morphinans and sinomenine

Autoimmune diseases (ADs), with significant effects on morbidity and mortality, are a broad spectrum of disorders featured by body's immune responses being directed against its own tissues, resulting in chronic inflammation and tissue damage. Sinomenine (SIN) is an alkaloid isolated from the root and stem of Sinomenium acutum which is mainly used to treat pain, inflammation and immune disorders for centuries in China. Its potential anti-inflammatory role for treating immune-related disorders in experimental animal models and in some clinical applications have been reported widely, suggesting an inspiring application prospect of SIN. In this review, the pharmacokinetics, drug delivery systems, pharmacological mechanisms of action underlying the anti-inflammatory and immunomodulatory effects of SIN, and the possibility of SIN as adjuvant to disease-modifying anti-rheumatic drugs (DMARDs) therapy were summarized and evaluated. This paper aims to reveal the potential prospects and limitations of SIN in the treatment of inflammatory and immune diseases, and to provide ideas for compensating its limitations and reducing the side effects, and thus to make SIN better translate to the clinic.

Topics: Animals; Anti-Inflammatory Agents; Immunity; Inflammation; Morphinans

Sinomenine (SIN) is the main pharmacologically active component of Sinomenii Caulis and protects against rheumatoid arthritis (RA). In recent years, many studies have been conducted to elucidate the pharmacological mechanisms of SIN in the treatment of RA. However, the molecular mechanism of SIN in RA has not been fully elucidated.. To summarize the pharmacological effects and molecular mechanisms of SIN in RA and clarify the most valuable regulatory mechanisms of SIN to provide clues and a basis for basic research and clinical applications.. We systematically searched SciFinder, Web of Science, PubMed, China National Knowledge Internet (CNKI), the Wanfang Databases, and the Chinese Scientific Journal Database (VIP). We organized our work based on the PRISMA statement and selected studies for review based on predefined selection criteria.. After screening, we identified 201 relevant studies, including 88 clinical trials and 113 in vivo and in vitro studies on molecular mechanisms. Among these studies, we selected key results for reporting and analysis.. We found that most of the known pharmacological mechanisms of SIN are indirect effects on certain signaling pathways or proteins. SIN was manifested to reduce the release of inflammatory cytokines such as Tumor necrosis factor-α (TNF-α), Interleukin-6 (IL-6), and IL-1β, thereby reducing the inflammatory response, and apparently blocking the destruction of bone and cartilage. The regulatory effects on inflammation and bone destruction make SIN a promising drug to treat RA. More notably, we believe that the modulation of α7nAChR and the regulation of methylation levels at specific GCG sites in the mPGES-1 promoter by SIN, and its mechanism of directly targeting GBP5, certainly enriches the possibilities and the underlying rationale for SIN in the treatment of inflammatory immune-related diseases.

Topics: Anti-Inflammatory Agents; Arthritis, Rheumatoid; Humans; Morphinans; Signal Transduction

Sinomenine is the main component of the vine

Topics: Arthritis, Rheumatoid; Humans; Medicine, Chinese Traditional; Morphinans; Neuralgia

To systemically evaluate the efficacy and safety of sinomenine combined with methotrexate(SIN+MTX) in the treatment of rheumatoid arthritis(RA). Literature databases of Wanfang, CNKI, VIP, SinoMed, PubMed, Cochrane Library and Web of Science were retrieved comprehensively for relevant clinical trials. The literature retrieval time was from database establishment to February 4, 2020. The quality of literatures was assessed by the Cochrane Evaluation Handbook 5.1.0, and qualified literature was reviewed and analyzed by using the RevMan 5.3 statistical software. Twenty randomized controlled trials met the inclusion criteria, and were enrolled in the Meta-analysis. The results showed that SIN+MTX remarkably reduced DAS28(MD=-0.85, 95%CI[-1.03,-0.67], P<0.000 01), and improved total efficiency(P<0.000 01). SIN+MTX could inhibit swollen joint count(MD=-1.19, 95%CI[-1.75,-0.63], P<0.000 1), tender joint count(MD=-1.58, 95%CI[-2.89,-0.28], P=0.02) and reduce morning stiffness time(MD=-8.44, 95%CI[-11.82,-5.07], P<0.000 01) compared with control group. The results showed that SIN+MTX was equal to control group in grip strength(SMD=0.20,95%CI[-1.11,1.51],P=0.77). SIN+MTX remarkably alleviated the erythrocyte sedimentation rate(MD=-9.87, 95%CI[-14.52,-5.22], P<0.000 1), C-reactive protein(SMD=-0.30, 95%CI[-0.51,-0.09], P=0.005), and rheumatoid factor(MD=-11.23,95%CI[-13.81,-8.65],P<0.000 01). The frequency of adverse reactions were reduced compared with that in the control group(P<0.000 01). Current clinical studies demonstrate that the efficacy and safety of SIN+MTX in the treatment of RA were superior to control group. However, due to the low quality and quantity of the included studies, high-quality randomized controlled trials are necessary to support the clinical evidences.

Topics: Antirheumatic Agents; Arthritis, Rheumatoid; Drugs, Chinese Herbal; Humans; Methotrexate; Morphinans

Purified from the roots of the plant

Topics: Analgesics; Animals; Chronic Pain; Humans; Morphinans; Neuroimmunomodulation

Sinomenine (SIN) is a bioactive alkaloid compound extracted from a Chinese medicinal plant Sinomenium acutum. It is a multitarget antitumor natural substance. Various mechanisms have been proposed for the antitumor effects of SIN, such as direct cytotoxicity, induction of apoptosis, sensitization attenuating radiotherapy and chemotherapy, reversal of drug resistance, resistance to distant metastasis, and antiangiogenesis. SIN can be used as a tumor cell killer and an adjuvant to radiotherapy and chemotherapy. However, recent studies are mostly limited to the basic experimental stage; no systematic clinical studies have yet been reported. Therefore, this paper aimed to review the mechanism underlying the antitumor effects of SIN by consulting relevant domestic and foreign studies and to provide a relevant reference for further development, use, and exploration of SIN.

Topics: Alkaloids; Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Chemotherapy, Adjuvant; Humans; Morphinans; Neoplasms; Plants, Medicinal; Radiotherapy, Adjuvant; Signal Transduction

Sinomenine is one of the most widely known alkaloids owing to different therapeutic properties including anti-inflammatory and immunosuppressive activities, as well as the potency in neurodegenerative disorders, such as Alzheimer's disease and Parkinson's disease. Sinomenine has been studied as a potential anti-inflammatory agent through appropriate structural modifications. Recently, a number of such approaches have been attempted in various laboratories with great success. Several series of derivatives have been synthesized with changes at rings A, B, C and D. Herein, we present an up to date review of modifications and bioactivities of important modified derivatives. These studies offer interesting knowledge on the bioactivity and structural specificity of sinomenine, providing better understanding of the structure-activity relationships (SAR) between designing and development of sinomenine derivative with better therapeutic and lower side effects.

Topics: Animals; Anti-Inflammatory Agents; Humans; Immunosuppressive Agents; Morphinans; Neurodegenerative Diseases

Cardio/cerebral-vascular diseases seriously threaten human health and are the leading cause of death. As such, there is great interest in identifying a potential mechanism that controls the development process of cardio/cerebral vascular diseases. Present studies demonstrate that inflammasomes play an important role in the process of ischemic cardio/cerebral vascular diseases (ICCVDs). Among the pathological process of ICCVDs, inflammasomes activated the sterile inflammatory response that accelerated the development of diseases and aggravated the acute lesion of tissue. As the most thoroughly studied inflammasome, the NLRP3 inflammasome has been proven to be a potential therapeutic target for ICCVDs. In this review, we summarized the mechanisms of Chinese herbal medicine which can affect ICCVDs via the regulation of the NLRP3 inflammasome. Our study discovers that active compounds of Chinese medicines have a negative effect on NLRP3 in different ICCVDs models. Astragaloside IV may influence the receptor of the cell membrane to inhibit NLRP3 activation. Resveratrol, colchicinesis, salvianolic acid B, chrysophanol and sulforaphane may directly damage the formation of NLRP3 by inhibiting ASC or Caspase-1. Most of the active natural compounds can negatively regulate the downstream products of NLRP3 inflammasome such as IL-18 and IL1

Topics: Cardiovascular Diseases; Cerebrovascular Disorders; Drugs, Chinese Herbal; Humans; Inflammasomes; Molecular Targeted Therapy; Morphinans; NLR Family, Pyrin Domain-Containing 3 Protein; Resveratrol; Spirostans

Sinomenine (SIN) is widely used in China to treat a variety of rheumatic diseases (RA), and has various pharmacological effects such as anti-inflammatory, analgesic, and anti-tumor effects. However, due to the histamine release characteristics of SIN, its adverse reactions such as allergic reactions, gastrointestinal reactions, and circulatory systemic reactions have been drawing increasing attention. We present here a systematic review of the chemical structure, pharmacological effects, clinical application, and adverse reactions of SIN, a detailed discussion on the relationship between histamine/histamine receptor and mechanism of action of SIN. In addition, we simulated the binding of SIN to four histamine receptors by using a virtual molecular docking method and found that the bonding intensity between SIN and receptors varied in the order shown as follows: H1R > H2R ~ H3R > H4R. The docking results suggested that SIN might exhibit dual regulatory effects in many processes such as cyclooxygenase-2 (COX-2) expression, NF-κB pathway activation, and degranulation of mast cells to release histamine, thereby exhibiting pro-inflammatory (adverse reactions)/anti-inflammatory effects. This study provides a theoretical basis for the clinical treatment of inflammations seen such as in RA using SIN, and also suggests that SIN has great potential in the field of cancer treatment and will have very important social and economic significance.

Topics: Cyclooxygenase 2; Hematologic Diseases; Histamine Release; Humans; Immunosuppressive Agents; Molecular Docking Simulation; Morphinans; Protein Binding; Receptors, Histamine; Rheumatic Diseases

To systematically evaluate the curative clinical efficacy and safety of sinomenine (SIN) in\ treatment of rheumatoid arthritis (RA) in comparison to methotrexate (MTX).. We searched the China National Knowledge Infrastructure Database, Chinese Biomedical\ Literature Database, China Science and Technology Journal Database, Wanfang Database, Pubmed and\ Cochrane Library electronically up to August 31, 2015, without language limitation. Only randomized\ controlled trials (RCTs) were included. Software Review Manager 5.3 was used for Meta-analysis.. A total of 16 eligible studies within 1500 RA patients were included. The meta-analysis indicated\ that on basis of MTX, SIN was more effective in total effective rate (P < 0.000 01). Besides, SIN\ alone versus MTX also showed advantages in RA therapy (P = 0.04) Taken together, adverse events\ occurred less frequently in combination of SIN and MTX than MTX alone (P < 0.0001), especially in digestive\ system (P < 0.000 01),while occurred more in dermato mucosal system with SIN treatment versus\ MTX (P = 0.02), and were similar for both remedies in nervous system (P = 0.12) and hematological\ system (P = 0.25).. Compared to MTX, SIN had better clinical efficacy and relatively fewer adverse events\ in treatment of RA, especially when it was used together with MTX. Due to the poor methodological\ quality, well-designed, multiple-center RCTs are still required to further confirm the findings.

Topics: Antirheumatic Agents; Arthritis, Rheumatoid; China; Drugs, Chinese Herbal; Humans; Methotrexate; Morphinans; Randomized Controlled Trials as Topic; Treatment Outcome

The current review emphasizes on the herbal bioenhancers which themselves do not possess inherent pharmacological activity of their own but when co-administered with Active Pharmaceutical Ingredients (API), enhances their bioavailability and efficacy. Herbal bioenhancers play a crucial role in enhancing the bioavailability and bioefficacy of different classes of drugs, such as antihypertensives, anticancer, antiviral, antitubercular and antifungal drugs at low doses. This paper highlights various natural compounds that can be utilized as an efficient bioenhancer. Several herbal compounds including piperine, quercetin, genistein, naringin, sinomenine, curcumin, and glycyrrhizin have demonstrated capability to improve the pharmacokinetic parameters of several potent API. This article also focuses on various United States patents on herbal bioenhancers, which has proved to be beneficial in improving oral absorption of nutraceuticals like vitamins, minerals, amino acids and certain herbal compounds. The present paper also describes proposed mechanism of action, which mainly includes absorption process, drug metabolism, and action on drug target. The herbal bioenhancers are easily available, safe, free from side effects, minimizes drug toxicity, shortens the duration of treatment, lowers the drug resistance problems and minimizes the cost of treatment. Inspite of the fact that herbal bioenhancers provide an innovative concept for enhancing the bioavailability of several potent drugs, there are numerous bioenhancers of herbal origin that are yet to be explored in several vital areas. These bioenhancers must also be implied to enhance the bioavailability and bioefficacy through routes other than the oral route of drug delivery. There is a vast array of unexploited plants which can be investigated for their drug bioenhancing potency. The toxicity profiles of these herbal bioenhancers must not be overlooked. Researches must be carried out to solve these issues and to deliver a safe and effective dose of drugs to attain desired pharmacological response.

Topics: Adjuvants, Pharmaceutic; Alkaloids; Benzodioxoles; Biological Availability; Carum; Cuminum; Curcumin; Ergolines; Flavanones; Genistein; Glycyrrhizic Acid; Humans; Morphinans; Piperidines; Plant Extracts; Polyunsaturated Alkamides; Quercetin; Zingiber officinale

Sinomenium acutum (Thumb.) Rehd. et Wils. (Menispermaceae, SA) has been used as a traditional Chinese medicine in the treatment of various diseases for hundreds of years; it possesses favorable effects against autoimmune diseases, especially rheumatoid arthritis (RA). A great number of investigations have been done on SA in the last decade, but they are usually scattered across various publications.. The purpose of this article is to summarize and review the published scientific information about the chemical constituents, pharmacological effects, pharmacokinetics, and clinic applications of this plant since 2000.. The information for 89 cases included in this review was compiled. The SA contains alkaloids, sterols, phospholipids, and some other components. A great deal of pharmacological and clinic research has been done on sinomenine, a main compound from SA, which mainly focuses on the immune system, cardiovascular system, and nervous system.. Previous studies strongly support its potential as an effective adaptogenic herbal remedy. There is no doubt that SA is being widely used now and will have extraordinary potential for the future.

Topics: Alkaloids; Animals; Anti-Inflammatory Agents; Antirheumatic Agents; Cardiovascular Agents; Drugs, Chinese Herbal; Ethnopharmacology; Humans; Morphinans; Neurotransmitter Agents; Sinomenium

Osteoarthritis (OA) is a common chronic progressing disease in the elderly. In current there are many methods of drug treatment for knee osteoarthritis, the intra-articular injection belongs to local application, can direct effect on arthritis, is the most used and most effective methods in osteoarthritis treatment. A single medication may exist that anti-inflammatory effects produce a marked effect but articular cartilage injury is induced, or articular cartilage repairing are promoted but anti-inflammatory effects is not ideal. Therefore,the best method of treatment should be a comprehensive analysis of articular structure, function and possible pathogenic factor to understand the expectations of level of recovery and use in patients to design an individualized treatment plan.

Topics: Glucocorticoids; Humans; Injections, Intra-Articular; Morphinans; Osteoarthritis, Knee

Sinomenine (SN), a pure compound extracted from the Sinomenium acutum plant, has been found to inhibit T- and B-lymphocyte activation, proliferation and function and to interfere with the differentiation, recruitment and function of several other cell types, such as dendritic cells (DC). SN has demonstrated its potential anti-inflammatory role for treating immune-related disorders in experimental animal models and in some clinical applications. This review will summarize its potential effects, mechanisms and applications.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Humans; Immunosuppressive Agents; Lymphocyte Activation; Lymphocytes; Morphinans; Sinomenium

The clinical or experimental study proves that Chinese medicine such as Tripteryglum wilfordii, Lignum Sappan, Caulis Sinomenii, Radix Trichosanthis and Herba Artemisiae Annuae have good immunosuppressive activity. Further researches on the immunosuppressive active components from Chinese medicine have been the main direction in recent years. The recent researches on immunosuppressive effect and possible mechanisms for the monomers such as triperine, triptolide, bazilein, potosappanin A, sinomenine, trichosanthin and artemisinin extracted from those Chinese medicine are introduced in this review.

Topics: Animals; Artemisinins; Diterpenes; Drugs, Chinese Herbal; Epoxy Compounds; Humans; Immunosuppressive Agents; Morphinans; Phenanthrenes; Plants, Medicinal; Trichosanthin

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Rheumatoid; Humans; Interleukin-1beta; Morphinans; Phytotherapy; Tumor Necrosis Factor-alpha

Sinomenine (SIN), an alkaloid isolated from CAULIS SINOMENII, has been used in the treatment of rheumatoid arthritis (RA) clinically. This study aimed to systematically evaluate the efficacy and safety of SIN by a comparison between SIN and non-steroidal anti-inflammatory drugs (NSAIDs). Forty-three electronic databases were systematically searched. The quality of eligible trials was assessed by Jadad's scale. Revman 5.0 software was used for data syntheses and meta-analyses. The results showed that (i) of the 121 potential studies identified, 10 clinical trials involving 1185 patients met the inclusion criteria; (ii) improved patients and rheumatoid factor disappearance patients after SIN treatments were significantly more than those treated by NSAIDs ( P < 0.00001 and P = 0.008); (iii) compared with NSAIDs, SIN was more effective in amelioration of morning stiffness ( P < 0.00001), painful joints ( P = 0.03), and erythrocyte sedimentation rate ( P < 0.00001), but there was no significant difference between the two remedies in the treatment of swollen joints, grip strength, and C-reactive protein ( P > 0.05); and (iv) adverse events occurred less frequently in the digestive system during SIN treatment than during NSAID treatment ( P = 0.0003) but occurred more frequently in the dermatomucosal system with SIN treatment ( P = 0.03), while adverse events of the nervous system were similar for both treatments ( P = 0.31). In conclusion, SIN may be a valuable remedy to treat RA clinically, although current evidence needs to be further verified by more high-quality trials.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Rheumatoid; Controlled Clinical Trials as Topic; Databases, Factual; Humans; Morphinans; Randomized Controlled Trials as Topic; Software; Treatment Outcome

To further understand sinomenine, this paper has introduced the abstract technology, assaying, pharmaceutical chemistry, pharmacological action, pharmacotoxicology, pharmacokinetics and clinical application of sinomenine based on the important and significant contents of reference which have been consulted in the past ten years. Sinomenine is a kind of non-steroidal anti-inflammatory drugs with very effective and little side effect and expected it as a good new drug withdrawal medicine in the future.

Topics: Animals; Anti-Arrhythmia Agents; Anti-Inflammatory Agents, Non-Steroidal; Antirheumatic Agents; Humans; Morphinans; Plant Roots; Plants, Medicinal; Sinomenium; Technology, Pharmaceutical

This article reviewed the progress in the study of the pharmacognosy, chemical compositions, pharmacological actions and clinical practices of Sinomenium acutum (Thunb.) Rehd. et Wils. An expectation for the further development and utilization of this plant was put forward.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Rheumatoid; Drugs, Chinese Herbal; Humans; Morphinans; Pharmacognosy; Phytotherapy; Plant Stems; Plants, Medicinal; Sinomenium

Topics: Administration, Cutaneous; Animals; Drugs, Chinese Herbal; Lactates; Morphinans; Plants, Medicinal; Pyrazines; Skin Absorption

The root and stem decoctions of Sinomenium acutum Rehd. et Wils. (formerly Sinomenium diversifolius Diels, one type of Fang-chi (Chinese)) have been used as a folk remedy for neuralgia and rheumatoid arthritis in many areas of the Far East. In Japan and China various viny plants have been identified as Fang-chi (Boi in Japanese) since antiquity. This uncertain nomenclature has made it difficult to evaluate the efficacy of the Fang-chi described in the classic literature. Among traditional Fang-chi plants only Sinomeniumacutum has been demonstrated to contain the alkaloid sinomenine, which is now known to be effective in neuralgia and rheumatic diseases. Sinomenine is a unique plant alkaloid, as it potently releases histamine in association with degranulation of tissue mast cells in mammalian tissues. This action occurs preferentially in the skin and joint capsules. The released histamine is responsible for the dominant pharmacological actions of sinomenine, such as vasodilatation, increased vascular permeability, acceleration of the thoracic and peripheral lymph flow, contraction of plain muscles, increased peristalsis of the intestines, and stimulation of gastric acid secretion. At toxic doses of sinomenine, convulsive central excitation was observed in most laboratory animals. Clinical side effects encountered with high doses of injected sinomenine or of decocted Sinomenium acutum were: injection site flare, pruritus in the head and upper part of the body, edema around the lips and eyelids, and temporary cephalalgia. Most of these side effects were reduced by classical antihistamines (H1-receptor antagonists). Daily subcutaneous injections of sinomenine for more than one week produced an analgesic effect in mice. Granulation tissue growth and adjuvant arthritis induced in rats were both inhibited by daily injections of a small dose of sinomenine hydrochloride or histamine dihydrochloride. These inhibitory effects were mediated through histamine H2-receptors probably on fibroblasts (for granulation tissue growth) and on T-cells (for adjuvant arthritis), since these effects were clearly inhibited by the H2-antagonist burimamide but not by the H1-antagonist mepyramine. The anti-rheumatic effect on Sinomenium acutum are probably genuine and can probably be attributed to the histamine-releasing properties of sinomenine.

Topics: Alkaloids; Analgesics; Animals; Burimamide; Cardiovascular System; Chemical Phenomena; Chemistry; Connective Tissue; Dogs; Haplorhini; Histamine; Histamine Release; Humans; Japan; Methyl Ethers; Mice; Morphinans; Plants, Medicinal; Pyrilamine; Rabbits; Rats; Rheumatic Diseases

Diabetic nephropathy (DN) is one of the main complications of diabetes. It is closely associated with the dysfunction of glomerular endothelial cells (GECs) under hyperglycemia. Severe inflammation is an important inducer for the development of GECs dysfunction, and it contributes to the disruption of tight junctions in GECs and the increased endothelial permeability. Sinomenine, an alkaloid monomer extracted from the rhizome of Sinomenium acutum, is recognized for its multiple pharmacological functions, including an anti-DN property. The present study aimed to explore the potential functional mechanism of Sinomenine against DN. Animals were randomly divided into Sham, DN, DN + Sinomenine (20 mg/kg), and DN + Sinomenine (40 mg/kg) groups. The Sinomenine or vehicle was administered every day for 6 weeks, followed by collecting renal tissues for further detection. Increased body weights, elevated blood glucose levels and UAE values, aggravated renal tissue pathology, higher concentrations of IL-18 and IL-1β in renal tissues, and reduced claudin-5 expression were observed in DN rats. However, the administration of Sinomenine significantly alleviated all these DN-related changes. Furthermore, human renal glomerular endothelial cells (HrGECs) were treated with high glucose (HG, 30 mM) with or without Sinomenine (50, 100 μM) for 24 h. We found that Sinomenine treatment ameliorated the elevated production of IL-18 and IL-1β, increased fluorescence intensity of FITC-dextran, declined trans-endothelial electrical resistance (TEER) value, and reduction of claudin-5 and C/EBP-α in HG-treated HrGECs. Moreover, the regulatory effect of Sinomenine on endothelial monolayer permeability in HG-treated HrGECs was abolished by the knockdown of C/EBP-α, indicating C/EBP-α is required for the effect of Sinomenine. We concluded that Sinomenine alleviated diabetic nephropathy-induced renal glomerular endothelial dysfunction via activating the C/EBP-α/claudin-5 axis.

Topics: Animals; CCAAT-Enhancer-Binding Protein-alpha; Claudin-5; Diabetic Nephropathies; Endothelial Cells; Interleukin-18; Morphinans; Permeability; Rats; Signal Transduction

The aim of the research was to investigate the anti-endotoxin and anti-inflammatory effects of Sinomenine, an agent commonly found in Chinese herbal medicines. Endotoxin (i.e., 1 mg lipopolysaccharide (LPS)/kg)) was administered via intraperitoneal (IP) injection to piglets in high-, middle-, and low-dose sinomenine groups. Piglets were then treated with 1, 5 or 10 mg/kg sinomenine, intramuscularly (i.m.), 3 hr after LPS. Vehicle was administered, as above, to drug control group piglets followed 3 hr later by 10 mg/kg sinomenine i.m.. LPS control group piglets were challenged with 1 mg/kg LPS IP, followed by vehicle i.m., and naïve control piglets were treated with normal saline IP, followed by normal saline i.m., as above. Temperatures were measured, and blood samples were collected from the precaval veins of piglets at 12, 24, and 48 hr post-LPS or vehicle injection. Clinical signs were recorded, and index levels were analyzed via ELISA. Sinomenine was found to reduce the incidence and severity of LPS-induced toxicities, including body temperature elevation, cell adhesion, and systemic inflammation. These data suggest that sinomenine may be effective for regulating inflammatory responses and has the potential for use as an anti-endotoxin therapy.

Topics: Acute-Phase Proteins; Animals; Carrier Proteins; Dose-Response Relationship, Drug; Gene Expression Regulation; L-Selectin; Lipopolysaccharides; Macrophage-1 Antigen; Male; Membrane Glycoproteins; Morphinans; Swine

A combination of conventional disease-modifying anti-rheumatic drugs improves the treatment of rheumatoid arthritis but with high side-effects. Methotrexate (MTX) combination therapy that with high therapeutic efficacy and low toxicity is in demand in many countries to replace the use of expensive biological agents.. This study was an open-label, 24-week, parallel randomized controlled trial conducted between November 2015 and December 2017.. Patients were randomly assigned at a 3:2 ratio to receive MTX combined with sinomenine (SIN) at a dose of 120 mg twice daily, or leflunomide (LEF) at a dose of 20 mg once daily. Efficacy and safety were assessed at weeks 4, 12 and 24. The primary efficacy endpoint was the proportion of patients achieving an American College of Rheumatology (ACR)50 response and a European League Against Rheumatism (EULAR) good response at week 24.. A total of 101/120 (84.2%) patients completed 24 weeks of observation. In the intention-to-treat (ITT) analysis, 65.3% of patients treated with MTX + SIN showed improved disease activity as determined by the ACR50 response at week 24 compared to 69.6% of patients treated with MTX + LEF. A similar insignificant pattern was found for the ACR20 and ACR70 responses, as well as the clinical disease activity index, EULAR response, and remission and low disease activity rates between these two treatment groups. The per-protocol analysis showed results consistent with those of the ITT analysis. Notably, significant reductions in gastrointestinal adverse reactions and liver toxicity were found in patients treated with MTX + SIN compared to patients treated with MTX + LEF (p < 0.05).. Considering the balance of efficacy and toxicity, the current study provides evidence that MTX + SIN combination therapy is probably one of the choices for treating patients with active rheumatoid arthritis in addition to MTX + LEF combination therapy.

