats, Sprague-Dawley; RNA, Messenger

Geniposide was prepared on a large-scale using a selective two-phase liquid-liquid extraction. The aqueous residue from the fruit of Gardenia jasminoides Ellis was treated with sodium carbonate and extracted with n-butanol several times. The n-butanol extracts were treated with activated granular charcoal to remove pigments and were then concentrated to produce a residue with a high solid content. The residue was crystallized to obtain geniposide with 98% purity. For large-scale synthesis, the residue (solid content 45%, geniposide 5.5%) was extracted to generate 70g of geniposide with 98% purity and 84.8% recovery using 1500g residue.

Topics: Butanols; Fruit; Gardenia; Hydrogen-Ion Concentration; Iridoids; Liquid-Liquid Extraction; Molecular Structure

Refined Qing-Kai-Ling (QKL), a modified Chinese medicine, consists of three main ingredients (Baicalin, Jasminoidin and Desoxycholic acid), plays a synergistic effect on the treatment of the acute stage of ischemic stroke. However, the rules of the combination and synergism are still unknown. Based on the ischemic stroke mice model, all different kinds of combination of Baicalin, Jasminoidin, and Desoxycholic acid were investigated by the methods of neurological examination, microarray, and genomics analysis. As a result, it confirmed that the combination of three drugs offered a better therapeutical effect on ischemic stroke than monotherapy of each drug. Additionally, we used Ingenuity pathway Analysis (IPA) and principal component analysis (PCA) to extract the dominant information of expression changes in 373 ischemia-related genes. The results suggested that 5 principal components (PC1-5) could account for more than 95% energy in the gene data. Moreover, 3 clusters (PC1, PC2+PC5, and PC3+PC4) were addressed with cluster analysis. Furthermore, we matched PCs on the drug-target networks, the findings demonstrated that Baicalin related with PC1 that played the leading role in the combination; Jasminoidin related with PC2+PC5 that played a compensatory role; while Desoxycholic acid had the least performance alone which could relate with PC3+PC4 that played a compatible role. These manifestations were accorded with the principle of herbal formulae of Traditional Chinese Medicine (TCM), emperor-minister-adjuvant-courier. In conclusion, we firstly provided scientific evidence to the classic theory of TCM formulae, an initiating holistic viewpoint of combination therapy of TCM. This study also illustrated that PCA might be an applicable method to analyze the complicated data of drug combination.

Topics: Animals; Cluster Analysis; Computational Biology; Deoxycholic Acid; Disease Models, Animal; Drug Combinations; Drugs, Chinese Herbal; Enzyme Inhibitors; Flavonoids; Gene Expression Profiling; Gene Expression Regulation; Iridoids; Ischemia; Male; Mice; Models, Statistical; Oligonucleotide Array Sequence Analysis; Polymerase Chain Reaction; Principal Component Analysis; Stroke

Both geniposide (Ge) and borneol (Bo) are bioactive substances derived from traditional Chinese medicine. Injections containing co-compound of Gardenia-Borneol are widely used for stroke treatment in China, such as "Xingnaojing" multi-component injection. As more and more adverse reactions (especially drug allergy) were reported, it is urgent to find more effective and safer routes of administration for such kinds of medicines. In this paper, bioavailabilities and brain-target effects of geniposide in Gardenia-Borneol co-compound through different administration routes in mice were investigated. Geniposide concentrations in plasma and in brain of mice were determined by reversed-phase high-performance liquid chromatography. The pharmacokinetics parameters of intranasal (i.n.) and intragastric (i.g.) administration were compared with intravenous (i.v.) administration. The bioavailabilities of Ge were 85.38% and 28.76% for i.n. and i.g. while T(max) were 1 min and 30 min. C(max) were 21.881 ± 5.398, 1.914 ± 0.327 and 42.410 ± 6.268 μg/mL for i.n., i.g. and i.v., respectively. The AUC of Ge in brain were 32413.6 ± 4573.9, 6440.1 ± 863.7 and 37270.5 ± 4160.6 ng/g ·min for i.n., i.g. and i.v., respectively. The drug target indexes (DTI) were 1.02 and 0.60 for i.n. and i.g. The results demonstrated that geniposide could be absorbed promptly and thoroughly by i.n. administration in mice and basically transported into the brain though blood vessel passways.

Topics: Animals; Biological Availability; Brain; Camphanes; Drug Administration Routes; Drugs, Chinese Herbal; Gardenia; Iridoids; Male; Medicine, Chinese Traditional; Mice; Plant Extracts

To establish a method for determination of geniposide in Beagle dogs plasma by high performance liquid chromatography (HPLC), and study the pharmacokinetics and bioavailability of geniposide in Beagle dogs after oral administration Xingnaojing.. To determine the geniposide in Beagle dogs plasma by HPLC after oral administration or intravenous injection Xingnaojing, and the pharmacokinetic parameters were calculated by the software of Kinetica.. The good linearity range of geniposide was 1.24 - 158.88 mg x L(-1). The main pharmacokinetic parameters after oral administration was as follows: Cmax (11.8 +/- 0.6) mg x L(-1), Tmax (52.0 +/- 4.5) min, AUC(1280.8 +/- 172.0) mg x min x L(-1), MRT(118.7 +/- 25.4) min, and these parameters after intravenous injection was follows: Cmax 107.4 +/- 6.3) mg x L(-1), AUC(7930.1 +/- 670.0) mg x min x L(-1), MRT(92.4 +/- 5.1) min. The bioavailability of geniposide in Beagle dogs after oral administration Xingnaojing was (6.46 +/- 0.87)%.. The HPLC method had good applicability. The extract recovery, method recovery, intra-day precision and inter-day precision of the method were all met the requirements. The stability in conditions of room temperature and freeze-thaw cycle was good. The results indicated that the oral administration bioavailability of geniposide was in low degree.

Topics: Administration, Oral; Animals; Biological Availability; Chromatography, High Pressure Liquid; Dogs; Drugs, Chinese Herbal; Iridoids

We reported previously that geniposide showed neurotrophic and neuroprotective activities with the activation of glucagons-like peptide 1 receptor (GLP-1R) in neurons. The current study was designed to further investigate the protective effect of geniposide on β-amyloid (Aβ)-induced cytotoxicity. Our results showed that pre-incubation with geniposide prevented Aβ₁₋₄₂-induced cell injury in primary cultured cortical neurons. Geniposide also induced the expression of insulin-degrading enzyme (IDE), a major degrading protease of Aβ, in a dose-dependent manner. Moreover, bacitracin, an inhibitor of IDE, and RNAi on Glp-1r gene decreased the neuroprotection of geniposide in Aβ₁₋₄₂-treated cortical neurons. Our findings indicated that geniposide activating GLP-1R to against Aβ-induced neurotoxicity involved in its regulation on the expression of IDE in cortical neurons, which provided an additional mechanistic insight into the role of GLP-1R in neuroprotection.

Topics: Amyloid beta-Peptides; Animals; Cerebral Cortex; Dose-Response Relationship, Drug; Female; Gene Expression Regulation, Enzymologic; Insulysin; Iridoids; Male; Neurons; Peptide Fragments; Rats; Rats, Sprague-Dawley

To optimize the extraction process for Qingrejiedu oral liquid with synthesizing multiple guidelines grading method.. Used the extraction rate of baicalin and geniposide comprehensive contribution rate variance as index, extraction time and added water were as factors, central composite design was used for optimization of extraction process, and forecasting analysis parameters.. The optimal extraction process was as follows: extraction time 139 min, added water 13 times, grading index was 0.768.. Synthesizing multiple guidelines grading method is ideal for multi-component extraction experiment design, its easy application, the forecast is good and so on.

Topics: Administration, Oral; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavonoids; Gardenia; Iridoids; Plants, Medicinal; Quality Control; Reproducibility of Results; Scutellaria baicalensis; Technology, Pharmaceutical

To optimize the process for preparing genipin with enzymolyzed geniposide by an orthogonal experiment.. The optimal enzymolysis process was selected by an orthogonal experiment, with the concentration of geniposide as the index as well as enzyme quantity, pH of enzymolysis solution, enzymolysis time and enzymolysis temperature as considerations.. The optimal hydrolysis conditions were as follow: rough genipin samples at the concentration of 40 g x L(-1) were selected and shook on a table concentrator at a speed of 100 r x min(-1), added with beta-glucosidase-geniposide 1 : 1 (weight proportion), with pH of enzymolysis solution at 4.5, hydrolyzation temperature at 50 degrees C, the conversion rate of genipin could reach 85.8%.. The process is so stable and feasible that it can provide theoretical basis for the preparation of genipin with enzymolyzed geniposide.

Topics: Hydrolysis; Iridoids; Technology, Pharmaceutical

Optimized-SopungSunkiwon (OSS) is a multi-herbal formula that contains six medicinal herbs from SopungSunkiwon, a traditional medicine used for neurodegenerative disorders. In this study, we investigated the anti-amnesic effects of OSS in a dementia model. Acetylcholinesterase (AChE) inhibition assay was performed to investigate the cholinergic antagonistic effect of OSS. In addition, a step-through passive-avoidance test was performed with scopolamine-induced memory impairment in mice, and immunohistochemistry was analyzed to investigate synaptic formation with synaptic proteins. OSS inhibited AChE activity, resulting in significant improvement of memory functions. In the passive-avoidance test, the latency time of OSS-treated mice was significantly longer than that of either the control or scopolamine-treated group. In the immunohistochemical analysis, synaptic proteins such as synaptophysin and PSD-95 were significantly increased in OSS-treated mice. These results demonstrate that OSS may affect impairment and enhancement of memory and increase synaptophysin and PSD-95 facilitating acetylcholine release and synaptic growth.

Topics: Acetylcholinesterase; Animals; Anthraquinones; Avoidance Learning; Brain; Carotenoids; Chromatography, High Pressure Liquid; Immunohistochemistry; Iridoids; Male; Maze Learning; Medicine, Korean Traditional; Memory; Memory Disorders; Mice; Mice, Inbred ICR; Neuroprotective Agents; Phytotherapy; Plant Extracts; Plants, Medicinal; Senna Extract; Sennosides

Both geniposide (Ge) and natural borneol (NB) are bioactive substances derived from traditional Chinese herbs. The effect of NB on the pharmacokinetics of Ge in rat via intranasal administration was investigated. The concentrations of Ge in plasma were determined by reversed-phase high-performance liquid chromatography (HPLC) after intranasal administration of Ge (4 mg/kg) alone and combined with different doses (0.08, 0.8, and 8 mg/kg) of NB. The intravenous administration was given as a reference (4 mg/kg of Ge and 8 mg/kg of NB). Compared with the intravenous administration, the absolute bioavailability of Ge was 76.14% through intranasal administration combined with NB. Compared with the intranasal administration of Ge alone, Ge could be absorbed rapidly in the nasal cavity combined with NB; the peak time of Ge in the plasma became shorter (3-5 min vs. 40 min); the peak concentration became higher (1.32-4.25 μg/ml vs. 0.67 μg/ml); and, the relative bioavailability of Ge combined with NB was 90.3%-237.8%. The enhancing effect was attenuated as the dose of NB decreased. The results indicated that NB can accelerate the absorption of Ge dose-dependently in the nasal cavity.

Topics: Absorption; Administration, Intranasal; Animals; Biological Availability; Camphanes; Drug Synergism; Gardenia; Injections, Intravenous; Iridoids; Male; Nasal Mucosa; Plants, Medicinal; Rats; Rats, Sprague-Dawley

Nonalcoholic steatohepatitis (NASH), a metabolic disorder of the liver, may gradually evolve into fibrosis or cirrhosis. Recent studies have suggested that geniposide can effectively inhibit experimental liver fibrosis. Therefore, the aim of this study was to determine whether geniposide can influence the early phase of fibrogenesis in an animal model of NASH.. Male Sprague-Dawley rats were given a high fat diet alone or the same diet combined with geniposide at doses of 25, 50 or 100 mg/kg for six weeks. Ten rats received corresponding solvent as a normal control.. Treatment with geniposide could improve liver histology through reducing the elevated liver index (liver weight/body weight), serum alanine aminotransferase and aspartate aminotransferase. Total cholesterol, triglycerides and free fatty acids in serum and liver decreased in geniposide-treated rats. Furthermore, geniposide increased serum insulin levels but reduced serum tumour necrosis factor-α level in high-fat diet rats. In addition, geniposide suppressed expression of CYP2E1 and increased peroxisome proliferator-activated receptor-α (PPARα) expression. These benefits may be associated with increased superoxide dismutase and decreased malondialdehyde in liver.. Geniposide exerts protective effects against hepatic steatosis in rats fed with a high fat diet; the underlying mechanism may be associated with its antioxidant actions or regulation of adipocytokine release and expression of PPARα.

Topics: Alanine Transaminase; Animals; Anti-Inflammatory Agents; Aspartate Aminotransferases; Cytochrome P-450 CYP2E1; Dietary Fats; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Fatty Liver; Insulin; Iridoids; Lipid Metabolism; Male; Malondialdehyde; Non-alcoholic Fatty Liver Disease; PPAR alpha; Rats; Rats, Sprague-Dawley; Superoxide Dismutase; Tumor Necrosis Factor-alpha

Genipin is the bioactive compound of geniposide and a natural cross-linking agent. In order to improve the preparation process of genipin, the hydrolysis of geniposide to genipin by immobilized β-glucosidase in an aqueous-organic two-phase system was studied. β-glucosidase was immobilized by the crosslinking-embedding method using sodium alginate as the carrier. The optimum reaction temperature, pH value and time were 55 °C, 4.5 and 2.5 h, respectively. To reduce genipin hydrolysis and byproduct production the reaction was carried out in an aqueous-organic two-phase system comprising ethyl acetate and sodium acetate buffer. The product was analyzed by HPLC, UV, IR, and NMR. The yield of genipin was 47.81% and its purity was over 98% (HPLC).

Topics: beta-Glucosidase; Enzyme Stability; Hydrogen-Ion Concentration; Hydrolysis; Iridoid Glycosides; Iridoids; Organic Chemistry Phenomena; Temperature; Time Factors

Combination therapies have recently been shown to be more effective than monotherapies that may provide synergistic effects in the treatment of stroke, but its selective mechanism still remains unclear. Based on the median-effect method, the combination therapy of jasminoidin and ursodeoxycholic acid had a synergic effect on reducing the infarct volume. The numbers of up- or down-regulated genes by at least 1.5-fold in the vehicle, jasminoidin, ursodeoxycholic acid, and the combination of jasminoidin and ursodeoxycholic acid treatment groups were 228, 95, 136, and 101, respectively. According to clustering and principal component analysis, the pattern of gene expression in the combination group was similar to that of jasminoidin group rather than ursodeoxycholic acid group. Based on these nine top sequences in the combination group excluding four overlapping pathways (MAPK-ERK, Kitlg, Icam1-Ap1, and prolactin), the jasminoidin group had four (PRLR-STAT1, AcvR2-AcvR1B, ACVR1/2A-SMAD1, GHR-NF-κB) contributing pathways, and the ursodeoxycholic acid group had one (IL-6) contributing pathway. Based on the multiple-pathway-dependent comparison analysis (MPDCA), it may lead to the conclusion that jasminoidin possibly contributes more important pharmacological effect in the combined treatment as jasminoidin regulated 80% of the pathways that the combination group mediated. The study reveals a horizontal synergistic effect by optimizing the fusion of more pathways from the compounds with more contribution to the combination therapy. Rather than selecting compounds only based on experience in the past, this study would give a new insight into the systematic strategies for designing synergistic combination therapies.

Topics: Animals; Brain Ischemia; Cluster Analysis; Drug Combinations; Drug Synergism; Gene Expression Regulation; Hippocampus; Iridoids; Male; Mice; Principal Component Analysis; Ursodeoxycholic Acid

Iridoids are one of the most widely distributed secondary metabolites in higher plants. They are pharmacologically active principles in various medicinal plants and key intermediates in the biosynthesis of monoterpenoid indole alkaloids as well as quinoline alkaloids. Although most iridoids are present as 1-O-glucosides, the glucosylation step in the biosynthetic pathway has remained obscure. We isolated a cDNA coding for UDP-glucose:iridoid glucosyltransferase (UGT85A24) from Gardenia jasminoides. UGT85A24 preferentially glucosylated the 1-O-hydroxyl group of 7-deoxyloganetin and genipin but exhibited only weak activity toward loganetin and no activity toward 7-deoxyloganetic acid. This suggests that, in the biosynthetic pathway of geniposide, a major iridoid compound in G. jasminoides, glucosylation occurs after methylation of 7-deoxyloganetic acid. UGT85A24 showed negligible activity toward any acceptor substrates other than iridoid aglycones. Thus, UGT85A24 has a remarkable specificity for iridoid aglycones. The mRNA level of UGT85A24 overlaps with the marked increase in genipin glucosylation activity in the methyl jasmonate-treated cell cultures of G. jasminoides and is related to iridoid accumulation in G. jasminoides fruits.

Topics: Base Sequence; DNA, Complementary; Fruit; Gardenia; Glycosyltransferases; Iridoids; Methylation; Molecular Sequence Data; Plant Proteins; Substrate Specificity

To optimize the process for purification of geniposide in the extract fluid of Gardenia jasminoides with macroporous absorption resin.. By comparing the content and transfer ratio of geniposide during the process of purification, we optimized the process for purification of geniposide.. The optimal process for purification of geniposide with D301R macroporous absorption resin included the diameter height ratio 1:7.5, the concentration of the extract fluid of Gardenia jasminoides 2:1, the flow rate 1BV/h (1BV represented one column volume), the sample volume 1/3BV. We loaded the sample and left it for 2 hours, afterwards, rinsed the macroporous absorption resin using 2BV water until the solution became colourless. Then we rinsed the macroporous absorption resin with 20% alcohol,and the volume of elution solvent was 2BV. We collected 20% alcohol elution solvent and recycle alcohol using rotating evaporation and dried the rest solution in a vacuum to get the light yellow powder which was the purified geniposide.. This process is simple and affordable, can be used to refine and purify geniposide in the extract fluid of Gardenia jasminoides Ellis. It provides a guidance for industrial production basis.

Topics: Absorption; Chromatography, High Pressure Liquid; Fruit; Gardenia; Iridoids; Reproducibility of Results; Resins, Synthetic; Solvents; Technology, Pharmaceutical

6,7-Dimethylesculetin (D), geniposide (G) and rhein (R) are the three major active ingredients of Yin-Chen-Hao-Tang (YCHT), a famous Chinese herbal formula, which has been shown to be clinically effective for treating hepatic injury (HI) syndrome. The present study was conducted to investigate the therapeutic and synergistic effects of COC (combination of D, G and R) on HI rats by combining pharmacokinetic with biochemical analysis strategy. Plasma was analyzed by using reversed-phase high performance liquid chromatography (RP-HPLC). Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) models were built to evaluate the therapeutic and synergistic effects of COC at the biochemical level. Here, we report that the COC combination could increase the plasma level, slow elimination rate, exert a more robust therapeutic effect than any one or two of the three individual compounds by hitting multiple targets in a rat model of HI. Overall, this beneficially accounts for the popular view that traditional Chinese medicine (TCM) formula usually takes multi-component to exert their therapeutic effects. We suggest that dissecting the mode of action of clinically effective formula to be capable of producing a sufficient effect at low doses.