Topics: Adult; Antirheumatic Agents; Arthritis, Rheumatoid; Chemical and Drug Induced Liver Injury; Drug Therapy, Combination; Female; Gastrointestinal Diseases; Humans; Leflunomide; Male; Methotrexate; Middle Aged; Morphinans; Treatment Outcome

Chemotherapy induced nausea and vomiting (CINV) remains a major problem that seriously impairs the quality of life (QoL) in cancer patients receiving chemotherapy regimens. Complementary medicines, including homeopathy, are used by many patients with cancer, usually alongside with conventional treatment. A randomized, placebo-controlled Phase III study was conducted to evaluate the efficacy of a complex homeopathic medicine, Cocculine, in the control of CINV in non-metastatic breast cancer patients treated by standard chemotherapy regimens.. Chemotherapy-naïve patients with non-metastatic breast cancer scheduled to receive 6 cycles of chemotherapy including at least three initial cycles of FAC 50, FEC 100 or TAC were randomized to receive standard anti-emetic treatment plus either a complex homeopathic remedy (Cocculine, registered in France for treatment of nausea and travel sickness) or the matching placebo (NCT00409071 clinicaltrials.gov). The primary endpoint was nausea score measured after the 1st chemotherapy course using the FLIE questionnaire (Functional Living Index for Emesis) with 5-day recall. Secondary endpoints were: vomiting measured by the FLIE score, nausea and vomiting measured by patient self-evaluation (EVA) and investigator recording (NCI-CTC AE V3.0) and treatment compliance.. From September 2005 to January 2008, 431 patients were randomized: 214 to Cocculine (C) and 217 to placebo (P). Patient characteristics were well-balanced between the 2 arms. Overall, compliance to study treatments was excellent and similar between the 2 arms. A total of 205 patients (50.9%; 103 patients in the placebo and 102 in the homeopathy arms) had nausea FLIE scores > 6 indicative of no impact of nausea on quality of life during the 1st chemotherapy course. There was no difference between the 2 arms when primary endpoint analysis was performed by chemotherapy stratum; or in the subgroup of patients with susceptibility to nausea and vomiting before inclusion. In addition, nausea, vomiting and global emesis FLIE scores were not statistically different at any time between the two study arms. The frequencies of severe (Grade ≥ 2) nausea and vomiting were low in our study (nausea: P: 17.6% vs C: 15.7%, p=0.62; vomiting: P: 10.8% vs C: 12.0%, p=0.72 during the first course).. This double-blinded, placebo-controlled, randomised Phase III study showed that adding a complex homeopathic medicine (Cocculine) to standard anti-emetic prophylaxis does not improve the control of CINV in early breast cancer patients.

Topics: Adult; Aged; Antiemetics; Antineoplastic Agents; Breast Neoplasms; Double-Blind Method; Female; Humans; Middle Aged; Morphinans; Plant Extracts; Quality of Life; Vomiting; Young Adult

The rhizome of Chinese medical plant QingTeng (scientific name: Sinomenium acutum (Thunb.) Rehd. et Wils.) is widely used by traditional medical doctors for anti-inflammation and immunoregulatory in China and other Asian countries.. The purpose of this study was to evaluate the effects and possible mechanisms of sinomenine resistance against DSS-induced inflammation in vitro and in vivo.. The UC model was induced by treating female mice with 3% DSS in vivo and human colonic epithelial cells (Hcoepic) with 0.8 mg/ml DSS in vitro. The mice and Hcoepic were then treated with sinomenine. Inflammatory factors were detected using ELISA and qRT-PCR. Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 and 14-3-3θ were analyzed by bioinformatic analysis and verified by western blotting, immunofluorescent staining or immunohistochemistry.. DSS-induced Hcoepic underwent high inflammation and oxidative stress conditions, whereas sinomenine reduced the uncontrolled immune microenvironment by suppressing NF-κB signaling and targeting 14-3-3θ. Knockdown of 14-3-3θ decreased the protective effect of sinomenine against DSS-induced inflammation in vitro. Moreover, sinomenine promoted 14-3-3θ protein expression and inhibited NF-κB p65 signaling in DSS-induced mice.. These findings suggest that 14-3-3θ plays an important role in sinomenine against DSS treatment, and sinomenine could be considered a potential drug for the treatment of UC.

Topics: Animals; Colitis, Ulcerative; Dextran Sulfate; Disease Models, Animal; Humans; Inflammation; Mice; Mice, Inbred C57BL; Morphinans; NF-kappa B; Signal Transduction

Sinomenine (SIN) is a major component contained in extracts of the Chinese medicinal herb Sinomenium acutum. SIN has various pharmacological properties, including cytoprotection, immunosuppression and anti-inflammation effects. Furthermore, recent studies have reported that SIN has anti-tumor and antidepressant effects, which has created a strong need for SIN kinetic studies. This paper reports a simple and sensitive competitive enzyme-linked immunosorbent assay (ELISA) for the pharmacokinetic evaluation of SIN. Anti-SIN serum was obtained by immunizing mice with an antigen conjugated with bovine serum albumin and carboxylic modified SIN using the N-succinimidyl ester method. Enzyme labeling of SIN with horseradish peroxidase was similarly performed using carboxylic modified SIN. Under optimized conditions, this ELISA shows a linear detection range from 40 to 5000 pg/mL, and a limit of detection of 12.1 pg/mL for 50-µL samples. This assay was specific for SIN and showed very slight cross-reactivity with dextromethorphan (0.45%), dimemorfan (0.22%) and codeine (0.01%), but no cross-reactivity with 2-methoxycyclohex-2-enone (<0.001%). Using this ELISA, SIN levels were easily determined in the blood of mice after oral administration of Kampo medicine, Boiogito. The ELISA may be a valuable tool for studies of the biological and pharmacological properties of SIN.

Topics: Animals; Antigens; Enzyme-Linked Immunosorbent Assay; Kinetics; Mice; Morphinans

Haematocarpus validus (Miers) Bakh. f. ex Forman, a lesser-known fruit and medicinal plant of high nutraceutical and medicinal value, is used as anti-arthritic, hepatoprotective, and anti-inflammatory agents in ethnomedicine. Metabolome studies in H. validus are a virgin area of research and here we report the spectra of non-volatiles present in the methanolic leaf and fruit extract, using high-resolution liquid chromatography-mass spectrometry. Furthermore, the alkaloid sinomenine was quantified using high-performance thin layer chromatography spectrodensitometric analysis owing to its pharmacological importance as anti-arthritic and anti-inflammatory drug. Electrospray ionization with protonation in positive mode was selected for the analysis and the spectral data was interrogated using MassHunter software. A total of 40 compounds were identified from leaf and fruit samples and the major classes of compounds identified were alkaloids, terpenoids, steroids, tripeptides, vitamins, and related compounds. For separation and quantitation of sinomenine, chloroform:methanol:water (60:30:6.5, v/v) was used as the mobile phase and sinomenine hydrochloride as reference compound. The analysis confirmed the presence of sinomenine in both non-defatted and defatted methanolic leaf extract with quantities 45.73 and 26.02 mg/100 g dry weight, respectively. H. validus is a non-conventional source of sinomenine, the anti-arthritic and anti-inflammatory alkaloid. Sinomenine detected in this study supports the ethnomedicinal uses of H. validus as an anti-arthritic agent. Further study is needed to elucidate the underlying molecular mechanism of its anti-arthritic attributes as well as the corresponding structure-activity relationships.

Topics: Alkaloids; Anti-Inflammatory Agents; Antineoplastic Agents; Fruit; Methanol; Morphinans; Plant Extracts

Topics: Animals; Apoptosis; Chemical and Drug Induced Liver Injury; Inflammation; Liver; Male; Mice; Mice, Inbred BALB C; Morphinans; Oxidative Stress

Knee osteoarthritis (KOA) is a degenerative disease in the knee joint, with chronic joint pain, swelling, stiffness, and dysfunction as the primary manifestations. Sinomenine hydrochloride injection is a proprietary Chinese medicine injection of sinomenine, the main active component of traditional Chinese medicine (TCM). Clinical studies show that Sinomenine hydrochloride injection has a good effect on the treatment of KOA. At present, there is still a lack of systematic reviews and meta-analyses to evaluate the efficacy and safety of sinomenine hydrochloride injection in the treatment of KOA. Our purpose is to supplement this deficiency.. Randomized controlled trials of sinomenine hydrochloride injection in the treatment of KOA were searched for Eight electronic resource databases. We will use Review Manager 5.3 software for heterogeneity assessment, meta-analysis, and subgroup analysis. We will use the Cochrane Manual to assess the quality of the included studies, and use reporting biases assessment and sensitivity analysis to evaluate the reliability and stability of the results.. This study will provide a high-quality synthesis to assess the efficacy and safety of sinomenine hydrochloride injection in the treatment of KOA.. This systematic review evaluates the efficacy and safety of sinomenine hydrochloride injection in the treatment of KOA.. INPLASY2021110057.

Topics: Humans; Meta-Analysis as Topic; Morphinans; Osteoarthritis, Knee; Reproducibility of Results; Systematic Reviews as Topic

Sinomenine (SIN) is an effective anti-inflammatory agent, but its therapeutic efficacy is limited by its short half-life and the high dosage required. Tissue-specific strategies have the potential to overcome these limitations. The synovial homing peptide (CKSTHDRLC) was identified to have high synovial endothelium targeting affinity. In this work, two peptide-drug conjugates (PDCs), conjugate (L) and conjugate (C), were synthesized, in which SIN was covalently connected to the linear and cyclic synovial homing peptide, respectively, via a 6-aminocaproic acid linker. An evaluation of biostability showed that conjugate (C) was more stable in mouse serum and inflammatory joint homogenate than conjugate (L). The two conjugates gradually released free SIN. Interestingly, conjugate (L) self-cyclized via a disulfide bridge in a biological environment, which significantly impacted its biostability. It had an almost equipotent half-life in serum but faster degradation in the inflammatory joint than conjugate (C). Therefore, conjugate (C) exhibited better therapeutic efficacy and tissue targeting. All the results indicated that PDCs particularly in its cyclic form might be more efficient for targeted deliver and represent a potential strategy for the treatment of rheumatoid arthritis.

Topics: Animals; Anti-Inflammatory Agents; Arthritis, Rheumatoid; Mice; Morphinans; Peptides

Overexpression of thioredoxin reductase (TrxR) has been linked to tumorigenesis and phenotypic maintenance of malignant tumors. Thus, targeting TrxR with natural molecules is a promising strategy for developing anticancer drugs. Sinomenine is a naturally occurring alkaloid isolated from Sinomenium acutum. The drug, Zhengqing Fengtongning made from sinomenine, has been universally applied in rheumatoid arthritis treatment in China as well as other Asian countries for decades. Recently, increasing evidence indicates that sinomenine appears to be a promising therapeutic agent against various cancer cells. However, the exact mechanism underlying the anticancer activity of sinomenine remains unclear. In this study, we identified sinomenine as a kind of new inhibitor for TrxR. Pharmacological inhibition of TrxR by sinomenine results in the decrease of thiols content, increases the levels of reactive oxygen species, and finally facilitates oxidative stress-mediated cancer cell apoptosis. It is vital that knockdown in TrxR1 by shRNA can increase cell sensitivity to sinomenine. Treatment with sinomenine in vivo leads to a decrease in TrxR activity and tumor growth, and an increase in apoptosis. Our findings provide a novel action mechanism related to sinomenine and presents an insight on how to develop sinomenine as a chemotherapeutic agent for cancer therapy.

Topics: Antirheumatic Agents; Apoptosis; Drug Repositioning; Female; Humans; Morphinans; Reactive Oxygen Species; Thioredoxin-Disulfide Reductase; Uterine Cervical Neoplasms

Topics: Acupuncture Points; Antirheumatic Agents; Arthritis, Rheumatoid; Humans; Morphinans; Powders

Sinomenine (SIN) is an anti-inflammatory drug that has been used for decades in China to treat arthritis. In a previous study, SIN acted on α7 nicotinic acetylcholine receptor (α7nAChR) to inhibit inflammatory responses in macrophages, which indicates a new anti-inflammatory mechanism of SIN. However, the level of α7nAChR was increased in the inflammatory responses and was downregulated by SIN in vitro, so the underlying mechanisms of SIN acting on α7nAChR remain unclear.. To analyze the role of α7nAChR in inflammation and the effect and mechanism of SIN regulation of α7nAChR.. The effects of SIN on α7nAChR in endotoxemic mice and LPS-stimulated macrophages were observed. Nicotine (Nic) was used as a positive control, and berberine (Ber) was used as a negative control targeting α7nAChR. The antagonists of α7nAChR, α-bungarotoxin (BTX) and mecamylamine (Me), were used to block α7nAChR. In RAW264.7 macrophage cells in vitro, α7nAChR short hairpin RNA (shRNA) was used to knock down α7nAChR. Macrophage polarization was analyzed by the detection of TNF-α, IL-6, iNOS, IL-10, Arg-1, and Fizz1. U0126 was used to block ERK phosphorylation. The cytokines α7nAChR, ERK1/2, p-ERK1/2 and Egr-1 were detected.. SIN decreased the levels of TNF-α, IL-6 and the expression of α7nAChR increased by LPS in endotoxemic mice. The above effects of SIN were attenuated by BTX. In the α7nAChR shRNA transfected RAW264.7 cells, compared with the control, α7nAChR was knocked down, and M1 phenotype markers (including TNF-α, IL-6, and iNOS) were significantly downregulated, whereas M2 phenotype markers (including IL-10, Arg-1, and Fizz1) were significantly upregulated when stimulated by LPS. SIN inhibited the expression of p-ERK1/2 and the transcription factor Egr-1 induced by LPS in RAW264.7 cells, and the above effects of SIN were attenuated by BTX. The expression of α7nAChR was suppressed by U0126, which lessened the expression of p-ERK1/2 and Egr-1.. SIN acts on α7nAChR to inhibit inflammatory responses and downregulates high expression of α7nAChR in vivo and in vitro. The increase of α7nAChR expression is correlated with inflammatory responses and participates in macrophage M1 polarization. SIN downregulates α7nAChR via a feedback pathway of α7nAChR/ERK/Egr-1, which contributes to inhibiting macrophage M1 polarization and inflammatory responses.

Topics: alpha7 Nicotinic Acetylcholine Receptor; Animals; Anti-Inflammatory Agents; Feedback; Interleukin-10; Interleukin-6; Lipopolysaccharides; Macrophages; Mice; Morphinans; RNA, Small Interfering; Tumor Necrosis Factor-alpha

Prostaglandin E

Topics: Anti-Inflammatory Agents; Dinoprostone; Methylation; Morphinans; Promoter Regions, Genetic; Prostaglandin-E Synthases

The work aims to explore the feasibility of Raman mapping in predicting the dissolution profiles of solid oral dosage form. In this study, N = 36 batches of representative sinomenine hydrochloride sustained-release tablets were prepared, using a D-optimal design, to introduce adequate variability, and the Raman mapping data of each tablet were acquired. The partial least squares regression models were established using three kinds of different modes, named single point mode, average mode and multi-point mode, to predict the dissolution profiles based on Raman mapping data. The percent dissolutions at specific time points and the parameters of an exponential function, which was employed to fit the dissolution profiles, were predicted, and the accuracy and precision of prediction were tested. The results showed that the multi-point mode displayed the best accuracy and precision in the prediction of both the dissolutions at the specific time points and the function parameters. In summary, the established method based on Raman mapping avoids the shortcomings of traditional dissolution testing protocols, such as complex operation, time-consuming and high analysis cost, thus has great potential of application and popularization.

Topics: Delayed-Action Preparations; Morphinans; Solubility; Tablets

Hepatocellular carcinoma (HCC) is one of the most frequent cancers. Sinomenine (SIN) is a compound derived from Sinomenium acutum. Our previous investigations have found that SIN inhibited protein kinase B (AKT) signaling to induce autophagic death of tumor cells. However, whether inhibition of this pathway by SIN could impact the proliferation of HCC cells is unknown. Thus, we applied SIN to SK-Hep-1 cells and used cell counting kit 8 (CCK8), lactate dehydrogenase (LDH), colony formation and 5-ethynyl-20-deoxyuridine (EdU) incorporation experiments to detect cell viability. Then, staining with annexin V/propidium iodide (PI) coupled with terminal deoxynucleotidyl transferase-mediated biotinylated uridine 5'-triphosphate (UTP) nick end labeling (TUNEL) staining were utilized to monitor apoptosis. Changes in cell mitochondrial membrane capacity were explored via 5,5',6,6'-Tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide (JC-1) staining, whilst Western blot or immunohistochemistry was applied to evaluate the expression levels of key proteins, consisting of Cleaved Caspase 3, AKT1, B-cell leukemia/lymphoma 2 (BCL-2), phosphatidylinositol 3-kinase (PI3K) p85α, and Cleaved Caspase 9 etc. The Balb/c nude mice were utilized to establish HCC xenograft tumor model, administered by SIN. After treatments, the tumor volume along with weight were measured. The results illustrated that SIN suppressed SK-Hep-1 HCC cells' proliferation, enhanced the collapse of potential of the mitochondrial membrane, triggered cell apoptosis, down-regulated PI3K p85α, AKT1, BCL-2, Pro-Caspase 9, Pro-Caspase 3 expressions, and up-regulated Cleaved Caspase 9 and Cleaved Caspase 3 expressions in vitro and in vivo. Meanwhile, SIN reduced the tumor volume along with weight of mice. In addition, insulin-like growth factor-1 (IGF-1), a powerful activator of the PI3K/AKT pathway, could reverse the high apoptosis of SK-Hep-1 HCC cells induced by SIN. Overall, inhibition of PI3K/AKT1 signaling cascade by SIN induced HCC cells apoptosis.

Topics: Animals; Apoptosis; Carcinoma, Hepatocellular; Caspase 3; Caspase 9; Cell Line, Tumor; Humans; Liver Neoplasms; Mice; Mice, Nude; Morphinans; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; Signal Transduction

Topics: Administration, Cutaneous; Drug Delivery Systems; Microdialysis; Morphinans; Skin; Skin Absorption

Sinomenine is the main bioactive ingredient of the medicinal plant Sinomenium acutum with neuroprotective potential. This study was designed to assess beneficial effect of sinomenine in alleviation of trimethyltin (TMT)-induced cognitive dysfunction. TMT was administered i.p. (8 mg/kg, once) and sinomenine was daily given p.o. 1 h after TMT for 3 weeks at doses of 25 or 100 mg/kg. Cognitive performance was assessed in various behavioral tests. In addition, oxidative stress- and inflammation-associated factors were measured and histochemical evaluation of the hippocampus was conducted. Sinomenine at a dose of 100 mg/kg significantly and partially increased discrimination index in novel object recognition (NOR), improved alternation in short-term Y maze, increased step-through latency in passive avoidance paradigm, and also reduced probe trial errors and latency in the Barnes maze task. Moreover, sinomenine somewhat prevented inappropriate hippocampal changes of malondialdehyde (MDA), reactive oxygen species (ROS), protein carbonyl, nitrite, superoxide dismutase (SOD), tumor necrosis factor α (TNFα), interleukin 6 (IL 6), acetylcholinesterase (AChE) activity, beta secretase 1 (BACE 1) activity, and mitochondrial membrane potential (MMP) with no significant effect on glutathione (GSH), catalase, glutathione reductase, glutathione peroxidase, and myeloperoxidase (MPO). In addition, lower reactivity (IRA) for glial fibrillary acidic protein (GFAP) as an index of astrocyte activity was observed and loss of CA1 pyramidal neurons was attenuated following sinomenine treatment. This study demonstrated that sinomenine could lessen TMT-induced cognitive dysfunction which is partly due to its attenuation of hippocampal oxidative stress and neuroinflammation.

Topics: Acetylcholinesterase; Animals; Cognitive Dysfunction; Glutathione; Hippocampus; Maze Learning; Morphinans; Neuroinflammatory Diseases; Neuroprotective Agents; Oxidative Stress; Rats; Rats, Wistar; Trimethyltin Compounds

Sinomenine is a bioactive alkaloid isolated from the Chinese medicinal plant of Sinomenium acutum (Thunb.) Rehd.et Wils. Currently, sinomenine hydrochloride (SIN) preparations, classified as a natural disease-modifying anti-rheumatic drug (nDMARD), have been used for therapy of rheumatoid arthritis (RA); however, the efficacy of SIN was seriously limited by its short half-life, low bioavailability, and dose-dependent adverse reactions. In this study, a biomimetic nanocomplex based on Prussian blue nanoparticles (PB NPs) was developed for overcoming clinical limitations of SIN and accordingly improving its efficacy. In vitro studies showed that the nanocomplexes significantly inhibited abnormal proliferation of fibroblast-like synoviocytes (FLSs) by scavenging reactive oxygen species (ROS) and inhibiting secretion of proinflammatory cytokines. In vivo imaging demonstrated that the improved immune-escape properties of the nanocomplexes resulted in markedly increased half-life of circulation and levels of accumulated drugs at arthritic sites of adjuvant-induced arthritis (AIA) rats. Notably, the nanocomplexes significantly suppressed joint inflammation and protected against bone destruction of AIA rats by inhibiting inflammatory cytokine secretion of the synovial macrophages and FLSs. These results indicate that the nanocomplexes provide an excellent carrier for controlled release and targeted accumulation of SIN within the arthritic sites, which consequently achieve disease-remitting effects of SIN on RA.

Topics: Animals; Arthritis, Rheumatoid; Cytokines; Morphinans; Multifunctional Nanoparticles; Rats

Topics: Antineoplastic Agents; Apoptosis; Cell Proliferation; Drug Screening Assays, Antitumor; Humans; Interleukin-6; MCF-7 Cells; Molecular Structure; Morphinans; NF-kappa B; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction; Structure-Activity Relationship

Mycoplasma pneumoniae (MPP) induced pneumonia is a common disease of children. Sinomenine (SIN) is an isoquinoline mainly sequestered from Sinomenium acutum. It is a promising drug for treating arthritis, lung, colon, liver and gastric cancer. Hence, the present study investigated the role and mechanism of SIN treatment in MPP induced pneumonia in experimental in-vivo mice model. The BALB/c male mice were separated into four groups (n = 6 mice/group): normal, MPP, MPP + SIN (20 mg/kg bw), and SIN (20 mg/kg bw) alone. Results were expressed as mean ± SD. Data were analyzed using one way Analysis of Variance (ANOVA) with the Dunnett's post hoc test using SPSS v 18.0. P value < 0.05 was considered significant. The total protein, cell count, inflammatory cytokines, MP-IgM, Monocyte chemo attractant protein-1 (MCP-1), and MP-DNA were measured. The protein expressions of Bax/Bcl-2, ERK, JNK, NF-κB were analyzed and histopathology of lungs was examined. SIN treatment significantly (p < 0.05) reduced the total proteins, cell counts in BALF, inflammatory cytokines, MP-IgM, MCP-1, MP-DNA and reversed the histological alterations. SIN attenuated the apoptotic pathway through the modulation of Bax/Bcl-2 expression. SIN alleviated pulmonary inflammatory mediators and apoptosis in MPP-infected mice via suppression of ERK/JNK/NF-κB signaling. SIN administration diminished inflammation and lung fibrosis by inhibiting apoptosis in MPP mice. Hence, SIN is a potential natural protective remedy for MPP.

Topics: Animals; bcl-2-Associated X Protein; Cytokines; Immunoglobulin M; Inflammation; Male; MAP Kinase Signaling System; Mice; Mice, Inbred BALB C; Morphinans; Mycoplasma pneumoniae; NF-kappa B; Pneumonia, Mycoplasma

Sinomenine hydrochloride (SH) is usually applied to treat rheumatoid arthritis (RA) with severe side effects due to oral administration. Cinnamaldehyde (CA) as essential oil possesses an anti-RA effect and can facilitate transdermal penetration. Hence, this study developed hexagonal liquid crystalline (HII) gels to deliver two components (SH and CA) across the skins. HII gels were prepared and characterized by polarized light microscopy (PLM), small-angle X-ray scattering (SAXS) and rheology. Moreover,

Topics: Acrolein; Administration, Cutaneous; Gels; Lipid Bilayers; Liquid Crystals; Morphinans; Oils, Volatile; Scattering, Small Angle; Skin; X-Ray Diffraction

To investigate the effects of sinomenine on orthodontic tooth movement and root resorption in rats, as well as the effect of sinomenine on the osteogenesis of periodontal ligament stem cells (PDLSCs).. Fifty-four male Wistar rats were randomly divided into 3 groups: control group, 20 mg/kg sinomenine group and 40 mg/kg sinomenine group. Fifty-gram orthodontic force was applied to all groups. Each group was injected intraperitoneally with corresponding concentration of sinomenine every day. After 14 days, all rats were sacrificed. Micro-computed tomography (micro-CT) scan was used to analyze tooth movement, root resorption and alveolar bone changes. The effect on periodontal tissue was analyzed by Masson, tartrate-resistant acid phosphatase (TRAP) and immunohistochemical staining. In vitro, PDLSCs were extracted and identified. The effect of sinomenine on proliferation was determined by cell-counting kit-8. The effect of sinomenine on osteogenesis was investigated by alkaline phosphatase (ALP) activity and alizarin red staining. qPCR and Western blotting were performed to explore the effects of sinomenine on the expression levels of ALP, runt-related transcription factor 2 (RUNX2), receptor activator of nuclear factor kappaB ligand (RANKL) and osteoprotegerin (OPG).. The tooth movement and root resorption of sinomenine groups were reduced. Sinomenine decreased trabecular spacing on compression side and increased alveolar bone volume and trabecular thickness on tension side. TRAP-positive cells in sinomenine groups decreased significantly. The expressions of TNF-α and RANKL were decreased, while the expressions of OPG, RUNX2 and osteocalcin were up-regulated. In vitro, 0.1 M and 0.5 M sinomenine enhanced ALP activity, mineral deposition and the expression of ALP, RUNX2 and OPG, and reduced the expression of RANKL.. Sinomenine could inhibit tooth movement, reduce root resorption, and exert a positive effect on bone formation in rats. Moreover, sinomenine promoted the osteogenesis of PDLSCs.