Topics: Animals; Anthraquinones; Artemisia; Carbon Tetrachloride; Chemical and Drug Induced Liver Injury; Chromatography, High Pressure Liquid; Drug Combinations; Drug Synergism; Drugs, Chinese Herbal; Gardenia; Iridoids; Least-Squares Analysis; Magnoliopsida; Male; Phytotherapy; Principal Component Analysis; Rats; Rats, Wistar; Rheum; Umbelliferones

To develop a HPLC- ELSD method for determination the contents of geniposide, crocin and crocetin in different processing products of Fructus Gardeniae.. The separation was performed in the HyperClone ODS C18 column (250 mm x 4. 6 mm, 5 microm) with linear gradient elution using methanol-water and 0.05% phosphoric acid in water, the flowing rate was 0.8 mL/min, the column temperature was 30 degrees C, and the ELSD parameter was as follow: 70 degrees C as atomization temperature and 2.0 L/min as the gas flowing rate.. The contents of geniposide and crocin in raw, yellowish, carbocoal and scorched Fructus Gardeniae decreased with the deepening of processing degree. However, the content of crocetin in carbocoal and scorched Fructus Gardeniae increased comparing with the raw one.. This is a simple and credible quality control method, and can be used for the quality control and comprehensive evaluation for different processed products of Fructus Gardeniae.

Topics: Carotenoids; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Fruit; Gardenia; Iridoids; Quality Control; Reproducibility of Results; Technology, Pharmaceutical; Vitamin A

Accumulation of visceral fat induces various symptoms of metabolic syndrome such as insulin resistance and abnormal glucose/lipid metabolism and eventually leads to the onset of ischemic cerebrovascular diseases. Geniposide, which is iridoid glycoside from the fruit of Gardenia jasminoides ELLIS, is recognized as being useful against hyperlipidemia and fatty liver. In order to clarify the effect of geniposide on metabolic disease-based visceral fat accumulation and the relevant molecular mechanism, experiments were performed in spontaneously obese Type 2 diabetic TSOD mice and the free fatty acid-treated HepG2 cells. In the TSOD mice, geniposide showed suppression of body weight and visceral fat accumulation, alleviation of abnormal lipid metabolism and suppression of intrahepatic lipid accumulation. In addition, geniposide alleviated abnormal glucose tolerance and hyperinsulinemia, suggesting that geniposide has an insulin resistance-alleviating effect. Next, in order to investigate the direct effect of geniposide on the liver, the effect on the free fatty acid-treated HepG2 fatty liver model was investigated using genipin, which is the aglycone portion of geniposide. Genipin suppressed the intracellular lipid accumulation caused by the free fatty acid treatment and also significantly increased the intracellular expression of a fatty acid oxidation-related gene (peroxisomal proliferator-activated receptor: PPARα). From these results, it was confirmed that geniposide has an anti-obesity effect, an insulin resistance-alleviating effect and an abnormal lipid metabolism-alleviating effect, and the metabolite genipin shows a direct effect on the liver, inducing expression of a lipid metabolism-related gene as one of its molecular mechanisms.

Topics: Adipose Tissue; Animals; Anti-Obesity Agents; Body Weight; Diabetes Mellitus, Type 2; Drug Evaluation, Preclinical; Fatty Acids, Nonesterified; Fatty Liver; Gardenia; Glucose Intolerance; Hep G2 Cells; Humans; Hyperinsulinism; Hypoglycemic Agents; Insulin Resistance; Iridoids; Lipid Metabolism; Liver; Male; Metabolic Diseases; Metabolic Syndrome; Mice; Mice, Obese; Obesity; Phytotherapy; Plant Preparations

To explore the network control mechanism of the calcium signaling pathway in cerebral ischemic injury after intervention by the main components of Qingkailing (see text), i.e. Baicalin, Jasminoidin and their combination.. Thirty mice were randomly divided into 5 groups, a baicalin group, a Jasminoidin group, a baicalin plus Jasminoidin group, a nimodipine group, and a model group (n = 6). The global cerebral ischemia-reperfusion mouse model was established. The mice were administrated respectively by injection of baicalin, Jasminoidin, mixture of baicalin and Jasminoidin, and nimodipine into the caudal vein, with the model group given no any drug. Three hours after operation, the brain was removed and sectioned. After calculation of cerebral ischemic area by 2,3,5-triphenyltetrazolium staining, the percentage of infarct volume was calculated. The total RNA of the mouse brain tissue was extracted to obtain the whole genome expression profile, and the differentially expressed genes related to the calcium signaling pathway was analyzed with Bayesian network structures.. Compared with the model group, the ischemic area was significantly reduced in the baicalin group, the Jasminoidin group, the Baicalin plus Jasminoidin group (all P < 0.05). The ischemic area in the baicalin plus Jasminoidin group was smaller than the other three groups (all P < 0.01). In the gene regulatory network structures of calcium signaling pathway, the average length and equitability were the highest in the baicalin plus Jasminoidin group, followed by the nimodipine group.. Compared with a single component, combination of Baicalin and Jasminoidin can more obviously intervene in the overall expression of calcium signaling pathway, and the mechanism is related with the aggregation characteristic of the gene expression network.

Topics: Animals; Brain Ischemia; Calcium Signaling; Drugs, Chinese Herbal; Flavonoids; Iridoids; Male; Mice; Nimodipine

Geniposide, an iridoid glycoside, is an important and characteristic compound in the fruits of Gardenia jasminoides Ellis, a commonly used medicinal herb in Chinese traditional and folk medicine for the treatment of inflammation and jaundice. However, few studies have been carried out on the metabolism of geniposide. In this study, we have established a rapid and sensitive method using ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UPLC/ESI-QTOF-MS) for analysis of the metabolic profile of geniposide in rat urine after oral administration. A total of ten metabolites were detected and identified by comparing their fragmentation patterns with that of geniposide using Metabolynx™ and MassFragment™ software tools. The results revealed that the principal metabolism pathways of geniposide in rat occurred after deglycosylation of the irdoid glycoside take place and this is followed by glucuronidation and the pyran-ring cleavages. The major metabolite, the glucuronic acid conjugate of genipin as observed in vivo, was further confirmed by the in vitro enzymatic study. The results of this work have demonstrated the feasibility of the UPLC/ESI-QTOF-MS approach for rapid and reliable characterization of metabolites from iridoid compounds.

Topics: Animals; Chromatography, High Pressure Liquid; Iridoids; Male; Metabolic Networks and Pathways; Rats; Rats, Wistar; Signal-To-Noise Ratio; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

Gardeniae Fructus is a traditional medicine used for the treatment of contusion such as ankle sprain. Geniposide is one of the main components of Gardeniae Fructus with diverse biological activities. In order to gain further insight into the therapeutic action of Gardeniae Fructus extract (GFE) and geniposide on ligament injuries, a new in vitro model was developed in the present study. Rat hind ankle ligament fibroblasts (RHALFs) derived from Sprague-Dawley rats were cultured, and the cell proliferation and collagen content were examined by MTT and a Sirius Red-based colorimetric assay after stimulating with each drug. The cell growth of RHALFs was promoted by culturing with 37.5-150 microg/mL of GFE and 25-200 microM of geniposide. The content of collagen in the RHALFs was significantly increased up to 131.4% and 124.2% of the control value by culturing with the GFE and geniposide, respectively. By contrast, both cell growth and collagen content were impaired by adding 25-200 microM of diclofenac, one of the common medications for ligament injuries. The findings suggest that GFE and geniposide may ameliorate the treatment of ligament injuries by proliferating ligament fibroblasts and promoting the synthesis of collagen. However, the use of diclofenac to treat acute ligament injuries should be reassessed although it possesses a potential effect on relieving symptoms.

Topics: Animals; Cell Proliferation; Cells, Cultured; Collagen; Diclofenac; Fibroblasts; Gardenia; Iridoids; Ligaments, Articular; Male; Plant Extracts; Rats; Rats, Sprague-Dawley

The major components of gardenia fruits are geniposide and water soluble pigment crocins. In this study, we investigate crocins and geniposide profiles of gardenia fruits from different cultivars and at the various stages of maturation. DPPH scavenging activity of gardenia fruits from different cultivars and at the various stages of fruit maturation was also assayed. Quantitative determination of crocins in the gardenia at the various stages of maturation revealed a significant increase when ripening. However, geniposide content was negatively correlated with ripening stages. A significant difference was observed when comparing crocin content of different gardenia from various cultivars and geniposide content also showed marked variety. Current study indicated no relationship between crocin and geniposide content in gardenia fruits at the various stages of maturation and DPPH radical scavenging activity. Data showed that, although crocins feature markedly less DPPH scavenging activity than gardenia ethanol extract, total crocin content of gardenias collected in various cultivars correlate, to a certain degree, with radical scavenging effects of the Chinese traditional medicine (r=0.75).

Topics: Antioxidants; Biphenyl Compounds; Carotenoids; Free Radical Scavengers; Fruit; Gardenia; Genotype; Iridoids; Molecular Structure; Picrates; Plant Extracts

The aim of this study was to investigate the inhibitory effect of geniposide on lipopolysaccharide (LPS)-induced interleukin-6 (IL-6) and interleukin-8 (IL-8) production in human umbilical vein endothelial cells (HUVECs).. Primary HUVECs were used. The mRNA/protein levels of IL-6 and IL-8 was determined by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). LPS-induced HUVEC migration and adhesion of monocytes to HUVECs were studied by monolayer wound healing experiments and monocytic cell adhesion assay, respectively. Expression of nuclear factor kappaB (NF-kappaB), inhibitory factor kappaB-alpha (IkappaB-alpha), p38 mitogen-activated protein kinase (MAPK) and ERK1/2 were determined by Western blot analysis.. Geniposide effectively inhibited LPS-induced expression of IL-6 and IL-8 in HUVECs at the transcription and translation levels. Additionally, geniposide suppressed LPS-induced HUVEC migration and U937 monocyte adhesion to HUVECs. Signal transduction studies indicate that geniposide blocked the activation of NF-kappaB, degradation of IkappaB-alpha, and phosphorylation of p38 MAPK and ERK1/2 in HUVECs challenged by LPS.. The results show that geniposide can inhibit LPS-induced IL-6 and IL-8 production in HUVECs by blocking p38 MAPK and ERK1/2 signaling pathways.

Topics: Blotting, Western; Cell Adhesion; Cell Movement; Cell Survival; Endothelial Cells; Enzyme-Linked Immunosorbent Assay; Extracellular Signal-Regulated MAP Kinases; Humans; I-kappa B Proteins; Indicators and Reagents; Interleukin-6; Interleukin-8; Iridoids; Lipopolysaccharides; Monocytes; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; U937 Cells; Umbilical Veins; Wound Healing

Eucommia ulmoides Oliver is a traditional medicinal plant of China, and it is one of the main sources of chlorogenic acid. Chlorogenic acid is an ester of caffeic acid, quinic acid, and a phenolic compound that has antibacterial, antifungal, antioxidant, and antitumor activities. The purpose of this study was to determine whether endophytic fungi isolated from Eucommia ulmoides Oliver had the same ability to produce chlorogenic acid. Primary screening was done by antibacterial and antifungal reactions, and the strain reselection was done with high-performance liquid chromatography (HPLC) to identify the fermentation products of the selected strains. Extracts of the leaf and cortex of Eucommia ulmoides Oliver were also deteted by HPLC, then positive results of HPLC were analyzed by GC-MS and LC-MS. In this study, 29 strains were isolated from Eucommia ulmoides Oliver. Most of them had antibacterial activity, and a few of them had antifungal activity. One ingredient of the B5 extract had a retention time identical to that of authentic chlorogenic acid. With GC-MS, other ingredients, isocoumarin and p-chlorocinnamide, were found. With LC-MS, chlorogenic acid and geniposide related to Eucommia ulmoides Oliver were found. The strain B5 was identified as Sordariomycete sp. Thus, endophytic fungi may produce the bioactive compound chlorogenic acid, as their host plant does, and could be used for the production of chlorogenic acid by fermentation in the future.

Topics: Anti-Bacterial Agents; Antifungal Agents; Antineoplastic Agents; Ascomycota; China; Chlorogenic Acid; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Eucommiaceae; Iridoids; Plant Leaves

To study the phenomenon of subsidence emergence in the process of Huanglian Jiedu Tang decoction extraction, and the mechanism of subsidence emergence.. UV was applied to determine the concentration of total alkaloids and total flavones; Simultaneous determination of berberine, jatrorrhizine, palmatine, baicalin and geniposide were carried out by HPLC; The solid holdup and precipitation rate were calculated by the formula. The relativity among these parameters was analyzed by the SPSS software program.. The contains of berberine, jatrorrhizine, palmatine in total alkaloids showed a good correlation with total alkaloids. The correlation between baicalin and total flavones was lower than that between geniposide and total flavones. Compared to precipitation rate, solided hold up shows a larger relevance with index component. With the change of time, the total alkaloids represented by berberine alkaloids and baicalin at a certain concentration can be regarded as the equilibrium point, or one generated by the critical point of precipitation, the reaction can generate "sediments"; because "precipitation objects" generated, reducing the concentration of the above-mentioned components, destroy the" balance". The relevant components of herbal medicine increased dissolution rate, at the same time might partly dissolved sediment, reaching a new equilibrium state.

Topics: Alkaloids; Berberine; Berberine Alkaloids; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavones; Flavonoids; Fractional Precipitation; Iridoids; Plants, Medicinal

Zhongtong Caji, a kind of liniment, is a traditional Chinese medicinal formula that is widely used for clinical treatment of inflammation and sprains. In this study, the principal effective compound of this formula, geniposide, was used as a criterion to represent the transdermal permeability of the whole formula. A passive diffusion of Zhongtong Caji through the stratum corneum was discovered by an in vitro experiment. The dosage-content relationship detected in subcutaneous tissue after in vivo drug administration was further evidence of its permeation. Blood analysis after different dosages showed that the geniposide could be absorbed and accumulated by subcutaneous tissue within 1h after drug administration, and it would be eliminated by blood circulation 1h after drug treatment.

Topics: Administration, Cutaneous; Animals; Chromatography, High Pressure Liquid; Dose-Response Relationship, Drug; Epidermis; In Vitro Techniques; Iridoids; Liniments; Male; Mice; Mice, Inbred ICR; Plant Preparations; Skin Absorption; Tissue Distribution

The traditional Chinese medicine formula Fuling Decoction (FD) has been clinically used for eczema treatment, but the unclear chemical distribution and the lack of quality control have strongly restricted its application. In this study, an analytical method incorporating ultra-fast liquid chromatography (UFLC) with MS and UV detection was developed for rapid profiling of the chemical constitutes from FD. Fourteen constitutes were identified by UFLC-ESI-MS, while four major components including genipingentiobioside, geniposide, paeoniflorin and liquiritin were quantified simultaneously by UFLC-DAD. The UFLC-based method was fully validated and can be applied to screening and determination of principal components in commercially FD prescriptions.

Topics: Benzoates; Bridged-Ring Compounds; Chromatography, Liquid; Drugs, Chinese Herbal; Flavanones; Glucosides; Iridoids; Monoterpenes; Quality Control; Spectrometry, Mass, Electrospray Ionization

To explore the role of activation of glucagon-like peptide 1 receptor (GLP-1R) and its relative cell signaling pathway in the cytoprotection of geniposide.. Cell viability was determined by MTT assay. Knockdown of the Glp-1r gene was carried out with shRNA. The levels of HO-1 protein and cAMP response element binding protein (CREB) phosphorylation were measured by Western blotting.. Geniposide protected PC12 cells from oxidative damage induced by 3-morpholinosydnonimine hydrochloride (SIN-1) by enhancing the expression of heme oxygenase 1 (HO-1) via the cAMP-PKA-CREB signal pathway. After transfecting PC12 cells with the AB1 enhancer from the HO-1 gene, luciferase activity induced by geniposide increased in a dose-dependent manner, but not in the PC12 cells whose Glp-1r gene was disrupted. Additionally, inhibition of HO-1 activity by Sn-protoporphyrin IX (SnPP) or shRNA-mediated knockdown of Glp-1r decreased the neuroprotection of geniposide in PC12 cells.. GLP-1R plays a critical role in geniposide-induced HO-1 expression to attenuate oxidative insults in PC12 cells.

Topics: Animals; Cell Survival; Cyclic AMP Response Element-Binding Protein; Cyclic AMP-Dependent Protein Kinases; Enhancer Elements, Genetic; Gene Expression; Gene Knockdown Techniques; Glucagon-Like Peptide-1 Receptor; Heme Oxygenase-1; Iridoids; Neuroprotective Agents; Oxidative Stress; PC12 Cells; Rats; Receptors, Glucagon

To observe the influence of ingredients in Xingnaojing,such as moschus, borneol and radix curcumae on intestine absorption kinetics of gardenia extract.. An in situ intestinal perfusion model of rats was employed to investigate the absorption of geniposide in gardenia extract.. While gardenia extract was administered solely, the absorptive rate constant (K) of geniposide was (0.055 +/- 0.006) h(-1); But while the extract was co-administered with radix curcumae,moschus and borneol, the K were (0.060 +/- 0.001), (0.066 +/- 0.008), (0.072 +/- 0.010) h(-1), respectively. The K was (0.076 +/- 0.011) h(-1) while the extract in total formulation for Xingnaojing.. The K, while the extract is co-administered with borneol or total formulation is significantly higher than solely used. Borneol and complex prescription can significantly increase the intestinal absorption of geniposide in gardenia extract.

Topics: Animals; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Gardenia; Intestinal Absorption; Iridoids; Linear Models; Male; Plant Extracts; Rats; Rats, Wistar

The objective of this research was to study the in situ and in vivo nasal absorption of Geniposide (Ge) co-administered with borneol. A rat in situ nasal perfusion technique with a novel volumeadjusted calculation was used to examine the absorption rate and extent of Ge. The influence of different experimental conditions such as purity of extract, drug concentration, co-administration with synthetic borneol or natural borneol were also investigated. Results indicated nasal absorption of Ge was primarily by passive diffusion that resembled first order kinetics. Following co-administration with borenol, the drug absorption was increased by 1.4 and 1.7 folds for natural borneol and synthetic borneol, respectively. However, the effect of other factors on drug absorption was not significant. In addition, it was also observed that there is a positive correlation between the absorption of water and Ge by the nasal route. In vivo studies carried out in rats where Ge was co-administered with NB and the pharmacokinetic profile obtained following intranasal administration were compared with those after intravenous administration. The bioavailability of Ge by intranasal was 101.5% and T(max) was 2.04 +/- 0.64 min. MRT was 218.7 +/- 74.1 min and 44.4 +/- 8.9 min for intranasal and intravenous, respectively. Combined with the borneol, Ge can be promptly and thoroughly absorbed intranasally in rats.

Topics: Absorption; Administration, Intranasal; Animals; Camphanes; Drug Interactions; Injections, Intravenous; Iridoids; Male; Nasal Mucosa; Random Allocation; Rats; Rats, Sprague-Dawley

To study the effect of geniposide on treating experimental CP rats.. The animal model of CP was made with rats by injecting hemorrhoid injection. Rats in experiment group were randomly devided into model group, Qianliekang tablets group (2 g x kg(-1)) and geniposide high, middle, low dose groups (20, 10, 5 mg x kg(-1)). Subsequently, the state of all rats, prostate index, WBC and lecithine corpuscle, LDH5/LDH1, and prostatic histopathological changes were observed. Count of total cellular score (TCS) and quantitation of inflammatory cell, fibroblasts, glandular organ, calculation of glandular cavity area, and their changes of morphology were analyzed.. Compared with model group, the prostate index, WBC and LDHS/LDH1 of the rats in Qianliekang tablets group, high dose geniposide group and middle dose geniposide group were significantly decreased, while the quantities of lecithine corpuscle were remarkably increased (P < 0.01 or P < 0.05). Compared with model group, the number of inflammatory cells and fibroblasts in Qianliekang tablets group, high dose geniposide group were decreased, and the quantity of glandular organ and area of glandular cavity in these groups were increased (P < 0.05 or P < 0.01).. Geniposide of high and middle dose can reduce leucocytes infiltration, restrain the hyperplasia of fibrous tissue, and recover the secretion function of prostate. It show that geniposide is significantly potential to cure rats which are exposed to chronic prostatitis.

Topics: Animals; Body Weight; Chronic Disease; Disease Models, Animal; Dose-Response Relationship, Drug; Iridoids; Isoenzymes; L-Lactate Dehydrogenase; Lactate Dehydrogenase 5; Leukocyte Count; Male; Prostate; Prostatitis; Rats; Rats, Sprague-Dawley

Lipopolysaccharide (LPS/endotoxin) is a key pathogen recognition molecule for sepsis. Currently, one of the therapeutic approaches for severe sepsis is focusing on the neutralization of LPS, and clinical trials have shown a lot of traditional Chinese herbs possess anti-sepsis function. Herein, to elucidate the bioactive components of traditional Chinese herbs that can neutralize LPS, the lipid A-binding abilities of sixty herbs were tested using affinity biosensor technology. The aqueous extract of Gardenia jasminoides Ellis, traditionally used to treat inflammation in Asian countries for centuries, was further investigated. Subsequently, a monomer, identified as geniposide, was isolated. In vitro, geniposide was found to directly bind LPS and neutralize LPS. It dose-dependently inhibited cytokines release from RAW264.7 cells induced by LPS without affecting the cell viability, and inhibited TNF-α mRNA expression up-regulated by LPS. However, geniposide did not decrease TNF-α release induced by CpG DNA, Poly I:C or IL-1β. Significantly, geniposide dose-dependently down-regulated TLR4 mRNA expression up-regulated by LPS, and suppressed the phosphorylations of p38 MAKP induced by LPS but not by IL-1β. In vivo, geniposide (40mg/kg) could significantly protect mice challenge with lethal heat-killed E. coli, and dose-dependently decreased the level of serum endotoxin which was tightly associated with the cytokine levels in endotoxemia mice. In summary, we successfully isolated geniposide from G. jasminoides Ellis. Geniposide directly bound LPS and neutralized LPS in vitro, and significantly protected sepsis model mice. Therefore, geniposide could be as a useful lead compound for anti-sepsis drug development.

Topics: Animals; Cell Line; Cytotoxicity Tests, Immunologic; Endotoxemia; Female; Gardenia; Gene Expression Regulation; Iridoids; Lipid A; Lipopolysaccharides; Macrophages; Male; Mice; Mice, Inbred Strains; Random Allocation; RNA, Messenger; Toll-Like Receptor 4

To investigate whether geniposide, an iridoid glucoside extracted from gardenia jasminoides ellis fruits, inhibits cell adhesion to human umbilical vein endothelial cells (HUVECs) induced by high glucose and its underlying mechanisms.. HUVECs were isolated from human umbilical cords and cultured. The adhesion of monocytes to HUVECs was determined using fluorescence-labeled monocytes. The mRNA and protein levels of vascular cell adhesion molecule-1 (VCAM-1) and endothelial selectin (E-selectin) were measured using real-time RT-PCR and ELISA. Reactive oxygen species (ROS) production was measured using a fluorescent probe. The amounts of nuclear factor-kappa B (NF-kappaB) and inhibitory factor of NF-kappaB (IkappaB) were determined using Western blot analysis. The translocation of NF-kappaB from the cytoplasm to the nucleus was determined using immunofluorescence.. Geniposide (10-20 mumol/L) inhibited high glucose (33 mmol/L)-induced adhesion of monocytes to HUVECs in a dose-dependent manner. This compound (5-40 mumol/L) also inhibited high glucose-induced expression of VCAM-1 and E-selectin at the gene and protein levels. Furthermore, geniposide (5-20 micromol/L) decreased ROS production and prevented IkappaB degradation in the cytoplasm and NF-kappaB translocation from the cytoplasm to the nucleus in HUVECs.. Geniposide inhibits the adhesion of monocytes to HUVECs and the expression of CAMs induced by high glucose, suggesting that the compound may represent a new treatment for diabetic vascular injury. The mechanism underlying this inhibitory effect may be related to the inhibition of ROS overproduction and NF-kappaB signaling pathway activation by geniposide.

Topics: Cell Adhesion; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Gardenia; Glucose; Humans; Iridoids; Monocytes; NF-kappa B; Reactive Oxygen Species; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Umbilical Veins

To develop an HPLC method for the determination of plasma concentration of jasminoidin and study the pharmacokinetics of jasminoidin in rabbits administered Xingnaojing naristillae by nasal medication.. After sampling blood from the left arteria carotis of rabbits which were administered Xingnaojing naristillae medication by nasal by 12 mg x kg(-1) (counted by gardenia extract) at 1, 3, 5, 10, 20, 30, 45, 60, 90, 120, 240 min, the plasma samples were dealt with acetonitrile precipitation and HPLC was used to determine the plasma concentration of jasminoidin. The pharmacokinetic parameters were computed by Kinetica software.. The calibration curve was linear (r = 0.999 6) within the range of 0.136 5-2.73 mg x L(-1) for jasminoidin in plasma. The average recovery was (97.14 +/- 3.78)%, (95.06 +/- 2.95)%, (91.50 +/- 1.82)%. The within-day and between-day precision met the requirements, because the RSD were both less than 4%. Jasminoidin was fitted to a two-compartment open pharmacokinetic model in rabbits. The mainly pharmacokinetic parameters were: C(max) = (2.013 +/- 0.563) mg x L(-1), T(max) = (6.405 +/- 1.764) min, K(e) = (0.032 5 +/- 0.013 3) min(-1), CL = (0.059 3 +/- 0.0246) L x min(-1) x kg(-1), AUC = (116.89 +/- 50.19) mg x min(-1) x L(-1), MRT = (84.447 +/- 19.420) min.. The method can be used to determine the concentration and to investigate the pharmacokinetics of jasminoidin in rabbits. Jasminoidin was absorbed rapidly by nasal medication and has a good perspective.

Topics: Administration, Intranasal; Animals; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Iridoids; Male; Rabbits

Oxidative stress in brain is emerging as a potential causal factor in aging and age-related neurodegenerative disorders. A large body of evidence shows that induction of endogenous antioxidative proteins seems to be a reasonable strategy for delaying the progression of cell injury. In this study, geniposide upregulates the expression of heme oxygenase-1 (HO-1) to attenuate the cell apoptosis induced by 3-morpholinosydnonimine hydrochloride (SIN-1) in primary cultured hippocampal neurons. Furthermore, geniposide induces the nuclear translocation of nuclear factor-E2-related factor 2 (Nrf2) and activation of phosphatidylinositol 3'-kinase (PI3K) in the presence of oxidative stress, and both LY294002 (a specific inhibitor of PI3K) and Zinc protoporphyrin (ZnPP, an inhibitor of HO-1) decrease the cytoprotective action of geniposide in hippocampal neurons. Taken together, the novel cytoprotective mechanism of geniposide to antagonize oxidative stress may be involved in PI3K- and Nrf2-mediated upregulation of the antioxidative enzyme HO-1.

Topics: Antioxidants; Apoptosis; Biological Transport; Cell Line; Cell Nucleus; Fruit; Gardenia; Heme Oxygenase-1; Hippocampus; Humans; Iridoids; Molsidomine; NF-E2-Related Factor 2; Oxidative Stress; Phosphatidylinositol 3-Kinases; Plant Extracts; Protoporphyrins; Signal Transduction; Up-Regulation

To determinate geniposide and croein-1 in Fructus Gardeniae from different breeds.. RP-HPLC method was adopted. The chromatographic separation was performed on an Eclipse XDB-C18 column (250 mm x 4.6 mm, 5 microm, Agilent). The mobile phase was a mixture of acetonitrile-0.1% phosphoric acid aqueous solution in gradient elution. The wavelength of 238 nm and 440 nm was selected to determinate geniposide and crocins simultaneously.. The contents of geniposide and croein-1 in different breeds of Fructus Gardeniae are different.. This method is simple and repeatable, and could be used for the quality control of Fructus Gardeniae.

Topics: Carotenoids; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Fruit; Gardenia; Iridoids; Reproducibility of Results

To study the chemical constituents from the roots of Tinospora sagittata var. yunnanensis.. Various chromatographic techniques were used to isolate and purify the constituents. The structures of these compounds were elucidated on the basis of spectral analysis.. Seven compounds were isolated from the plant and their structures were identified as tinophylloloside (1), epitinophylloloside (2), 2-deoxycrustecdysone (3), polypodine B (4) , (+)-5'-methoxyisolariciresinol 3alpha-O-beta-D-glucopyranoside (5), geniposide (6), adenine (7).. All compounds are isolated from the plant for the first time,and compounds 4, 5, 6 and 7 are isolated firstly from the genus.

Topics: Adenine; Ecdysterone; Iridoids; Magnetic Resonance Spectroscopy; Plant Roots; Plants, Medicinal; Tinospora

Hepatic glycogen phosphorylase (GP) and glucose-6-phosphatase (G6Pase) play an important role in the control of blood glucose homeostasis and are proposed to be potential targets for anti-diabetic drugs. Geniposide is an iridoid glucoside extracted from Gardenia jasminoides Ellis fruits and has been reported to have a hypoglycemic effect. However, little is known about the biochemical mechanisms by which geniposide regulates hepatic glucose-metabolizing enzymes. The present study investigates whether the hypoglycemic effect of geniposide is mediated by GP or G6Pase.. Type 2 diabetic mice, induced by a high-fat diet and streptozotocin injection, were treated with or without geniposide for 2 weeks. Blood glucose levels were monitored by a glucometer. Insulin concentrations were analyzed by the ELISA method. Total cholesterol (TC) and triglyceride (TG) levels were measured using Labassay kits. Activities of hepatic GP and G6Pase were measured by glucose-6-phosphate dehydrogenase-coupled reaction. Real-time RT-PCR and Western blotting were used to determine the mRNA and protein levels of both enzymes.. Geniposide (200 and 400 mg/kg) significantly decreased the blood glucose, insulin and TG levels in diabetic mice in a dose-dependent manner. This compound also decreased the expression of GP and G6Pase at mRNA and immunoreactive protein levels, as well as enzyme activity.. Geniposide is an effective hypoglycemic agent in diabetic mice. The hypoglycemic effect of this compound may be mediated, at least in part, by inhibiting the GP and G6Pase activities.

Topics: Animals; Blood Glucose; Cholesterol; Diabetes Mellitus, Experimental; Diet; Dietary Fats; Glucose-6-Phosphatase; Glycogen Phosphorylase; Hypoglycemic Agents; Iridoids; Liver; Male; Mice; Mice, Inbred C57BL; Molecular Structure; Random Allocation; Triglycerides

Oxidative stress plays a critical role in the pathogenic cascade leading to neuronal degeneration in AD. Consequently, the induction of endogenous antioxidative proteins by antioxidants seems to be a very reasonable strategy for delaying the disease's progression. In previous work, we identified the neurotrophic and neuroprotective effects of geniposide, which result from the activation of glucagon-like peptide 1 receptor (GLP-1R). In this study, we explore the role of PI3 kinase signaling pathway in the neuroprotection of geniposide in PC12 cells.. Cell viability was determined by MTT assay. Apoptosis was detected by Hoechst and PI double staining. The protein expression of Bcl-2 and phosphorylation of Akt308, Akt473, GSK-3beta, and PDK1 was measured by Western blot.. Geniposide induced the expression of the antiapoptotic protein Bcl-2, which inhibited apoptosis in PC12 cells induced by H(2)O(2), and this effect could be inhibited by preincubation with LY294002, a selective inhibitor of PI3K. Furthermore, geniposide enhanced the phosphorylation of Akt308, Akt473, GSK-3beta and PDK1 under conditions of oxidative stress.. These results demonstrate that the PI3K signaling pathway is involved in the neuroprotection of geniposide in PC12 cells against the oxidative damage induced by H(2)O(2) in PC12 cells.

Topics: Alzheimer Disease; Animals; Glucagon-Like Peptide 1; Glucagon-Like Peptide-1 Receptor; Humans; Hydrogen Peroxide; Iridoids; Neurons; Neuroprotective Agents; Oxidants; Oxidative Stress; PC12 Cells; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Proto-Oncogene Proteins c-bcl-2; Rats; Receptors, Glucagon; Signal Transduction

Phosphine-catalyzed [3 + 2] cycloaddition of ethyl-2,3-butadienoate with enone (S)-3b occurs with high levels of regio- and stereocontrol to deliver the cis-fused cyclopenta[c]pyran 4 characteristic of the iridoid family of natural products. Cycloadduct 4 was converted to the iridoid glycoside (+)-geniposide in 10 steps.

Topics: Catalysis; Iridoids; Molecular Structure; Phosphines; Stereoisomerism

To screen effective principles from traditional Chinese medicine, a method named hepatocyte extraction combined with HPLC (HE-HPLC) was establish in this study. The active principles in the fruits of Gardenia jasminoides Ellis extract were combined with the hepatocytes under imitated physiological environments. Then the unattached substances were washed off by PBS with pH 7.4. After that the conjugated compounds were eluted by PBS with pH 4.0. These compounds released from target sites were collected and handled through SPE to be condensed, and analyzed by HPLC. The results indicated that two characteristic active compounds in the fruits of G. jasminoides extract binded to the hepatocytes. One of them is geniposide. The other is continued to be identified. It is showed that active principles which could bind to hepotocyte (through receptors, Channels, enzymes, etc.) could be detected, at least partly, by HE-HPLC analysis. There was a significant correlation between the retention properties of the active compounds which was obtained by HE-HPLC and their pharmacological effects on hepotocytes.

Topics: Animals; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Fruit; Gardenia; Hepatocytes; Iridoids; Phytotherapy; Plant Extracts; Rats

To investigate the regulation of geniposide nasal absorption.. We used rat in situ nasal recirculation method as experimental model. The effects of perfusion volume, flow rate, perfusion concentration on nasal absorption of geniposide were studied.. Geniposide was stable in rats nasal lavage solution. At the perfusion volume 5 mL and flow rate 2.5 mL x min(-1), the absorption rate of geniposide remains constant in a dose-independent manner.. The mechanism of geniposide nasal absorption is passive diffusion following first order kinetics and the absorption rate constant is 4.20 x 10(-3).

Topics: Absorption; Administration, Intranasal; Animals; Diffusion; Drugs, Chinese Herbal; Iridoids; Kinetics; Linear Models; Male; Nasal Mucosa; Perfusion; Rats; Sensitivity and Specificity; Solutions

Seven new iridoid glucosides, 6''-O-trans-sinapoylgenipin gentiobioside (1), 6''-O-trans-p-coumaroylgenipin gentiobioside (2), 6''-O-trans-cinnamoylgenipin gentiobioside (3), 6'-O-trans-p-coumaroylgeniposide (4), 6'-O-trans-p-coumaroylgeniposidic acid (5), 10-O-succinoylgeniposide (6), and 6'-O-acetylgeniposide (7), two new monoterpenoids, 11-(6-O-trans-sinapoylglucopyranosyl)gardendiol (8) and 10-(6-O-trans-sinapoylglucopyranosyl)gardendiol (9), and three known ones, 6'-O-trans-sinapoylgeniposide (10), geniposide (11), and 10-O-acetylgeniposide (12), were isolated from the fruit of Gardenia jasminoides. The structures of these compounds were elucidated on the basis of 1D and 2D NMR spectra analyses. Furthermore, short-term memory assays on an Abeta transgenic drosophila model showed that compounds 4 and 6-12 can improve the short-term memory capacity to varying degrees, with compounds 4 and 7 being the most active ones, suggesting that these compounds may have a potential antagonism effect against Alzheimer's disease.

Topics: Alzheimer Disease; Animals; Animals, Genetically Modified; Disease Models, Animal; Drosophila; Fruit; Gardenia; Iridoids; Memory, Short-Term; Molecular Structure; Nuclear Magnetic Resonance, Biomolecular; Stereoisomerism

To observe the influence of natural borneol and synthetic borneol on mucosal permeability of Gardenia extract.. Taken frog skin as a vitro model to study the vitro mucosal permeation the impacts of the natural borneols and synthetic borneols on the P(app) of the Jasminoidin were studied, and the effect of different borneols on the stability of Jasminoidin were investigated. Compared the 10 h accumulated infiltration rate of each group the effects of influence factors,such as C(Ge), C(B) and rotation speed on P(app) were investigated by using response surface method.. The P(app) of Jasminoidin of natural borneol and synthetic borneol group were 1.44 fold and 1.77 fold of control group (P < 0.01). For two borneol groups, the results also showed a significant difference too (P < 0.05). Jasminoidin began to degrade about 8 h after the effect of frog skin for control group and synthetic borneol group, but was stable within 12 h for natural borneol group. The accumulated permeation rate of 10 h was same for different borneol groups. It was about 1.3 fold of control group. The C(Ge) had a salinence influence on the P(app) (P < 0.01) and C(B) had a salience influence on time-lag (P < 0.01).. Both the natural borneol and synthetic borneol can accelerate the permeation of Jasminoidin and the synthetic borneol has stronger effect on the P(app). Both two different borneol can reduce the degradation effect of frog skin to Jasminoidin, but the natural borneol has a better protect effect on it. By using more natural borneol, the mucosal permeability of Gardenia extract can be increased, the time-lag can be reduced, and Jasminoidin has better stability.

Topics: Administration, Cutaneous; Camphanes; Dosage Forms; Drugs, Chinese Herbal; Gardenia; Iridoids; Mucous Membrane; Nasal Mucosa; Permeability; Skin; Skin Absorption

To investigate the effect of geniposide on vascular dementia (VaD) in rats.. VaD rat model was established by permanent ligation of bilateral common carotid arteries. Morris water maze performance was assessed after 4 weeks of treatment with geniposide, followed by pathological examinations of hippocampus and cerebral cortex.. The VaD rats had significantly longer escape latency and lower percentage of activities in platform quadrant than the rats in the sham surgery group (P<0.05). The VaD rats treated with geniposide [50 mg/(kg x d), 75 mg/(kg x d)] had significantly shorter escape latency (except the first day of the test) and significantly higher percentage of activities in platform quadrant than the VaD rats without treatment (P<0.05). No significant differences were found between the geniposide treated group and the Donepezil treated group and the sham surgery control group. Geniposide significantly alleviated neurons,apoptosis and necrosis induced by chronic cerebral hypoperfusion of cortex and hippocampus.. Chronic cerebral hypoperfusion can induce cognitive impairment. Geniposide can improve cognitive ability of Vascular Dementia in rats. This may represent a potential treatment strategy for vascular dementia.

Topics: Animals; Behavior, Animal; Cognition; Dementia, Vascular; Hippocampus; Iridoids; Male; Random Allocation; Rats; Rats, Sprague-Dawley

Gardenia jasminoides E. (Rubiaceae) methanol extracts showed the highest level of antifungal activity against Pleurotus ostreatus, a wood-rotting fungus, compared to five other methanol plants extracts; [Thuja orientalis L. (Cupressaceae), Datura innoxia (Solanaceae), Ligustrum japonicum T. (Oleaceae), Juniperus chinensis var. procumbens (Cupressaceae) and Mallotus japonica M. (Euphorbiaceae)] and selected for further analysis. Two antifungal compounds were isolated from n-butanol and ethyl acetate solubles in the methanol extracts of Gardenia jasminoides leaves and stems by bioassay-guided fractionation, using Pleurotus ostreatus. The antifungal compounds found for the first time in Gardenia jasminoides against Pleurotus ostreatus were identified as genipin and geniposide based on instrumental analyses. Both also had potent inhibitory effects on two plant pathogenic fungi; Fusarium oxysporum and Corynespora cassiicola.