Topics: Animals; Core Binding Factor Alpha 1 Subunit; Male; Morphinans; Osteogenesis; Periodontal Ligament; Rats; Rats, Wistar; Root Resorption; Stem Cells; Tooth Movement Techniques; X-Ray Microtomography

Radioactive iodine (RAI) plays an important role in the diagnosis and treatment of papillary thyroid cancer (PTC). The curative effects of RAI therapy are not only related to radiosensitivity but also closely related to the accumulation of radionuclides in the lesion in PTC. Sinomenine hydrochloride (SH) can suppress tumor growth and increase radiosensitivity in several tumor cells, including PTC. The aim of this research was to investigate the therapeutic potential of SH on PTC cell redifferentiation. In this study, we treated BCPAP and TPC-1 cells with SH and tested the expression of thyroid differentiation-related genes. RAI uptake caused by SH-pretreatment was also evaluated. The results indicate that 4 mM SH significantly inhibited proliferation and increased the expression of the thyroid iodine-handling gene compared with the control group (p < 0.005), including the sodium/iodide symporter (NIS). Furthermore, SH also upregulated the membrane localization of NIS and RAI uptake. We further verified that upregulation of NIS was associated with the activation of the thyroid-stimulating hormone receptor (TSHR)/cyclic adenosine monophosphate (cAMP) signaling pathway. In conclusion, SH can inhibit proliferation, induce apoptosis, promote redifferentiation, and then increase the efficacy of RAI therapy in PTC cells. Thus, our results suggest that SH could be useful as an adjuvant therapy in combination with RAI therapy in PTC.

Topics: Adenosine Monophosphate; Humans; Iodides; Iodine; Iodine Radioisotopes; Morphinans; Receptors, Thyrotropin; Sodium; Symporters; Thyroid Cancer, Papillary; Thyroid Neoplasms; Thyrotropin

The present study designed and prepared near-infrared responsive sinomenine hydrochloride(SIN) reservoir microneedles and evaluated the feasibility of this type of microneedles in increasing the drug loading and transdermal absorption by characterizing their mechanical properties and in vitro release characteristics.SIN was selected as the model drug, and methoxy poly(ethylene glycol) poly(caprolactone)(mPEG-PCL) copolymers and indocyanine green(ICG) were employed as amphiphilic block copolymers and light inductor to prepare near-infrared responsive nanoparticles.Based on the preparation principle of bubble microneedles, near-infrared responsive SIN reservoir microneedles were designed and prepared.The features of the near-infrared responsive SIN reservoir microneedles were characterized by measuring the morphology, length, mechanical properties, and skin penetration of microneedles.Meanwhile, the drug release performance of reservoir microneedles was evaluated by in vitro release assay.The results showed that the prepared SIN microneedles were conical, with an exposed tip height of about 650 μm.Each needle could load about 0.5 mg of drugs per square centi-meter, and this type of microneedle showed good mechanical properties and performance in skin penetration.The results of the in vitro release assay showed that the 24 h cumulative release per unit area and release rate of the microneedle were 825.61 μg·cm~(-2) and 74.3%, respectively, which indicated that its release kinetics was in line with the first-order kinetic model.This study preliminarily proved that the reservoir microneedle could effectively increase the drug loading with good mechanical properties and release perfor-mance.

Topics: Drug Delivery Systems; Drug Liberation; Indocyanine Green; Morphinans; Needles; Polyethylene Glycols

Ischemia-reperfusion (IR) injury is induced by an interrupted blood flow and succeeding blood restoration, which is common in the operation of liver transplantation. Serious IR injury is a major reason leading to transplant failure. Hepatic IR is featured by excessive inflammatory response, oxidative stress, and apoptosis. Sinomenine (SIN) is derived from the herb Sinomeniumacutum and shows properties of anti-inflammation and antiapoptosis in multiple IR-induced organ injuries. However, the effect of SIN in hepatic IR has not been investigated.. This study aims to investigate impacts of SIN on hepatic IR and the involved signaling pathway. An in vivo rat model of syngeneic orthotopic liver transplantation was constructed to induce the hepatic IR injury.. Results showed that SIN pretreatment provided a significant prevention against IR-induced hepatic injury as manifested by the downregulated activities of serum alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase, the alleviatedoxidative stress as shown by increased activities of serum superoxide dismutase and glutathione peroxidase, and decreased serum level of malondialdehyde, the suppressed inflammatory responses as shown by downregulated serum tumor necrosis factor-α, interleukin (IL)-6, IL-8 levels, and upregulated IL-10 level, as well as attenuated apoptosis as shown by decreased protein expression of cleaved caspase-3 and -9. In line with these results, SIN pretreatment also alleviatedthe hepatic histopathological changes in IR rats and induced Nrf-2/HO-1 activation. The use of brusatol, a selective inhibitor for Nrf-2, effectively reversed SIN-induced above effects.. Altogether, our results demonstrate that SIN might be a useful therapeutic drug for preventing hepatic IR-induced injury during clinical liver transplantation.

Topics: Alanine; Alanine Transaminase; Animals; Aspartate Aminotransferases; Caspase 3; Glutathione Peroxidase; Interleukin-10; Interleukin-8; Lactate Dehydrogenases; Liver Diseases; Malondialdehyde; Morphinans; Rats; Reperfusion Injury; Superoxide Dismutase; Tumor Necrosis Factor-alpha

Gestational diabetes mellitus (GDM) is a dangerous complication of pregnancy which is induced via dysfunction in glucose metabolism during pregnancy. Sinomenine (SM) has already proved an antidiabetic effect against streptozotocin (STZ)-induced diabetes mellitus (DM) in rats. In this protocol, we examined the protective effect of SM against STZ-induced GDM in rats. Wistar rats were divided into three groups and STZ (40 mg/kg) was used to induce GDM. At the end of the experimental protocol, bodyweight, pub weight, and survival rate were estimated. Blood glucose level (BGL), fasting insulin (FINS), free fatty acid (FFA), Hemoglobin A1C (HbA1c), and C-peptide were measured. Lipid, antioxidant, inflammatory cytokines, and inflammatory mediators were also determined. RT-PCR was used for estimation of the role of TLR4/MyD88/NF-κB signaling pathway. SM treatment significantly (p < .001) reduced BGL, hepatic glycogen, and improved the levels of FINS, C-peptide, FFA, and HbA1c. SM significantly (p < .001) suppressed the levels of total cholesterol (TC), low-density lipoprotein (LDL), triglycerides (TG), coronary artery index (CAI), very low-density lipoprotein (VLDL), atherogenic index (AI), and boosted high-density lipoprotein (HDL) levels. SM significantly (p < .001) decreased the lipid peroxidation (LPO) level and enhanced glutathione peroxidase (GPx), total antioxidant capacity (TAC), glutathione S-transferase (GST), superoxide dismutase (SOD), respectively. It reduced the levels of inflammatory cytokines including interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and inflammatory mediators viz., nuclear kappa B factors (NF-κB). SM significantly (p < .001) reduced the mRNA expression of Myd88, NLRP3, TLR4, and NF-κB, which were boosted in the GDM group rats. These findings suggest that SM could be a probable drug to be used for treating GDM via inhibition of the TLR4 signaling pathway. PRACTICAL APPLICATIONS: It is well known that gestational diabetes mellitus (GDM) is a dangerous health problem during the pregnancy. SM reduced the glucose level; boosted the level of fasting insulin (FINS) and bodyweight. SM significantly improved the number of pubs and their survival rates. SM suppressed oxidative stress and inflammation via activation of TLR4/MyD88/NF-κB signaling pathway. According to our research, SM can be used as a preventive drug in the treatment of GDM during pregnancy.

Topics: Animals; Diabetes, Gestational; Female; Inflammation; Morphinans; Myeloid Differentiation Factor 88; NF-kappa B; Oxidative Stress; Pregnancy; Rats; Rats, Wistar; Signal Transduction; Toll-Like Receptor 4

Sepsis is a systemic inflammatory response syndrome caused by a host's immune response to infection. Acute lung injury (ALI) is one of the most common complications of sepsis with high mortality and morbidity. Recent evidence demonstrated that the 'gut-lung axis' was related to the progression of septic acute lung injury, which regarded gut microbiota and intestinal barrier as two critical factors correlated with acute lung injury. Sinomenine is an isoquinoline alkaloid component extracted from Sinomenium acutum Rehd，et Wils, which has been already reported to have significant anti-inflammatory, immunosuppressive, and anti-arthritis properties. In this research, we observed that sinomenine could repair the lung injury and alleviate inflammatory response induced by cecum ligation and puncture (CLP). Illumine sequencing of 16S rDNA revealed that sinomenine could improve the richness of gut microbiota and modulate the composition of intestinal flora in cecum ligation and puncture mice. Meanwhile, sinomenine could reduce the colon pathological damage and improve the intestine barrier integrity in cecum ligation and puncture mice. We also found that the molecular mechanism of sinomenine's protective effect on intestinal tract was related to the activation of aryl hydrocarbon receptor/nuclear factor erythroid-2 related factor 2（Nrf2）pathway both in vivo and vitro experiments. Collectively, the prevention of septic acute lung injury by sinomenine might be mediated by modulating gut microbiota and restoring intestinal barrier via aryl hydrocarbon receptor/Nrf2-dependent pathway.

Topics: Acute Lung Injury; Animals; Anti-Inflammatory Agents; Basic Helix-Loop-Helix Transcription Factors; Cecum; Cell Line, Tumor; Cytokines; Disease Models, Animal; Gastrointestinal Microbiome; Gastrointestinal Tract; Homeostasis; Humans; Inflammation; Ligation; Male; Mice, Inbred ICR; Morphinans; NF-E2-Related Factor 2; Permeability; Protective Agents; Punctures; Receptors, Aryl Hydrocarbon

To study the effects of sinomenine on conditioned place preference (CPP) zebrafish induced by morphine and expression levels of intracephalic tyrosine hydroxylase (TH), NMDA receptor subunit 2B (NR2B), μ-opioid receptor (zfmor) and δ-opioid receptors (zfdor1 and zfdor2), morphine (40mg/kg) was administrated to zebrafish and the effect of CPP was detected in these zebrafish treated with sinomenine. The expression of TH and NR2B was detected by immunohistochemistry; and the mRNA expression of opioid receptors zfmor, zfdor1 and zfdor2 in the zebrafish brain was assayed by RT-qPCR. In the CPP test, morphine induced significant behavioral alteration, while pretreatment with sinomenine or methadone, resulted in decreased activity time in the morphine-paired compartment significantly. Morphine also increased the integral optical density value of TH- and NR2B-positive cells in the zebrafish brain, and reduced the amount of opioid receptors. However, the compound sinomenine could attenuate these effects. These findings demonstrate that sinomenine (80mg/kg) decreased the CPP effects of zebrafish induced by morphine significantly, downregulated expression of TH and NR2B, and upregulated μ-opioid (zfmor) and δ-opioid (zfdor1 and zfdor2) receptor expression in the CPP zebrafish brains.

Topics: Animals; Gene Expression Regulation; Morphinans; Morphine; Receptors, N-Methyl-D-Aspartate; Receptors, Opioid; Tyrosine 3-Monooxygenase; Zebrafish

Topics: Airway Remodeling; Animals; Anti-Asthmatic Agents; Asthma; Cell Line; Cell Movement; Cell Proliferation; Disease Models, Animal; Epithelial Cells; Epithelial-Mesenchymal Transition; Female; Humans; Interleukin-4; Lung; Mice, Inbred BALB C; Morphinans; Ovalbumin; Signal Transduction; Smad3 Protein; Transforming Growth Factor beta1

Atherosclerosis (AS) is a cardiovascular disease that arise due to dysfunction of lipid deposition and metabolism. AS is causes the mortality and morbidity worldwide. Sinomenine isolated from the Sinomenium acutum is used extensively against the various cardiac diseases in China. However, the anti-atherosclerosis effect of sinomenine still not explore. In this study, we explore the cardioprotective and anti-atherosclerosis effect of sinomenine against Vitamin D3 and High fat induced atherosclerosis in rats. Sprague Dawley (SD) rats were used in this study. The rats were received the vitamin D (60000) and High fat diet to induce the atherosclerosis and divided into groups and received the oral administration of sinomenine (2.5, 5 and 10 mg/kg) and simvastatin (5 mg/kg). Body weight, organ weight and biochemical parameters were estimated. The mRNA expression of MyD88, TLR4, NF-κB and IκB were estimated. Sinomenine treated rats significantly (p<0.001) suppressed the body weight and modulated the organ weight (hepatic, renal and heart). Sinomenine significantly (p<0.001) decreased the level of triacylglycerols (TG), low density lipoprotein cholesterol (LDL-c), total cholesterol (TC), very low-density lipoprotein cholesterol (VLDL-c) and augmented the level of high-density lipoprotein cholesterol (HDL-c). Sinomenine treatment also reduced the level of atherogenic index (TC/HDL-c and LDL-c/HDL-c). Sinomenine treatment decrease the ratio of HMG CoA/Mevalonate and level of collagen and total protein. Sinomenine significantly (p<0.001) altered the level of heart parameters, antioxidant parameters and inflammatory cytokines. Sinomenine significantly (p<0.001) reduced the expression of MyD88, TLR4, NF-κB and IκB. Taken together, sinomenine exhibited the protective effect against the atherosclerosis via alteration of TLR4/NF-κB signaling pathway.

Topics: Administration, Oral; Animals; Antioxidants; Atherosclerosis; Cholecalciferol; Cytokines; Diet, High-Fat; Inflammation; Inflammation Mediators; Lipid Metabolism; Male; Morphinans; NF-kappa B; Oxidative Stress; Phytotherapy; Rats, Sprague-Dawley; Signal Transduction; Sinomenium; Toll-Like Receptor 4

To study the effect of sinomenine (Sin) on isoproterenol (Iso, β-agonist)-induced cardiac hypertrophy (CH), we set up four mouse groups: control, Iso model, Iso+metoprolol (Met, β blocker) 60 mg/kg and Iso+Sin 120 mg/kg. CH was induced by Iso (s.c. for 28 days) in mice, and Sin or Met were orally administered by gavage for 28 days in total. Left ventricular diastolic anterior wall thickness (LVAWd), left ventricular diastolic posterior wall thickness (LVPWd), left ventricular ejection fraction (LVEF), and short axis shortening (FS) were measured by echocardiography. Malondialdehyde (MDA) and total superoxide dismutase (T-SOD) were measured by commercial kits. Lactate dehydrogenase (LDH), tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β) were measured by ELISA kits. Histological changes were observed using hematoxylin-eosin (HE) and Masson staining. Protein level of nuclear transcription factor-kappa B (NF-κB) was detected by immunohistochemistry. Compared with the control group, LVAWd, Left ventricular weight index (LVWI) and myocardial fibrosis of the Iso model group significantly increased, as well as NF-κB, LDH, MDA, TNF-α, and IL-1β levels. However, the activity of T-SOD decreased. Compared with the Iso model group, LVWI of Iso model+Sin or Iso model+Met group was improved, LVAWd, LVPWd and myocardial fibrosis decreased, and NF-κB, LDH, MDA, TNF-α and IL-1β levels decreased. T-SOD activity also increased. This study reveals that Sin inhibits the activation of NF-κB, lowers the levels of TNF-α and IL-1β, has anti-oxidative stress effect and inhibits myocardial inflammation in mouse heart, thereby demonstrating its efficacy in preventing Iso induced CH.

Topics: Animals; Cardiomegaly; Fibrosis; Isoproterenol; Mice; Morphinans; NF-kappa B; Stroke Volume; Superoxide Dismutase; Tumor Necrosis Factor-alpha; Ventricular Function, Left

A novel structure of sinomenine isoxazole derivatives is synthesised from sinomenine hydrochloride and aromatic aldehydes and requires six steps. 19 target compounds have been obtained in good yields. The sinomenine hydrochloride transforms to 4-alkynyl sinomenine, which is a key intermediate product to synthesise the target sinomenine isoxazole compounds, after a neutralisation reaction with ammonia and substitution reaction with 3-chloropropyne. Another key intermediate product is 1,3-dipole, which can be obtained from aromatic aldehyde. After treatment with hydroxylamine hydrochloride and then sodium carbonate solution, aromatic aldehyde is converted to aldehyde oxime, which reacts with N-chlorosuccinimide (NCS) to afford aryl hydroximino chloride. 1,3-Dipole is eventually formed

Topics: Aldehydes; Carbon-13 Magnetic Resonance Spectroscopy; Cycloaddition Reaction; Isoxazoles; Morphinans; Proton Magnetic Resonance Spectroscopy; Spectroscopy, Fourier Transform Infrared

Lung cancer is the leading cause of cancer deaths worldwide, with a high morbidity and less than 20% survival rate. Therefore, new treatment strategies and drugs are needed to reduce the mortality of patients with lung cancer. α7 nicotinic acetylcholine receptor (α7 nAChR), as a receptor of nicotine and its metabolites, is a potential target for lung cancer treatment. Our previous studies revealed that sinomenine plays anti-inflammation roles via α7 nAChR and down-regulates the expression of this receptor, thus increasing the inflammatory response. Hence, sinomenine is possibly a natural ligand of this receptor. In the present study, the effects of sinomenine on lung cancer A549 cells and tumor-bearing mice were determined to investigate whether this alkaloid has an inhibitory effect on lung cancer via α7 nAChR. CCK-8 assay, wound-healing test, and flow cytometry were performed for cell proliferation, cell migration, and apoptosis analysis in vitro, respectively. Xenograft mice were used to evaluate the effects of sinomenine in vivo. Results showed that sinomenine decreased cell proliferation and migration abilities but increased the percentage of apoptotic cells. Tumor volume in tumor-bearing mice was significantly reduced after sinomenine treatment compared with that in the vehicle group mice (p < 0.05). Furthermore, the effects of sinomenine were abolished by the α7 nAChR antagonist mecamylamine and the allosteric modulator PNU-120596, but no change occurred when the mice were pretreated with the muscarinic acetylcholine receptor antagonist atropine. Meanwhile, sinomenine suppressed α7 nAChR expression in vitro and in vivo, as well as the related signaling molecules pERK1/2 and ERK1/2 and the transcription factors TTF-1 and SP-1. By contrast, sinomenine up-regulated the expression of another transcription factor, Egr-1. These effects were restricted by mecamylamine and PNU but not by atropine. Results suggested that sinomenine can inhibit lung cancer via α7 nAChR in a negative feedback mode.

Topics: A549 Cells; alpha7 Nicotinic Acetylcholine Receptor; Animals; Antineoplastic Agents; Humans; Lung Neoplasms; Male; Mice, Inbred C57BL; Morphinans; Signal Transduction; Transcription Factors

Topics: Animals; Astrocytes; Body Weight; Cytokines; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Encephalomyelitis, Autoimmune, Experimental; Female; Inflammasomes; Mice; Mice, Inbred C57BL; Microglia; Morphinans; Myelin-Oligodendrocyte Glycoprotein; NLR Family, Pyrin Domain-Containing 3 Protein; Peptide Fragments; Pyroptosis; Random Allocation; Specific Pathogen-Free Organisms; Spinal Cord

To investigate the therapeutic effects and potent mechanism of sinomenine (SIN) nanoliposomes on nephropathy in diabetic rats.. Cell viabilities of HK-2 cells with oxidative injury were obviously improved by incubating with SIN at 320 μg/mL for 92.9%. Significantly up-regulated GPX1, SOD2 and GSH contributed to the down-regulated ROS content in SIN-treated groups. Moreover, 6-week administration of SIN improved renal functions and worsening nephropathy morphology of DKD rats. SIN also ameliorated gradually increased renal cell apoptosis, suppressed expression levels of fibrosis-related proteins as well as IL-6 and ICAM-1, and regulated JAK2/STAT3/SOCS1 pathway, thereby exhibited protective effects on renal tissues of DKD rats.. SIN protects nephrocytes and decreases renal tissue injury via inhibiting oxidative stress, reducing renal cell apoptosis and fibrosis, regulating the JAK2/STAT3/SOCS1 pathway in DKD rats.

Topics: Animals; Apoptosis; Blood Glucose; Cell Line; Cell Survival; Diabetes Mellitus, Experimental; Diabetic Nephropathies; Dose-Response Relationship, Drug; Fibrosis; Humans; Hydrogen Peroxide; Janus Kinase 2; Male; Morphinans; Oxidative Stress; Rats; Rats, Sprague-Dawley; STAT3 Transcription Factor; Streptozocin; Suppressor of Cytokine Signaling 1 Protein

BACKGROUND This study aims to explore the effect of Sinomenine (SIN) on pregnancy outcomes of recurrent spontaneous abortion (RSA) in a mouse model. MATERIAL AND METHODS Thirty female CBA/J mice were allocated into 3 groups randomly, then mated with BALB/c mice (CBA/J×BALB/c) as normal-pregnancy group (n=10), or mated with DBA/2 mice (CBA/J×DBA/2) as RSA model (n=10), or CBA/J×DBA/2 mice treated with SIN as RSA+SIN group (n=10). The number of surviving and reabsorbed embryos in each group were counted on day 13.5 of gestation. The mouse serum was collected to determine the levels of interferon-γ (IFN)-γ and IL-4 by ELISA. Immunohistochemistry, qRT-PCR and immunoblotting were used to determine the location, mRNA and protein expressions of IFN-γ, IL-4, T-bet and GATA3 in the decidual and placental tissue. RESULTS In the RSA group, the amount of reabsorbed embryo was significantly higher than that in the normal-pregnancy group. However, SIN treatment showed a rescue effect on spontaneous abortion in RSA mice. IFN-γ, IL-4, T-bet, and GATA3 were all expressed in placental tissues and mainly located in the cytoplasm. The RSA group demonstrated higher expression levels of IFN-γ and T-bet than in the RSA+SIN and normal-pregnancy groups. Although RSA and RSA+SIN groups showed lower expression levels of IL-4 and GATA3 than in the normal-pregnancy group, there was no significant difference between RSA and RSA+SIN groups regarding IL-4 and GATA expression levels. CONCLUSIONS SIN treatment demonstrates a therapeutic effect on spontaneous abortion in RSA mice, possibly through regulating the balance of Th1/Th2 in maternal circulation and decidual tissues.

Topics: Abortion, Spontaneous; Animals; Decidua; Disease Models, Animal; Embryo, Mammalian; Female; GATA3 Transcription Factor; Interferon-gamma; Interleukin-4; Male; Mice; Morphinans; Placenta; Pregnancy; T-Box Domain Proteins; Th1-Th2 Balance

Topics: Animals; Antirheumatic Agents; Arthritis, Experimental; Cells, Cultured; Cytokines; Fibroblasts; Humans; Inflammation Mediators; Joints; Kelch-Like ECH-Associated Protein 1; Male; Mice, Inbred DBA; Mice, Knockout; Morphinans; NF-E2-Related Factor 2; Phosphorylation; Reactive Oxygen Species; Sequestosome-1 Protein; Signal Transduction

Sinomenine is an alkaloid derived from Chinese medicinal plant Sinomenium acutum. Our previous studies suggested that sinomenine can inhibit the metastasis of breast cancer. However, whether sinomenine can inhibit the metastasis characteristics of breast cancer side population (SP) cells is still unknown. In present study, we isolated the side population (SP) cells from MDA-MB-231 cells by fluorescence-activated cell sorting (FACS). MDA-MB-231 SP cells were treated with different concentrations of sinomenine at the absence or presence of hypoxia, and cell viability were measured by CCK-8 assay. The transwell invasive assay were conducted to assess of the effect of sinomenine on the invasion of hypoxic MDA-MB-231 SP cells. The protein expression was detected by Western blot assay. Sinomenine inhibited the cell viability and invasion of hypoxic MDA-MB-231 SP cells. Western blot assay results showed that the upregulation of MMP-2 and MMP-9 by hypoxia was inversed by sinomenine. Additionally, it was found that sinomenine suppressed the activation of PI3K/Akt/mTOR pathway under hypoxia in MDA-MB-231 SP cells. Moreover, the inhibiton of sinomenine on metastasis of hypoxic MDA-MB-231SP cells and PI3K/Akt/mTOR pathway could be rescued by PI3K activator IGF-1. Our study suggested that sinomenine inhibits invasion of breast cancer SP cells under hypoxia through PI3K/Akt/mTOR pathway.

Topics: Antineoplastic Agents; Breast Neoplasms; Cell Hypoxia; Cell Proliferation; Cell Survival; Dose-Response Relationship, Drug; Drug Screening Assays, Antitumor; Female; Humans; Molecular Structure; Morphinans; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Structure-Activity Relationship; TOR Serine-Threonine Kinases; Tumor Cells, Cultured

The objective of study was to investigate the inhibitory effect of sinomenine on neuropathic pain on dorsal root ganglia (DRG). The DRG cell line and spinal nerve ligation (SNL) model were used in this study. The effect of sinomenine on the cell viability was examined by MTT assay. The expression of p38 MAPK, NF-κB, c-fos, SP and TNF-α was detected by using immunofluorescence and immunohistochemistry assay. We also assessed the level of p-CaMKII, COX-2, p-CREB, IL-17A, TLR4 and IL-1β via western blotting and RT-qPCR. Compared to the controls, sinomenine showed a protective effect on TNF-α-induced apoptosis on DRG cells in a dose-dependent manner, with an increase of cell viability and a decrease of reactive oxygen species level as well as LDH release. Parallelly, sinomenine treatment significantly reduced the expression of various factors related to stress and inflammation, including p38 MAPK, NF-κB, c-fos, p-CAMKII, COX-2, p-CREB, TLR4 and IL-17A in DRG cells in vitro. Furthermore, we found that administration of sinomenine significantly reduced mechanical withdrawal threshold and thermal withdrawal latency and inhibited the inflammation and activation of p38 signaling in SNL rats. It is noting that combined therapy of sinomenine and pulsed radiofrequency exhibited higher efficacy of dorsal root ganglia inflammation than single treatment as well as the combination of oxycodone and pulsed radiofrequency. Sinomenine inhibited the apoptosis of DRG cell by regulating p38 MAPK/CREB signalling pathway, which provides evidence to alleviate neuropathic pain in clinic.