Topics: Antifungal Agents; Cholagogues and Choleretics; Fungi; Gardenia; Iridoid Glycosides; Iridoids; Microbial Sensitivity Tests; Molecular Structure; Plant Extracts

To determine the O/W partition coefficient of geniposide in fructus gardeniae extract and investigate the absortion kinetics of geniposide in whole small intestine and different intestinal segments of rats.. The shake-flask method was employed to determine the O/W partition coefficient of geniposide; an in situ intestinal perfusion model was employed to investigate the absorptive kinetics of geniposide.. The partition coefficient (P) of geniposide was 0.1077, logP was -0.9678; the absorptive rate constants (K) of geniposide at the concentration of 0.078, 0.311, 0.780 g x L(-1) were (0.130 +/- 0.007), (0.056 +/- 0.003), (0.031 +/- 0.006) h, respectively. The K of geniposide were (0.019 +/- 0.003), (0.015 +/- 0.002), (0.012 +/- 0.002) h at duodenum, jejunum, ileum, respectively.. According to the P and the logP, it could be indicated that the absorption of geniposide at small intestine was poor absorption; The absorption rate was increased with the decrease of the extract concentration; Their absorption was first-order process besides the passive diffusion mechanism, and facilitated diffusion and active transport may also take part in the transport process. Geniposide was absorbed at all smallintestinal segments of rats, but the duodenal absorption ratewas higher more than other section.

Topics: Animals; Drugs, Chinese Herbal; Gardenia; Intestinal Absorption; Intestines; Iridoids; Kinetics; Male; Random Allocation; Rats; Rats, Sprague-Dawley

Phytochemical study of the aqueous extract of the flowering tops of Lamium album led to identification of the antiviral iridoid isomers lamiridosins A and B (1, 2). These compounds were found to significantly inhibit hepatitis C virus entry (IC(50) 2.31 muM) in vitro. Studies of 14 iridoid analogues showed that, while the parent iridoid glucosides demonstrated no anti-HCV entry activity, the aglycones of shanzhiside methyl ester (4), loganin (5), loganic acid (6), geniposide (10), verbenalin (12), eurostoside (15), and picroside II (17) exhibited significant anti-HCV entry and anti-infectivity activities.

Topics: Antiviral Agents; Hepacivirus; Iridoids; Lamiaceae; Molecular Structure; Plants, Medicinal; Stereoisomerism

A number of studies have shown that oxidative stress and mitochondrial involvement are major triggering factors in the development of neurodegenerative diseases. Cobalt chloride (CoCl(2))-induced cell death in PC12 cells may serve a simple and convenient in vitro model of hypoxia-induced neuronal cytotoxicity. To explore the effect of geniposide on CoCl(2) which induced cytotoxicity and mitochondrial function in rat pheochromocytoma PC12 cells, we analyzed the influence of geniposide on the expression of apoptosis-related proteins.. PC12 cells and RNAi PC12 cells were treated with 0, 12.5, 25, 50, 100 micromol/L geniposide for 12 hours and then exposure to 400 micromol/L CoCl(2) for 12 hours. Cell viability, cell morphology, and expression of Bcl-2, Bax, P53 and caspase-9 were determined using Western blotting.. Pretreatment with geniposide markedly improved the cells viability and morphology, decreased the expression of Bax, P53 and caspase-9, and increased the expression of Bcl-2 in PC12 cells challenged by CoCl(2)2. However, in the RNAi PC12 cells, geniposide had no significant effect on the expression of these proteins.. Geniposide protects PC12 cells from CoCl(2) involved in mitochondrial mediated apoptosis, and GLP-1R might play a critical role in the neuroprotection of geniposide in PC12 cells.

Topics: Animals; Apoptosis; bcl-2-Associated X Protein; Cobalt; Glucagon-Like Peptide-1 Receptor; Iridoids; Mitochondria; Neuroprotective Agents; PC12 Cells; Proto-Oncogene Proteins c-bcl-2; Rats; Receptors, Glucagon; Signal Transduction

Geniposide, an iridoid glucoside, is a major constituent in the fruits of Gardenia jasminoides (Gardenia fruits), a popular Chinese herb. Genipin, the aglycone of geniposide, is used to prepare blue colorants in food industry and also a crosslinking reagent for biological tissue fixation. In this study, we investigated the metabolism and pharmacokinetics of genipin and geniposide in rats. Blood samples were withdrawn via cardiopuncture and the plasma samples were assayed by HPLC method before and after hydrolysis with sulfatase and beta-glucuronidase. The results indicated that after oral administration of genipin or Gardenia fruit decoction, genipin sulfate was a major metabolite in the bloodstream, whereas the parent forms of genipin and geniposide were not detected. Importantly, oral administration of 200mg/kg of genipin resulted in a mortality of 78% (7/9) in rats.

Topics: Animals; Calibration; Chemical Phenomena; Chemistry, Physical; Cholagogues and Choleretics; Chromatography, High Pressure Liquid; Data Interpretation, Statistical; Gardenia; Hydrolysis; Injections, Intravenous; Iridoid Glycosides; Iridoids; Male; Plant Extracts; Rats; Rats, Sprague-Dawley; Reproducibility of Results

Geniposide is an iridoid glycoside isolated from the fruit of Gardenia jasminoides Ellis used as a Chinese traditional medicine for treatment of generalized vitiligo. Stem cell factor from keratinocyte recognizes and activates its receptor c-kit carried by melanocyte to potent enhance melanocytic melanogenesis that can be suppressed by norepinephrine. This study addresses the action and mechanism of geniposide enhancing melanogenesis in norepinephrine-exposed normal human epidermal melanocyte. Flow cytometry results from this study exhibited the augmentation effect of geniposide on production of c-kit receptor by norepinephrine-exposed normal human epidermal melanocyte. However, geniposide did not affect the production of stem cell factor by norepinephrine-exposed normal human epidermal keratinocyte assessed by cellular enzyme-linked immunosorbent assay (ELISA). ELISA indicated that at the presence of stem cell factor, geniposide was capable of elevating the level of extracellular signal-regulated kinase 1/2 phosphorylation within norepinephrine-exposed normal human epidermal melanocyte, which is known to be involved in stem cell factor/c-kit downstream signalling. And inhibition of c-kit with inhibitory antibody K44.2 completely blocked the increase in geniposide-stimulated extracellular signal-regulated kinase 1/2 phosphorylation. In addition, spectrophotometry demonstrated the enhancement effect of geniposide on melanogenesis (tyrosinase activity and melanin production) in norepinephrine-exposed normal human epidermal melanocyte at the presence of stem cell factor, which was blocked by c-kit inhibitory antibody K44.2. Data from this study suggest that geniposide can enhance melanogenesis by stem cell factor/c-kit signalling in which the expression of c-kit receptor is augmented in norepinephrine-exposed normal human epidermal melanocyte.

Topics: Cell Line; Enzyme-Linked Immunosorbent Assay; Epidermal Cells; Extracellular Signal-Regulated MAP Kinases; Flow Cytometry; Humans; Iridoids; Keratinocytes; Melanins; Melanocytes; Norepinephrine; Oxidoreductases; Phosphorylation; Proto-Oncogene Proteins c-kit; Signal Transduction; Stem Cell Factor

A method for the rapid and simultaneous determination of 6,7-dimethylesculetin (CAS 120-08-1) and geniposide (CAS 24512-63-8) in rat plasma has been developed, using validated high performance liquid chromatography (HPLC) with solid phase extraction (SPE). The HPLC analysis was performed on a commercially available column (200 mm x 4.6 mm, 5 microm) with acetonitrile-methanol-0.1% aqueous formic acid as mobile phase and the UV detection at 343 nm and 238 nm for 6,7-dimethylesculetin and geniposide, respectively. The calibration curves for 6,7-dimethylesculetin and geniposide were linear over the range 0.4-25.6 microg/mL and 1.12-71.68 microg/mL, respectively. The lower limits of quantitation were 0.40 microg/ mL and 1.12 microg/mL, and the lower limits of detection were 0.06 microg/mL and 0.09 microg/ mL, respectively. The intra-day and inter-day precision for 6,7-dimethylesculetin and geniposide were < 5%, whereas the absolute recovery percentages were > 74%. A successful application of the developed HPLC analysis was demonstrated for the pharmacokinetic study of a Traditional Chinese Medicine formula of Yin Chen Hao Tang preparation.

Topics: Algorithms; Animals; Calibration; Chromatography, High Pressure Liquid; Coumarins; Drugs, Chinese Herbal; Half-Life; Iridoids; Male; Rats; Rats, Wistar; Reproducibility of Results

The conditions to determine geniposide and genipin using gradient liquid chromatography-tandem mass spectrometry (LC-MS/MS) via electrospray ionization were obtained using fractional factorial experimental design approaches, guided with Taguchi orthogonal arrays to enhance peak intensity. Geniposide, the major iridoid glycoside component of Gardenia herbs, which has been recognized to have choleretic effects, is transformed to genipin in animals. In this paper, the gradient establishment times, ionization source temperatures, and the concentrations of volatile additive ammonium acetate were investigated under the guidance of experimental designs to obtain LC-MS/MS signals of the highest peak intensity. Using geniposide and genipin standards, the methods are validated at the concentration ranges of 0.5-1000ng/mL and 10-5000ng/mL using ammonium adducts. The correlation coefficients of geniposide and genipin standard curves are greater than 0.999. Compared with the sensitivities of previously published LC-MS/MS methods, the methods developed in this work provide 6-fold sensitivity improvement. The lowest concentrations of geniposide and genipin, 0.19 and 2ng/mL, respectively, to generate detectable LC-MS/MS signal peaks are one order of magnitude lower than the repoered values in previous publications. The measurement accuracy and precision of geniposide are within 23% and 15%, respectively. The accuracy and precision of genipin are within 16% and 12.5%, respectively. When the validated calibration curves of geniposide and genipin are used to determine spiked control samples in rat blood dialysates, the geniposide determination errors are within 15% accuracy and within 5.8% precision, respectively, and the genipin determination errors are within 23% accuracy and within 3.6% precision, respectively.

Topics: Animals; Calibration; Chromatography, Liquid; Gardenia; Iridoid Glycosides; Iridoids; Rats; Reference Standards; Sensitivity and Specificity; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

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 investigate the different permeation enhancers on the transdermal permeation of Xiao'er Niuhuang tuire cataplasms (XNTC).. Using improved franz-type diffusion cell with excised rat skin in vitro as the transdermal barrier, the content of permeated geniposide was determined by HPLC to study the kinetic parameters such as cumulative permeation quantity and permeation rate.. The result showed that the process of penetrating of geniposide in XNTC through skin could be in accordance with zero-rade releasing equation and XNTC was stable during the course of experiment.. 5% Propylene glycol (PG)-azone (2:3) has the best permeation-enhancing effect, and the results provided a primary basis for the future research on Xiao'er Niuhuang tuire cataplasms.

Topics: Animals; Azepines; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; In Vitro Techniques; Iridoids; Pharmaceutical Vehicles; Propylene Glycol; Rats; Rats, Sprague-Dawley; Skin; Skin Absorption

To investigate the protective mechanism of geniposide, baicalin and berberine on hypoxia and reoxygenation injury in cultured rat cerebral microvascular endothelial cells.. A model of four hours hypoxia and twelve hours reoxygenation injury in rat cerebral microvascular endothelial cells in vitro was established. The injured cells were treated with geniposide (0.128, 0.064, 0.032 mmol x L(-1)), baicalin (0.028, 0.014, 0.007 mmol L(-1)) and berberine (0.024, 0.012, 0.006 mmol L(-1)), respectively. The immunocytochemical method and techniques of image quantitative analysis were used to detect the mean optical density and mean area in order to match the protein expression of VCAM-1. The method of RT-PCR was adopted to observe and match the mRNA expression of VCAM-1.. As compared with the normal group, the mean optical density, the mean area and the mRNA expression of VCAM-1 of model group were significant increased (P < 0.01, P < 0.01, P < 0.01). As compared with the model group, both the mean optical density and the mean area of all treated groups were decreased, and there was significant difference between them (P < 0.01, P < 0.01). As compared with normal group, the mean optical density of baicalin (0.007 mmol x L(-1)) and berberine (0.012, 0.006 mmol x L(-1)) were significant decreased (P < 0.05, P < 0.01, P < 0.01), but there was no significant difference between the other groups and the normal group. As compared with normal group, the mean area of baicalin (0.0014 mmol x L(-1)) was significant decreased (P < 0.05), but there was significant difference between the other groups and the normal group. The mRNA expression of all treated groups was not only lower than that of the model group but also higher than that of the normal group (P < 0.05, P < 0.05).. The results suggest that geniposide, baicalin and berberine, which are effective compositions of huanglian jiedu decoting, can protect hypoxia-reoxygenation injuried rat cerebral microvascular endothelial cells. One of the protected mechanisms is that they can inhibit the expression of VCAM-1.

Topics: Animals; Berberine; Cell Hypoxia; Cells, Cultured; Cerebrum; Drugs, Chinese Herbal; Endothelium, Vascular; Flavonoids; Gene Expression; Humans; Hypoxia; Iridoids; Male; Oxygen; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Vascular Cell Adhesion Molecule-1

Yin Chen Hao Tang preparation (YCHTP) is a classic traditional Chinese medicine formula, which is commonly used for clinical treatment of hepatological diseases. In this study, a rapid and validated high-performance liquid chromatography (HPLC) method was developed to simultaneously identify 6,7-dimethylesculetin and geniposide in rat plasma. This assay was performed on a Dikma Diamonsil RP(18) column (200 mm×4.6 mm, 5 μm) with acetonitrile-methanol-water (0.1% formic acid) as the mobile phase, showing acceptable linearity, intra- and inter-day precision and accuracy (R.S.D.=5%), and absolute recovery for two analytes (74%); the limits of quantitation were 0.4 and 1.12 μg/ml, and the limits of detection were 0.06 and 0.09 μg/ml for two analytes. The developed method was successfully applied to study the effect of formula compatibility on the pharmacokinetics of 6,7-dimethylesculetin and geniposide in YCHTP when orally administrating an effective human daily dose of YCHTP to rats. We surmise that formula compatibility can significantly influence the pharmacokinetics of YCHTP, and we have elucidated and validated the compatible administration of YCHTP.

Topics: Administration, Oral; Animals; Calibration; Chromatography, High Pressure Liquid; Coumarins; Drugs, Chinese Herbal; Iridoids; Limit of Detection; Male; Rats; Rats, Wistar; Reproducibility of Results

When cultivated with Aspergillus niger, geniposide, an important drug, is transformed into genipin and genipinine. A simple and rapid HPLC method for simultaneous determination of geniposide and its two metabolites in broth of A. niger is described. The chromatographic separation was achieved on a C18 ODS column (250 x 4.6 mm) by gradient elution with 0.1% formic acid in water and 0.1% formic acid in acetonitrile as the gradient mixtures. The flow rate was 1 mL/min, the detection wavelength was 238 nm and the column temperature was kept at 28 degrees C. The retention times of geniposide, genipin and genipinine were 10.9, 13.8 and 21.5 min, respectively. The mean absolute recoveries of three analysts were over 98%. Quantification limits were 0.01 microg/mL for geniposide and 0.02 microg/mL for the two metabolites. The method was applied for the quantification of geniposide, genipin and genipinine during fermentation and the evaluation of the bioavailabilities of these three compounds in Caco-2 monolayer.

Topics: Aspergillus niger; Biological Availability; Caco-2 Cells; Chromatography, High Pressure Liquid; Humans; Iridoid Glycosides; Iridoids; Linear Models; Pyridines; Sensitivity and Specificity

Dehydroandrographolide, andrographolide and geniposide are the main active constituents of many herbal medicines, e.g., Fructus gardeniae, Common Andrographis Herb. They are used as the markers to control the quality of such herbal medicines and their herbal preparations. In this paper, a simple and sensitive high-performance liquid chromatographic (HPLC) method coupled with photodiode array detection (DAD) and electrospray mass spectrometry (ESI/MS) were developed to determine the three compounds simultaneously in extracts of medicinal herbs and herbal preparations produced by different companies. The extracts were separated on a C(18) reversed phase HPLC column, with a gradient solvent system, the time for the separation of the three target analytes was 1 0min. The abundance ions were recorded using selected ion monitoring (SIM) mode with m/z 297.3, 297.3 and 411.1 for dehydroandrographolide, andrographolide and geniposide, respectively. The limit of detection for dehydroandrographolide, andrographolide and geniposide were 20, 30 and 150 ng mL(-1), respectively. The proposed method was successfully applied to the determination of the contents of the compounds in related to medicinal herbs and preparations.

Topics: Andrographis; Chromatography, High Pressure Liquid; Diterpenes; Iridoids; Plant Extracts; Plants, Medicinal; Pyrans; Quality Control; Spectrometry, Mass, Electrospray Ionization

Geniposide (GP) as an agonist of glucagon-like peptide-1 receptor (GLP-1R) is an iridoid glycoside from the fruit of Gardenia jasminoides Ellis used as a Chinese traditional medicine for treatment of vitiligo vulgaris. Interaction of c-kit receptor with its ligand-SCF potent enhances the melanocytic melanogenesis, which can be repressed by norepinephrine (NE). To discover economic and efficient drug against vitiligo vulgaris, this paper addresses the action and mechanism of GP abrogating the NE-induced hypopigmentation in melanocyte. Flow cytometry exhibited the up-regulation effect of GP on NE-suppressed production of c-kit by normal human epidermal melanocyte (HEMn) in a concentration-dependent manner, and exendin-(9-39) (selective GLP-1R antagonist) appeared to alleviate the GP-stimulated expression of c-kit. However, neither NE nor GP affected the production of SCF by normal human epidermal keratinocyte (HEKn) assessed by cellular enzyme-linked immunosorbent assay. Spectrophotometry documented that GP abrogated the repression effect of NE on tyrosinase activity and melanin production in HEMn in the presence of recombination SCF significantly. The response of melanocytic melanogenesis to GP was blocked by exendin-(9-39) or K44.2 antibody (c-kit inhibitory antibody). Data from this paper provide the evidence that GP abrogates the NE-induced hypopigmentation by the activation of GLP-1R-dependent c-kit receptor signaling in which c-kit expression is augmented in HEMn.

Topics: Cell Line; Flow Cytometry; Gardenia; Glucagon-Like Peptide-1 Receptor; Humans; Hypopigmentation; Iridoids; Keratinocytes; Medicine, Chinese Traditional; Melanocytes; Norepinephrine; Proto-Oncogene Proteins c-kit; Receptors, Glucagon; Signal Transduction; Up-Regulation; Vitiligo

To investigate the protective effect of geniposide, baicalin and berberine for the rat cerebral microvascular endothelial cell.. The model of hypoxia and reoxygenation injury in rat cerebral microvascular endothelial cells in vitro was established. Both normal and model cells were treated with geniposide (1.024, 0.512, 0.256, 0.128, 0.064, 0.032, 0.016, 0.008 micromol x mL(-1)), baicalin (0.224, 0.112, 0.056, 0.028, 0.014, 0.007, 0.003 micromol x mL(-1)) and berberine (0.192, 0.096, 0.048, 0.024, 0.012, 0.006, 0.003 micromol x mL(-1)). Cell activity was measured by methyl thiazolyl tetrazolium (MTT) test.. After hypoxia/hypoglycemia cultures for 4 hour and reoxygenation for 12 hour, geniposide (0.128, 0.064, 0.032 micromol x mL(-1)), baicalin (0.028, 0.014, 0.007 micromol x mL(-1)) and berberine (0.024, 0.012, 0.006 micromol x microL(-1) could protect the injuried cerebral microvascular endothelial cells.. Appropriate concentration of geniposide, baicalin and berberine, which are effective components of Huanglian Jiedu decoction, could protect the injuried cerebral microvascular endothelial cells.

Topics: Animals; Berberine; Cell Hypoxia; Cell Survival; Cells, Cultured; Cerebral Cortex; Dose-Response Relationship, Drug; Drug Combinations; Drugs, Chinese Herbal; Endothelial Cells; Flavonoids; Iridoids; Male; Neuroprotective Agents; Oxygen; Plants, Medicinal; Pyrans; Rats; Rats, Sprague-Dawley

Gardenia yellow powders A, B and C, containing geniposide at 0.284%, 0.938% and 2.783%, respectively, were administered orally to male and female SD rats as 3% feed admixtures for 13-weeks to evaluate any potential toxicity. Mean geniposide intake values were 5.72, 18.9 and 56.3mg/kg/day in groups receiving these feed admixtures, respectively. All animals survived the duration of the study. The following findings were evident in the gardenia yellow C group: chromatouria, slightly increased plasma total bilirubin, blackish brown discoloration of the kidneys and liver, brown pigments in the proximal tubular epithelium of the kidneys. Slightly increased plasma total bilirubin was considered to be due to interference of metabolite of geniposide with the system of measurement and not to be a toxic effect since there were no related changes in histopathology of the liver or in any blood chemistry parameters. Other findings were limited to pigmentations or discolorations attributable to metabolites of geniposide. No treatment-related effects were evident on body weight, food consumption, ophthalmology, hematology or organ weights in any group. Therefore, it was concluded that 3-month ingestion of the gardenia yellow powder containing geniposide at 2.783% (approximately 60 mg/kg/day as geniposide intake) does not cause any severe toxic effects.

Topics: Administration, Oral; Animals; Blood Chemical Analysis; Body Weight; Eating; Female; Food Coloring Agents; Gardenia; Histocytochemistry; Iridoids; Male; Plant Extracts; Pyrans; Rats; Rats, Sprague-Dawley; Statistics, Nonparametric; Survival Analysis

A sensitive and specific method was developed and validated for the determination of paeoniflorin in rat brain with liquid chromatography-tandem mass spectrometry. Sample pretreatment involved protein precipitation following solid-phase extraction. Paeoniflorin and geniposide (internal standard) were separated isocratically on a Waters Symmetry C18 column (150 mm x 2.1 mm i.d., 5 microm), using a mobile phase of methanol/water with 0.1% formic acid (50:50, v/v) at a flow-rate of 200-300 microL/min in 4min. A Finngan LTQ tandem mass spectrometer equipped with electrospray ionization source was operated in the positive ion mode. Selective reaction monitoring was performed to quantify paeoniflorin and the internal standard at m/z transitions of 503-->381 and 411-->231, respectively. A good linearity was found in the range of 2-500 ng/mL (R(2)=0.9939). The intra- and inter-batch assay precisions (coefficient of variation, CV) at 5, 50 and 400 ng/mL (n=5) ranged from 6.3% to 9.7% and 1.2% to 7.2%, respectively, and the accuracies were from 95.9% to 101.6% and 99.4% to 102.9%, respectively. The mean recoveries of paeoniflorin were 81.2%, 80.9% and 82.3% at 5, 50 and 400 ng/mL (n=5), respectively, and the mean recovery of the internal standard was 76.7% with a concentration of 50 ng/mL (n=5). Stability studies showed that paeoniflorin was stable in different conditions. Finally, the method was successfully applied to the pharmacokinetic study of paeoniflorin in rat brain following a single subcutaneous administration (10 mg/kg) to rats.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Benzoates; Brain; Brain Chemistry; Bridged-Ring Compounds; Calibration; Chromatography, Liquid; Drugs, Chinese Herbal; Glucosides; Injections, Subcutaneous; Iridoids; Male; Molecular Structure; Monoterpenes; Paeonia; Plant Roots; Pyrans; Rats; Rats, Sprague-Dawley; Reference Standards; Reproducibility of Results; Sensitivity and Specificity; Solid Phase Extraction; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry; Tissue Distribution

A new method for the simultaneous quantitative determination of geniposide, baicalin and berberine hydrochloride in Angong Niuhuang pill using reversed phase high performance liquid chromatographic method had been developed.. The optimum chromatographic conditions were as follows: Agilent Zorbax SB - C18 column (4.6 mm 250 mm, 5 m), acetonitrile-H2O (containing 6 mmol L(-1) KH2PO4, pH 4.6) gradient elution; as a detection wavelength of 343 nm.. The calibration curves of geniposide, baicalin and berberine were linear at the ranges of 4.50-110.00, 5.00-153.00, 6.40-191.00 mg L(-1), respectively. The limits of detection of the method were 0.77 ng for geniposide, 1.53 ng for baicalin and 1.43 ng for berberine hydrochloride. The recoveries of the method were 104.44% (RSD 1.79% ) for geniposide, 96.98% (RSD 1.76%) for baicalin, 101.08% (RSD 3.1%) for berberine hydrochloride.. This method had been successfully applied to determine the content of geniposide, baicalin and berberine hydrochloride in Angong Niuhuang pill.

Topics: Berberine; Chromatography, High Pressure Liquid; Drug Combinations; Drugs, Chinese Herbal; Flavonoids; Iridoids; Materia Medica; Plants, Medicinal; Pyrans

Nine novel monoterpene alkaloid derivatives (3(a)-(c), 4(a)-(c), 5(a)-(c)) were prepared as a reaction of genipin 2 with L-amino acid methyl hydrochlorides utilized the reaction of reductive amination. Genipin was obtained by beta-glucosidase, catalyzed hydrolysis of the iridoid glucoside geniposide 1. The chemical structures were confirmed by 1D-, 2D-NMR and MS.

Topics: Alkaloids; Amino Acids; Fruit; Gardenia; Iridoid Glycosides; Iridoids; Magnetic Resonance Spectroscopy; Monoterpenes; Pyrans; Spectrometry, Mass, Electrospray Ionization

To establish a RP-HPLC method for fast and simultaneous determination of four index contents, which are Genpioside, Baicalin, Berberine Hydrochloride and Ammonium Glyeyrrhizinate, in Qingwei Huanglian tablets.. A separation was well achieved by gradient elution on a Hypersil C18 (4.6 mm x 250 mm, 5 microm) column with the mobile phases of acetontrile -0.5% triethylamine water-solution (pH 3.0) at a flow rate of 1.0 mL x min(-1), detection wavelength of 230 nm, and room temperature.. The calibration curve were linear in the ranges of 4.82-77.2, 5.80-92.8, 1.63-26.1 and 6.40-102 microg x mL(-1) (r = 0.9997-0.9999) for genpioside, baicalin, berberine hydrochloride and ammonium glyeyrrhizinate, respectively. The average recoveries of four index components were not less than 98.0%.. The method is convenient, rapid and accurate. It is suitable for the quality control of Qingwei Huanglian tablets.

Topics: Berberine; Chromatography, High Pressure Liquid; Coptis; Drug Combinations; Drugs, Chinese Herbal; Flavonoids; Gardenia; Glycyrrhiza; Glycyrrhizic Acid; Iridoids; Plants, Medicinal; Quality Control; Scutellaria baicalensis; Tablets

Gardenia fruit has been traditionally used as a folk medicine for centuries in Asian countries. Extraction with ethanol was used to obtain an extract (GFE) that contains two known constituents, geniposide and genipin, which were subsequently evaluated for anti-inflammatory activity. GFE, genipin, and geniposide showed acute anti-inflammatory activities in carrageenan-induced rat paw edema. In a dose-dependent manner, GFE also inhibited vascular permeability induced by acetic acid. Both genipin and geniposide inhibited production of exudate and nitric oxide (NO) in the rat air pouch edema model. However, genipin possessed stronger anti-inflammatory activity than geniposide, as demonstrated by the results with carrageenan-induced rat paw edema, carrageenan-induced air pouch formation, and measurement of NO content in the exudates. GFE caused a dose-dependent inhibition of acetic acid-induced abdominal writhing in mice. Collectively, genipin, rather than geniposide, is the major anti-inflammatory component of gardenia fruit.

Topics: Animals; Anti-Inflammatory Agents; Carrageenan; Dose-Response Relationship, Drug; Edema; Female; Fruit; Gardenia; Inflammation; Iridoid Glycosides; Iridoids; Male; Mice; Nitrates; Plant Extracts; Pyrans; Rats; Rats, Sprague-Dawley

A new HPLC method for the determination of geniposide in rat serum with solid-phase extraction (SPE) for preconcentration is described. Geniposide and an internal standard (paeoniflorin) were extracted from serum by SPE using C18 cartridges. Analysis of the extract was then performed on a reversed-phase C18 column using acetonitrile-water (16:84, v/v) as the eluting solvent system, and UV detection at 238 nm was used to measure the analyte with a limit of quantitation about 0.1 microg/mL. The calibration curve for geniposide was linear (r = 0.9993) in the concentration range 0.1-16.0 microg/mL. The intra- and inter-day precision of the geniposide were determined and their RSD did not exceed 10%. The validated method has been successfully applied for pharmacokinetic studies of geniposide from rat serum after oral administration of Yin-Zhi-Ku decoction.

Topics: Administration, Oral; Animals; Chemical Fractionation; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Iridoids; Male; Pyrans; Rats; Rats, Sprague-Dawley

The effect of ultrasound on the leaching process, in which Geniposide is leached from the Gardenia fruit by deionized water at 20 degrees C, was investigated. The phase equilibrium and the dynamics were measured at static, stirring, and ultrasonically assisted conditions, respectively. The experimental results show that the extraction yield of Geniposide with ultrasound at 0.1533 W cm(-2), is increased by 16.5%, in comparison with that without ultrasound when the ratio of the solvent volume to the fruit weight is 40 ml/g. A model for mass transfer, based on the intraparticle diffusion and the external mass transfer, was developed. And the dynamic curves calculated by the model are in a good agreement with the experimental data. The external mass transfer coefficient k(f)/R and intraparticle diffusion coefficient D(e)/R2 were obtained by fitting of the experiment data. The external mass transfer coefficient with ultrasound at 0.1533 W cm(-2) is 1.63 times higher than that in static process, and the intraparticle diffusion coefficient with ultrasound at 0.1533 W cm(-2) is 3.25 times higher than that in static process.

Topics: Chemistry; Diffusion; Fruit; Gardenia; Iridoids; Kinetics; Plant Extracts; Pyrans; Spectrophotometry; Temperature; Ultrasonics; Ultraviolet Rays

The effects of pulse ultrasound with different pulse parameter on the adsorption isotherm and kinetics of Geniposide on Resin 1300 were studied. And the mass transfer model describing the adsorption process was constructed. Amount of Geniposide adsorbed on Resin 1300 in the presence of ultrasound is lower than that in the absence of ultrasound. At our experimental conditions, the adsorption equilibrium constant decreases with increasing ultrasonic intensity and pulse duty ratio, and with decreasing pulse period. In addition, pulse ultrasound can enhance both liquid film diffusion and intraparticle diffusion, and the intensification of liquid film diffusion with pulse ultrasound is stronger than that of intraparticle diffusion. The intraparticle diffusion coefficient D(e)/R2 increases with increasing ultrasonic intensity and pulse duty ratio, and with decreasing pulse period.

Topics: Adsorption; Chemistry; Diffusion; Iridoids; Kinetics; Plant Extracts; Polymers; Pyrans; Resins, Plant; Ultrasonics

Geniposide from Gardenia jasminoides protected neuronal cells from damage in oxygen and glucose deprivation-exposed hippocampal slice culture. Geniposide showed a greater protective effect on the granule cell layer than on the pyramidal cell layer including CA 1 and CA 3. On the basis of the experimental results, geniposide may be a therapeutic agent for ischemia in patients.

Topics: Animals; Cell Death; Gardenia; Glucose; Hippocampus; Hypoxia; Iridoids; Neurons; Organ Culture Techniques; Pyramidal Cells; Pyrans; Rats; Rats, Sprague-Dawley

Crocetin esters present in saffron (Crocus sativus L.) stigmas and in Gardenia jasminoides Ellis fruit are the compounds responsible for their color. Of the fifteen crocetin esters identified in this study, five new compounds were tentatively identified: trans and cis isomers of crocetin (beta-D-triglucoside)-(beta-D-gentibiosyl) ester, trans and cis isomers of crocetin (beta-D-neapolitanose)-(beta-D-glucosyl) ester, and cis crocetin (beta-D-neapolitanose)-(beta-D-gentibiosyl) ester. The most relevant differences between both species were a low content of the trans crocetin (beta-D-glucosyl)-(beta-D-gentibiosyl) ester, the absence of trans crocetin di-(beta-D-glucosyl) ester in gardenia, and its higher content of trans crocetin (beta-D-gentibiosyl) ester and cis crocetin di-(beta-D-gentibiosyl) ester. With the same chromatographic method it was possible to identify, in a single run, ten glycosidic compounds in saffron extracts with a UV/vis pattern similar to that of picrocrocin; among them, 5-hydroxy-7,7-dimethyl-4,5,6,7-tetrahydro-3H-isobenzofuranone 5-O-beta-D-gentibioside and 4-hydroxymethyl-3,5,5-trimethyl-cyclohexen-2-one 4-O-beta-D-gentibioside were tentatively identified for the first time in saffron. Of these ten glycosides, only the O-beta-D-gentibiosyl ester of 2-methyl-6-oxo-2,4-hepta-2,4-dienoic acid was found in gardenia samples, but it was possible to identify the iridoid glycoside, geniposide.

Topics: Carotenoids; Chromatography, Liquid; Crocus; Cyclohexenes; Esters; Flowers; Fruit; Gardenia; Glucosides; Glycosides; Iridoids; Pyrans; Spectrometry, Mass, Electrospray Ionization; Terpenes; Vitamin A

A high-performance liquid chromatography method with solid-phase extraction is introduced for the determination of geniposide in rat urine after oral administration of yin-zhi-ku decoction. Geniposide and an internal standard (paeoniflorin) are extracted from urine using Strata cartridges. Analysis of the extract is then performed on a reversed-phase C18 column using acetonitrile-water (14:86, v/v) as eluting solvent system. UV detection is set at 238 nm. The calibration curve for geniposide is linear (r = 0.9996) in the concentration range of 2.0-240 microg/mL. Both intra- and interday precision of the geniposide are determined, and their relative standard deviation does not exceed 10%. The validated method is successfully applied to determine geniposide from rat urine after oral administration of yin-zhi-ku decoction.

Topics: Administration, Oral; Animals; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Iridoids; Male; Pyrans; Rats; Rats, Sprague-Dawley; Reference Standards; Reproducibility of Results

To estimate the therapeutic effect of single or combined use of jasminoidin and cholalic acid on focal cerebral ischemia rat with magnetic resonance-diffusion-weighted imaging (MR-DWI) technique, ultra-microscopy, and neuro-behavior scoring.. The model of cerebral ischemia-reperfusion injury was induced by string method. Three hours after reperfusion, MR-DWI was applied with ultra-microscopy and neuro-behavior test to give evaluation on cerebral ischemic rats, and pathologic, ultramicroscopic observation of tissue were taken as adjuvant measures to comprehensively evaluate the pharmacological effect on ischemia-reperfusion rats and delimit the efficacy of the two different components and their combination.. Compared with the model group, ADC and DCavg values of the foci in all the treated groups had the incrensing trend. There was significant difference arund the foci in the group of combined use of jasminoidin and cholalic acid (P < 0.05).. Combined use of jasminoidin and cholalic acid had protective effects on nerve and brain. MR-DWI technique accompanied with ultramicroscopic observation of tissues and neuro-behavior test is an effective method for evaluating the effect of neuro-protective agent.

Topics: Animals; Brain Ischemia; Cholic Acids; Diffusion Magnetic Resonance Imaging; Drug Therapy, Combination; Drugs, Chinese Herbal; Gardenia; Iridoids; Male; Neuroprotective Agents; Phytotherapy; Pyrans; Random Allocation; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Reproducibility of Results; Treatment Outcome

A simple, rapid, and specific analytical method for simultaneous determination of geniposide, baicalin, cholic acid and hyodeoxycholic acid in 50 microL samples of rat serum was developed by high performance liquid chromatography-tandem mass spectrometry. The quantification of the target compounds was determined by multiple reaction monitoring (MRM) mode using electrospray ionization (ESI). The correlation coefficients of the calibration curves were better than 0.997. The intra- and inter-day accuracy, precision, and linear range had been investigated in detail. This method was subsequently applied to pharmacokinetic studies of geniposide, baicalin, cholic acid and hyodeoxycholic acid in rats successfully.

Topics: Animals; Calibration; Cholic Acid; Chromatography, High Pressure Liquid; Deoxycholic Acid; Flavonoids; Iridoids; Male; Pyrans; Rats; Rats, Wistar; Reference Standards; Spectrometry, Mass, Electrospray Ionization

In our previous studies, we demonstrated the apoptotic cascades protein kinase C (PKC) delta/c-Jun NH2-terminal kinase (JNK)/Fas/caspases induced by penta-acetyl geniposide [(Ac)5GP]. However, the upstream signals mediating PKCdelta activation have not yet been clarified. Ceramide, mainly generated from the degradation of sphingomyelin, was hypothesized upstream above PKCdelta in (Ac)5GP-transduced apoptosis. Furthermore, nerve growth factor (NGF)/p75 is supposed to be involved because(Ac)5GP-induced apoptosis was demonstrated previously in glioma cells. In the present study, (Ac)5GP was shown to activate neutral sphingomyelinase (N-SMase) immediately, with its maximum at 15 min. The NGF and p75 enhanced by (Ac)5GP was inhibited when added with GW4869, the N-SMase inhibitor, indicating NGF/p75 as the downstream signals of N-SMase/ceramide. To investigate whether N-SMase is involved in (Ac)5GP-transduced apoptotic pathway, cells were treated with (Ac)5GP added with or without GW4869. It showed that N-SMase inhibition blocked FasL expression and caspase 3 activation. Likewise, p75 antagonist peptide attenuated the FasL/caspase 3 expression. The PKCdelta translocation induced by (Ac)5GP was also eliminated by GW4869 and p75 antagonist peptide. To further confirm whether N-SMase activation plays an important role in (Ac)5GP-induced apoptosis, cells were analyzed the apoptotic rate by 4', 6-diamidino-2-phenylindole (DAPI) staining. (Ac)5GP-induced apoptosis was reduced 40 and 80% by 10 and 20 microM GW4869, respectively. It indicated that N-SMase activation is pivotal in (Ac)5GP-mediated apoptosis. In conclusion, SMase and NGF/p75 are suggested to mediate upstream above PKCdelta, thus transducing FasL/caspase cascades in (Ac)5GP-induced apoptosis.

Topics: Animals; Apoptosis; Caspase 3; Caspases; Enzyme Activation; Fas Ligand Protein; Glioma; Iridoids; Membrane Glycoproteins; Models, Biological; Nerve Growth Factor; Protein Kinase C-delta; Protein Transport; Pyrans; Rats; Receptor, Nerve Growth Factor; Signal Transduction; Sphingomyelin Phosphodiesterase; Tumor Cells, Cultured; Tumor Necrosis Factors

The emerging data show that the insulinotrophic hormone glucagon-like peptide-1(GLP-1) and its agonist extendin-4 have neurotrophic function to inducing neuronal differentiation of PC12 cells and prevent neurons damage challenged by oxidative stress. Here, with the model of high throughput screen for GLP-1 receptor agonists, we screen and identify that geniposide is a novel agonist for GLP-1 receptor. Furthermore, geniposide induces the neuronal differentiation of PC12 cells with resulting neurites outgrowth; we also observe an increase in expression of growth-associated protein-43. U0126, a selective MEK inhibitor, prevents neurites out growth and phosphorylation of mitogen-activated kinase proteins in PC12 cells induced by geniposide. All these results show that activation of GLP-1 receptor by geniposide to induce the neuronal differentiation of PC12 cells involves in MAPK signaling cascade.