Topics: Analgesics; Analgesics, Opioid; Animals; Anti-Inflammatory Agents; Apoptosis; Behavior, Animal; Cell Line; Combined Modality Therapy; Cyclic AMP Response Element-Binding Protein; Disease Models, Animal; Ganglia, Spinal; Inflammation; Inflammation Mediators; Male; Morphinans; Neuralgia; Oxycodone; p38 Mitogen-Activated Protein Kinases; Pain Threshold; Pulsed Radiofrequency Treatment; Rats, Sprague-Dawley; Signal Transduction

Sinomenine (SIN), isolated from Caulis sinomenii, is a benzyltetrahydroisoquinoline-type alkaloid with potent anti-inflammatory and analgesic effects. SIN-HCl has been used in the forms of tablets or enteric-coated tablets in the treatment of rheumatoid arthritis in China for years, while its short half-life leads to attenuated therapeutic effects and serious side effects. In the current study, three phenolic acids, including salicylic acid (SAA), 2,3-dihydroxybenzoic acid (23DHB), and 2,4-dihydroxybenzoic acid (24DHB), were firstly employed as coamorphous coformers to prepare three binary SIN-phenolic acid coamorphous systems. These new coamorphous systems were characterized by powder X-ray diffraction (PXRD), modulated temperature differential scanning calorimetry (mDSC), and Fourier transform infrared spectroscopy (FTIR). The formation of SIN-phenolic acid coamorphous systems are supported by the absence of diffraction peaks in their PXRD spectra, as well as the single T

Topics: Calorimetry, Differential Scanning; Delayed-Action Preparations; Drug Stability; Hydroxybenzoates; Morphinans; Solubility; Spectroscopy, Fourier Transform Infrared; X-Ray Diffraction

Topics: Animals; Anti-Inflammatory Agents; Cell Movement; Crk-Associated Substrate Protein; Enzyme Activation; Focal Adhesion Kinase 1; Macrophages; Mice; Morphinans; RAW 264.7 Cells; Signal Transduction; Sinomenium; src-Family Kinases

Natural product structure and fragment-based compound development inspire pseudo-natural product design through different combinations of a given natural product fragment set to compound classes expected to be chemically and biologically diverse. We describe the synthetic combination of the fragment-sized natural products quinine, quinidine, sinomenine, and griseofulvin with chromanone or indole-containing fragments to provide a 244-member pseudo-natural product collection. Cheminformatic analyses reveal that the resulting eight pseudo-natural product classes are chemically diverse and share both drug- and natural product-like properties. Unbiased biological evaluation by cell painting demonstrates that bioactivity of pseudo-natural products, guiding natural products, and fragments differ and that combination of different fragments dominates establishment of unique bioactivity. Identification of phenotypic fragment dominance enables design of compound classes with correctly predicted bioactivity. The results demonstrate that fusion of natural product fragments in different combinations and arrangements can provide chemically and biologically diverse pseudo-natural product classes for wider exploration of biologically relevant chemical space.

Topics: Biological Products; Cheminformatics; Chromones; Drug Discovery; Griseofulvin; Indoles; Morphinans; Quinidine; Quinine; Small Molecule Libraries

Sinomenii Caulis (QingFengTeng) and Ramulus Cinnamomi (GuiZhi) are traditional Chinese drugs that have been used for anti-inflammation. In this study, the team plans to find out the material basis of a Chinese herb combination composed of the two herbs with different ratios.. The extracts of the herbal compound with various ratios obtained from ethanol extraction were analyzed by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) and gas chromatography coupled mass spectrometry to identify the basic chemical compounds. Simultaneously, the contents of the eight main components (sinomenine, magnoflorine, laurifoline, dauricine, coumarin, cinnamyl alcohol, cinnamic acid and cinnamaldehyde) from herb formula were determined by gradient elution by high-performance liquid chromatography. Furthermore, the content of sinomenine and cinnamaldehyde were determined by isocratic elution, respectively.. Eighteen compounds in the herb formula were identified by UHPLC-Q-TOF-MS. The components in the GuiZhi are mostly volatile oils and the kinds of compounds isolated from the formula in the ratio of 4:1 were the most. Wherein eight compounds were identified as the main detection targets in the content determination.. The extraction rate of sinomenine in QingFengTeng was related to the proportion of GuiZhi in the drug pairs. Synchronously, the addition of sinomenine in different proportions also had some influence on the extraction of cinnamaldehyde in GuiZhi. Furthermore, the series of methods was successfully applied to the simultaneous determination of chemical compounds in different samples of QingFengTeng-GuiZhi decoction.

Topics: Acrolein; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Gas Chromatography-Mass Spectrometry; Morphinans; Oils, Volatile

Although sinomenine (SIN) has been used to treat several inflammation-related diseases in the clinic for decades, the detailed anti-inflammatory mechanism remains elusive. Here, we present a chemoproteomic study that supports a polypharmacological mode of action for SIN to inhibit inflammation. Notably, functional validation revealed multiple new protein regulators whose knockdown could significantly affect inflammation.

Topics: Animals; Anti-Inflammatory Agents; Cell Survival; Dose-Response Relationship, Drug; Inflammation; Lipopolysaccharides; Mice; Molecular Structure; Morphinans; Proteomics; RAW 264.7 Cells

Transarterial chemoembolization is the preferred treatment for patients with middle and advanced-stage hepatocellular carcinoma (HCC); however, most hepatic artery embolization agents have various disadvantages. The purpose of this study was to evaluate phytantriol-based liquid crystal injections for potential use in treatment of HCC.. Using sinomenine (SN) and 5-fluorouracil (5-FU) as model drugs, three precursor in situ liquid crystal injections based on phytantriol (P1, P2, and P3) were prepared, and their in vitro biocompatibility, anticancer activity, and drug release investigated, to evaluate their feasibility for use in treatment of HCC. The properties of the precursor injections and subsequent cubic liquid crystal gels were observed by visual and polarizing microscopy, in an in vitro gelation experiment. Biocompatibility was evaluated by in vitro hemolysis, histocompatibility, and cytotoxicity assays.. Precursor injections were colorless liquids that formed transparent cubic liquid crystal gels on addition of excess water. The three precursor injections all caused slight hemolysis, without agglutination, and were mildly cytotoxic. Histocompatibility experiments showed that P1 had good histocompatibility, while P2 and P3 resulted in strong inflammatory responses, which subsequently resolved spontaneously. In vitro anti-cancer testing showed that SN and 5-FU inhibited HepG2 cells in a time- and concentration-dependent manner and had synergistic effects. Further, in vitro release assays indicated that all three preparations had sustained release effects, with cumulative release of >80% within 48 h.. These results indicate that SN and 5-FU have synergistic inhibitory effects on HepG2 cells, which has not previously been reported. Moreover, we describe a biocompatible precursor injection, useful as a drug carrier for the treatment of liver cancer, which can achieve targeting, sustained release, synergistic chemotherapy, and embolization. These data indicate that precursor injections containing SN and 5-FU have great potential for use in therapy for liver cancer.

Topics: Animals; Antineoplastic Agents; Carcinoma, Hepatocellular; Cell Death; Drug Carriers; Drug Liberation; Drug Synergism; Fatty Alcohols; Fluorouracil; Gels; Hemolysis; Hep G2 Cells; Humans; Injections; Liquid Crystals; Liver Neoplasms; Morphinans; Rats, Sprague-Dawley; Water

Recent evidence has been demonstrated that Sinomenine (SIN) exerts antitumor activity in vitro. However, the clinical utility of SIN remains limited mainly because of its poor bioavailability. Exosomes are nanoscale vesicles that play crucial roles in intracellular communications through functionally active substances such as DNA and RNA. Exosomes have been utilized as nanocarriers for targeted drug delivery of different anticancer drugs.. The present study aimed to evaluate the effectiveness of combined Exosomes-SIN for the treatment of hepatocellular carcinoma (HCC) in a rat model. To do so, we prepared a mixture of SIN and exosomes (Exo-SIN) to improve the bioavailability of SIN to treat liver cancer. The in vitro releasing profile of the Exo-SIN was examined.. We observed a continuous, slow release of SIN from Exo-SIN in simulated body fluid as well as tumor microenvironment. In the cytotoxicity test, Exo-SIN exhibited a significantly stronger inhibition in HepG2 cells compared to free SIN. The flow cytometry assessments showed that Exo-SIN could suppress HepG2 cell migration in a Transwell assay and induce cell cycle arrest and cellular apoptosis. Western blotting showed that survivin, a crucial protein for the survival of living cells, was significantly downregulated after treatment with Exo-SIN.. In conclusion, our data suggested that Exo-SIN could serve as a potential, effective delivery platform for hepatic carcinoma therapy.

Topics: Animals; Antineoplastic Agents; Apoptosis; Carcinoma, Hepatocellular; Cell Movement; Cell Proliferation; Cells, Cultured; Drug Carriers; Drug Delivery Systems; Drug Screening Assays, Antitumor; Exosomes; Humans; Liver Neoplasms; Morphinans; Rats; Rats, Sprague-Dawley

Sinomenine is a morphinan alkaloid with a variety of biological activities. Its derivatives have shown significant cytotoxic activity against different cancer cell lines in many studies. In this study, two series of sinomenine derivatives were designed and synthesized by modifying the active positions C

Topics: A549 Cells; Antineoplastic Agents; Apoptosis; Cell Line; Cell Line, Tumor; Cell Proliferation; Dose-Response Relationship, Drug; Drug Design; Drug Screening Assays, Antitumor; HEK293 Cells; HeLa Cells; Hep G2 Cells; Humans; MCF-7 Cells; Molecular Docking Simulation; Morphinans

The new paradigm of precision medicine in oncology questions today the respective place of evidence-based medicine and doctor-patient relationship. Based on the results of a randomized study comparing the efficacy of a homeopathic molecule in the prevention of nausea and vomiting induced by chemotherapy in non-metastatic breast cancer, this article extends and develops the discussion of maintaining an unresolved tension between medical art and medical science, between care and cure. This tension sets a base for the authors of the therapeutic alliance in medicine, defined as a dialectic constantly adjourned between the alliance of the doctor with the patient and his therapy, and the therapeutic effect of this alliance. Because if a policy or a public opinion were to promote an exclusively rational medicine deprived of the field of relation to care, or on the contrary a medicine based only on clinical sense and intuition, then respectively the ethics of care and the progress of therapy would be threatened. It is advisable to be aware of erring from the truth, amplified today by social networks, as much due to a tide of scientific positivism, as an excess of the "good caring soul". Taking into account the therapeutic alliance makes it possible to no longer oppose scientific medicine and care relationship.

Topics: Breast Neoplasms; Delivery of Health Care; Evidence-Based Medicine; Female; Humans; Materia Medica; Medicine; Metaphor; Morphinans; Nausea; Online Social Networking; Physician-Patient Relations; Precision Medicine; Proof of Concept Study; Randomized Controlled Trials as Topic; Science; Therapeutic Alliance; Vomiting

Rheumatoid arthritis (RA) is a chronic autoimmune joint disorder that affects about 1% of the world population and may lead to severe disability and comorbidity. Despite breakthroughs in past decades to understand its pathogenesis and the development of transforming disease-modifying antirheumatic drugs, the symptoms of many patients are not substantially improved. Sinomenine (SIN), a natural alkaloid with poor solubility, has been used to treat RA in China for years because of its unique immunoregulative activity. However, its commercial hydrochloride form has a short half-time, which may cause huge fluctuations of blood drug concentration leading to severe adverse reactions. In this study, co-amorphous systems of SIN with three nonsteroidal anti-inflammatory drugs (NSAIDs), including indomethacin, naproxen, and sulindac, were prepared for the combination therapy, as well as the improvement of its aqueous solubility and controlled release. Each co-amorphous sample was characterized by powder X-ray diffraction (PXRD), temperature-modulated differential scanning calorimetry (mDSC), and Fourier transform infrared spectroscopy (FTIR). The CO

Topics: Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Rheumatoid; Calorimetry, Differential Scanning; Delayed-Action Preparations; Drug Combinations; Drug Stability; Humans; Morphinans; Solubility; Spectroscopy, Fourier Transform Infrared; X-Ray Diffraction

Sinomenine (SIN) is a clinical drug for treating rheumatoid arthritis (RA) in China. Our previous study found SIN inhibited inflammation via alpha7 nicotinic acetylcholine receptor (α7nAChR) in macrophages in vitro. Adenosine receptor A

Topics: alpha7 Nicotinic Acetylcholine Receptor; Animals; Antirheumatic Agents; Arthritis, Experimental; Arthritis, Rheumatoid; Male; Morphinans; NF-kappa B; Rats; Rats, Sprague-Dawley; Receptor, Adenosine A2A; Synoviocytes

Our present study focused on assessing whether Sinomenine (SIN) could attenuate sepsis-induced acute lung injury (ALI).. The mice were conditioned with SIN 1 h before intraperitoneal injection of lipopolysaccharide (LPS). Lung wet/dry (W/D) ratio, inflammatory level in bronchoalveolar lavage fluid (BALF), malondialdehyde (MDA) levels, superoxide dismutase (SOD) activity and inflammatory cytokines production were detected. The expression of nuclear factor erythroid 2-like 2 (Nrf2) and autophagy-related proteins were detected by Western blot and immunohistochemical analyses. In addition, the RAW264.7 cells were treated with SIN 1 h before treatment with LPS. Inflammatory cytokines, iNOS and COX2 were detected. The expression of Nrf2 and autophagy-related proteins were explored by Western blot analysis.. Experiments in vivo and in vitro discovered that LPS significantly increased the degree of injury, inflammatory cytokines production and oxidative stress. However, the increase was significantly inhibited by treatment of SIN. In addition, SIN was found to upregulate the expression of Nrf2 and autophagy-related proteins both in vivo and in vitro.. Our data suggested that SIN could attenuate septic-associated ALI effectively, probably due to the inhibition of inflammation and oxidative stress through Nrf2 and autophagy pathways.

Topics: Acute Lung Injury; Animals; Autophagy; Cytokines; Female; Inflammation; Kelch-Like ECH-Associated Protein 1; Lipopolysaccharides; Mice; Mice, Inbred ICR; Morphinans; NF-E2-Related Factor 2; Oxidative Stress; RAW 264.7 Cells; Sepsis

This study is designed to investigate the effects and mechanisms of sinomenine (Sin) in stress load-induced heart failure in mice.. We used aortic constriction (AB) to cause pressure overload as our heart failure model. Sin was received in mice as the treatment group. Cardiac function and structural changes were detected using echocardiography. Heart-lung mass ratios were measured. The serum levels of IL-10 and IL-17 proteins were detected by using ELISA, cardiac hypertrophy markers atrial natriuretic peptide (ANP), myocardial I and III collagen mRNA levels were detected by RT-PCR. Myocardial type I and III collagen protein levels were detected by Western blotting.. Sin significantly improved stress load-induced heart failure (P < 0.05), reduced the heart-lung mass ratio, ANP, collagen-I and -III mRNA and protein levels (P < 0.05); Sin can enhance the ratio of IL-10/IL-17.. Sin may be a promising drug target to improve heart failure. Its role is related to reduce serum ANP levels, inhibit the mRNA and protein level of type I and III collagen and enhance the ratio of IL-10/IL-17.

Topics: Animals; Atrial Natriuretic Factor; Cardiotonic Agents; Collagen Type I; Collagen Type III; Disease Models, Animal; Heart Failure; Interleukin-10; Interleukin-17; Male; Mice; Mice, Inbred C57BL; Morphinans

Plasma cell mastitis (PCM) is a special form of mastitis characterized by periductal inflammation and large-scale plasma cell infiltration. At present, the recurrence rate of PCM after excision is quite high, making PCM a major problem for mammary surgeons. However, no effective drug exists for the treatment of PCM. Numerous studies have demonstrated that Sinomenine hydrochloride (SH) has potent anti-inflammatory and immunoregulatory properties. However, the efficacy and the underlying mechanisms of SH in the treatment of PCM remain unclear. In the present study, we first investigated the therapeutic effects of SH in the PCM mouse model and clarified the possible mechanisms. We found that the levels of plasmocytes and lymphocytes infiltration were alleviated significantly in the 100 mg/kg SH group compared to the control group. In addition, few CD138+ plasma cells were found in the mammary glands of the 100 mg/kg SH group. The levels of Bcl-2 in the 100 mg/kg SH group were dramatically decreased compared with those in the saline group. Mechanistically, we demonstrated that SH inhibited the progression of PCM mainly through downregulating IL-6/JAK2/STAT3 levels. Collectively, our results suggested that SH could inhibit the progression of PCM by suppressing IL-6/JAK2/STAT3 cascades and ultimately achieve a therapeutic effect in PCM. This study provides theoretical support for the clinical application of SH in the treatment of PCM.

Topics: Animals; Anti-Inflammatory Agents; Disease Models, Animal; Disease Progression; Female; Humans; Interleukin-6; Janus Kinase 2; Mammary Glands, Animal; Mastitis; Mice; Morphinans; Plasma Cells; Signal Transduction; STAT3 Transcription Factor; Tyrphostins

The conventional medications are still facing a huge challenge for the treatment of rheumatoid arthritis (RA). Thus, looking for an effective therapy of RA has became an urgent issue nowadays. In this study, a novel thermosensitive liposome loaded with sinomenine hydrochloride (SIN-TSL) was developed by a pH gradient method. The SIN-TSL had a mean particle size of around 100 nm, and an high entrapment efficiency and drug loading capacity. The results also suggested that SIN-TSL had a thermosensitive drug release behaviour, with the drug release rate at 43 °C was much faster than the one at 37 °C. The SIN-TSL could be effectively taken up by lipopolysaccharide-activated HUVECs, without any cytotoxicity was observed. In addition, both in vitro and in vivo studies indicated that the SIN-TSL combined with microwave hyperthermia exhibited superior anti-rheumatoid arthritis effect. Overall, these results suggest that SIN-loaded thermosensitive liposomes combined with microwave hyperthermia could provide an optional strategy for alleviating the clinical symptoms of RA.

Topics: 1,2-Dipalmitoylphosphatidylcholine; Animals; Antirheumatic Agents; Arthritis, Rheumatoid; Cells, Cultured; Cholesterol; Combined Modality Therapy; Cytokines; Disease Models, Animal; Drug Compounding; Drug Liberation; Female; Human Umbilical Vein Endothelial Cells; Humans; Hyperthermia, Induced; Joints; Lipids; Liposomes; Microwaves; Morphinans; Particle Size; Rats, Wistar; Solubility

To explore the potential targets underlying the effect of sinomenine (SIN) on rheumatoid arthritis (RA) by utilizing a network pharmacology approach.. SIN and its drug targets were identified using network analysis followed by experimental validation. First, the Pharmmapper, UniProt and GeneCards databases were mined for information relevant to the prediction of SIN targets and RA-related targets. Second, the SIN-target gene and SIN-RA target gene networks were created in Cytoscape software followed by the collection of the candidate targets of each component by R software. Eventually, the key targets and enriched pathways were examined by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis.. Sixty-seven potential targets of SIN and 3797 related targets involved in RA were subjected to network analysis, and the 20 intersection targets indicated the principal pathways linked to RA. Additionally, 16 key targets, which were linked to more than three genes, were determined to be crucial genes. GO analysis showed that 14 biological processes, 5 cellular components and 2 molecular functions were identified, when corrected by a P value ≤ 0.01. Seven related signaling pathways were identified by KEGG analysis, when corrected according to a Bonferroni P value ≤ 0.05.. The present study explored the potential targets and signaling pathways of SIN during the treatment of RA, which may help to illustrate the mechanism (s) involved in the action of SIN and may provide a better understanding of its anti-rheumatoid arthritis effects in terms of inhibiting angiogenesis, synovial hyperplasia, and bone destruction.

Topics: Antirheumatic Agents; Arthritis, Rheumatoid; Computational Biology; Gene Regulatory Networks; Humans; Hyperplasia; Molecular Docking Simulation; Morphinans; Neovascularization, Pathologic; Protein Interaction Maps; Signal Transduction; Synovial Membrane

Premature and ill neonates undergo painful but medically necessary procedures while hospitalized. Although opiate drugs are administered as analgesics, problems associated with their side effects, tolerance, and potential dependence necessitate research into alternative pain-relieving medications. Here we test two plant-derived compounds in infant rats: sinomenine, which targets the Mas-related G-protein-coupled receptor member X2 opioid receptor; and salvinorin A, which is a κ opioid receptor agonist. In adult animals both sinomenine and salvinorin A are analgesic, but neither has been tested in infants.. We used the formalin and thermal plantar tests in rats 7 and 21 days of age (PN7 and PN21) for behavioral signs of pain. In addition, brain sections were stained using Fos immunohistochemistry to examine patterns of brain activation in the midbrain periaqueductal gray and the paraventricular nucleus of the hypothalamus.. Sinomenine was analgesic in both the formalin and thermal tests on animals 21 days of age. At PN7 only the highest dose elevated response latencies in the thermal test and there were no effects of sinomenine in the formalin test. Analysis of Fos expression in the sinomenine-treated animals showed no drug effect, in contrast to the behavioral results. Salvinorin A was analgesic in the formalin test only at the highest dose at 21 days of age but not in the thermal test at either age.. The increased modest effectiveness of sinomenine in older animals and the minimum salvinorin A drug effect suggest that the compounds act on sites that develop during the preweaning period (sinomenine) or after weaning (salvinorin A).

Topics: Analgesics; Analgesics, Opioid; Animals; Behavior, Animal; Diterpenes, Clerodane; Hot Temperature; Humans; Infant; Infant, Newborn; Morphinans; Pain; Pain Measurement; Phytotherapy; Plant Extracts; Rats, Long-Evans; Receptors, Opioid; Receptors, Opioid, kappa; Salvia; Sinomenium

Amyloid beta is a major constituent of the plaques found in the brains of patients suffering from Alzheimer's disease (AD). A growing body of research work suggests that neuroinflammation plays important roles in the development of AD. Thus, considerable efforts are directed towards identification of compounds that can reduce or inhibit neuroinflammation. Here, we show that sinomenine, a compound present in a Chinese medicinal plant, Sinomenium acutum, inhibits oligomeric amyloid beta-induced production of reactive oxygen species (ROS), nitric oxide (NO) and inflammation-related molecules from astrocytic cells. The conditioned medium from oligomeric amyloid beta-treated astrocytic cells induces cell death in the hippocampal neuronal cells. Importantly, sinomenine inhibits this cell death. In addition, this compound has inhibitory effects on the production of ROS, NO and inflammation-related factors from oligomeric amyloid-beta treated human astrocytes. Finally, the conditioned medium from oligomeric amyloid beta-treated human astrocytes induces cell death in the primary culture of human neurons, which is inhibited by sinomenine. Thus, sinomenine inhibits amyloid beta-induced production of toxic factors from astrocytes, and confers protection to hippocampal neuronal cells as well as human neurons against indirect toxicity. The results suggest that this compound could provide beneficial effects in AD and other neurodegenerative conditions by reducing inflammation and neuronal cell death.

Topics: Amyloid beta-Peptides; Animals; Astrocytes; Cell Line; Hippocampus; Humans; Inflammation; Mice; Morphinans; Neurons; Neuroprotective Agents; Nitric Oxide; Reactive Oxygen Species

Sinomenine (SN) has been used in the clinical treatment of systemic lupus erythematosus and rheumatoid arthritis for many years. Studies showed that SN held protective effects such as anti-inflammation, scavenging free radicals and suppressing immune response in many autoimmune diseases. The purpose of the present study is to explore the mechanism of anti-inflammation of SN on lipopolysaccharide (LPS)-induced macrophages activation and investigate whether the TLR4/NF-κB signaling pathway participated in. Macrophages isolated from mouse peritoneal cavity were stimulated by 1 µg/mL LPS for 24 h. And then the cells were treated with various concentrations of SN, TLR4 inhibitor respectively for additional 48 h. Drug toxicity was detected by MTT assay and Transwell experiment was used to assess chemotaxis. Furthermore, TLR4 and MyD88 mRNA levels were detected by real-time PCR. Western blotting was used to examine TLR4, MyD88 and phosphorylated IκB protein expression in macrophages. Immunofluorescence assay was applied to observe p65 NF-κB protein expression in macrophage nucleus. We extracted macrophages with high purity and activity from the abdominal cavity of mice. SN remarkably inhibited the chemotaxis and secretion function of LPS-stimulated macrophages. It also down-regulated both the protein levels of inflammatory cytokines (TNF-α, IL-1β and IL-6) and the RNA and protein levels of the key factors (TLR4, MyD88, P-IκB) in TLR4 pathway. The expression of p65 NF-κB protein in nuclei was down-regulated, which was correlated with a similar decrease in P-IκB protein level. In conclusion, SN can inhibit the LPS induced immune responses in macrophages by blocking the activated TLR4/NF-κB signaling pathway. These results may provide a therapeutic approach to regulate inflammatory responses.

Topics: Animals; Anti-Inflammatory Agents; Cell Survival; Cells, Cultured; Chemotaxis; Gene Expression Regulation; Lipopolysaccharides; Macrophages; Male; Mice; Morphinans; Myeloid Differentiation Factor 88; NF-kappa B; Signal Transduction; Toll-Like Receptor 4

Sinomenine (SIN) is an anti-inflammatory alkaloid derived from

Topics: Administration, Cutaneous; Animals; Antirheumatic Agents; Chondroitin Sulfates; Drug Delivery Systems; Drug Liberation; Morphinans; Needles; Permeability; Povidone; Rats; Rats, Sprague-Dawley; Skin; Skin Absorption

Traumatic brain injury (TBI) is a significant medical problem with limited treatment options and is one of the main causes of life-long disability. Neuroinflammation orchestrated by activated microglia/macrophages at the site of injury plays a critical role in the onset of many pathological events following TBI, leading to blood brain barrier (BBB) dysfunction, neuronal damage and long term neuronal and behavioral deficits. Current treatment involves intravenous administration of anti-inflammatory drugs which have limited clinical outcomes only when dosed within the early time window after injury. Hence there is an urgent need to develop improved drug delivery systems which have potential to cross impaired BBB, target and deliver drugs selectively to activated microglia/macrophages at the sites of injury, and suppress the detrimental effects of acute inflammation. In this study, we have used Sinomenine (Sino), a potent anti-inflammatory and antioxidant drug conjugated to hydroxyl terminated generation-4 PAMAM dendrimer (D-Sino) as a potential therapy for attenuating early inflammation in TBI. D-Sino conjugates were synthesized using highly robust copper-catalyzed click reaction with high purity. D-Sino conjugates enhanced the intracellular availability of Sino due to their rapid cellular uptake, significantly attenuated early/acute inflammation by suppressing pro-inflammatory cytokines (TNF-α, IL-1β, CCL-3 and IL-6), and reduced oxidative stress (iNOS and NO) in LPS activated murine macrophages (RAW 264.7) by inhibiting NF-κB activation and its nuclear translocation (the root cause for inflammation inception) significantly more as compared to the free drug. Upon systemic administration in a rabbit model of pediatric TBI, D-Sino conjugates specifically targeted activated microglia/macrophages at the site of injury in the brain. Single dose of D-Sino attenuated inflammation in the injured brain areas by suppressing inflammatory cytokines expression whereas free Sino treatment did not demonstrate a significant effect. Together, these results suggest that D-Sino conjugate may open up new avenues for increasing the therapeutic window in the treatment of early inflammation and for improving the efficacy of the drug in TBI. Moreover, this treatment can work in conjunction with current clinical practices such as therapeutic hypothermia and pharmacologically induced coma for many indications associated with TBI, where acute inflammation plays a critical role in di

Topics: Animals; Brain Injuries, Traumatic; Child; Dendrimers; Disease Models, Animal; Humans; Mice; Microglia; Morphinans; Rabbits

Topics: Antineoplastic Agents; Cell Line, Tumor; Cell Movement; Gene Expression Regulation; Humans; Lung Neoplasms; Matrix Metalloproteinase 9; Matrix Metalloproteinases; MicroRNAs; Morphinans; Tissue Inhibitor of Metalloproteinase-2

Oxidative stress, inflammation and pyroptosis are three of the most important mechanisms in the pathophysiology of temporal lobe epilepsy (TLE). Most people with TLE are refractory to the existing drugs. Sinomenine has shown neuroprotective effects through counteracting oxidative stress, inflammation and pyroptosis. In this study, we evaluated the effect of sinomenine on seizure behavior, oxidative stress, inflammation and pyroptosis markers in addition to its neuroprotective potential in intrahippocampal kainate-induced rat model of TLE. For this purpose, male rats (n = 60) were randomly divided into five groups, i.e., sham, kainate (lesion) with an intrahippocampal injection of kainate, kainate groups receiving sinomenine at doses of 30 or 50 mg/kg, and kainate group receiving valproic acid at a dose of 200 mg/kg (as the positive control). Our obtained data showed that sinomenine administration at a dose of 50 mg/kg can significantly decreases severity of seizures and incidence of status epilepticus (SE), hippocampal aberrant MFS and DNA fragmentation and prevents reduction of neuronal density. It also significantly restored level of ROS, MDA, HO-1 and SOD but its effect on GSH level was not significant. Additionally, sinomenine at a dose of 50 mg/kg partially counteracted the increase of NF-κB, TLR 4, TNFα, GFAP and caspase 1. These results suggest that sinomenine has anticonvulsant and neuroprotective effects by reducing hippocampal oxidative stress, inflammation, pyroptosis and apoptosis in intrahippocampal kainate model of TLE.