Topics: Animals; Blotting, Western; Butadienes; Cell Differentiation; Cell Line; Colforsin; Dose-Response Relationship, Drug; Drug Interactions; Enzyme Inhibitors; GAP-43 Protein; Gene Expression; Glucagon-Like Peptide 1; Glucagon-Like Peptide-1 Receptor; Humans; Iridoids; Luciferases; Mitogen-Activated Protein Kinase Kinases; Nerve Growth Factor; Neurons; Neuroprotective Agents; Nitriles; PC12 Cells; Pyrans; Rats; Receptors, Glucagon; Transfection

To study the hepatotoxicity effects in rats with different extract of Fructus Gardeniae.. Observe the change of appearance, behavior and weight of rats through oral gavage daily for 3 d. Weigh the liver and calculate the liver index. Detect the ALT, AST and TBIL. Observe the liver tissue by optical microscope.. The weight and index of liver were increased by 3.08 g x kg(-1) aqueous extract, 1.62 g x kg(-1) alcoholic extract and 0.28 g x kg(-1) geniposide, compared to those of the blank group (P < 0.005, P < 0.001) and the activities of ALT, AST and the content of TBIL were also increased, compared to those of the blank group (P < 0.05, P < 0.001). The liver cells were obviously swell, necrotic and changed with inflammatory infiltrate.. Aqueous extract, alcoholic extract and geniposide displayed hepatotoxicity, and the geniposide which was the main substance of the Fructus Gardeniae might be mainly responsible for the hepatotoxicity.

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; Bilirubin; Drugs, Chinese Herbal; Female; Fruit; Gardenia; Iridoids; Liver; Male; Organ Size; Plants, Medicinal; Pyrans; Random Allocation; Rats; Rats, Sprague-Dawley

To elucidate the therapeutic effect and the influence on PI3K-Akt-PKB-BAD-CREB-PCREB pathway in focal cerebral ischemia rat responses before and after treatment with baicalin and jasminoidin given alone or in combination.. Rat model of ischemia reperfusion was established with thread. Generally accepted methods were used, including TTC staining, behavior test, as well as micro and ultra microscopy which can dynamically and accurately monitor pathological and physiological changes after cerebral ischemia on earlier period, to evaluate the brain injury induced by ischemia and the attenuations by the drugs. The difference of PI3K-Akt-PKB-BAD-CREB-PCREB expression was detected by western-blot technology.. The combination of baicalin and jasminoidin composition can be potential neuroprotective agent. TTC staining technology combined with behavior grade and ultrmicro-structure observation on brain tissue is effective method to evaluate protective agent, which is related to signal transduction PI3K-Akt-PKB-BAD-CREB-PCREB pathway. The results provide benofical basis for revealing the complex of therapeutic mechanism of traditional Chinese medicine Qingkai Ling (QKL).

Topics: Animals; Behavior, Animal; Brain; Brain Ischemia; CREB-Binding Protein; Cyclic AMP Response Element-Binding Protein; Drug Combinations; Flavonoids; Gardenia; Injections; Iridoids; Male; Neuroprotective Agents; Plants, Medicinal; Proto-Oncogene Proteins c-akt; Pyrans; Random Allocation; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Scutellaria; Signal Transduction

To compare the individual effects of baicalin and jasminoidin with the combined effect of them on cerebral ischemia-reperfusion injury, and test whether the combined administration of baicalin and jasminoidin can improve the therapeutic effect. Male Sprague-Dawley rats underwent focal cerebral ischemia for 1.5 h and reperfusion for 24 h. Just before reperfusion, tested drugs (baicalin, jasminoidin, a drug combination consisting of baicalin and jasminoidin, or nimodipine) were intravenously treated. Diffusion weighted imaging (DWI) of magnetic resonance imaging (MRI), behavior examination, 2,3,5-triphenyltetrazolium chloride (TTC) staining, histological examination, and real-time PCR for BDNF and caspase-3 were performed. All of the drug treatments could significantly ameliorate the results of TTC and histological examination, and the baicalin/jasminoidin combination did so most prominently. This combination could also significantly ameliorate DWI of MRI and behavior examination results, and promote the expression of BDNF and inhibit the expression of caspase-3. On the whole, both baicalin and jasminoidin have a preventive effect against ischemic stroke, although their effects are not very strong. However, the combination of baicalin and jasminoidin can significantly improve their effectiveness. This may be related to its better regulation on the BDNF and caspase-3.

Topics: Analysis of Variance; Animals; Brain; Brain Ischemia; Brain-Derived Neurotrophic Factor; Caspase 3; Disease Models, Animal; Drug Combinations; Drug Therapy, Combination; Drugs, Chinese Herbal; Flavonoids; Iridoids; Male; Neuroprotective Agents; Nimodipine; Pyrans; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Statistics, Nonparametric

To determinate 3 kinds of effective components (5 compounds) in Huanglian Jiedu decoction at the same time, namely berberine, palmtine, jatrorrhizine, baicalin and geniposide.. A HPLC method detected by 3 different UV wavelengths 345 nm for berberine, palmtine and jatrorrhizine, 280 nm for baicalin, 238 nm for geniposide. Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column was used, the mixture of H2O (A) , CH3OH (B) and 0.05% H3PO4 solution (C) as mobile phase in gradient mode. The column temperature was 35 degrees C and the flow rate was 1 mL x min(-1).. This method was applied to determined 5 compounds in Huanglian Jiedu decoction rapidly and accurately.. The quality control method is convenient, reasonable and credible for Huanglian Jiedu decoction.

Topics: Berberine; Berberine Alkaloids; Chromatography, High Pressure Liquid; Drug Combinations; Drugs, Chinese Herbal; Flavonoids; Iridoids; Plants, Medicinal; Pyrans; Quality Control; Ultraviolet Rays

By using amplified fragment length polymorphism (AFLP) markers, this paper studied the genetic relationships among five wild or cultivated Gardenia jasminoides Ellis populations in Jiangxi Province. Chemical fingerprint was also built with HPLC method. The results showed that there was a great genetic difference among these samples. The UPGMA obtained with NTSYS-PC 2. 10e software suggested that there were seven branches of population, and the population from near geographical location clustered firstly. The geniposide content of these branches was not correlated with UPGMA. It could be concluded that the authenticity was resulted from the co-action of genotype and environmental change. The microelements content in G. jasminoides fruit measured by inductively coupled plasma showed that there was a negative correlation between Zn and geniposide contents.

Topics: Amplified Fragment Length Polymorphism Analysis; China; Chromatography, High Pressure Liquid; DNA, Plant; Drugs, Chinese Herbal; Ecology; Ecosystem; Gardenia; Iridoids; Pyrans

To study the effect of geniposide on serum IL-1beta and TNF-alpha levels of rheumatoid arthritis rats, as well as the mechanism of this drug.. To establish an experimental rat model of type II collagen-induced arthritic (CIA). The inhibitory effects on paw edema were observed, and serum IL-1beta and TNF-alpha levels were determined in experimental rats.. Compared with the model, geniposide delayed the starting time of right paw edema significantly, and the levels of serum IL-1beta and TNF-alpha were significantly decreased by geniposide at high dose or medium dose (P < 0.01).. Geniposide can lower serum IL-1beta and TNF-alpha levels in rheumatoid arthritis rats. The effect may be close related to inhibitory development of rheumatoid arthritis by the agent.

Topics: Animals; Arthritis, Rheumatoid; Collagen Type II; Dose-Response Relationship, Drug; Edema; Gardenia; Hindlimb; Interleukin-1; Iridoids; Male; Plants, Medicinal; Pyrans; Rats; Rats, Wistar; Tumor Necrosis Factor-alpha

Geniposide, an iridoid glycoside isolated from the fruit of Gardenia jasminoides Ellis, has biological capabilities of detoxication, antioxidation, and anticarcinogenesis. We have recently found that geniposide possesses a potential for detoxication by inducing GST activity and the expression of GST M1 and GST M2 subunits. In this study, the signaling pathway of geniposide leading to the activation of GSH S-transferase (GST) was investigated. Primary cultured rat hepatocytes were treated with geniposide in the presence or absence of mitogen-activated protein kinase (MAPK) inhibitors and examined for GST activity, expression of GST M1 and M2 subunits, and protein levels of MAPK signaling proteins. Western blotting data demonstrated that geniposide induced increased protein levels of GST M1 and GST M2 (approximately 1.76- and 1.50-fold of control, respectively). The effect of geniposide on the increased protein levels of GST M1 and GST M2 was inhibited by the MEK-1 inhibitor PD98059, but not by other MAPK inhibitors. The GST M1 and GST M2 transcripts as determined by RT-PCR and GST activity were also inhibited concurrently by the MEK-1 inhibitor PD98059. The protein levels of up- and down-stream effectors of the MEK-1, including Ras, Raf, and Erk1/2, and the phosphorylation state of Erk1/2 were found to be induced by geniposide, indicating a two-phase influence of geniposide. The results suggest that geniposide induced GST activity and the expression of GST M1 and GST M2 acting through MEK-1 pathway by activating and increasing expression of Ras/Raf/MEK-1 signaling mediators.

Topics: Animals; Enzyme Activation; Enzyme Inhibitors; Glutathione Transferase; Hepatocytes; Iridoids; Isoenzymes; Male; MAP Kinase Kinase 1; Mitogen-Activated Protein Kinases; Phosphorylation; Pyrans; Rats; Rats, Sprague-Dawley; Signal Transduction; Transcription, Genetic

High-speed counter-current chromatography (HSCCC) was applied to the isolation and purification of geniposide from Gardenia jasminoides Ellis. Analytical HSCCC was used for the preliminary selection of a suitable solvent system composed of ethyl acetate-n-butanol-water (2:1:3, v/v/v). According to the above solvent system, preparative HSCCC was successfully performed with the optimal solvent system composed of ethyl acetate-n-butanol-water (2:1.5:3, v/v/v) yielding 389 mg of geniposide at over 98% purity from 1g of the partially purified extract with 38.9% recovery in a one-step separation.

Topics: Countercurrent Distribution; Gardenia; Iridoids; Magnetic Resonance Spectroscopy; Pyrans; Spectrometry, Mass, Electrospray Ionization

The discovery of new topoisomerase I inhibitors is necessary since most of the antitumor drugs are targeted against type II and only a very few can specifically affect type I. Topoisomerase poisons generate toxic DNA damage by stabilization of the covalent DNA-topoisomerase cleavage complex and some have therapeutic efficacy in human cancer. Two iridoids, aucubin and geniposide, have shown antitumoral activities, but their activity against topoisomerase enzymes has not been tested. Here it was found that both compounds are able to stabilize covalent attachments of the topoisomerase I subunits to DNA at sites of DNA strand breaks, generating cleavage complexes intermediates so being active as poisons of topoisomerase I, but not topoisomerase II. This result points to DNA damage induced by topoisomerase I poisoning as one of the possible mechanisms by which these two iridoids have shown antitumoral activity, increasing interest in their possible use in cancer chemoprevention and therapy.

Topics: Camptothecin; DNA Damage; DNA Topoisomerases, Type I; DNA Topoisomerases, Type II; Enzyme Inhibitors; Glucosides; Humans; Iridoid Glucosides; Iridoids; Neoplasms; Pyrans; Topoisomerase I Inhibitors; Topoisomerase II Inhibitors; Tumor Cells, Cultured

A new iridoid, gardaloside (1), and a new safranal-type monoterpene, jasminoside G (2), together with 10 known compounds including nine iridoids and a second safranal-type monoterpene, were isolated from the fruits of Gardenia jasminoides. The structures of 1 and 2 were established on the basis of spectroscopic evidence. Of these compounds, geniposide (3), 6alpha-hydroxygeniposide (5), ixoroside (7), and shanzhiside (8) showed significant inhibition of IL-2 secretion by phorbol myristate acetate and anti-CD28 monoclonal antibody co-stimulated activation of human peripheral blood T cells.

Topics: Antibodies, Monoclonal; CD28 Antigens; Fruit; Gardenia; Humans; Immunosuppressive Agents; Interleukin-2; Iridoids; Molecular Structure; Monoterpenes; Plants, Medicinal; Pyrans; T-Lymphocytes; Taiwan; Tetradecanoylphorbol Acetate

A method was established for the simultaneous quantification of nine components of five different structural types in Qingkailing injection. High performance liquid chromatography coupled with a photo diode array detector and an evaporative light scattering detector (HPLC-DAD-ELSD) was employed in the determination. Four monitoring wavelengths of 240, 254, 280 and 330 nm were set to determine nucleosides (uridine and adenosine), iridoid glucoside (geniposide), flavone glycoside (baicalin) and organic acids (chlorogenic acid and caffeic acid) respectively, and a combined evaporative light scattering detector was used to detect three steroid compounds (cholic acid, ursodesoxycholic acid and hyodeoxycholic acid). The proposed method permitted the simultaneous separation and determination of five groups of compounds in Qingkailing injection, and acceptable validation results of the precision, repeatability, stability and accuracy tests were achieved. The method was applied to the analysis of 19 Qingkailing injection samples from three different plants, and the results indicated that the method could be used as a convenient and reliable method in the multi-component determination and quality control of traditional Chinese medicines.

Topics: Adenosine; Caffeic Acids; Chlorogenic Acid; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Flavonoids; Iridoids; Medicine, Chinese Traditional; Scattering, Radiation; Uridine

The aim of this study was to evaluate the in vitro antioxidant activity of the methanol extract of Plantago bellardii All. aerial parts. This was assessed by two different tests, scavenging of 1,1-diphenyl-2-picrylhydrazil (DPPH) radical, and inhibition of lipid peroxidation on liposomes prepared from bovine brain extract. In both tests the extract showed a potent antioxidant effect. The characterization of the major compounds in the extract as rutin, geniposide and verbascoside was performed by isolation and HPLC comparison with authentic samples. They were quantified by HPLC for the flavonoids and colorimetry for iridoids. The compounds that contribute most to the antioxidant activity were shown to be verbascoside and rutin.

Topics: Antioxidants; Glucosides; Iridoid Glucosides; Iridoids; Molecular Structure; Phenols; Plant Extracts; Plantago; Pyrans; Rutin

To estimate the uncertainty for quantitative analysis results of geniposide, chloregenic acid and crocinl I in Gardenia jasminoides Ellis.. HPLC method was employed to determine the amounts of geniposide, choloregenic acid and crocinl I in Gardenia jasminoides Ellis. Analyzing the uncertainty sources arising from the procedure of analysis, the standard uncertainty and combined uncertainty and expanded uncertainty were calculated according to the data of HPLC.. This method met the requirements of modern pharmaceutical analysis, and the expand uncertainties for the HPLC method of the three components are 0.1024, 0.2254, 0.1264, respectively.. Applying measurement uncertainty to the evaluation of quantitative analysis results of active components in Gardenia jasminoides Ellis. is an improvement to the actual error evaluation system.

Topics: Algorithms; Carotenoids; Chlorogenic Acid; Chromatography, High Pressure Liquid; Fruit; Gardenia; Iridoids; Models, Statistical; Plants, Medicinal; Pyrans; Quality Control; Uncertainty

Geniposide, an iridoid glycoside isolated from the fruit of Gardenia jasminoides Ellis, has the biological capabilities of detoxication, antioxidation, and anticarcinogenesis. In this study, the mechanism of geniposide affecting the GST (glutathione S-transferase) system was investigated. Primary cultured rat hepatocytes were treated with geniposide and examined for total GST activity and expression of GST subunits. The results showed that the geniposide-induced GST activity was dose and time dependent. Western blotting data demonstrated that geniposide induced increased protein levels of GSTM1 and GSTM2 (approximately 1.7- and 1.8-fold of control, respectively), but did not increase those of GSTA1. The corresponding transcripts levels were confirmed by RT-PCR. Using PD98059, the effect of geniposide was verified to be via the MEK pathway. The results suggest that geniposide possesses a potential for detoxication by inducing GST activity via increasing the transcription of GSTM1 and GSTM2.

Topics: Animals; Cells, Cultured; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Enzyme Induction; Flavonoids; Fruit; Glutathione Transferase; Glycosides; Hepatitis; Hepatocytes; Iridoids; Isoenzymes; Male; Plant Extracts; Pyrans; Rats; Rats, Sprague-Dawley; RNA, Messenger; Rubiaceae; Time Factors

Inchin-ko-to (ICKT), an herbal medicine, and its ingredients exert potent choleretic effects by a "bile acid-independent" mechanism. The current study was designed to determine whether ICKT or its ingredients potentiate multidrug resistance-associated protein 2 (Mrp2; Abcc2)-mediated choleresis in vivo. Biliary secretion of Mrp2 substrates and the protein mass, subcellular localization, and messenger RNA (mRNA) level of Mrp2 were assessed in rat liver after infusion of genipin, an intestinal bacterial metabolite of geniposide, a major ingredient of ICKT. The function of Mrp2 was also assessed by the adenosine triphosphate (ATP)-dependent uptake of Mrp2-specific substrates using canalicular membrane vesicles (CMVs) from the liver. Infusion of genipin increased bile flow by 230%. It also increased biliary secretion of bilirubin conjugates and reduced glutathione (GSH) by 513% and 336%, respectively, but did not increase bile acid secretion. The ATP-dependent uptake of estradiol 17-beta-D-glucuronide (E(2)17 beta G; by 265%), leukotriene C4 (LTC(4); by 161%), taurolithocholate-3-sulfate (TLC-3S; by 266%), and methotrexate (MTX; by 234%) was significantly stimulated in the CMVs from the liver. These effects were not observed in Mrp2-deficient rats. Under these conditions, genipin treatment increased the protein mass of Mrp2 in the CMVs but not the mRNA level. In immunoelectron microscopic studies, a marked increase in Mrp2 density in the canalicular membrane (CM) and microvilli was observed in the genipin-treated liver tissue sections when compared with the vehicle-treated liver tissue sections. In conclusion, genipin may enhance the bile acid-independent secretory capacity of hepatocytes, mainly by stimulation of exocytosis and insertion of Mrp2 in the bile canaliculi. ICKT may be a potent therapeutic agent for a number of cholestatic liver diseases.

Topics: Actin Cytoskeleton; Adenosine Triphosphate; Administration, Oral; Animals; Anions; ATP Binding Cassette Transporter, Subfamily B, Member 11; ATP-Binding Cassette Transporters; Bile; Cholagogues and Choleretics; Drugs, Chinese Herbal; Estradiol; Gene Expression; In Vitro Techniques; Iridoid Glycosides; Iridoids; Liver; Male; Membrane Transport Proteins; Multidrug Resistance-Associated Protein 2; Multidrug Resistance-Associated Proteins; Phalloidine; Pyrans; Rats; Rats, Sprague-Dawley; RNA, Messenger; Tritium

The EtOH extract of gardenia (Gardenia jasminoides Ellis) fruits was previously found to possess potent anti-angiogenic activity in the chick embryo chorioallantoic membrane (CAM) assay. Bioassay-guided fractionation and purification of the EtOH extract yielded an active anti-angiogenic compound, which was determined to be an iridoid glucoside, geniposide, by spectral analyses. Geniposide showed anti-angiogenic activity in a dose-dependent manner. It also exhibited an inhibitory effect in the range of 25-100 microM on the growth of the transformed NIH3T3 cell line.

Topics: Angiogenesis Inhibitors; Animals; Chickens; Choroidal Neovascularization; Dose-Response Relationship, Drug; Fruit; Gardenia; Iridoids; Phytotherapy; Plant Extracts; Pyrans

Inchin-ko-to (TJ-135) is an herbal medicine used in Japan for treatment of icteric patients with cirrhosis. Its efficacy as an anti-fibrogenic drug was evaluated in relation to stellate cell activation.. Liver fibrosis was induced in rats by repeated injections of carbon tetrachloride (CCl4) or pig-serum. Oral administration of TJ-135 improved the mortality of rats given CCl4 with reduced extents of liver necrosis and fibrosis. Similar improvement of liver fibrosis was found in rats given pig-serum showing no liver necrosis. DNA synthesis of stellate cells activated in vitro after isolation from normal rat liver was decreased by culture with TJ-135 in a dose-related manner, accompanied by decreased smooth muscle alpha actin expression and contractility. Such attenuation was not found in the cells cultured with geniposide, an iridoid compound of TJ-135, but genipin, an aglycone of geniposide formed in the gut by action of bacterial flora, markedly decreased stellate cell activation without affecting synthesis of proteins other than collagen.. TJ-135 may be useful for treatment of liver fibrosis and portal hypertension through suppression of activated hepatic stellate cell function by genipin, an absorbed form of its component.

Topics: Animals; Carbon Tetrachloride; Cells, Cultured; Cholagogues and Choleretics; Drugs, Chinese Herbal; Iridoid Glycosides; Iridoids; Liver; Liver Cirrhosis, Experimental; Necrosis; Pyrans; Rats; Rats, Inbred F344; Survival Analysis; Swine

To evaluate the pharmacological actions of herbal medicines, metabolic activities of herbal medicine components, ginsenoside Rb1, glycyrrhizin, geniposide and baicalin to their bioactive compounds compound K, 18beta-glycyrrhetic acid, genipin and baicalein by fecal specimens were measured. Their metabolic activities were 646.1+/-591.4, 29.4+/-51.7, 926.3+/-569.6 and 3884.6+/-1400.1 micromol/h/g, respectively. The profiles of these metabolic activities of baicalin and ginsenoside Rb1 were not significantly different to those of water extracts of Scutellariae Radix and Ginseng Radix. None of the metabolic activities tested were different between males and females, or between ages. However, the difference in these metabolic activities in individuals was significant. These results suggest that the human intestinal microflora enzymes that convert herbal components to their bioactive compounds may be used as selection markers of responders to traditional medicines.

Topics: Adult; Chromatography, Thin Layer; Feces; Female; Flavanones; Flavonoids; Ginsenosides; Glycyrrhetinic Acid; Glycyrrhizic Acid; Humans; In Vitro Techniques; Iridoid Glycosides; Iridoids; Male; Panax; Plant Extracts; Pyrans; Scutellaria baicalensis; Stereoisomerism

A high performance liquid chromatographic method has been developed for the determination of geniposide in Xinxue granules. Geniposide was extracted by ultrasonic extraction for 30 min. The chromatographic separation was carried out on a Diamonsil C18 column (200 mm x 4. 6 mm i.d., 5 microm) with a mobile phase consisting of an acetonitrile-water (15:85, v/v) mixture. The detection wavelength was set at 238 nm. The calibration curve was linear in the ranged of 25 - 400 mg/L for geniposide. The average recovery of geniposide was 101.2% with a relative standard deviation (RSD) of 1.6% and contents of geniposide in the granules ranged from 0.841 to 0.923 mg/g. This method is simple, reliable and suitable for quality control of Xinxue granules.

Topics: Chromatography, High Pressure Liquid; Drug Combinations; Drugs, Chinese Herbal; Iridoids; Pyrans

To study in vitro dissolution rate of geniposide in Huangqin Qingfei dispersible tablet.. A reversed-phase HPLC method was developed for determination of geniposide. In vitro dissolution rates were compared between Huangqin Qingfei dispersible tablet and conventional tablet in the dissolution medium of pH 1.0, 2.85, 4.5, 6.8, and 8.0 accordingly. Zero-class model, single-index model, logarithm normal school model, and Weibull distributing model were used to simulate the dissolution curve.. The dissolution rate of two tablets is not affected by pH so much, and they can dissolve within 5 to 10 minutes. Weibull distributing model is the best simulation for in vitro dissolution. Comparing with conventional tablet, dispersible tablet dissolve quickly and completely.. The in vitro dissolution rate of geniposide in Huangqin Qingfei dispersible tablet conforms to Weibull distributing model. The dispersible tablet is able to release rapidly.

Topics: Chromatography, High Pressure Liquid; Drug Combinations; Drugs, Chinese Herbal; Gardenia; Iridoids; Kinetics; Plants, Medicinal; Pyrans; Scutellaria; Solubility; Tablets; Time Factors

Gardenia fruit (Gardenia jasminoides ELLIS) is widely used as a natural food colorant and as a traditional Chinese medicine for treatment of hepatic and inflammatory diseases. "Gardenia yellow" is a natural food colorant which is extracted by ethanol from gardenia fruit. The purpose of this study was to evaluate the genotoxicity of gardenia yellow. Genotoxicity of gardenia yellow and its components, crocetin, gentiobiose (a component of crocin), geniposide and genipin (formed by hydrolysis of geniposide), was studied by Ames test, rec-assay, and sister chromatid exchange (SCE) using V79 cells. Gardenia yellow and its components were found not to be mutagenic in the Salmonella reverse mutation assay. Gardenia yellow and genipin caused damage of DNA in rec-assay. Gardenia yellow induced a significant dose-dependent increase of SCE frequency (8.6 times at 1000 microg/ml as the value for the solvent control). Only genipin induced SCEs significantly among the components of gardenia yellow. Moreover, genipin induced a significant increase of tetraploids at all doses tested (95% at 8 microg/ml). Gardenia yellow preparation was analyzed by capillary electrophoresis (CE), and geniposide was detected. However, genipin was not observed. In conclusion, we have shown that genipin possesses genotoxicity. Furthermore, there were unidentified genotoxicants in gardenia yellow.

Topics: Bacillus subtilis; Carotenoids; Coloring Agents; Disaccharides; DNA Damage; Electrophoresis, Capillary; Food Coloring Agents; Gardenia; Iridoid Glycosides; Iridoids; Mutagenicity Tests; Plant Extracts; Pyrans; Sister Chromatid Exchange; Vitamin A

A high-performance liquid chromatographic method was applied to the determination of the geniposide concentration in Gardenia fruit and preparations of traditional Chinese medicine using a mobile phase of acetonitrile-methanol-5 mM monosodium phosphate (pH 4.6) (5:15:80, v/v/v). Intra-assay and inter-assay accuracy and precision of the analyses were < or = 10% in the range of 0.1 through 50 microg/ml. The presence of geniposide in the medicinal herb and its preparations was ascertained by retention time, spiking with an authentic standard, change of detection wavelength and change of the composition of the mobile phase. The concentration of geniposide in the fruit of Gardenia jasminoides Ellis var. grandiflora Nakai is higher than that in Gardenia jasminoides Ellis. The concentration of geniposide in the traditional Chinese herbal medicine preparations, Huang-Lian-Jiee-Dwu-Tang (66.27 +/- 1.98 mg/g) and In-Chern-Hau-Tang (68.54 +/- 2.62 mg/g) was less than in the herb Gardenia jasminoides Ellis (73.44 +/- 2.62 mg/g) itself.

Topics: Chromatography, High Pressure Liquid; Gardenia; Iridoids; Medicine, Chinese Traditional; Pyrans; Reference Standards; Reproducibility of Results; Spectrophotometry, Ultraviolet

Gardenia blue dye was obtained through the reaction of methylamine with genipin, the aglycone of geniposide isolated from the fruits of Gardenia jasminoides. The resulting blue pigments were passed through Bio-Gel P-2 resin yielding five fractions, GM1-GM5. Four fractions (GM1-GM4) were all blue pigments, and the first eluted higher molecular weight fraction GM1 had a higher tinctorial strength than the later eluted lower molecular weight fractions, GM2-GM4. The last eluted GM5 fraction with lambda(max) of 292 nm was colorless and was confirmed as the true intermediate of the blue pigments on the basis of UV-vis spectrophotometric evidence. The GM5 fraction was composed of two epimeric isomers, and their structures were characterized by (1)H NMR, (1)H-(1)H COSY, (13)C NMR, and HMQC and HMBC spectral measurements.

Topics: Gardenia; Iridoids; Isomerism; Methylamines; Molecular Weight; Pigments, Biological; Pyrans; Solubility

Natural products are normally obtained by organic solvent extraction and many subsequent chromatographic separations. Compounds of interest are often isolated with very low yield and limited purity. An aqueous two-phase extraction process combined with a simple ethanol treatment, for removing excess inorganic salt, has been developed for preparation of geniposide from gardenia. The system was comprised of PE62, a random copolymer composed of 20% ethylene oxide and 80% propylene oxide, KH2PO4 and ethanol. To find optimal conditions, the partition behavior of geniposide under an aqueous two-phase system was investigated. Various factors were considered, including the concentration of salt, the concentration of polymer, the sample loading, and the addition of ethanol. The experimental results demonstrated that increasing salt concentration or decreasing PE62 concentration results in enhancement of the geniposide partition in the salt-rich phase. The addition of ethanol and higher sample loading also promoted the partition efficiency of geniposide. Based on this study, an optimized system containing 5% PE62, 7.5% KH2PO4, and 10% ethanol was tested on a large-scale extraction. A 39.0-g aliquot of final product (in powder form) with 77% purity of geniposide can be effectively extracted from 500 g of gardenia fruit. This process is proved to be useful for industrial application of geniposide preparation.

Topics: Chromatography, High Pressure Liquid; Gardenia; Iridoids; Pyrans; Spectrophotometry, Ultraviolet

Fruits of Gardenia jasminoides contain geniposide which can be transformed to blue pigments by a simple modification. Colorless geniposide obtained from gardenia fruits by charcoal and silica gel column chromatographies was hydrolyzed with beta-glucosidase to yield genipin. The resulting genipin was transformed to blue pigments by reaction with amino acids (glycine, lysine, or phenylalanine). The stability of the blue pigments against heat, light, and pH was studied to examine the blue dye for possible use as a value-added food colorant. Thermal degradation reactions at temperatures of 60-90 degrees C were carried out at different pH levels within the range 5.0-9.0 (pH 5.0, acetate buffer; pH 7.0, phosphate buffer; and pH 9.0, CHES buffer). The blue pigments remained stable after 10 h at temperatures of 60-90 degrees C, and in some cases, more new pigments formed. The pigments were more stable at alkaline pH than neutral and acidic pH. Similarly, the pigments were stable under light irradiance of 5000-20 000 lux. In this case, pH effect was not significant.

Topics: Drug Stability; Fruit; Hydrogen-Ion Concentration; Iridoids; Light; Pigments, Biological; Pyrans; Temperature

Geniposide is one of the constituents of Gardenia fruit (Gardenia jasminoides Ellis, Rubiaceae), which has been used in traditional medicine. Although its anti-inflammatory and antithrombotic effects have been reported, the way it acts is still unclear. We have investigated the effects of geniposide and its metabolite genipin on thrombogenesis and platelet aggregation. In an in vivo model, geniposide and genipin significantly (P < 0.05) prolonged the time required for thrombotic occlusion induced by photochemical reaction in the mouse femoral artery. In an in vitro study, both geniposide and genipin inhibited collagen-induced, but did not inhibit arachidonate-induced, mouse platelet aggregation. However aspirin, a cyclooxygenase inhibitor, inhibited arachidonate-induced platelet aggregation but only partially inhibited the collagen-induced one. We also showed, by measuring PLA(2)-catalyzed arachidonic acid release, that geniposide inhibited phospholipase A(2) (PLA(2)) activity. We conclude that geniposide showed an antithrombotic effect in vivo due to the suppression of platelet aggregation. PLA(2) inhibition by geniposide is one possible anti-platelet mechanism.

Topics: Animals; Disease Models, Animal; Fibrinolytic Agents; Fruit; Iridoid Glycosides; Iridoids; Male; Mice; Phospholipases A; Phytotherapy; Plant Extracts; Platelet Aggregation; Pyrans; Rubiaceae; Thrombosis

The primary objective of this study was to investigate the effect of geniposide, a potent anti-inflammatory, on ovalbumin-antigen-induced tracheal permeability and transepithelial electrical resistance in guinea pigs. Two weeks after sensitization with ovalbumin (100 mg/ml), the permeability of guinea-pig tracheas was evaluated by flux measurements using the transcellular tracer, [(14)C]estradiol, and the paracellular tracer, [(14)C]mannitol. The effect of extracellular Ca(2+) with geniposide was also studied, using deletion of Ca(2+) in the donor chamber. The in vivo treatment effect of aerosolized geniposide on tracheal permeability in the ovalbumin-sensitized guinea pigs was also evaluated. The results indicate that tight junction permeability of ovalbumin-sensitized trachea was significantly dose dependent and decreased by geniposide (1-10 mM), as evidenced by substantial recovery of transepithelial electrical resistance and decreased transepithelial permeability of [(14)C]mannitol at (1.32+/-0.12) x 10(-5) cm/s. The effect of combination of the removal of extracellular Ca(2+) with geniposide had no effect on tight junction permeability of ovalbumin-sensitized trachea and revealed that transepithelial electrical resistance and junction permeability did not recover. In addition, the cAMP levels and phosphodiesterase activity were not significantly influenced in ovalbumin-sensitized tracheal tissues after geniposide treatment. Inhaled geniposide (50 mM, 30 min after ovalbumin sensitization) significantly restored junction permeability induced by ovalbumin (100 mg/ml, 2 min). Junction permeability did not recover on pretreatment with geniposide (50 mM for 30 min over 16 days consecutive before ovalbumin sensitization) after exposure of conscious guinea pigs to aerosol ovalbumin. In conclusion, geniposide has inhibitory effects on ovalbumin-induced junction permeability and recovery of transepithelial electrical resistance in guinea pig trachea, showing its potential as anti-asthma therapy.

Topics: Aerosols; Animals; Anti-Inflammatory Agents; Calcium; Cyclic AMP; Guinea Pigs; Iridoids; Male; Mannitol; Ovalbumin; Permeability; Pyrans; Tight Junctions; Trachea

Topics: Administration, Oral; Animals; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Humans; Iridoid Glycosides; Iridoids; Male; Mice; Mice, Inbred Strains; Plant Extracts; Pyrans; Rubiaceae

We showed previously that a Kampo (Chinese/Japanese herbal) medicine, Inchin-ko-to (ICKT), inhibits hepatocyte apoptosis induced by transforming growth factor beta1 in vitro. The present study investigated whether ICKT or its ingredients inhibit Fas-mediated liver apoptosis in vivo.. Acute liver injury was induced by an intravenous injection of anti-Fas antibody, Jo2. The effects of ICKT and its ingredients on lethality, histology, apoptotic index, serum transaminase levels, caspase activation, mitochondrial membrane potential (Deltapsi(m)), and mitochondrial permeability transition (MPT) were analyzed. Apoptosis in mouse hepatocytes in vitro was also evaluated.. Pretreatment with ICKT rescued 75% of Jo2-treated mice and markedly suppressed liver apoptosis/injury. Genipin, an intestinal bacterial metabolite of geniposide that is a major ingredient of ICKT, was found to be an active principle of ICKT. Genipin also suppressed in vitro Fas-mediated apoptosis in primary-cultured murine hepatocytes. Activation of caspase 3 and 8 in the liver homogenate and rapid reduction of triangle uppsi(m) of hepatocytes isolated from Jo2-treated mice were inhibited by genipin preadministration. The resistance to Ca(2+)-induced MPT was enhanced in liver mitochondria of genipin-treated mice.. These results suggest that the antiapoptotic activity of genipin via the interference with MPT is a possible mechanism for therapeutic effects of ICKT.

Topics: Alanine Transaminase; Animals; Apoptosis; Aspartate Aminotransferases; Cells, Cultured; Cholagogues and Choleretics; fas Receptor; Female; Glycyrrhizic Acid; Intracellular Membranes; Iridoid Glycosides; Iridoids; Liver; Membrane Potentials; Mice; Mice, Inbred BALB C; Mitochondria, Liver; Permeability; Plants, Medicinal; Pyrans; Ursodeoxycholic Acid

The effect of a methanol extract of Eucommiae Cortex on collagen synthesis was investigated in false aged model rats. Granuloma formation and collagen synthesis were significantly increased by the administration of the methanol extract of Eucommiae Cortex. The effective component of Eucommiae Cortex was then discussed by fractionating the methanol extract of Eucommiae Cortex. Eucommiol, a main component in the water fraction of the methanol extract, was found to be an effective compound. In our previous paper, we reported the promoting effect of Eucommia ulmoides OLIVER leaf on collagen synthesis, and found geniposidic acid and aucubin were the main effective compounds in the leaf. Based on our data in this paper, we clarified that the main effective components of the Eucommia ulmoides OLIVER leaf and Eucommiae Cortex were different. Geniposidic acid and aucubin were reported to be contained at a high concentration in the fresh cortex of Eucommia ulmoides OLIVER, but during the drying process and storage, most of them were destroyed by enzymes in the cortex and very little remained in the Eucommiae Cortex. Therefore, we investigated the effect of the methanol extract of fresh cortex of Eucommia ulmoides OLIVER. A stronger effect than Eucommiae cortex was shown, and geniposidic acid, aucubin and geniposide were concluded to be the main effective components. Although geniposide was found to be an effective compound, when the dose was higher than 50 mg/kg/d, toxicity was shown. The pharmaceutical effect of eucommiol was reported for the first time.

Topics: Aging; Alcohols; Animals; Collagen; Cyclopentanes; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Iridoids; Plant Leaves; Plants, Medicinal; Pyrans; Rats; Rats, Wistar; Trees

Human uridinediphosphate-glucuronosyltransferase 1A1 (UGT1A1) was expressed in Salmonella typhimurium TA1535 cells by transfection of the cells with plasmids carrying the UGT1A1 cDNA. UGT1A1 cDNA was isolated by a polymerase chain reaction from human liver total RNA and was inserted into the pSE420 plasmid, linked to the trc promoter and terminator. The plasmid thus constructed was introduced into Salmonella TA1535 cells. The expression of human UGT1A1 protein was confirmed by Western blot analysis. The maximal expression was observed at 24 h after the addition of isopropyl-beta-D-thiogalactopyranoside, an inducer. However, the bilirubin conjugation activity of the membrane fraction from the Salmonella cells was not detectable. When a beta-glucuronidase inhibitor such as saccharic acid 1,4-lactone, glycyrrhizin or 1-naphtyl-beta-D-glucuronide was added to the reaction mixture, the bilirubin conjugation activity of the human UGT1A1 was detected. When geniposide was added to the reaction mixture, the bilirubin conjugation activity of UGT1A1 was not seen. Taking these results into account, the established Salmonella strain possesses the beta-glucuronidase activity. Since the beta-glucuronidase activity of the Salmonella was lower than that of E. coli, it was concluded that Salmonella seemed to be a good host to express UGT protein. This is the first study to demonstrate the establishment of a bacterial strain expressing native human UGT protein showing catalytic activity.