Topics: Animals; Anticonvulsants; Disease Models, Animal; Epilepsy, Temporal Lobe; Hippocampus; Inflammation; Kainic Acid; Male; Morphinans; Neuroprotective Agents; Oxidative Stress; Pyroptosis; Rats; Reactive Oxygen Species

Topics: Administration, Topical; Animals; Area Under Curve; Drug Carriers; Elasticity; Joints; Male; Microdialysis; Microscopy, Confocal; Monoterpenes; Morphinans; Polyethylene Glycols; Rabbits; Rats; Rats, Sprague-Dawley; ROC Curve; Thermodynamics; Transition Temperature

Liver disease is common and often life-threatening. Sinomenine (SIN) is an active ingredient extracted from . In vivo experiments, mice were randomly divided into six groups (n=10): control group, model group, SIN (25 mg/kg) group, SIN (50 mg/kg) group, SIN (100 mg/kg) group and SIN (100 mg/kg) + SRI-011381 group. Alanine transaminases (ALT), aspartate transaminases (AST) and alkaline phosphatase (ALP) were detected. The pathological lesion was measured by HE staining. Apoptosis was measured by TUNEL staining. In vitro experiments, BRL-3A cells were treated with APAP (7.5 mM) and then subjected to various doses of SIN (10, 50 and 100 μg/mL) at 37°C for 24 h. Inflammatory factors and oxidative stress index were measured by ELISA. The expression of proteins was detected by Western blot.. The results showed that compared with the control group, the levels of ALT, AST and ALP in the serum of APAP-induced mice were significantly increased, followed by liver histological damage and hepatocyte apoptosis. Besides, APAP reduced the activity of SOD and GSH-Px, while increasing the content of MDA and LDH. Notably, APAP also promoted the expression of NLRP3, ASC, caspase-1 and IL-1β. Interestingly, SIN treatment dose-dependently reduced APAP-induced liver injury and oxidative stress, inhibited the activation of NLRP3 inflammasomes, and reduced the levels of inflammatory cytokines. In vitro studies have shown that SIN treatment significantly reduced the viability of BRL-3A cells and oxidative stress and inflammation. In addition, the Western blotting analysis showed that SIN inhibited the activation of TGF-β/Smad pathway in a dose-dependent manner in vitro and in vivo. These effects were significantly reversed by TGF-β/Smad activator SRI-011381 or TGF-β overexpression.. The study indicates that SIN attenuates APAP-induced acute liver injury by decreasing oxidative stress and inflammatory response via TGF-β/Smad pathway in vitro and in vivo.

Topics: Acetaminophen; Animals; Apoptosis; Cells, Cultured; Chemical and Drug Induced Liver Injury; Dose-Response Relationship, Drug; Inflammation; Mice; Mice, Inbred C57BL; Morphinans; Oxidative Stress; Smad Proteins; Structure-Activity Relationship; Transforming Growth Factor beta

This study aimed to evaluate the effects of two different structural alcohol permeation enhancers (menthol and propylene glycol) on the internal structure and in vitro properties of the dual drug-loaded lyotropic liquid crystalline (LLC) gels. The LLC gels were prepared and characterized by polarized light microscopy, small-angle X-ray scattering, differential scanning calorimetry, attenuated total reflectance-Fourier transform infrared spectrum, and rheology. Based on the results, the inner structure of the gels was Q

Topics: Acrolein; Administration, Cutaneous; Animals; Drug Delivery Systems; Drug Liberation; Female; Gels; Liquid Crystals; Male; Menthol; Morphinans; Propylene Glycol; Rats; Rats, Sprague-Dawley

Sinomenine was found to play anti-cancer functions in different type of cancers, while the mechanisms underlying the anticancer effects of sinomenine in retinoblastoma (RB) remains unclear. The present study was designed to explore the impacts of sinomenine on cell proliferation and invasion ability of RB cells and the related mechanism. Human retinoblastoma cell line WERI-RB-1 and Y79 cells were cultured and treated by different concentration of sinomenine, and then the proliferation ability of the cells was determined via performing the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) proliferation assay. The cell apoptosis was examined via performing the flow cytometry assay. Then scratch wound healing analysis as well as and transwell invasion analysis have been performed to determine the effect of sinomenine on cell migration ability as well as invasion ability. The proteins level of phosphatidylinositol 3-kinase (PI3K)/AKT signaling molecules were determined with Western blot assay. We found that sinomenine was able to decrease the proliferation and promote the apoptosis of RB cells in a dose-dependent manner; moreover, sinomenine also significantly suppressed the migration as well as invasion ability of WERI-RB-1 and Y79 cells in vitro. Furthermore, sinomenine also de-activated PI3K/AKT signaling in WERI-RB-1 cells via inhibited the phosphorylation of PI3K and AKT proteins. Sinomenine can exert anti-tumor function on RB cells in vitro, therefore sinomenine might be a potential alterative medication for the treatment for RB.

Topics: Cell Movement; Cell Proliferation; Drug Screening Assays, Antitumor; Humans; Morphinans; Neoplasm Invasiveness; Phosphatidylinositol 3-Kinase; Phosphorylation; Proto-Oncogene Proteins c-akt; Retinal Neoplasms; Retinoblastoma; Signal Transduction

Sinomenium acutum stem is a popular traditional Chinese medicine used to treat bone and joint diseases. Sinomenine is considered the only chemical marker for the quality control of S. acutum stem in mainstream pharmacopeias. However, higenamine in S. acutum stem is a novel stimulant that was banned by the World Anti-Doping Agency in 2017. Therefore, enhancing the quality and safety control of S. acutum stem to avoid potential safety risks is of utmost importance. In this study, a fast, sensitive, precise, and accurate method for the simultaneous determination of 11 alkaloids in S. acutum stem by ultrahigh-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UHPLC-QQQ-MS/MS) was established. This method successfully analyzed thirty-five batches of S. acutum stem samples. The average contents of sinomenine, magnoflorine, coclaurine, acutumine, higenamine, sinoacutine, palmatine, magnocurarine, columbamine, 8-oxypalmatine, and jatrorrhizine were 24.9 mg/g, 6.35 mg/g, 435 μg/g, 435 μg/g, 288 μg/g, 44.4 μg/g, 22.5 μg/g, 21.1 μg/g, 15.8 μg/g, 9.30 μg/g, and 8.75 μg/g, respectively. Multivariate analysis, including principal component analysis (PCA), orthogonal partial least square method-discriminant analysis (OPLS-DA), and hierarchical cluster analysis (HCA), were performed to characterize the importance and differences among these alkaloids in S. acutum stem samples. As a result, sinomenine, magnoflorine, coclaurine, acutumine, and higenamine are proposed as chemical markers for quality control. Higenamine and coclaurine are also recommended as chemical markers for safety control. This report provides five alkaloids that can be used as chemical markers for improving the quality and safety control of S. acutum stem. It also alerts athletes to avoid the risks associated with consuming S. acutum stem.

Topics: Alkaloids; Aporphines; Chromatography, High Pressure Liquid; Cluster Analysis; Drugs, Chinese Herbal; Isoquinolines; Least-Squares Analysis; Morphinans; Plant Extracts; Plant Stems; Principal Component Analysis; Sinomenium; Solvents; Spiro Compounds; Tandem Mass Spectrometry; Tetrahydroisoquinolines

Matrix metalloproteinases (MMPs) are required for collagen degradation which play a key pathological role in arthritis progression. Herein, the effect of sinomenine (SN) on Interleukin 1 beta (IL-1β)-induced MMPs production and its underlying mechanism were explored in SW1353 cells. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that 200 and 400 µM SN significantly inhibited SW1353 cell proliferation, thus the lower dose of SN (25-100 µM) were used in the subsequent experiments. Notably, the increased mRNA and protein levels of suppressor of cytokine signaling (SOCS) 3 were dose-dependently induced by SN. SN significantly suppressed mRNA and protein levels of MMPs in IL-1β-induced SW1353 cells. Through Western blot analysis, SN showed inhibitory effect on IL-1β-induced TAK1 and p65 phosphorylation. Moreover, SN blocked the interaction of TRAF6 and TAK1 resulting in inactivation of IL-1β pathway. Mechanistically, the inhibitory effect of SN on MMPs levels alongside TRAF6 and TAK1 interactions was abrogated by silencing SOCS3. Moreover, SN did not inhibit TAK1 kinase activity. In TAK1 silencing cells, the levels of MMPs and p65 phosphorylation of SN-treatedcells were lower than dimethyl sulfoxide (DMSO)-treated cells, indicating that blocking interaction was not a unique way for SN to inhibit MMPs levels. Finally, SN significantly inhibited IL-6-induced Janus tyrosine kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) phosphorylation in SW1353 cells. The levels of JAK2 phosphorylation and MMPs did not show a significant difference between IL-6 + SOCS3-small interfering RNA (siRNA) + SN group and IL-6 + SOCS3-siRNA + DMSO group. These findings demonstrated that SOCS3 expression was increased by SN blocked IL-1β-induced interaction between TRAF6 and TAK1 as well as IL-6 pathway activation, thereby culminating in the inhibition of MMPs levels.

Topics: Arthritis; Cell Line, Tumor; Cell Proliferation; Chondrocytes; Humans; Interleukin-1beta; Matrix Metalloproteinase Inhibitors; Matrix Metalloproteinases; Morphinans; Suppressor of Cytokine Signaling 3 Protein; Up-Regulation

Sinomenine (SIN) is clinically used as an anti-rheumatic drug. However, the metabolic and pharmacological mechanisms of SIN combined with its metabolites are unclear. This study aims to explore the cyclic metabolic mechanism of SIN, the anti-inflammation effects of SIN and its major metabolites (N-demethylsinomenine (DS) and sinomenine-N-oxide (SNO)), and the oxidation property of SNO.. SIN was administrated to rats via gavage. Qishe pills (a SIN-containing drug) were orally administrated to humans. The bio-samples were collected to identify SIN's metabolites. Enzymatic and non-enzymatic incubations were used to reveal SIN's metabolic mechanism. Impacts of SIN, SNO and DS on the inflammation-related cytokine's levels and nuclear translocation of NF-κB were evaluated in LPS-induced Raw264.7 cells. ROS induced by SNO (10 μM) was also assessed.. CYP3A4 and ROS predominantly mediated the formation of SNO, and CYP3A4 and CYP2C19 primarily mediated the formation of DS. Noteworthily, SNO underwent N-oxide reduction both enzymatically, by xanthine oxidase (XOD), and non-enzymatically, by ferrous ion and heme moiety. The levels of IL-6 and TNF-α and nuclear translocation of NF-κB were ameliorated after pretreatment of SIN in LPS-induced Raw264.7 cells, while limited attenuations were observed after pretreatment of DS (SNO) even at 200 μM. In contrast, SNO induced ROS production.. This study elucidated that SIN underwent both enzymatic and non-enzymatic cyclic metabolism and worked as the predominant anti-inflammation compound, while SNO induced ROS production, suggesting more studies of SIN combined with SNO and DS are necessary in case of DDI and potential toxicities.

Topics: Animals; Anti-Inflammatory Agents; Female; Humans; Inflammation; Male; Mice; Microsomes, Liver; Morphinans; Oxidative Stress; Rats; Rats, Sprague-Dawley; RAW 264.7 Cells

In comparison with pristine sinomenine and carborane precursors, the calculations of molecular docking with matrix metalloproteinases (MMPs) and methylcarboranyl-

Topics: Animals; Antineoplastic Agents; Antirheumatic Agents; Boron; Boron Neutron Capture Therapy; Cell Line, Tumor; Drug Evaluation, Preclinical; Drug Screening Assays, Antitumor; Magnetic Resonance Spectroscopy; Matrix Metalloproteinase 1; Matrix Metalloproteinase 13; Molecular Docking Simulation; Morphinans; Rats; Spectroscopy, Fourier Transform Infrared; Synoviocytes

To investigate the effect of topical sinomenine (SIN) on ocular surface damage in experimental dry eye in mice.. Experimental dry eye was created using scopolamine hydrobromide in female C57BL/6 mice. Eye drops consisting of 0.05%, or 0.1% SIN or phosphate-buffered saline (PBS) were applied to the experimental dry eye in mice. Tear product and corneal staining scores were measured at 7 and 14 days after treatment. Interleukin (IL)-1β and tumour necrosis factor (TNF)-α levels in the SIN groups at 14 days after treatment were compared with those of other groups.. Mice treated with 0.05% or 0.1% SIN showed a significant improvement in tear product and corneal irregularity compared to the control and PBS-treated groups. A significant decrease in the levels of IL-1βand TNF-α was observed in the 0.05% and 0.01% SIN-treated groups.. Topical SIN eye drop application can effectively improve clinical signs and decrease inflammation in the ocular surface, and alleviate ocular surface damage in dry eye.

Topics: Animals; Cornea; Disease Models, Animal; Dry Eye Syndromes; Female; Interleukin-1beta; Mice, Inbred C57BL; Morphinans; Ophthalmic Solutions; Scopolamine; Tears; Tumor Necrosis Factor-alpha

Sinomenine (Sino) is diffusely applied in heal rheumatoid arthritis and neuralgia. Howbeit, the activities of Sino in breast cancer cells remain confused. The research attempted to probe the anti-tumor function of Sino in breast cancer cells and divulge the feasible molecular mechanism. Sion at the 1-16 μM concentrations was exploited for the exposure of MDA-MB-231 or MCF7 cells, and cell growth, migration, invasion, cell cycle-relevant and apoptosis-correlative factors were estimated. Micro RNA (miR)-29 expression was evaluated via enforcing qRT-PCR, and the actions of miR-29 in MDA-MB-231 cells growth, migration and invasion were appraised after the overexpressed or suppressed vectors transfection. The functions of PDCD-4 in JNK and MEK/ERK pathways were estimated by employing western blot. We found that, Sino exposure impeded cell proliferation, provoked cell apoptosis and barricaded cell migration and invasion in MDA-MB-231 and MCF7 cells. Enhancement of miR-29 was observed in Sino-managed cells, and miR-29 overexpression further potentiated the activities of Sino in MDA-MB-231 cells. Additionally, Sino remarkably enhanced PCDC-4 expression via adjusting miR-29 in MDA-MB-231 cells. Beyond that, overexpressed PCDC-4 obstructed JNK and MEK/ERK pathways in MDA-MB-231 cells. Taken together, the explorations unveiled that Sino restrained MDA-MB-231 cells proliferation, migration, invasion, and provoked apoptosis through modulation of miR-29/PDCD-4 axis. Highlight Sino inhibits MDA-MB-231 and MCF7 cells proliferation and provokes apoptosis; Sino restrains MDA-MB-231 and MCF7 cells migration and invasion; Sino ascends miR-29 expression in MDA-MB-231 and MCF7 cells; Sino adjusts cell growth, migration and invasion via modulating miR-29; Sino up-regulates PDCD-4 expression through mediating miR-29; PDCD-4 obstructs JNK and MEK/ERK pathways in MDA-MB-231 cells.

Topics: Antineoplastic Agents; Apoptosis; Apoptosis Regulatory Proteins; Breast Neoplasms; Cell Movement; Cell Proliferation; Humans; JNK Mitogen-Activated Protein Kinases; MAP Kinase Signaling System; MCF-7 Cells; MicroRNAs; Morphinans; Neoplasm Invasiveness; RNA-Binding Proteins

Sinomenine (SIN) has been reported its antitumor effects on various types of human cancers, but there is no available information regarding the antitumor effects of SIN and cisplatin on gastric cancer. Here, we examined the antitumor effects of SIN combined with cisplatin on gastric cancer cells as well as the underlying biological mechanisms. CCK-8 assay and Calcusyn 2.0 software analysis, Hoechst 33258 staining and flow cytometry, transwell assay showed that SIN and cisplatin synergistically inhibited growth, induced apoptosis, and suppressed invasion than did either drug alone in gastric cancer cells. Interestingly, no change in the AKT level was found, whereas SIN and cisplatin led to a dramatic decrease in p-AKT level compared with either alone treatment. SIN and cisplatin further decreased the Bcl-2, procaspase-3, and β-catenin, but increased Bax, cleaved dcaspase 3, MMP9, and MMP2 in combined group than in either alone group. Immunofluorescence staining showed again a significant decrease in nucleus β-catenin was found in combined group. These data suggested that SIN sensitizes human gastric cancer cells to cisplatin through negative regulation of PI3K/AKT/Wnt signaling pathway. In conclusion, SIN and cisplatin exerted synergistic antitumor effects in gastric cancer cells and might constitute a promising therapeutic approach for gastric cancer.

Topics: Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Cell Line, Tumor; Cell Proliferation; Cisplatin; Drug Synergism; Humans; Morphinans; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Stomach Neoplasms; Wnt Signaling Pathway

The goal of this paper was to develop and evaluate dual component-loaded with the hydrophilic sinomenine hydrochloride (SH) and lipophilic cinnamaldehyde (CA) cubic liquid crystal gels for transdermal delivery. The gels was prepared with a vortex method using phytantriol/water (70:30, w/w) and characterized by polarized light microscopy, small-angle X-ray scattering and rheology. The inner structure of the gels were Pn3m cubic phase and exhibited a pseudoplastic fluid behavior. Furthermore, the

Topics: Acrolein; Administration, Cutaneous; Animals; Antirheumatic Agents; Arthritis, Rheumatoid; Drug Carriers; Drug Combinations; Drug Compounding; Drug Evaluation, Preclinical; Drug Liberation; Fatty Alcohols; Gels; Hydrophobic and Hydrophilic Interactions; Liquid Crystals; Male; Morphinans; Rats; Skin; Water

This study aimed to explore the effect of sinomenine on the proliferation and differentiation of MC3T3-E1 cells and the related mechanism. Mouse preosteoblastic cell line MC3T3-E1 cells were divided into four groups: control group, treatment of sinomenine with the concentrations of 100, 500 or 1000 μM. The proliferation and apoptosis of the cells were determined by MTT assay at 0, 24, 48 and 72 h, and flow cytometry method was used to determine the effect of sinomenine on the apoptosis of MC3T3-E1 cells at 72 h; furthermore, after 72 h, the cell culture supernatants were collected, and the levels of alkaline phosphatase and osteocalcin were examined by enzyme-linked immunosorbent assay (ELISA); next, cells were collected, and the expression of type I collagen (COL1A1), osteopontin (OPN), protein kinase B (Akt), runt-related transcription factor 2 (Runx2), B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) were examined by RT-qPCR and Western Blot methods. It was observed that treatment of 500 and 1000 μM sinomenine has induced significant increase in the proliferation and decrease in the apoptosis of MC3T3-E1 cells; furthermore, sinomenine also lead to increased secretion of osteocalcin and alkaline phosphatase in MC3T3-E1 cells; finally, sinomenine induced marked increase in the expression of type I collagen, osteopontin and also induced the activation of the Akt/Runx2 signaling pathway. To sum up, we observed that sinomenine may promote the proliferation and differentiation of MC3T3-E1 cells via activating the Akt/Runx2 signaling pathway.

Topics: Animals; Apoptosis; Cell Line; Cell Proliferation; Cell Survival; Collagen Type I; Core Binding Factor Alpha 1 Subunit; Mice; Morphinans; Osteoblasts; Osteocalcin; Osteopontin; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2

Traumatic spinal cord injury (SCI) is a devastating condition with few efficacious drugs. Sinomenine, a bioactive alkaloid extracted from medicinal herb, has been used as a treatment of rheumatoid diseases. This present study explored the therapeutic effects of sinomenine on locomotor dysfunction and neuropathology in SCI. Our findings revealed that sinomenine mitigated neurological deficits and enhanced neuronal preservation, paralleled with a reduction of apoptosis. Also, sinomenine significantly reduced inflammatory cytokines and oxidative stress factors. We further examined erythroid-2-related factor 2 (Nrf2) nuclear translocation, which mainly controls the coordinated expression of important antioxidant and detoxification genes. An increase in Nrf2 translocation from cytoplasm to nucleus and Nrf2-mediated transactivation was observed after sinomenine administration. Knocking down Nrf2 by siRNA could counteract sinomenine-mediated anti-oxidant stress and anti-inflammation following H

Topics: Animals; Female; Hydrogen Peroxide; Inflammation; Morphinans; NF-E2-Related Factor 2; Oxidative Stress; PC12 Cells; Rats; Rats, Sprague-Dawley; Signal Transduction; Spinal Cord Injuries; Thoracic Vertebrae

Sinomenine, an alkaloid originally isolated from the roots and the rhizome of Sinomenium acutum is used as a traditional Chinese herbal medicines for rheumatoid arthritis and neuralgia. The aims of this study were to investigate the effects of oral administration of shinomenine on formalin-induced nociceptive behavior in mice and the opioid receptor subtypes involved in the antinociceptive effects of sinomenine. Our findings showed that a single dose of oral-administrated sinomenine inhibited the formalin induced licking and biting responses in a dose-dependent manner. Intraperitoneal pretreatment with naloxone hydrochloride, an opioid receptor antagonist, and β-funaltrexamine hydrochloride (β-FNA), a selective μ-opioid receptor antagonist, significantly attenuated sinomenine induced antinociception, but not by naltrindole, a nonselective δ-opioid receptor antagonist and nor-binaltorphimine, a selective κ-opioid receptor antagonist. Furthermore, in western blot analysis, oral administration of sinomenine resulted in a significant blockage of spinal extracellular signal-regulated protein kinase (ERK1/2) activation induced by formalin. Naloxone hydrochloride and β-FNA significantly reversed the blockage of spinal ERK1/2 activation induced by sinomenine. These results suggest that sinomenine-induced anti nociceptive effect and blockage of spinal ERK1/2 activation may be triggered by activation of μ-opioid receptors.

Topics: Administration, Oral; Analgesics; Animals; Dose-Response Relationship, Drug; Formaldehyde; Male; Mice; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Morphinans; Naloxone; Naltrexone; Nociception; Receptors, Opioid, mu; Spinal Cord

Background Prostate cancer is an extremely common disease in males, and the mortality of prostate cancer has been rising year by year. Sinomenine has been reported to exhibit anti-tumor effect on various cancers, but the function of Sinomenine in prostate cancer remains unclear. The study aimed to explore the effect of Sinomenine on proliferation, apoptosis, migration and invasion in prostate cancer cells. Methods PC3 cells were stimulated by different concentrations of Sinomenine, then cell proliferation, migration, invasion and apoptosis were examined by CCK-8, BrdU, Transwell and flow cytometry, respectively. Subsequently, miR-23a mimic and miR-23a inhibitor were transfected into PC3 and LNCaP cells, cell proliferation, apoptosis, migration and invasion were reassessed in these transfected cells. The related factors of cell cycle, apoptosis, metastasis and PI3K/AKT and JAK/STAT signal pathways were measured by western blot. Results Sinomenine significantly suppressed cell proliferation, promoted apoptosis and inhibited migration and invasion, as well as down-regulated Cyclin D1, CDK4, Bcl-2, MMP-2, MMP-9, Vimentin protein levels and up-regulated p16 and Bax protein levels in PC3 cells. Additionally, Sinomenine decreased the expression level of miR-23a, and overexpression of miR-23a reversed the effects of Sinomenine on cell proliferation, apoptosis, migration and invasion in PC3 and LNCaP cells. Further, Sinomenine inactivated PI3K/AKT and JAK/STAT signal pathways by regulation of miR-23a. Conclusions The data suggested that Sinomenine could suppress cell proliferation, migration, invasion, and promote apoptosis of prostate cancer cells through regulation of miR-23a. These findings might provide a possible strategy for the clinical treatment of prostate cancer.

Topics: Apoptosis; Cell Movement; Cell Proliferation; Humans; Male; MicroRNAs; Morphinans; Neoplasm Invasiveness; Prostatic Neoplasms

Sinomenine (SIN) is an anti-inflammatory and antiarthritic alkaloid derived from Sinomenium acutum, and the product Zhengqing Fengtongning produced from SIN has been marketed in China for treating rheumatoid arthritis (RA). Interestingly, we recently found that SIN could significantly ameliorate bone destruction induced by breast cancer cells in mice. Micro-CT examination showed that bone loss of the trabecular bones in tumor-bearing mice was markedly decreased by i.p. treatment of SIN at 150 mg/kg body weight. A mechanistic study demonstrated that SIN could suppress osteoclast formation and bone absorption induced by both MDA-MB-231 cells and MDA-MB-231 cell-conditioned medium (MDA-MB-231 CM) in preosteoclastic RAW264.7 cells. The MDA-MB-231 CM-induced osteoclast-related genes TRAP and OSCAR were obviously downregulated by SIN. In addition, mRNA expression of c-Fos and NFATc1 and nuclear translocation of c-Fos and NFATc1 protein were inhibited by SIN during MDA-MB-231 CM-induced osteoclastogenesis, while NF-κB signaling was not impacted by SIN. More interestingly, SIN was demonstrated to decrease hIL-8 mRNA expression in cultured MDA-MB-231 cells and to inhibit hIL-8 protein expression in MDA-MB-231 cells cocultured with preosteoclastic RAW264.7 cells while simultaneously downregulating CXCR1, the ligand of IL-8 related to bone destruction, during MDA-MB-231 CM-induced osteoclastogenesis. Previously, IL-8/CXCR1 was reported to be associated with the pathogenesis and progression of RA, and SIN was observed to markedly ameliorate bone erosion of RA patients. Our current findings may extend the utilization of SIN to preventing osteoclastogenesis and bone destruction in breast cancer patients and may enable IL-8/CXCR1 to serve as new targets for both anticancer and antiarthritic drug discovery.

Topics: Animals; Cell Line, Tumor; Female; Humans; Interleukin-8; Mammary Neoplasms, Experimental; Mice; Mice, Inbred BALB C; Morphinans; NFATC Transcription Factors; Osteoclasts; Osteogenesis; Osteolysis; Proto-Oncogene Proteins c-fos; RAW 264.7 Cells; Receptors, Interleukin-8A; Signal Transduction

Sinomenine (SIN) has been shown to protect against IL-1β-induced chondrocyte apoptosis in vitro. However, the role of SIN in the anterior cruciate ligament transection (ACLT)-induced osteoarthritis (OA) mouse model and its underlying molecular mechanisms remain unclear. In the present study, the protective effect of SIN on ACLT-induced articular cartilage degeneration and IL-1β-induced chondrocyte apoptosis miR-223-3p/NLRP3 signaling regulation was investigated. Safranin O staining was performed to evaluate the pathological changes of articular cartilage. Chondrocyte apoptosis was measured with Annexin V-fluorescein isothiocyanate/polyimide (annexin V-FITC/PI) staining using flow cytometry. Gene and protein expression were detected by RT-qPCR and Western blotting, respectively. SIN administration markedly improved articular cartilage degradation in mice undergoing ACLT surgery. In addition, SIN treatment downregulated the levels of inflammatory cytokines and the protein expression of NLRP3 inflammasome components and upregulated the expression of miR-223-3p in OA mice and IL-1β-stimulated chondrocytes. In vitro, we found that NLRP3 was a direct target of miR-223-3p, and overexpression of miR-223-3p blocked IL-1β-induced apoptosis and the inflammatory response in chondrocytes. These findings indicate that miR-223-3p/NLRP3 signaling could be used as a potential target of SIN for the treatment of OA.

Topics: Animals; Antirheumatic Agents; Cartilage Diseases; Cartilage, Articular; Gene Expression Regulation; Inflammasomes; Mice; MicroRNAs; Morphinans; NLR Family, Pyrin Domain-Containing 3 Protein; Protective Agents; Signal Transduction

The objective of the present study is to investigate the inhibitory effects of sinomenine (SIN) on angiogenesis in a collagen-induced arthritis (CIA) mouse model.. Arthritis assessments for all mice were recorded. The histopathological assessments were performed following haematoxylin and eosin (HE) staining. Immunohistochemistry and enzyme-linked immunosorbent assay (ELISA) analyses were used to detect the expression of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and angiopoietin 1 (ANG-1) in the serum and in the membrane. Immunohistochemistry was employed to detect the synovium microvessel density (MVD).. Compared with the CIA model group, SIN significantly ameliorated swelling and erythema extension, decreased the arthritis index, reduced inflammation, cartilage damage and bone erosion, and lessened the number of CD31 positive cells on the synovium. Moreover, the levels of HIF-1α, VEGF and ANG-1 in the synovium and in the peripheral serum were increased in the untreated CIA model group but were significantly reduced in the 30 mg/kg, 100 mg/kg and 300 mg/kg SIN treatment groups.. SIN could mitigate CIA by inhibiting angiogenesis, and the mechanism may associate with the HIF-1α-VEGF-ANG-1 axis. Additionally, our study provides a referable experimental basis for the use of SIN for the treatment of rheumatoid arthritis.

Topics: Angiogenesis Inhibitors; Angiopoietin-1; Animals; Antirheumatic Agents; Arthritis, Experimental; Arthritis, Rheumatoid; Collagen; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Hypoxia-Inducible Factor 1, alpha Subunit; Mice; Mice, Inbred C57BL; Morphinans; Neovascularization, Pathologic; Synovial Membrane; Vascular Endothelial Growth Factor A

Intra-articular injection of sinomenine (SN) is an effective treatment method for knee osteoarthritis (OA), however, SN could be eliminated quickly in vivo. To extend the residence time of SN in the joint cavity, the SN-hyaluronic acid (HA) conjugate was prepared previously. This study was performed to evaluate the pharmacokinetics and pharmacodynamics of SN-HA conjugate after intra-articular administration for the treatment of OA.. A high performance liquid chromatography-mass spectrometry (HPLC-MS/MS) method was established to determine the SN content in rat synovial fluid. One hundred and twenty rats were randomly divided into two groups, the SN-HA group and SN group. The concentration of SN in articular cavity washings was determined by HPLC-MS/MS. The protective effect on the cartilage was evaluated by histological evaluation in a model of papain induced rabbit knee osteoarthritis.. The method was validated with respect to sensitivity, specificity, linearity, precision, accuracy and especially the stability of analytes under various conditions, and was successfully applied in evaluating the pharmacokinetic profiles of SN in the joint cavity. Compared to the SN injection, the drug exposure in joint cavity was significantly increased following SN-HA injection administration, and AUC(0-12h) was 2.9 times of SN injection, mean residence time (MRT) was 1.88 times of SN injection. In the pharmacodynamic study, there was no significant difference between the SN-HA twice-treated group and SH/HA five-times mixture-treated group.. The local bioavailability of SN in joint cavity was improved significantly after conjugated with HA. The SN-HA conjugate showed good synergism effect of OA inhibition. The results indicated that the SN-HA conjugate seemed to be an effective therapeutic means for the treatment of OA.

Topics: Animals; Hyaluronic Acid; Injections, Intra-Articular; Male; Morphinans; Osteoarthritis, Knee; Rats; Rats, Sprague-Dawley

Management of chronic pain is restricted by the lack of effective tools. This study evaluated the efficacies of sinomenine combined gabapentin or ligustrazine hydrochloride in treating peripheral and central chronic neuropathic pain. The study was conducted in mice with photochemically induced sciatic nerve injury, and in rats with photochemically induced spinal cord injury. For assessing the effectiveness of combined therapy, sinomenine, gabapentin or ligustrazine hydrochloride was injected intraperitoneally (i.p.), and pain behavioral tests were performed. At sub-effective dosages, pre-administration of sinomenine (for 60 min) plus gabapentin or ligustrazine hydrochloride, generated significant anti-allodynic effects in mice or rats with peripheral or central neuropathic pain. However, these effects were abolished when gabapentin or ligustrazine hydrochloride was pre-administered, and then sinomenine was given 60 min later. The combined efficacies of sinomenine and gabapentin or ligustrazine hydrochloride, cannot be blocked or reversed by the naloxone, suggesting the underlying mechanism is not mediated by opioid receptors. Moreover, following repeated treatments, sinomenine and gabapentin combination increased the baseline mechanical threshold, while generating prolonged analgesia, without introducing notable side effects. Sinomenine can enhance the efficacy of gabapentin or ligustrazine hydrochloride in rodent models of peripheral or central neuropathic pain, without introducing tolerance or other notable side effects. Findings of current study suggest that combing sinomenine and gabapentin or ligustrazine hydrochloride could be highly beneficial in neuropathic pain therapies.

Topics: Analgesics; Animals; Disease Models, Animal; Drug Synergism; Gabapentin; Male; Mice; Mice, Inbred C57BL; Morphinans; Neuralgia; Peripheral Nerve Injuries; Pyrazines; Spinal Cord Injuries

Sinomenine (SIN) is an isoquinoline derived from Caulis Sinomenii that has been used to treat rheumatoid arthritis and osteoarthritis for several decades in China. This study aims to reveal the effects of SIN on mouse chondrogenic ATDC5 cells growth and inflammation. SIN was used to treat ATDC5 cells injured by lipopolysaccharides (LPS). The following parameters were determined for evaluating the treatment effects of SIN: cell viability, apoptosis, reactive oxygen species generation, and pro-inflammatory cytokines release. Besides, the expression of LPS-sensitive miRNA (miR-192) and the activation of NF-κB and MAPK signaling were studied to explain SIN's function. SIN with concentration of 30 μM significantly attenuated LPS-induced cell damage via increasing cell viability, inhibiting apoptosis and reactive oxygen species generation, and declining IL-6 and TNF-α release. miR-192 was downregulated by SIN treatment. Restoration of miR-192 expression by miRNA transfection could significantly impede SIN's protective action. Besides, the inhibitory effects of SIN on the activation of NF-κB and MAPK signaling were attenuated by miR-192 overexpression. Furthermore, GDF11 was found to be a target gene of miR-192. LPS-mediated injury to chondrogenic ATDC5 cells can be relieved by SIN via downregulating miR-192 and subsequently repressing the activation of NF-κB and MAPK signaling.

Topics: Animals; Cell Line; Chondrogenesis; Down-Regulation; Lipopolysaccharides; Mice; MicroRNAs; Mitogen-Activated Protein Kinases; Morphinans; NF-kappa B

Topics: Administration, Cutaneous; Animals; Antioxidants; Arthritis, Rheumatoid; Bone and Bones; Dialysis; Drug Carriers; Drug Delivery Systems; Liposomes; Male; Morphinans; Permeability; Rabbits; Rats, Sprague-Dawley; Skin; X-Ray Microtomography

The aim of this study was to investigate the release behaviors of sinomenine hydrochloride loaded via in situ hexagonal liquid crystal (ISH), and its potential to improve the local bioavailability in knee joints of sinomenine hydrochloride (SMH) after intra-articular administration. The ISH was prepared by a liquid precursor mixture containing phytantriol (PT), Vitamin E acetate (VEA), ethanol (ET), and water. The in vitro release profiles revealed a sustained release of SMH from the optimized ISH formula (PT/VEA/ET/water, 60.8:3.2:16.0:20.0, w/w/w/w), which was selected for the in vivo pharmacokinetics and preliminary pharmacodynamics studies. In both healthy and adjuvant-induced arthritis (AA) rats, the SMH loaded ISH showed significantly smaller SMH AUC

Topics: Animals; Anti-Inflammatory Agents; Drug Liberation; Fatty Alcohols; Injections, Intra-Articular; Interleukin-1beta; Joints; Liquid Crystals; Morphinans; Rats; Skin Irritancy Tests; Synovial Membrane

Gastric carcinoma (GC) is a pernicious neoplasm with high morbidity and mortality. Sinomenine (SIN) has long been exploited to heal rheumatoid arthritis. Recently, SIN has been discovered to exert the antitumour functions in diverse cancers. However, the impacts of SIN on GC remain indistinct. We attempted to expose the antitumour effect of SIN on GC. MKN45 and SGC-7901 cells were administered with SIN for 24 hours, cell viability, proliferation, apoptosis, migration, invasion and the associated proteins in the above processes were examined via exploiting CCK-8, BrdU, flow cytometry, Transwell and Western blot. MiR-204 expression in GC tumour tissues, different GC cell lines and SIN-stimulated GC cells was investigated by executing RT-qPCR. The above cell biological processes were reassessed after transfection with miR-204 inhibitor. The latent mechanisms were probed by examining AMPK and Wnt/β-catenin pathways. We found that SIN memorably repressed cell proliferation, evoked apoptosis and affected CyclinD1, Bcl-2, Bax and cleaved-caspase-3 expression in MKN45 and SGC-7901 cells. Cell migration, invasion and expression of MMP-9 and Vimentin were all restrained by SIN stimulation. The increase of miR-204 was discovered in GC tissues and SIN-treated MKN45 and SGC-7901 cells. But suppression of miR-204 was observed in AGS, MKN28, MKN45 and SGC-7901 cells. Suppression of miR-204 overturned the inhibitory functions of SIN in MKN45 and SGC-7901 cells. Besides, SIN prohibited AMPK and Wnt/β-catenin pathways via enhancement of miR-204. In conclusion, these findings suggested that SIN exerted the antitumour activity in GC cells by hindering AMPK and Wnt/β-catenin pathways via enhancement of miR-204.

Topics: Adult; AMP-Activated Protein Kinases; Apoptosis; Carcinoma; Cell Movement; Cell Proliferation; Drug Screening Assays, Antitumor; Female; Gene Expression Regulation, Neoplastic; Humans; Male; MicroRNAs; Middle Aged; Morphinans; Stomach; Stomach Neoplasms; Up-Regulation; Wnt Signaling Pathway

To clarify the inner framework and relative properties in vitro of Lyotropic liquid crystal (LLC) based on various prescriptions by using hydrophilic sinomenine hydrochloride (SH) and lipophilic cinnamaldehyde (CA) as model drugs.. Phase structures were checked by polarized light microscopy (PLM) and small-angle X-ray scattering (SAXS). Rheological studies and Attenuated Total Reflectance Fourier Transform Infrared (ATR-FTIR) analysis were carried out to reveal their molecular interactions. In vitro release and skin permeation were conducted by Franz diffusion cell.. PLM and SAXS showed double diamond cubic crystal. All the samples displayed characteristics of non-Newtonian fluid, and the molecular interactions increased with the reducing water. ATRFTIR showed that the strongest strength of hydrogen bond emerged in the formulation with 32% water. Released SH of S2 and S3 arrived over 80%, while S1 only reached 45%, and that of CA was about 23%. Water-rich prescription gave higher percutaneous penetration for hydrophilic drugs, whereas no significant difference existed in CA permeation.. Proportion of Phytantriol to water determined the LLC assembling and affected the dissolving status of hydrophilic substance, thereby impacting on the location sites of guest molecular interactions among the substances, rheology properties, and finally the release and penetration behavior in vitro. Adjusting the basic prescription was the key to obtain satisfactory percutaneous delivery and stability for LLC carrying multi-therapeutic agents.

Topics: Acrolein; Hydrophobic and Hydrophilic Interactions; Liquid Crystals; Microscopy, Polarization; Morphinans; Rheology; Scattering, Small Angle; Spectroscopy, Fourier Transform Infrared; X-Ray Diffraction

Pathological changes, such as articular cartilage degeneration, destruction, and hyperosteogeny, are regarded as the main features of osteoarthritis (OA). Sinomenine (SIN) is a monomeric component purified from the plant Sinomenium acutum which has been found to have anti-inflammatory effects, however, the mechanism of action of SIN on OA is not clear. In this study, we evaluated whether SIN could regulate the inflammatory response induced by interleukin (IL)-1β and improve outcomes in the instability model of OA (medial meniscus mice (DMM)) by acting on the Nrf2/HO-1 and NF-κ B signaling pathways in chondrocytes. From our experiments, which include Griess reaction, ELISA, Western blot, and immunofluorescence, we found that SIN not only down-regulated the expression of pro-inflammatory factors induced by IL-1β, including; inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nitricoxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6), but also decreased the production of IL-1β-induced cartilage matrix catabolic enzymes including; ADAMTS-5 and MMPs, in mouse chondrocytes. In addition, the degradation of aggrecan and type II collagen protein in the extracellular matrix (ECM) stimulated by IL-1β was reversed. Most importantly, we have revealed for the first time that in OA, SIN inhibited the inflammatory response and ECM degradation by activating the Nrf2/HO-1 signaling pathways and inhibiting NF-κB activity in mouse-cartilage cells. In in vivo experiments, SIN treatment helped to improve the cartilage destruction in OA model mice. In conclusion, this study has demonstrated that SIN inhibits the IL-1β-induced inflammatory response and cartilage destruction by activating the Nrf2/HO-1 signaling pathway and inhibiting the NF-κB signaling pathway in mouse chondrocytes, suggesting a new use for SIN in the treatment of OA.

Topics: Animals; Anti-Inflammatory Agents; Cartilage, Articular; Cells, Cultured; Chondrocytes; Heme Oxygenase-1; Knee Joint; Male; Membrane Proteins; Mice, Inbred C57BL; Morphinans; NF-E2-Related Factor 2; NF-kappa B; Osteoarthritis; Signal Transduction

The initiation of pressure ulcers is accompanied by inflammation. Sinomenine emerges as a potential anti-inflammation agent. The aim of this study was to corroborate its anti-inflammatory property in skin keratinocyte HaCaT cells. Long non-coding RNA colon cancer associated transcript-1 (CCAT1)-associated mechanisms were also investigated.. HaCaT cells were stimulated with lipopolysaccharide (LPS) for 6 h after sinomenine pre-administration. Transfection was carried out to induce CCAT1 overexpression or silence it in HaCaT cells. Viability and apoptosis of HaCaT cells were determined by MMT and observed using flow cytometry, respectively. Protein expression was quantified using Western blot or ELISA. CCAT1 was measured by qRT-PCR.. LPS notably decreased cell viability and exaggerated apoptosis with the cleavage of caspase-3/-9. The secretion of inflammatory factors was promoted. Sinomenine pre-administration maintained cell viability, blocked apoptosis and relieved inflammation with the decrease in cleaved caspase-3/-9 and inflammatory factors. LPS-induced phosphorylation of p65, IκBα and p38MAPK and overexpression of CCAT1 were precluded by sinomenine. CCAT1 overexpression, which per se induced inflammatory lesions, negated the positive effects of sinomenine with the restored phosphorylation of p65, IκBα, and p38MAPK.. Sinomenine played a protective role against LPS-induced inflammation. The anti-inflammatory activity of sinomenine might be mediated by CCAT1 down-regulation.

Topics: Anti-Inflammatory Agents; Antirheumatic Agents; Cells, Cultured; Down-Regulation; Gene Expression Regulation; Humans; Inflammation; Keratinocytes; Lipopolysaccharides; Morphinans; RNA, Long Noncoding; Skin

Sinomenine is a bioactive alkaloid extracted from Sinomenium acutum. Here, we investigated the antidepressant effects of sinomenine in mice. The antidepressant actions of sinomenine were first examined in the forced-swim test and the tail-suspension test, and then assessed in the chronic social defeat stress (CSDS) model of depression. Changes in the brain-derived neurotrophic factor (BDNF) signaling pathway after CSDS and sinomenine treatment were also investigated. A tryptophan hydroxylase inhibitor and a BDNF signaling inhibitor were also used to determine the pharmacological mechanisms of sinomenine. It was found that sinomenine induced antidepressant-like effects in the forced-swim test and tail-suspension test without affecting the locomotor activity of mice. Sinomenine also prevented the CSDS-induced depressive-like symptoms. Moreover, sinomenine fully restored the CSDS-induced decrease in the hippocampal BDNF signaling pathway, whereas a BDNF signaling inhibitor, but not a tryptophan hydroxylase inhibitor, blocked the antidepressant effects of sinomenine. In conclusion, sinomenine exerts antidepressant effects in mice by promoting the hippocampal BDNF signaling pathway.

Topics: Animals; Antidepressive Agents; Brain-Derived Neurotrophic Factor; Depression; Depressive Disorder; Disease Models, Animal; Hippocampus; Male; Mice; Mice, Inbred C57BL; Morphinans; Neurogenesis; Serotonin; Signal Transduction; Stress, Psychological

A series of new sinomenine derivatives were designed, synthesized, and evaluated in tumor inhibitory activity, such as human triple negative breast cancer cell line (MDA-MB-231), glioma cell line (A172), human lung cancer cell line (A549), human colon cancer cell line (HCT-8). The modifications were carried out on rings A and C of the sinomenine by esterificating on phenolic hydroxyl with good yields. The highlight of this work was that the synthetic procedures were concise and sinomenine derivatives demonstrated promising antitumor activities.

Topics: Antineoplastic Agents; Cell Line, Tumor; Cell Proliferation; Cell Survival; Drug Design; Drug Screening Assays, Antitumor; Humans; Molecular Structure; Morphinans; Nuclear Magnetic Resonance, Biomolecular; Structure-Activity Relationship

Renal cell carcinoma (RCC) is a heterogeneous histological disease and the most common kidney cancer. The mortality rate of RCC remains high despite the improved treatment. Sinomenine is an isoquinoline extracted from Chinese medicinal plant Sinomenium acutum, famous for its ability to suppress several cancer cell types. Our research aimed to explore the anti-cancer potential of sinomenine in RCC. Results showed that sinomenine reduced the viability by reducing sphere-forming ability and enhancing pro-apoptosis effect in ACHN cells in a dose dependent manner. The expression levels of proliferation/apoptosis markers further validated the result. In addition, sinomenine significantly regulated the level of autophagy-related proteins with decreased expression of p62, and increased Beclin1 and LC3 II/LC3 I. Furthermore, phosphatidylinositol-3 kinase (PI3K)/AKT/mechanistic target of rapamycin (mTOR), the negatively regulated cell autophagy signaling pathway, was inhibited by sinomenine with decreased membrane translocation of AKT in ACHN cell lines. All in all, our study demonstrated that sinomenine promoted apoptosis in RCC via enhancing autophagy through PI3K/AKT/mTOR pathway.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Autophagy; Carcinoma, Renal Cell; Cell Line, Tumor; Cell Proliferation; Cell Survival; Humans; Kidney Neoplasms; Morphinans; Phosphatidylinositol 3-Kinase; Proto-Oncogene Proteins c-akt; Signal Transduction; TOR Serine-Threonine Kinases

The surface molecular imprinting technique has been proposed as a prospective strategy for template molecule recognition and separation by devising the recognition sites on the surface of imprinted materials. The purpose of this study was to establish a novel drug delivery system which was developed by surface molecular imprinting method using β-cyclodextrin (β-CD)-grafted chitosan (CS) (CS-g-β-CD) microspheres as matrix and sinomenine hydrochloride (SM) as the template molecule. By adjusting the amount of functional monomer and cross-linking agent, we got the more excellent adsorption of CS-g-β-CD molecularly imprinted polymers (MIPs-CS-g-β-CD). When the amount of functional monomer was 6 mmol and cross-linking agent was 20 mmol, the maximum binding capacity of MIPs and non-imprinted polymers (NIPs) was 55.9 mg/g and 37.2 mg/g, respectively. The results indicated that the recognition of SM with MIPs was superior to NIPs. The adsorption isotherms of MIPs-CS-g-β-CD indicated that the adsorption behavior fitted better to the Langmuir model, which showed that the adsorption process of polymer was monomolecular layer. In in vitro drug release studies, the accumulative release amount of MIPs-CS-g-β-CD was up to 78% within 24 h. MIPs exhibited an excellent controlled SM release profile without burst release and the mechanism of SM release was shown to conform to non-Fick diffusion. Therefore, MIPs-CS-g-β-CD were successfully applied to extraction of SM and used as the materials for drug delivery system.

Topics: beta-Cyclodextrins; Chitosan; Delayed-Action Preparations; Microspheres; Morphinans

Endotoxin is a significant contributing factor underlying the occurrence of fever, diarrhea, inflammation, edema, coagulation, shock and other syndromes associated with gram-negative bacterial infections. To date, there is no effective treatment for endotoxemia.. The aim of this study was to characterize differentially expressed genes in sinomenine-treated and lipopolysaccharide (LPS)-induced endothelial cells by microarray analysis and to determine the potential pharmacological activity of sinomenine.. The cultured cells of five treatment groups (n = 3) were collected. Participants: total RNA was extracted and subjected to Agilent Porcine 4 × 44 K whole genome microarray.. Kyoto encyclopedia of genes and genomes and gene ontology software were applied to screen and analyze differentially regulated genes.. The results showed that 723 differentially regulated genes were identified including 410 up-regulated genes and 313 down-regulated genes in therapy group vs. LPS group. Ten genes may be key controlled genes in the pathogenesis of LPS, including five up-regulated genes (ARG1, TLR2, IL1A, VCAM1, DKK3) and five down-regulated genes (HABP2, ID1, CHDH, GPX3, PTGFR), which primarily contribute to biological processes such as inflammatory response, vascular lesion, metabolic process and cell cycle. IL1A and FMO3 were considered as potent target genes.. Global gene expression profile analysis showed that sinomenine might effectively be useful to regulate inflammatory responses as part of future anti-endotoxin therapies.

Topics: Animals; Anti-Inflammatory Agents; Cells, Cultured; Down-Regulation; Drug Evaluation, Preclinical; Endothelial Cells; Gene Expression Profiling; Lipopolysaccharides; Morphinans; Oligonucleotide Array Sequence Analysis; Up-Regulation

In this study, the suppressive effect of sinomenine on the activation of SHh and the progression of breast cancer metastasis in vitro and in vivo was investigated. MDA-MB-231 breast cancer cells were treated with sinomenine and/or cyclopamine a proven SHh inhibitor. Sinomenine and cyclopamine both suppressed cell proliferation and migration, but sinomenine had a stronger suppressive effect in MDA-MB-231. In addition, sinomenine could suppress the activation of NF-κB and SHh signaling pathways, but cyclopamine could not suppress the activation of NF-κB. Subsequently, a mouse breast cancer-lung metastasis model was established. Our data on tissue examination and gene detection showed that SHh signaling was markedly activated in the metastatic model mice. The progression of lung metastasis was suppressed when mice were fed sinomenine and/or cyclopamine, while sinomenine had a stronger suppressive effect than cyclopamine in the model mice. In conclusion, sinomenine has a better effect than cyclopamine on the inhibition of breast cancer metastasis to lung in vivo and vitro, and inhibits NF-κB activation and NF-κB-mediated activation of SHh signaling pathway.