Topics: Animals; Bilirubin; Blotting, Western; Cell Fractionation; DNA Primers; DNA, Complementary; Enzyme Inhibitors; Escherichia coli; Flavonoids; Gene Expression; Glucuronates; Glucuronidase; Glucuronosyltransferase; Glycyrrhizic Acid; Humans; Iridoids; Microsomes, Liver; Plasmids; Polymerase Chain Reaction; Pyrans; RNA; Salmonella typhimurium; Transfection

To determine the iridoid contents in Gardenia sootepensis.. Using RP-HPLC with geniposidic acid, geniposide and scandoside methyl ether as standards.. According to the selected chromatographic conditions, the linear ranges of geniposidic acid, geniposide and scandoside methyl ether were 0.097-0.606 microgram, 0.0638-3.990 micrograms and 0.198-1.239 micrograms, respectively. The average recoveries of these three compounds were 100.0%, 99.9% and 100.1%, and RSD 1.23%, 1.38% and 1.53% (n = 5) respectively.. The method is easy, rapid, and reproducible.

Topics: Chromatography, High Pressure Liquid; Fruit; Gardenia; Glucosides; Iridoid Glucosides; Iridoids; Plants, Medicinal; Pyrans

To demonstrate the chemical constituents of the fruits of Gardenia sootepensis.. Column chromatographic technology was employed to separate the water soluble part, and the compounds obtained were elucidated by spectral and chemical analysis.. Ten compounds have been isolated, seven of which elucidated respectively as D-mannitol, beta-sitosterol, deacetylasperulosidic acid methyl ester, geniposidic acid, geniposide, scandoside methyl ester and dimeric molecule of quinide.. All the compounds were obtained from this plant for the first time.

Topics: Drugs, Chinese Herbal; Fruit; Gardenia; Glucosides; Iridoid Glucosides; Iridoids; Mannitol; Plants, Medicinal; Pyrans; Sitosterols

The suppressing effect of crude extracts of Tochu tea, an aqueous extract of Eucommia ulmoides leaves and a popular beverage in Japan, on the induction of chromosome aberrations in CHO cells and mice was studied. When CHO cells were treated with Tochu tea crude extract after MMC treatment, the frequency of chromosome aberrations was reduced. Out of 17 Tochu tea components, 5 irridoids (geniposidic acid, geniposide, asperulosidic acid, deacetyl asperulosidic acid, and asperuloside) and 3 phenols (pyrogallol, protocatechuic acid, and p-trans-coumaric acid) were found to have anticlastogenic activity. Since the anticlastogenic irridoids had an alpha-unsaturated carbonyl group, this structure was considered to play an important role in the anticlastogenicity. The anticlastogenic effect of Tochu tea extracts was examined in mice using a micronucleus assay. When mice received 1.0 ml 4% Tochu tea extract by oral gavage 6 h before intraperitoneal injection of MMC, a decrease in the frequency of micronuclei was observed. This decrease was not due to a delay in the maturation of micronucleated reticulocytes.

Topics: Animals; Antimutagenic Agents; Beverages; CHO Cells; Chromatids; Chromosome Aberrations; Cricetinae; Cyclopentane Monoterpenes; Glucosides; Iridoid Glucosides; Iridoids; Japan; Male; Mice; Mice, Inbred ICR; Micronucleus Tests; Mitomycin; Mutagens; Phenols; Plant Extracts; Pyrans

The antitumor effects of two iridoid compounds, geniposidic acid (GA) and geniposide (GP), were investigated in mice along with their possible effects on radioprotection after sublethal X-irradiation. Decreases in the growth of the implanted tumor by ascitic cells were a result of intraperitoneal administration of GA and GP at high concentrated levels. This result was achieved by exerting the levels of dosage in a dose-dependent manner. Except on the 12th day after treatment by the dosage of 500 mg/kg, reduced radiation effects of mice treated with the drugs in the 30 min preirradiated period by GA and GP on peripheral leukocytes were not observed significantly by the sublethal whole-body X-irradiation. And except on the 7th day after treatment, when these two compounds were administered i.p. to mice 30 min before 4 Gy irradiation, neither GA nor GP enhanced significantly the postirradiation responses of splenic blastogenesis by PHA. In addition, GA might be a more potent tumor growth inhibitor than GP when combined with the X-irradiation, though there was no significant synergetic effect on their combined antitumor activity. The preliminary results of GA and GP on hematological and blastogenic observations in this study suggested that they may very well, partially, play a role in an effective anticancer product with the ability to decrease undesirable radiation damage to the hematologic tissue after high dose irradiation.

Topics: Animals; Antineoplastic Agents; Carcinoma, Ehrlich Tumor; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Glucosides; Iridoid Glucosides; Iridoids; Leukocyte Count; Lymphocyte Activation; Male; Mice; Mice, Inbred ICR; Pyrans; Radiation-Protective Agents; Time Factors; Whole-Body Irradiation

The geniposide and paeoniflorin in Jiaowei Xiaoyao Pills were separated and determined by HPLC, with a C18 column and a mixture of CH3CN-0.1% H3PO4(13:87) as mobile phase. The method is simple, rapid and accurate. The components have a good linearity, and the average recovery with RSD(n = 5) is 101.81% + 2.38% for geniposide and 98.75% + 1.95% for paeoniflorin.

Topics: Benzoates; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Drug Combinations; Drugs, Chinese Herbal; Glucosides; Iridoids; Monoterpenes; Pyrans; Quality Control

We examined effect of iridoid glucosides, aucubin, catalpol, geniposide and gardenoside, and their enzymic hydrolysates on neurite outgrowth of PC12h cells. Except for aucubin, these glucosides induced neurite outgrowth at 0.1 microgram/ml and above in medium after 3 d of treatment. Hydrolysates of the four glucosides all caused neuritogenesis. Geniposide hydrolysate enhanced responses of cells to carbachol and KCl-induced depolarization in terms of cytoplasmic free-calcium concentration. The aglucone of geniposide, genipin, also promoted neurite outgrowth in a dose-dependent manner (ED50 = 0.7 microM). The neuritogenic effect of genipin was partially or considerably inhibited in the presence of H-89 and genistein. All the results presented suggest that certain iridoid compounds can induce neuronal differentiation in PC12h cells through activation of components of the intracellular signal transduction pathway.

Topics: Animals; beta-Glucosidase; Calcium; Glucosides; Hydrolysis; Iridoid Glucosides; Iridoid Glycosides; Iridoids; Neurites; Neurons; PC12 Cells; Protein Kinase Inhibitors; Pyrans; Rats; Signal Transduction

A geniposide-hydrolyzing beta-glucosidase was discovered in Eubacterium sp. A-44, a human intestinal anaerobe. The enzyme was intracellularly distributed in the bacterium, and purified to homogeneity from the extract using Butyl-Toyopearl 650M, Sephacryl S-300, hydroxyapatite and chromatofocusing column chromatography. The enzyme was a single polypeptide chain with the molecular weight of 90 kDa and the N-terminal amino acid sequence initiated from methionine up to the 29th residue did not show more than 50% homology against known protein sequences. A broad substrate specificity was shown for the beta-glucosidase to hydrolyze aryl beta-D-glucosides (p-nitrophenyl beta-D-glucopyranoside-pNPG, esculin and salicin), alkyl beta-D-glucosides (geniposide and amygdalin) and cellobiose. The Km values (mM) for various beta-D-glucosides were 0.068 for geniposide, 0.10 for pNPG, 0.21 for esculin, 0.22 for salicin, 2.9 for amygdalin, and 0.91 for cellobiose. The pH optimum with pNPG and geniposide as the substrates was 6.0. The enzyme was inhibited by sulfhydryl reagents, Cu2+, and nojirimycin bisulfite.

Topics: Amino Acid Sequence; beta-Glucosidase; Eubacterium; Feces; Humans; Hydrolysis; Iridoids; Molecular Sequence Data; Molecular Weight; Pyrans

Geniposide, a main iridoid glucoside of Gardenia fruit, is transformed to genipin, a genuine choleretic, in vivo in rats (Aburada et al., J. Pharmacobio-Dyn., 1, 81 (1978)). As geniposide was not hydrolyzed to any metabolite by rat liver homogenate, which has beta-D-glucosidase and esterase activities, beta-D-glucosidases in intestinal bacteria seem to be required for an exhibition of its choleretic action. The crude extract of Eubacterium sp. A-44, a human intestinal anaerobe, hydrolyzed geniposide, but that of Ruminococcus sp. PO1-3, another human anaerobe, did not, though both extracts had beta-D-glucosidase activities for p-nitrophenyl beta-D-glucopyranoside. Only one of three beta-D-glucosidases from E. sp. A-44 and none of two from R. sp. PO1-3 hydrolyzed geniposide to genipin. However, carboxylesterases from E. sp. A-44 and pig liver were unable to hydrolyze geniposide to geniposidic acid, but hydrolyzed genipin to an aglycone of geniposidic acid, indicating that geniposide is first hydrolyzed to genipin by beta-D-glucosidases and subsequently to the aglycone of geniposidic acid by esterases. Thus, when geniposide is orally administered, genipin seems to be effectively produced in the intestine and then absorbed to act as a genuine choleretic.

Topics: Animals; Bacteria; beta-Glucosidase; Chromatography, Thin Layer; Esterases; Eubacterium; Feces; Humans; In Vitro Techniques; Intestines; Iridoids; Liver; Male; Peptococcaceae; Pyrans; Rats; Rats, Wistar; Subcellular Fractions

The paper reports the content determination of geniposide in Fructus Gardeniae and its different processed products by HPLC. The method is simple, accurate and reproducible, with an average recovery of 101.97%, RSD 1.01%.

Topics: Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Hot Temperature; Iridoids; Pyrans; Technology, Pharmaceutical

Four iridoid constituents: geniposide, gardenoside, geniposidic acid and genipin-1-beta-gentiobioside, have been separated by using an ODS (7 microns) column with gradient elution. The iridoid contents of the crude drug were quantified by peak height ratio. Thirty-one specimens from various sources were analyzed.

Topics: Drugs, Chinese Herbal; Glucosides; Iridoid Glucosides; Iridoids; Pyrans

Six oligosaccharides (I-VI) and six iridoid glucosides (VII-XII) isolated from the ethanolic extract of Lantana camara roots were identified as stachyose (I), verbascose (II), ajugose (III), verbascotetracose (IV), alpha-D-galac-(1-[-6)-alpha-D-galac(-1](3)-6-D-gluc(V ) , alpha-D-galac-(1-6)-alpha-D-galac(-1]-(4)6-D-)gluc(VI) , theveside (VII), 8-epiloganin (VIII), shanzhsid methyl ester (IX), theviridoside (X), lamiridoside (XI) and geniposide (XII), on the basis of spectral analysis (1H-NMR, 13C-NMR, FD-MS, GC-MS), physico-chemical constants and preparation of derivatives. V and VI were new compounds named lantanose A and lantanose B, respectively. The others were isolated from this plant for the first time.

Topics: Drugs, Chinese Herbal; Iridoids; Molecular Structure; Oligosaccharides; Pyrans; Trisaccharides

We have previously demonstrated that geniposide (GP) inhibits the aflatoxin B1 (AFB1) induced-hepatotoxicity and hepatic DNA binding in rats. To address the mechanism of action, the effects of GP on AFB1-induced DNA repair synthesis and AFB1 biotransformation in cultured rat hepatocytes were investigated. By evaluation of unscheduled DNA synthesis (UDS), GP reduced AFB1-induced DNA repair synthesis in a dose-dependent manner in hepatocyte cultures. GP elevates the metabolism of AFB1 and decreases the formation of AFM1. The enzyme activities of glutathione S-transferase (GST) and GSH-peroxidase (GSH-Px) in AFB1-treated hepatocyte cultures are enhanced in the presence of GP. GP reduces AFB1-induced DNA repair synthesis through an increased AFB1 detoxication metabolism. It provides one possible mechanism for the chemopreventive activity of GP.

Topics: Aflatoxin B1; Animals; Cells, Cultured; DNA Repair; DNA Replication; Dose-Response Relationship, Drug; Female; Iridoids; Kinetics; Liver; Pyrans; Rats; Rats, Inbred Strains

The effects of geniposide pretreatment on both hepatic aflatoxin B1 (AFB1)-DNA binding and AFB1 hepatotoxicity in rats has been examined. For these studies, male Sprague-Dawley rats were treated with AFB1 (2 mg/kg) by i.p. administration, and the different degrees of hepatic damage were revealed by the elevations of levels of serum marker enzymes such as aspartate aminotransferase (AST), alanine amino-transferase (ALT) and gamma-glutamyltranspeptidase (gamma-GT). After pretreatment of animals with geniposide (10 mg/kg) daily for 3 consecutive days, the enzyme elevations were significantly suppressed. This suggested that the geniposide possessed chemopreventive effects on the early acute hepatic damage induced by AFB1. Under these experimental conditions, consistent elevation of the activities of glutathione S-transferase (GST) and gamma-glutamylcysteine synthetase but not glutathione peroxidase (GSH-Px) and gamma-glutamyltranspeptidase were observed. Treatment of rats with geniposide significantly lowered hepatic GSH and GSSG levels, but the ratio of GSH to GSSG was not changed. Geniposide treatment also decreased AFB1-DNA adduct formation in AFB1-treated animals. From these results, we suggest that the protective effect of geniposide on AFB1 hepatotoxicity in rats might be due to the hepatic tissues' defense mechanisms that involve the enhanced GST activity for AFB1 detoxication and induction gamma-glutamylcysteine synthetase for GSH biosynthesis.

Topics: Aflatoxin B1; Alanine Transaminase; Animals; Aspartate Aminotransferases; DNA; Dose-Response Relationship, Drug; Drug Antagonism; gamma-Glutamyltransferase; Glutamate-Cysteine Ligase; Glutathione; Glutathione Peroxidase; Iridoids; Liver; Male; Organ Size; Pyrans; Rats; Rats, Inbred Strains

This paper reports the study of six fractions and one chemical constituent isolated from the traditional Chinese herb Gardenia jasminoides. The results showed that two fractions (G5.G6,), alcohol extract (G1) and genipoiside(A) had obvious anti-inflammatory effects and were comparatively effective in treating soft tissue injuries in animals.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Buttocks; Drugs, Chinese Herbal; Iridoids; Male; Mice; Pyrans; Rabbits; Rats; Wounds and Injuries

During the course of our studies on the metabolism of iridoid glycosides by human intestinal bacteria, we found that geniposide (1) and gardenoside (4) were transformed to new nitrogen-containing compounds, genipinine (3) and gardenine (6), respectively, along with the known aglycones. Although the amounts of new metabolites were somewhat lower than those of the aglycones, they were quantitatively analyzed by means of liquid chromatography/mass spectrometry (LC/MS). Of 25 strains of human intestinal bacteria, Peptostreptococcus anaerobius, Klebsiella pneumoniae, Fusobacterium nucleatum, and Bacteroides fragilis ssp. thetaotus produced appreciable amounts of 3, while a bacterial mixture of human feces produced 10 times or more higher amounts of 3, as compared to the individual strains.

Topics: Bacteria; Biotransformation; Bridged Bicyclo Compounds; Feces; Humans; Intestines; Iridoids; Male; Pyrans; Pyridines

The hepatotoxic effects of geniposide were investigated in rats. Increases in serum alanine aminotransferase and aspartate aminotransferase activities as a result of oral administration of 320 mg geniposide/kg body weight were suppressed when geniposide was administered ip or when the rats were pretreated with chloramphenicol. The non-protein sulphydryl content of the liver 4 hr after oral administration of geniposide decrease in a dose-dependent manner. Genipin, the aglycone of geniposide, had a marked reactivity with sulphydryl groups of glutathione and cysteine in vitro. The hepatotoxic effects of ip administration of genipin at a dose of 80 mg/kg body weight were comparable with those of oral administration of geniposide at a dose of 320 mg/kg. Buthionine sulphoximine pretreatment enhanced the toxicity of geniposide, while cysteine pretreatment completely suppressed it. These results suggest that the conversion of geniposide to genipin is causally related to the hepatotoxicity of geniposide and that hepatic non-protein sulphydryls are important in modulating the toxicity.

Topics: Administration, Oral; Alanine Transaminase; Animals; Aspartate Aminotransferases; Bilirubin; Buthionine Sulfoximine; Chloramphenicol; Cysteine; Injections, Intraperitoneal; Iridoid Glycosides; Iridoids; Liver; Male; Methionine Sulfoximine; Molecular Structure; Pyrans; Rats; Rats, Inbred Strains; Sulfhydryl Compounds

A method for the determination of geniposide in Fructus Gardeniae was established by HPLC. HPLC conditions: column: TSK GEL LS-410, mobile phase: 0.05 mol/L Na2 HPO4-MeOH (80:20) (pH = 6.86). Quantitative determination was made using peak area. Samples of various parts of the plant and of different habitats together with their pharmaceutical preparations were analyzed.

Topics: Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Iridoids; Pyrans

Acute toxicity by gardenia yellow color was studied in rats. Oral administration of the colorant at doses of 800 mg/kg up to 5000 mg/kg caused diarrhea and increases in serum alanine aminotransferase and aspartate aminotransferase activities in a dose-dependent manner. After 24 h of oral treatment with 2000 mg/kg of the colorant, liver showed partially hemorrhagic changes and the intestinal tract, especially the duodenum, appeared blue. The toxicity induced by the colorant was stronger by oral administration than by intraperitoneal administration. The content of geniposide, an iridoid compound, was estimated to be 28% of the colorant, and this iridoid accounted for almost all the hepatotoxic activity of the colorant.

Topics: Administration, Oral; Alanine Transaminase; Animals; Aspartate Aminotransferases; Coloring Agents; Diarrhea; Gardenia; Injections, Intraperitoneal; Iridoids; Liver; Male; Plant Extracts; Pyrans; Rats; Rats, Inbred Strains

Topics: Chromatography, Thin Layer; Drugs, Chinese Herbal; Fruit; Hot Temperature; Iridoids; Pyrans

Topics: Administration, Oral; Animals; Biotransformation; Intestinal Mucosa; Iridoids; Pyrans; Rats

Topics: Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Glucosides; Glycosides; Iridoid Glucosides; Iridoids; Pyrans

The intravenous administration of 50 and 100 mg/kg of genipin (GP), gardenogenins (GAR-G), deacetylasperulosidic acid methylester genins (DAM-G), and scandoside methylester genin (SSM-G) exhibited the bile acid-independent choleretic actions. The action of DAM-G was stronger than the actions of other compounds tested. The choleretic action of SSM-G was milder, but longer lasting than those of GAR-G and DAM-G.

Topics: Animals; Bile; Bile Acids and Salts; Cholagogues and Choleretics; Electrolytes; Glucosides; Glycosides; Iridoids; Male; Pyrans; Rats; Rats, Inbred Strains; Time Factors

Topics: 1-Naphthylisothiocyanate; Alanine Transaminase; Animals; Aspartate Aminotransferases; Bilirubin; Iridoids; Jaundice; Male; Plants, Medicinal; Pyrans; Rats; Thiocyanates

Topics: 1-Naphthylisothiocyanate; Animals; Cholestasis, Intrahepatic; Iridoids; Male; Medicine, Chinese Traditional; Medicine, East Asian Traditional; Plant Extracts; Plants, Medicinal; Pyrans; Rats; Rats, Inbred Strains

Topics: Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Dosage Forms; Iridoids; Medicine, Chinese Traditional; Medicine, East Asian Traditional; Plants, Medicinal; Pyrans

Topics: Animals; Dietary Carbohydrates; Iridoids; Male; Metabolism; Plants; Pyrans; Rats; Rats, Inbred Strains

Topics: Carotenoids; Glucosides; Glycosides; Iridoids; Plants, Medicinal; Pyrans