Topics: Animals; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Hedgehog Proteins; Humans; Lung Neoplasms; Mice; Morphinans; Neoplasm Metastasis; NF-kappa B; Signal Transduction; Veratrum Alkaloids

Fibroblast like synoviocyte (FLS) is a crucial in the pathogenesis of rheumatoid arthritis (RA), and involved in inflammation and joint destruction. Sinomenine (SIN), an alkaloid derived from the plant Sinomenium acutum, has anti-inflammatory and analgesic effect and been used for RA treatment in China. Alpha 7 nicotinic acetylcholine receptors (α7nAChR), as the key receptor in cholinergic anti-inflammatory pathway (CAP) to inhibit inflammation, has been detected in RA patients synovium, but its role is still unclear. Here we investigated the association between the aggressive proliferation of FLS and α7nAChR expression and the effect of sinomenine. FLS was isolated from synovial tissues of adjuvant-induced-arthritis (AIA) rat. Tumor necrosis factor(TNF)-α was used to induce the aggressive proliferation of FLS. MTT assay was applied to evaluate the proliferation of FLS. The messenger RNA (mRNA) and protein levels of α7nAChR and early growth response gene-1 (Egr-1) were measured. The results showed that TNF-α induced FLS proliferation in vitro (P < .01) and increased the phosphorylation of ERK1/2 and the expression of Egr-1 and α7nAChR (P < .05 or P < .01). U0126, the inhibitor of ERK1/2 inhibited α7nAChR expression and FLS proliferation significantly (P < .05 or P < .01). Specific short interference RNA(siRNA) of α7nAChR decreased α7nAChR expression and inhibited FLS proliferation as well. SIN inhibited the proliferation of FLS and decreased the phosphorylation of ERK1/2, and the expression of Egr-1 and α7nAChR induced by TNF-α (P < .05). In conclusion, the expression of α7nAChR involved in the aggressive proliferation of FLS induced by TNF-α and was regulated by ERK/Egr-1 signal pathway. SIN inhibited FLS proliferation and α7nAChR expression through inhibiting ERK/Egr-1 signal pathway, this may contribute to the anti-inflammatory and anti-arthritic effect of SIN.

Topics: alpha7 Nicotinic Acetylcholine Receptor; Animals; Anti-Inflammatory Agents; Arthritis, Experimental; Arthritis, Rheumatoid; Cell Proliferation; Cells, Cultured; Disease Models, Animal; Early Growth Response Protein 1; Extracellular Signal-Regulated MAP Kinases; Gene Expression Regulation; Humans; Male; Morphinans; Rats; Rats, Sprague-Dawley; RNA, Small Interfering; Signal Transduction; Sinomenium; Synoviocytes

Sinomenine (SIN) has been shown to have protective effects against brain damage following traumatic brain injury (TBI). However, the mechanisms and its role in these effects remain unclear. This study was conducted to investigate the potential mechanisms of the protective effects of SIN.. The weight-drop model of TBI in Institute of Cancer Research (ICR) mice were treated with SIN or a vehicle via intraperitoneal administration 30 min after TBI. All mice were euthanized 24 h after TBI and after neurological scoring, a series of tests were performed, including brain water content and neuronal cell death in the cerebral cortex.. The level of cytochrome. SIN protected neuronal cells by protecting them against apoptosis via mechanisms that involve the mitochondria following TBI.

Topics: Animals; Apoptosis; Apoptosis Regulatory Proteins; Brain Edema; Brain Injuries, Traumatic; Cerebral Cortex; Cytochromes c; Cytoprotection; Disease Models, Animal; Dose-Response Relationship, Drug; Glutathione Peroxidase; Male; Malondialdehyde; Mice, Inbred ICR; Mitochondria; Morphinans; Nerve Degeneration; Neurons; Neuroprotective Agents; Oxidative Stress; Signal Transduction; Superoxide Dismutase-1

Radiation therapy is one of the most important treatments for unresectable and locally advanced esophageal squamous cell carcinoma (ESCC), however, the response to radiotherapy is sometimes limited by the development of radioresistance. Sinomenine hydrochloride (SH) has anticancer activity, but its effect on the radiosensitivity of ESCC is unclear. We determined the effect of SH on the radiosensitivity of ESCC cells and elucidated its potential radiosensitization mechanisms in vitro and in vivo. ESCC cells were subjected to SH and radiation, both separately and in combination. Untreated cells served as controls. The CCK‑8 assay was used to evaluate cell proliferation, and the clonogenic assay to estimate radiosensitization. Flow cytometry was used to investigate cell cycle phases and cell apoptosis. Bcl‑2, Bax, cyclin B1, CDK1, Ku86, Ku70, and Rad51 expression was evaluated using western blotting. In vivo, tumor xenografts were created using BALB/c nude mice. Tumor‑growth inhibition was recorded, and Ki‑67 and Bax expression in the tumor tissues was assessed using immunohistochemistry. SH inhibited ESCC cell growth and markedly increased their radiosensitivity by inducing G2/M phase arrest. SH combined with radiation therapy significantly increased ESCC cell apoptosis. The molecular mechanism by which SH enhanced radiosensitivity of ESCC cells was related to Bcl‑2, cyclin B1, CDK1, Ku86, Ku70, and Rad51 downregulation and Bax protein expression upregulation. SH combined with radiation considerably delayed the growth of tumor xenografts in vivo. Immunohistochemical analysis showed that in the SH combined with radiation group, the expression of Bax was significantly higher while that of Ki‑67 was lower than the expressions in the control groups. Taken together, our findings showed that SH could improve the sensitivity of radiation in ESCC cells by inducing G2/M phase arrest, promoting radiation‑induced apoptosis and inhibiting DSB‑repair pathways. SH appears to be a prospective radiosensitizer for improving the efficacy of radiotherapy for ESCC.

Topics: Animals; Apoptosis; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Esophageal Neoplasms; Esophageal Squamous Cell Carcinoma; Flow Cytometry; Gene Expression Regulation, Neoplastic; Humans; Mice; Morphinans; Neoplasm Proteins; Radiation Tolerance; Xenograft Model Antitumor Assays

Sinomenine, a major bioactive ingredient isolated from traditional Chinese medicine Sinomenium acutum, has been reported to have analgesic effects in various pain animal models. N-demethylsinomenine, the N-demethylated product of sinomenine, has been identified to be the major metabolite of sinomenine and is also a natural component extracted from Sinomenium acutum. This study examined the anti-allodynic effects of N-demethylsinomenine in a mouse model of postoperative pain. A significant and sustained mechanical allodynia that lasted for 4 days was induced by making a surgical incision on the right hind paw in mice. Acute treatment with N-demethylsinomenine (10-40 mg/kg, s.c.) relieved the mechanical allodynia in a dose-dependent manner. Although there was no difference in maximal analgesic effect between N-demethylsinomenine (40 mg/kg, s.c.) and sinomenine (40 mg/kg, s.c.), the onset of action of N-demethylsinomenine was quicker than sinomenine. Repeated treatment with N-demethylsinomenine (10-40 mg/kg/day, s.c.) also dose-dependently exerted sustained antinociception against postoperative allodynia and did not produce analgesic tolerance and carry-over effect. The anti-allodynia induced by N-demethylsinomenine (40 mg/kg, s.c.) was attenuated by bicuculline, a selective γ-aminobutyric acid type A (GABA

Topics: Analgesics; Animals; Bicuculline; Disease Models, Animal; Dose-Response Relationship, Drug; GABA-A Receptor Antagonists; Hyperalgesia; Male; Mice; Mice, Inbred ICR; Morphinans; Pain, Postoperative; Receptors, GABA-A

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 “This paper belongs to a set of over 400 papers (as per February 2020) that share very similar Western blots with tadpole-like shaped bands, the same background pattern, and striking similarities in title structures, paper layout, bar graph design, and - in a subset - flow cytometry panels”, 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; Apoptosis Regulatory Proteins; Cell Survival; Down-Regulation; Dual Specificity Phosphatase 1; Female; Guinea Pigs; Humans; Inflammation; Interleukin-6; Interleukin-8; Keratinocytes; Lipopolysaccharides; Male; MAP Kinase Signaling System; MicroRNAs; Morphinans; Signal Transduction; Tumor Necrosis Factor-alpha; Up-Regulation

Topics: Antineoplastic Agents; Brain Neoplasms; Cell Line, Tumor; Cell Movement; Epithelial-Mesenchymal Transition; Glioblastoma; Humans; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Morphinans

BACKGROUND In the present study, we aimed to investigate the effects of sinomenine (SIN) on chronic intermittent hypoxia (CIH)- induced lung injury in rats, and to explore the underlying mechanisms. MATERIAL AND METHODS To perform the investigation, a CIH rat model was established. ELISA assay was applied to detect the level of inflammatory cytokines. Oxidative stress bio-markers (MDA, SOD, and CAT) were determined in lung tissues. In addition, the expression level of NADPH oxidase 2 (Nox2) was analyzed by Western blotting and qRT-PCR, respectively. RESULTS The results showed that compared with other groups, more obvious pulmonary pathological changes were observed in the CIH group. The level of inflammatory cytokines in the CIH group was markedly higher than that in the control and Con-S groups. Compared with the control and Con-S groups, oxidative stress was notably increased in the CIH group. Expression of Nox2 was also increased in the CIH group. The effects caused by CIH in rats were attenuated by SIN treatment. CONCLUSIONS SIN can reverse chronic intermittent hypoxia-induced lung injury through inhibiting inflammation and oxidative stress.

Topics: Animals; Chronic Disease; Cytokines; Down-Regulation; Hypoxia; Inflammation; Inflammation Mediators; Lung; Lung Injury; Male; Mice; Morphinans; NADPH Oxidases; NF-E2-Related Factor 2; Oxidative Stress; Rats, Sprague-Dawley; Signal Transduction

BACKGROUND The purpose of this study was to investigate the effects of sinomenine (SIN) on CFA-induced inflammatory pain in rats, and to explore the underlying molecular mechanisms. MATERIAL AND METHODS To determine the potential influences of SIN in the pathogenesis of inflammatory pain, an inflammatory pain (IP) mouse model was established and rats were treated with SIN (30 mg/kg). Behavioral tests were used to assess the MWT and TWL of the rats. ELISA assay was used to detect the level of inflammation cytokines. Western blotting and qRT-PCR were carried out to measure the related protein and mRNA expression level, respectively. RESULTS We found that the MWT and TWL of the CFA-treated rats were markedly lower than that of the control rats, and they were significantly increased by SIN administration. The results suggest that IP rats had higher levels of TNF-α, IL-1β and IL-6 compared with the control rats. SIN administration decreased the levels of TNF-α, IL-1β, and IL-6. In addition, we found that p-p65 and p-p38 expression notably decreased after SIN treatment in IP rats. Moreover, the results showed that SIN inhibited Cox-2 and PGE2 expression in IP rats. CONCLUSIONS The data indicate that SIN had a protective role in inflammatory pain through repressing inflammatory mediators via preventing the p38MAPK-NF-κB pathway.

Topics: Animals; Cyclooxygenase 2; Cytokines; Freund's Adjuvant; Inflammation; Interleukin-1beta; Male; Morphinans; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Pain; Rats; Rats, Sprague-Dawley; Signal Transduction; Tumor Necrosis Factor-alpha

This study developed a new transdermal delivery system for the improved delivery of sinomenine hydrochloride (SH). The delivery system utilized the advantages of lyotropic liquid crystals (LLC) creating an adaptable system that offers a variety of options for the field of transdermal delivery. The formulation was prepared, characterized, and evaluated for its skin penetration in vitro. In the study, the appearance of samples was characterized by visual observation, and these LLC gels were colorless and transparent. Polarizing light microscopy (PLM) and small-angle X-ray diffraction (SAXS) were used to analyze the internal structures of gels, and the gels displayed a cubic double-diamond (P

Topics: Administration, Cutaneous; Animals; Drug Delivery Systems; Liquid Crystals; Morphinans; Permeability; Rats; Scattering, Small Angle; Skin

Rheumatoid arthritis (RA) is an autoimmune disease, which is characterized by inflammatory synovitis, and the subsequent destruction of articular cartilage and bone. Sinomenine is a traditional Chinese medicine, which has been employed as a clinical treatment for RA for several years in China. The present study investigated the anti‑arthritic effects of sinomenine on Sprague‑Dawley rats with collagen‑induced arthritis (CIA). The differentially expressed proteins in serum were measured by proteomic analysis in order to generate a differentially expressed protein network. A total of 320 differentially expressed proteins were detected in the drug‑treated group compared with in the control group. In the sinomenine‑treated group, 79 differentially expressed proteins were detected compared with in the model group, and among these, 46 proteins were upregulated. Gene ontology analysis revealed that five functions were affected by sinomenine treatment of CIA rats compared with in the model group. In addition, Ingenuity® Pathway Analysis was used to measure enriched signaling pathways, which revealed nuclear factor‑κB, histones, heat shock proteins and protein kinase B as core proteins, generating ~60 pair associations in the network. To the best of our knowledge, the present study is the first to perform proteomic analysis in sinomenine‑treated CIA rats, and the results revealed that numerous targets were involved in the process. In addition, the present study provided a novel approach and evidence for exploring the biological effects of sinomenine. Therefore, the findings of the present study may provide a novel insight into the anti‑RA mechanisms of sinomenine, and may justify further exploration into its function in other relevant diseases.

Topics: Animals; Antirheumatic Agents; Arthritis, Experimental; Autoimmune Diseases; Female; Gene Expression Profiling; Gene Expression Regulation; Morphinans; Rats; Rats, Sprague-Dawley

Sinomenine, a bioactive alkaloid isolated from the traditional Chinese herb Sinomenium acutum, possesses antiinflammatory, antinociceptive, antifibrotic, and antitumorigenic properties. In this work, we sought to explore the biological effects of sinomenine on glioma cells. It was found that sinomenine caused a concentration-dependent inhibition of viability in both U87 and U251 glioma cells. Sinomenine at 16 μmol/L caused 55% to 60% reduction in the proliferation of U87 and U251 cells. Moreover, sinomenine treatment induced a G0/G1 cell cycle arrest and apoptosis. Mechanistically, sinomenine promoted p53 expression and acetylation and reduced the expression of sirtuin 1. Ectopic expression of sirtuin 1 significantly prevented sinomenine-induced p53 acetylation and growth suppression in glioma cells. Moreover, sinomenine inhibited the growth of U87 xenograft tumors in vivo and raised the p53 protein expression. Collectively, sinomenine shows antiproliferative effects against glioma cells which is mediated through downregulation of sirtuin 1 and induction of p53 activity.

Topics: Acetylation; Animals; Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Cell Proliferation; Cell Survival; Disease Models, Animal; G1 Phase Cell Cycle Checkpoints; Glioma; Humans; Mice; Morphinans; Sirtuin 1; Tumor Suppressor Protein p53; Xenograft Model Antitumor Assays

Epilepsy is a common neurological disorder and is not well controlled by available antiepileptic drugs (AEDs). Inflammation is considered to be a critical factor in the pathophysiology of epilepsy. Sinomenine (SN), a bioactive alkaloid with anti-inflammatory effect, exerts neuroprotective activity in many nervous system diseases. However, little is known about the effect of SN on epilepsy.. The chronic epilepsy model was established by pentylenetetrazole (PTZ) kindling. Morris water maze (MWM) was used to test spatial learning and memory ability. H.E. staining and Hoechst 33258 staining were used to evaluate hippocampal neuronal damage. The expression of nucleotide oligomerization domain (NOD)-like receptor protein 1 (NLRP1) inflammasome complexes and the level of inflammatory cytokines were determined by western blot, quantitative real-time PCR and enzyme-linked immunosorbent assay (ELISA) kits.. SN (20, 40, and 80 mg/kg) dose-dependently disrupts the kindling acquisition process, which decreases the seizure scores and the incidence of fully kindling. SN also increases the latency of seizure and decreases the duration of seizure in fully kindled rats. In addition, different doses of SN block the hippocampal neuronal damage and minimize the impairment of spatial learning and memory in PTZ kindled rats. Finally, PTZ kindling increases the expression of NLRP1 inflammasome complexes and the levels of inflammatory cytokines IL-1β, IL-18, IL-6, and TNF-α, which are all attenuated by SN in a dose- dependent manner.. SN exerts anticonvulsant and neuroprotective activity in PTZ kindling model of epilepsy. Disrupting the kindling acquisition, which inhibits NLRP1 inflammasome-mediated inflammatory process, might be involved in its effects.

Topics: Analysis of Variance; Animals; Anticonvulsants; CARD Signaling Adaptor Proteins; Caspase 1; Convulsants; Cytokines; Disease Models, Animal; Epilepsy; Kindling, Neurologic; Male; Maze Learning; Morphinans; Nerve Tissue Proteins; Pentylenetetrazole; Proto-Oncogene Proteins c-bcl-2; Rats; Rats, Sprague-Dawley; RNA, Messenger

BACKGROUND This study aimed to investigate the effect and underlying molecular mechanism of sinomenine (SIN) on ankylosing spondylitis (AS). MATERIAL AND METHODS To study the potential role of SIN in the pathogenesis of AS, an AS mouse model was established and mice were treated with different concentrations of SIN (10, 30, and 50 mg/kg, administered intraperitoneally). Markers of inflammation and oxidative stress were determined by ELISA assay. Western blot analysis and qRT-PCR were used to quantify the levels of related proteins and gene mRNA expression. RESULTS The results suggest that AS mice has higher levels of TNF-α, IL-1β, and IL-6 (p<0.01 for all), and lower levels of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-PX) (p<0.01 for all). SIN treatment reduced the level of TNF-α, IL-1β, and IL-6 in a dose-dependent manner, and the levels of SOD, CAT, and GSH-PX were dose-dependently increased (p<0.05 for all). The results also revealed that NF-κBp65 expression decreased, while the level of IkB increased, in a dose-dependent manner, after SIN treatment in AS mice (p<0.05 for all). The level of p-p38 was dose-dependently reduced in AS mice by SIN treatment (p<0.05). Moreover, SIN inhibited Cox-2 expression in AS mice in a dose-dependent manner (p<0.05). CONCLUSIONS SIN has a beneficial role in AS through suppressing inflammatory mediators and by down-regulating oxidative stress via inhibiting the MAPKp38/NF-kB pathway and Cox-2 expression.

Topics: Animals; Antioxidants; Catalase; Disease Models, Animal; Glutathione Peroxidase; Inflammation; Interleukin-1beta; Interleukin-6; Mice; Mice, Inbred BALB C; Morphinans; NF-kappa B; Oxidative Stress; Signal Transduction; Spondylitis, Ankylosing; Superoxide Dismutase; Tumor Necrosis Factor-alpha

Three different kinds of sinomenine in situ liquid crystal were prepared for different prescriptions, to investigate the rheological properties before and after in situ treatment and evaluate its feasibility for embolization. Rheological experiments were carried out with cone plate fixtures. Both the steady-state rheological and non-steady-state rheological properties of in-situ gels and the swelling gels were studied and compared. Steady-state rheological study results showed that all the three liquid embolic agents were non-newtonian fluid before and after in situ treatment, which would become less ropy when they were pressed with shear stress; their viscosities differed by 2-5 orders of magnitude. It had a yield value of about 10 Pa before in situ treatment and about 4 500 Pa after in situ treatment. All the six systems had thixotropy while their dynamic viscosities were not influenced by the shear rate, all less than 0.3 Pa·s before in situ treatment more than 1 Pa·s after in situ treatment, differing by an order of magnitude. The results of temperature sweeping showed a slight decrease with a steady rate in viscosity within the range of 10-50 °C, differing by 3-4 orders of magnitude. The results of unsteady rheology showed that there was no obvious linear viscoelastic region in the three kinds of agents, indicating the properties of liquid. After in situ treatment, their linear viscoelastic range γ<1% (No.3 was 5%), and their elastic modulus G' was larger than the viscous modulus G", indicating the properties of solid. Frequency scanning results showed that for the systems at low frequencies, G">G', system viscosity in a dominant position; while at high frequencies, G'>G", system elasticity in a dominant position. The results of compound viscosity test also proved that the liquid embolic agent in situ can form a cubic liquid crystal (the structure of No. 3 was destroyed after in situ treatment). The DHR-2 rheometer was used to investigate the rheological properties of in situ gels with three different prescriptions. The method is simple and the result is reliable, which can provide more theoretical reference for the

Topics: Elasticity; Liquid Crystals; Morphinans; Rheology; Viscosity

Studies have shown that the inflammatory activation of miroglia (MG) and nuclear factor kappa B ( NF-κB ) play a dominant role in inflammatory response. Previous studies have shown that sinomenine, an anti-inflammatory agent extracted from Sinomenium acutum, can directly protect neurons against cerebral ischemia injury. However, there are no reports on its effect on ischemia/reperfusion-induced inflammatory activation of MG. In the present study, an in vitro ischemia/reperfusion model was developed with mouse BV-2 microglia cells, a model of oxygen-glucose deprivation/reperfusion (OGD/R), and the inhibitory effect of sinomenine pretreatment on inflammatory activation was confirmed through measurement of inflammatory indicators. Mechanistically, sinomenine suppressed OGD/R-induced inflammatory activation through the SP1/miRNA-183-5p/IκB-α pathway. In conclusion, this study shows that sinomenine effectively inhibits OGD/R-induced inflammatory activation in MG by suppressing the activation of transcription specificity protein 1 (SP 1). This finding is of significance for the clinical use of sinomenine in treating cerebral ischemia/reperfusion injury.

Topics: Animals; Anti-Inflammatory Agents; Cell Line; Glucose; Inflammation; Mice; Microglia; MicroRNAs; Morphinans; NF-KappaB Inhibitor alpha; Oxygen; Reperfusion Injury; Signal Transduction; Sinomenium; Sp1 Transcription Factor

Sinomenine, an anti-rheumatoid arthritis drug used in China for decades, is usually co-administered with cardiovascular (CV) drugs to reduce arthritis-related risk of cardiovascular diseases. This study was to investigate whether and how CV drugs affect the pharmacokinetic profile of sinomenine.. In rat liver microsomes (RLMs), the key metabolic enzymes of sinomenine were identified by using specific inhibitors. The influences of CV drugs, including propranolol, verapamil, warfarin, atorvastatin, simvastatin, and lovastatin, on the metabolism of sinomenine were examined. Cocktail probe, RT-qPCR, and western blotting were performed to unveil the underlying mechanism of the drug-drug interaction.. The key metabolic enzymes of sinomenine were identified to be CYP3A1/2 and CYP2D1 in RLMs. Among the CV drugs screened, simvastatin and lovastatin were shown to inhibit the liver metabolism of sinomenine with Ki values of 13.00 and 25.83 μM, respectively. Single administration of simvastatin or lovastatin in rats increased the AUC value of sinomenine to 1.40- or 1.50-fold, and decreased the CLz/F value to 68.19% or 65.44%, respectively. In contrast, multiple administrations of simvastatin, but not lovastatin, increased the CLz/F value of sinomenine to 1.38-fold and decreased the AUC value to 71.59%. Further studies showed that the long-term administration of simvastatin could up-regulate the expression of CYP3A1/2 to account for the effect.. This study demonstrated the potential effect of simvastatin and lovastatin on the metabolism of sinomenine for the first time. The findings provide guidelines for the co-administration of sinomenine with simvastatin or lovastatin in clinic.

Topics: Animals; Antirheumatic Agents; Cytochrome P-450 Enzyme System; Drug Combinations; Drug Interactions; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Lovastatin; Male; Microsomes, Liver; Morphinans; Rats; Rats, Sprague-Dawley; Simvastatin; Tissue Distribution

Sinomenine (SIN), a pure alkaloid isolated from Sinomenium acutum, has been widely used in arthritis for its anti‑inflammatory effect, but little is known about the effect of SIN on human ulcerative colitis (UC). In the present study, the effect and mechanism of SIN was examined in a dextran sulfate sodium (DSS)‑induced murine colitis model, which mimics human UC. Oral administration of SIN significantly suppressed the elevated disease activity index and ameliorated colonic histological damage in a DSS‑induced colitis model. Tumor necrosis factor‑α, interleukin‑6 and inducible nitric oxide synthase levels were also reduced as detected by reverse transcription‑quantitative polymerase chain reaction. In addition, SIN reversed the decreased colon length and colonic superoxide dismutase activity. Furthermore, western blot analysis revealed that nuclear factor‑erythroid 2‑related factor 2 (Nrf2) and its downstream genes, heme oxygenase‑1 and NADP(H) quinone oxidoreductase 1 (NQO‑1), were markedly activated by SIN. The current results indicated that SIN alleviated DSS‑induced colitis in mice, which may be due to its antioxidant properties and was at least in part dependent on the Nrf2/NQO‑1 signaling pathway. Therefore, SIN may have potential applications as a protective drug for patients with UC.

Topics: Animals; Antioxidants; Colitis; Dextran Sulfate; Female; Mice, Inbred C57BL; Morphinans; NAD(P)H Dehydrogenase (Quinone); NF-E2-Related Factor 2; Oxidative Stress; Signal Transduction; Sinomenium

Stephania epigaea Lo is an important herbal medicine used as antiphlogistic and analgesic drugs. Its major components are dicentrine (1) and sinomenine (2). In the present study, a rapid, accurate, and precise method for simultaneous quantitation of dicentrine (1) and sinomenine (2) in S. epigeae using

Topics: Aporphines; Magnetic Resonance Spectroscopy; Morphinans; Stephania

Melanoma is a common skin tumor in adults with high metastasis and mortality rates. Thus, finding a better effective approach to treat melanoma has become very urgent. Sinomenine (SIN), the major active compound of. This study aimed to explore the effects of SIN on melanoma in vitro and in vivo, in addition to exploring the underlying mechanism.. Mouse melanoma cell B16-F10 treated by SIN was analyzed by CCK8 assay and flow cytometry. Melanoma xenograft model was then established by subcutaneously injection with B16-F10 cells. Tumor growth was measured by immunohistochemistry. To further investigate the relative mechanism, the autophagy and PI3K/Akt/mTOR pathway were examined by immunofluorescence and Western blot.. Our results revealed that SIN dose dependently inhibited the proliferation of B16-F10 cells in vitro and attenuated melanoma growth in vivo. In addition, SIN treatment promoted the apoptosis of B16-F10 cells in a dose-dependent manner, as demonstrated by the increase in apoptotic cells, Bax/Bcl-2 ratio, and caspase-3 activity. Moreover, preconditioning with SIN dramatically enhanced autophagy activity by increasing Beclin-1 and LC3II/LC3I expression, in addition to decreasing p62 expression and augmenting the number of LC3 puncta, in B16-F10 cells. More importantly, autophagy inhibitor chloroquine partly abolished SIN's effects on cell growth and apoptosis. Furthermore, our results showed that SIN-triggered activation of autophagy was mediated by PI3K/Akt/mTOR signaling pathway.. Our study has identified a novel function of SIN and provided a molecular basis for potential applications of SIN in the treatment of melanoma and other cancers.

Topics: Animals; Apoptosis; Autophagy; Cell Line, Tumor; Humans; Melanoma, Experimental; Mice; Mice, Inbred BALB C; Morphinans; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Proto-Oncogene Proteins c-akt; Signal Transduction; TOR Serine-Threonine Kinases

Acute lung injury (ALI) is a common disease characterized by pulmonary inflammation and oxidative stress. Sinomenine (SIN) is an alkaloid originally extracted from the Chinese medicinal plant Sinomenium acutum. It has been shown to have anti-inflammatory and anti-oxidative effect. However, it's unclear whether SIN can alleviate ALI. In this study, we assessed the effect of SIN on Escherichia coli (E.coli)-induced ALI mouse model. Mice were conditioned with SIN or placebo 1 h before intratracheally instilled with E.coli. Lung water content, malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, Myeloperoxidase (MPO) levels and inflammatory cytokines production were measured. Immunohistochemistry and western blot were performed to measure target protein expression. E.coli induced histological changes indicating tissues damage and increased W/D ratio, MPO activity, MDA content, and inflammatory cytokines production in the Lung. Whereas in mice pretreated with SIN, these changes were absent. E.coli-induced NF-κB activation was also inhibited by SIN. In addition, SIN increased the expression of HO-1, NQO1 and Nrf2 in lung tissues. Our results suggest that SIN attenuates ALI through the inhibition of inflammation and oxidative stress.

Topics: Acute Lung Injury; Animals; Bronchoalveolar Lavage Fluid; Cytokines; Escherichia coli; Female; Inflammation; Lung; Malondialdehyde; Mice, Inbred ICR; Morphinans; NF-E2-Related Factor 2; NF-kappa B; Oxidative Stress; Protective Agents; Protein Kinase C; Signal Transduction; Superoxide Dismutase

The use of plant‑based compounds derived from traditional medicine to improve human diseases has been gaining momentum, due to their high bioavailability and moderate adverse effects. Sinomenine is one such biomonomer alkali compound derived from Sinomenium acutum and is known for its anti‑inflammatory and antitumor effects. However, the molecular mechanism(s) of its antitumor properties are not fully characterized. In the present study, we evaluated the radiosensitizing effects of the water‑soluble sinomenine, sinomenine hydrochloride (SH) in human cervical cancer cell line (HeLa). SH sensitized HeLa cells to ionizing radiation (IR) by promoting accumulation of IR‑induced DNA double‑strand breaks (DSBs) and also by interfering with DNA damage checkpoint activation. We then investigated the molecular mechanisms underlying the SH‑mediated cellular sensitization to IR and found that SH inhibited the expression of DNA damage response (DDR) factors Ku80 and Rad51 at the transcription level. Finally, the radiosensitizing activity of SH was confirmed in a cervical cancer mouse xenograft model. The combinatorial treatment of SH and IR significantly slowed the tumor growth rate compared with IR alone. Collectively, our study not only provides molecular insights into the novel role of SH in cellular response to IR, but also suggests a therapeutic potential of SH as a radiosensitizer in cervical cancer therapy.

Topics: Animals; Cell Cycle Checkpoints; DNA Breaks, Double-Stranded; DNA Damage; Female; HeLa Cells; Humans; Mice; Morphinans; Radiation Tolerance; Radiation-Sensitizing Agents; Radiation, Ionizing; Sinomenium; Uterine Cervical Neoplasms; Xenograft Model Antitumor Assays

BACKGROUND This study aimed to investigate the effects of SIN on ankle fracture and the underlying mechanisms in MG-63 cells. MATERIAL AND METHODS qRT-PCR and ELISA assay were used to detect the mRNA and protein levels of cytokines in peripheral blood of children with or without ankle fracture. The expression and activity of antioxidant and detoxifying enzymes were detected by ELISA assay. Pretreated MG-63 cells with/without SIN were stimulated with 1 μg/ml bradykinin (BK). A CCK-8 kit was used to detect the cell viability. The cytokines produced from MG-63 cells were detected by Western blotting and qRT-PCR. Moreover, Western blotting was used to detect the levels of p-p38 and p-NF-κB (p65), and the activation level of the Nrf2 signaling pathway was examined by qRT-PCR and Western blotting. RESULTS In this study, we found that compared with the healthy children, the mRNA and protein levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor-alpha (TNF-α) were significantly upregulated in children with ankle fracture. In addition, the expression and activity of antioxidant and detoxifying enzymes were imbalanced in children with ankle fracture. SIN treatment did not have a cytotoxic effect on MG-63 cells. SIN dose-dependently suppressed BK-induced upregulation of IL-1β, IL-6, TNF-α, p-p38, and p-NF-κB (p65). Furthermore, SIN dramatically inhibited oxidative stress induced by BK via balancing the expression and activity of antioxidant and detoxifying enzymes and inhibited the activation of Nrf2 signaling. CONCLUSIONS SIN might be a potential agent for the treatment of ankle fracture through reducing inflammatory response and oxidative stress.

Topics: Ankle Fractures; Ankle Joint; Cell Line, Tumor; Child; Child, Preschool; Cytokines; Female; Humans; Inflammation; Interleukin-1beta; Interleukin-6; Male; Morphinans; NF-E2-Related Factor 2; NF-kappa B; Oxidative Stress; Phosphorylation; Reactive Oxygen Species; Signal Transduction; Tumor Necrosis Factor-alpha

To investigate the effect of sinomenine on lipopolysaccharide (LPS)-induced inflammation in RAW264.7 macrophages and the underlying mechanisms.
 Methods: The mouse RAW264.7 macrophages were treated with sinomenine and/or LPS with or without heme oxygenase-1 (HO-1) inhibitor Znpp. Real-time PCR, ELISA, immunofluenscence, and Western blot were used to detect the mRNA expression of TNF-α and IL-6, the release of TNF-α and IL-6, the protein expression of HO-1 and autophagy, respectively.
 Results: Compared with the control group, the mRNA expression and release of inflammatory cytokines TNF-α and IL-6 were increased, the green fluorescence of autophagy-related protein LC3 was accumulated and the protein expression of HO-1 was increased in RAW264.7 cells after LPS treatment (P<0.05). Compared with the LPS group, sinomenine treatment could reduce the mRNA expression and release of TNF-α and IL-6, accompanied by increasess in green fluorescence aggregation of LC3 and HO-1 production (P<0.05). HO-1 inhibitor Znpp could weaken the ability of sinomenine through suppressing TNF-α and IL-6 expression and decreasing the aggregation of LC3 green fluorescence (P<0.05).
 Conclusion: Sinomenine could alleviate LPS-induced inflammation in RAW264.7 macrophages, which might be related to HO-1 mediated autophagy. This study provides an experimental and theoretical basis for the clinical application of sinomenine in prevention and treatment of inflammation.. 目的：探讨青藤碱对脂多糖诱导的RAW264.7巨噬细胞炎症的影响和机制。方法：以小鼠RAW264.7巨噬细胞为研究对象，在有或无血红素氧合酶-1(heme oxygenase-1，HO-1)抑制剂Znpp处理下，采用青藤碱和/或脂多糖(lipopolysaccharide，LPS)处理RAW264.7巨噬细胞。应用Real-time PCR检测细胞炎症因子TNF-α和IL-6 mRNA表达，ELISA检测细胞炎症因子TNF-α和IL-6水平，免疫荧光试验分析细胞自噬情况，Western印迹检测细胞HO-1蛋白表达。结果：与对照组相比，LPS作用后RAW264.7细胞炎症因子TNF-α和IL-6表达和释放增多，自噬相关蛋白LC3绿色荧光聚集，HO-1表达水平升高(P<0.05)；与LPS组相比，加用青藤碱处理能减少TNF-α和IL-6表达和释放，进一步促进LC3绿色荧光聚集，增加HO-1水平(P<0.05)；用HO-1抑制剂Znpp预处理后，青藤碱对LPS诱导TNF-α和IL-6表达和释放的抑制作用减弱，LC3绿色荧光聚集减少(P<0.05)。结论：青藤碱能减轻LPS诱导的RAW264.7巨噬细胞炎症，其机制可能与HO-1介导的自噬激活有关，这为青藤碱应用于炎症反应的防治提供了实验基础和理论依据。.

Topics: Animals; Anti-Inflammatory Agents; Autophagy; Gene Expression Regulation, Enzymologic; Heme Oxygenase-1; Inflammation; Lipopolysaccharides; Macrophages; Mice; Morphinans

BACKGROUND Sinomenine (SIN) is an extract of the Chinese medicinal herb Sinomenium acutum; it has various pharmacological properties, including immunosuppression and anti-inflammation. The present study aimed to investigate whether SIN has an anti-depressant-like effect in a mouse model of depression induced by chronic unpredictable mild stress (CUMS), and to explore the underlying molecular mechanisms. MATERIAL AND METHODS A mouse model of depression was established and treated with different concentrations of SIN (30, 100, or 300 mg/kg). Then, behavioral tests, including sucrose preference test (SPT), forced swimming test (FST), and the tail suspension test (TST), were performed. The levels of norepinephrine (NE), 5-hydroxytryptamine (5-HT), and proinflammatory cytokines (interleukin-1β [IL-1β] interleukin-6 [IL-6], and tumor necrosis factor-α [TNF-α]) in the hippocampus of mice were detected by ELISA assay. The levels of p-p38, p-p65, NLRP3, ASC, and caspase-1 were measured by Western blot or/and qRT-PCR. RESULTS The results showed that SIN significantly relieved CUMSinduced depressive-like behaviors. Compared with the model mice, SIN treatment significantly increased the sucrose preference of the mice, and the immobility time in the forced swimming and the tail suspension test were shortened. In addition, SIN decreased CUMS-induced reduction in the concentrations of NE and 5-HT in the hippocampus of mice. SIN reduced CUMS-induced increases in the levels of IL-1β, IL-6, and TNF-α in the hippocampus of mice. Furthermore, activation of the p38MAPK-NF-κB pathway and the nucleotide binding and oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome induced by CUMS were inhibited by SIN treatment. CONCLUSIONS In conclusion, our results indicate the antidepressantlike effects of SIN on chronic unpredictable mild stress-induced depression in a mouse model.

Topics: Animals; Antidepressive Agents; Behavior, Animal; China; Depression; Depressive Disorder; Disease Models, Animal; Hippocampus; Inflammation; Interleukin-1beta; Interleukin-6; Male; Mice; Mice, Inbred ICR; Morphinans; Norepinephrine; Serotonin; Stress, Psychological; Tumor Necrosis Factor-alpha

Rheumatoid arthritis (RA), a chronic systemic inflammatory disorder affects many adults. Sinomenine, a natural product, has been clinically available for the treatment of RA in China. SND-117, a sinomenine derivative with much more potent activity, might serve as a candidate for anti-arthritis. The aim of the present study was to develop a sensitive and rapid high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method for quantification of SND-117 in rat plasma and to understand its absolute bioavailability. The HPLC-MS/MS method was developed and fully validated for determination of SND-117 in rat plasma, and the pharmacokinetic differences were investigated after different administration routes. The pharmacokinetics parameters were calculated by non-compartment model with DAS 3.0 software. After the oral or intravenous administration of different doses of SND-117, the time to peak is 1.5h, half-life time is 8-10h. The absolute oral bioavailability of SND-117 in rats was 9.60%. The results showed that SND-117 in rats was quickly absorbed, slowly eliminate, and the kinetics were linear. This method was suitable for pharmacokinetic studies of SNA-117 in rats.

Topics: Administration, Intravenous; Administration, Oral; Animals; Antirheumatic Agents; Area Under Curve; Arthritis, Rheumatoid; Chromatography, High Pressure Liquid; Limit of Detection; Morphinans; Rats; Rats, Sprague-Dawley; Tandem Mass Spectrometry

With the advent of aging, the morbidity rates of such diseases as osteoarthritis, rheumatoid arthritis, and osteoporosis has witnessed a significant increase. As a common rattan drug, sinomenine (SIN) has been widely applied for the treatment of various arthritic diseases in traditional Chinese medicine (TCM) clinics. Given that SIN has been reported to inhibit the expression of Prostaglandin E2 (PGE2) in several types of cells, in this study, the influence of SIN treatment on PGE2 expression in mesenchymal stem cells (MSCs), thereby changing the osteoprotegerin (OPG) receptor/activator for the nuclear factor-κ B ligand (RANKL) ratio, was investigated. Our results showed that, when compared with the untreated cells, treatment with 0.25mM SIN can down-regulate the mRNA and protein expression levels of the Prostaglandin E synthase 3 (PTGES3) or PGE2 and RANKL, while the OPG was up-regulated. After being cultured with SIN treated MSC-conditioned medium (stMSC-CM), the amount of TRAP-positive multinucleated osteoclasts differentiated from RAW264.7 cells was reduced. Also, the expression levels of specific markers for active osteoclasts were decreased when incubated with stMSC-CM. Moreover, these changes were able to be recovered when the exogenous RANKL was added to the MSC-CM culture. This indicates that the increased OPG/RANKL ratio can reduce the osteoclastogenesis of RAW264.7 cells. Our results demonstrated that SIN has an inhibitory effect on osteoclast differentiation through mechanisms involving the inhibition of the PGE2-induced OPG/RANKL ratio. SIN can also serve as a proinflammatory mediator to regulate the MSC immunosuppressive effects. Our findings suggest that SIN can be useful for the treatment of bone diseases associated with over-activity of osteoclasts.

Topics: Animals; Cell Differentiation; Cell Line; Cells, Cultured; Dinoprostone; Down-Regulation; Humans; Membrane Glycoproteins; Mesenchymal Stem Cells; Mice; Morphinans; Osteoblasts; Osteoclasts; Osteogenesis; Prostaglandin-E Synthases; RANK Ligand; RAW 264.7 Cells; RNA, Messenger; Up-Regulation

Sinomenium acutum has been used in traditional medicine to treat a painful disease such as rheumatic arthritis and neuralgia. Sinomenine, which is a main bioactive ingredient in Sinomenium acutum, has been reported to have an analgesic effect in diverse pain animal models. However little is known about the detailed mechanisms underlying peripheral analgesic effect of sinomenine. In the present study, we aimed to elucidate its cellular mechanism by using formalin-induced acute inflammatory pain model in mice. We found that intraperitoneal (i.p.) administration of sinomenine (50mg/kg) suppressed formalin-induced paw licking behavior in both the first and the second phase. Formalin-induced c-Fos protein expression was also suppressed by sinomenine (50mg/kg i.p.) in the superficial dorsal horn of spinal cord. Whole-cell patch-clamp recordings from small-sized dorsal root ganglion (DRG) neurons revealed that sinomenine reversibly increased the spike threshold and the threshold current intensity for evoking a single spike and decreased firing frequency of action potentials evoked in response to a long current pulse. Voltage-gated sodium currents (I

Topics: Animals; Antirheumatic Agents; Cells, Cultured; Disease Models, Animal; Dose-Response Relationship, Drug; Formaldehyde; Ganglia, Spinal; Inflammation; Male; Mice; Mice, Inbred C57BL; Morphinans; Neuralgia; Pain; Pain Measurement; Patch-Clamp Techniques; Proto-Oncogene Proteins c-fos; Sensory Receptor Cells; Sodium; Voltage-Gated Sodium Channels

Sinomenine hydrochloride (SH) is an ideal drug for the treatment of rheumatoid arthritis and osteoarthritis. However, high plasma concentration of systemically administered SH can release histamine, which can cause rash and gastrointestinal side effects. Topical delivery can increase SH concentration in the synovial fluid without high plasma level, thus minimizing systemic side effects. However, passive diffusion of SH was found to be inefficient because of the presence of the stratum corneum layer. Therefore, an effective method is required to compensate for the low efficiency of SH passive diffusion. In this study, transdermal experiments in vitro and clinical tests were utilized to explore the optimized parameters for electroporation of topical delivery for SH. Fluorescence experiment and hematoxylin and eosin staining analysis were performed to reveal the mechanism by which electroporation promoted permeation. In vitro, optimized electroporation parameters were 3 KHz, exponential waveform, and intensity 10. Using these parameters, transdermal permeation of SH was increased by 1.9-10.1 fold in mice skin and by 1.6-47.1 fold in miniature pig skin compared with passive diffusion. After the electroporation stimulation, the intercellular intervals and epidermal cracks in the skin increased. In clinical tests, SH concentration in synovial fluid was 20.84 ng/mL after treatment with electroporation. Therefore, electroporation with optimized parameters could significantly enhance transdermal permeation of SH. The mechanism by which electroporation promoted permeation was that the electronic pulses made the skin structure looser. To summarize, electroporation may be an effective complementary method for transdermal permeation of SH. The controlled release of electroporation may be a promising clinical method for transdermal drug administration.

Topics: Administration, Cutaneous; Aged; Aged, 80 and over; Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis; Delayed-Action Preparations; Drug Delivery Systems; Electroporation; Female; Humans; Male; Mice; Middle Aged; Morphinans; Pain; Swine; Swine, Miniature; Synovial Fluid; Transcutaneous Electric Nerve Stimulation

Sinomenine is a pure alkaloid with immunosuppressive effects that is extracted from the Chinese medicinal plant Sinomenium acutum. We studied the therapeutic effects of sinomenine on inflammatory bowel disease. In this study, we randomly divided mice into the following ten groups: Control group; DSS-induced colitis group; Salicylazosulfapyridine (SASP)-treated group; Chitosan-treated group; low-, medium-, and high-dose sinomenine-treated and sinomenine enteric-coated microspheres-treated groups. We recorded changes in colon length, disease activity index (DAI), and colon pathology, measured TLR4, MyD88, SIGIRR, NF-κB p65 protein levels and inflammatory serum cytokine levels. Except for the Control group, the weight of mice in each group decreased, the DAI of the DSS-induced colitis group was significantly higher than the other groups, and the DAIs of the sinomenine- and sinomenine enteric-coated microspheres-treated groups were significantly lower than that of the SASP-treated group. TLR4, MyD88, NF-κB p65 and proinflammatory cytokine expressions decreased dose dependently in the sinomenine and sinomenine enteric-coated microspheres-treated groups and were generally lower in the sinomenine enteric-coated microspheres groups. However, SIGIRR and anti-inflammatory IL-10 expressions exhibited the opposite pattern. Based on the superior therapeutic effect, sinomenine enteric-coated microspheres might regulate TLR/NF-κB signaling and would be beneficial for an effective and safe therapy of inflammatory bowel disease.

Topics: Animals; Anti-Inflammatory Agents; Colitis; Colon; Dextran Sulfate; Disease Models, Animal; Disease Progression; Down-Regulation; Drug Delivery Systems; Female; Inflammatory Bowel Diseases; Medicine, Chinese Traditional; Mice; Mice, Inbred BALB C; Microspheres; Morphinans; Myeloid Differentiation Factor 88; NF-kappa B; Sinomenium; Toll-Like Receptor 4

Recently, microsomal prostaglandin E synthase 1 (mPGES-1) has attracted much attention from pharmacologists as a promising strategy and an attractive target for treating various types of diseases including rheumatoid arthritis (RA), which could preserve the anti-inflammatory effect while reducing the adverse effects often occur during administration of non-steroidal anti-inflammatory drugs (NSAIDs). Here, we report that sinomenine (SIN) decreased prostaglandin (PG)E

Topics: A549 Cells; Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Experimental; Cell Culture Techniques; Cell Survival; Edema; Female; Gene Expression; Macrophages, Peritoneal; Male; Mice, Inbred DBA; Morphinans; Prostaglandin-E Synthases; Rats, Sprague-Dawley; Transfection

The aim of this study was to investigate in piglets, the anti-endotoxin and anti-inflammatory effects of sinomenine, an agent commonly found in Chinese herbal medicines. In high-, middle- and low-dose sinomenine groups, piglets were initially challenged with endotoxin (i.e., 1 mg lipopolysaccharide (LPS)/kg) by intraperitoneal (IP) injection and, 3 h later, intramuscularly (IM) with sinomenine at 1, 5, or 10 mg/kg. In a drug control group, piglets were dosed IP with vehicle and 3 h late IM with 10 mg/kg sinomenine while those in an LPS control group were challenged with 1 mg LPS/kg (IP) and then vehicle 3 h later; naïve control piglets were administered normal saline IP and then IM only. At 12, 24, and 48 h post-LPS/vehicle injection, blood samples were collected from the precaval vein of piglets. Clinical signs were recorded during the trial and index levels were analyzed by ELISA kits. The results revealed sinomenine could reduce the incidence/severity of certain LPS-induced toxicities, e.g., cell adhesion, systemic inflammation, and multiple organ dysfunction. Taken together, the data suggested to us that sinomenine might effectively be useful to regulate inflammatory responses as part of future anti-endotoxin therapies.

Topics: Animals; Animals, Newborn; Anti-Inflammatory Agents; Endotoxins; Inflammation; Injections, Intramuscular; Lipopolysaccharides; Morphinans; Severity of Illness Index; Swine; Treatment Outcome

The immunosuppressive mechanism of sinomenine in organ allotransplantation was investigated, especially its effect of blocking dendritic cell (DC) maturation, which might influence the frequency of regulatory T cells (Tregs).. Bone marrow cells from male donor Wistar rats were induced to differentiate into DCs in vitro in the presence or absence of sinomenine, and characterized by flow cytometry. These two groups of DCs were respectively injected into male recipient Sprague-Dawley rats via the tail vein, at both high and low doses. Sprague-Dawley rats receiving saline injection were used as controls. Seven days later, renal transplantation was performed from donor Wistar rats to the recipient Sprague-Dawley rats. Seven days after transplantation, spleens were collected from the recipients. The proportions of Tregs and Foxp3. With sinomenine treatment, the frequency of mature DCs was reduced, as indicated by lower expression of the surface markers CD80, CD86, and RT1B. In recipient Sprague-Dawley rats that received sinomenine-treated DCs before renal allotransplantation, the proportions of splenic Tregs and Foxp3. Pre-transplant infusion of donor-derived sinomenine-induced maturation arrested DCs could result in the increase of Foxp3

Topics: Adoptive Transfer; Animals; Antirheumatic Agents; Cell Differentiation; Dendritic Cells; Forkhead Transcription Factors; Kidney Transplantation; Male; Morphinans; Rats; Rats, Sprague-Dawley; Rats, Wistar; Spleen; T-Lymphocytes, Regulatory; Transplantation, Homologous

The purpose of this study was to investigate the feasibility of delivering sinomenine hydrochloride (SH) transdermally using composite dissolving microneedles (DM) which integrated with liquid crystals (H

Topics: Administration, Cutaneous; Animals; Drug Delivery Systems; Liquid Crystals; Morphinans; Needles; Permeability; Rats; Rats, Wistar; Skin; Skin Absorption

Zhengqing Fengtongning (ZQFTN), the pharmaceutical preparation of sinomenine (SIN) derived from the medicinal plant Sinmenium acutum, is well-known in China as an effective treatment for rheumatoid arthritis (RA). However, its histamine-release anaphylactoid reactions (HRARs) occur often in some patients. Therefore, it is desirable to establish effective clinical protocols to manage such HRARs. In the study, rat models with systemic HRARs and local HRARs of the skin were established. The level of vascular permeability and mast cell numbers was determined by quantitative analysis using Evans blue dye and histological assays. The levels of histamine, leukotriene B4 (LTB4) and IL-33 in plasma were detected by UHPLC-SPE-MS, ELISA and immunohistochemistry assays, respectively. The results demonstrated that SIN significantly induced both systemic and local HRARs in rats, showing significant decrease of body temperature, increases in vascular permeability in skin, injury of lung tissues and mast cell infiltration and IL-33 expression in skin and lung tissues. Mechanistic study showed that tranilast could prevent SIN-triggered HRARs via inhibition of H1 receptor gene expression and NF-κB signaling. Our findings provide evidence that mast cell membrane stabilizers and H1 receptor blockers effectively prevent SIN-induced HRARs, and cromolyn, cetirizine and tranilast can be used in the clinic for the management of HRARs induced by ZQFTN.

Topics: Acute Lung Injury; Anaphylaxis; Animals; Anti-Allergic Agents; Cetirizine; Cromolyn Sodium; Female; Histamine; Histamine H1 Antagonists; Histamine Release; Interleukin-33; Leukotriene B4; Lung; Mast Cells; Morphinans; NF-kappa B; ortho-Aminobenzoates; Rats, Sprague-Dawley; Receptors, Histamine H1; Signal Transduction; Skin

BACKGROUND Sinomenine hydrochloride is an alkaloid that is extracted from the Chinese herbal plant Sinomenium acutum, and is used as a herbal medicine in the treatment or rheumatic disease. This report is the first to describe a case of sinomenine hydrochloride-induced agranulocytosis. CASE REPORT A 44-year-old woman with systemic lupus erythematosus (SLE) and systemic sclerosis (SSc) was treated with sinomenine hydrochloride and developed agranulocytosis with a neutrophil count of 0.01×10^9/L. She subsequently developed an opportunistic bacterial infection. Bone marrow aspiration showed a severe reduction in the proportion of mature granulocytes. The patient discontinued sinomenine hydrochloride therapy and was treated with granulocyte colony-stimulating factor (G-CSF) and antibiotics. The patient showed a return to normal granulocyte levels within ten days of discontinuing treatment with sinomenine hydrochloride. CONCLUSIONS The findings of this case report show that monitoring of bone marrow function and granulocyte levels should be performed during treatment with sinomenine hydrochloride.

Topics: Adult; Agranulocytosis; Antirheumatic Agents; Drugs, Chinese Herbal; Female; Humans; Morphinans

Glioblastoma is the most common malignant primary brain tumor, and it is one of the causes of cancer fatality in both adult and pediatric populations. Patients with glioblastoma require chemotherapy after surgical resection and radiotherapy. Therefore, chemotherapy constitutes a viable approach for the eradication of glioblastoma cells. In this study, the anti-tumor activity of sinomenine hydrochloride (SH) was evaluated in U87 and SF767 cells in vitro and in vivo. The results showed that SH potently inhibited U87 and SF767 cell viability and did not cause caspase-dependent cell death, as demonstrated by the absence of significant early apoptosis and caspase-3 cleavage. Instead, SH activated an autophagy-mediated cell death pathway, as indicated by the accumulated microtubule-associated protein light chain 3B (LC3B)-II, triggered autophagic flux and enhanced cell viability after pretreatment with autophagy inhibitors. SH-mediated autophagy in the two cell lines was implicated in reactive oxygen species (ROS) generation, protein kinase B (Akt)-mammalian target of rapamycin (mTOR) pathway suppression and c-Jun NH2-terminal kinase (JNK) pathway activation. The ROS antioxidant

Topics: Animals; Antineoplastic Agents; Apoptosis; Autophagy; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; Brain Neoplasms; Cell Line, Tumor; Glioblastoma; Humans; JNK Mitogen-Activated Protein Kinases; Lysosomes; Male; Mice; Mice, Inbred BALB C; Microtubule-Associated Proteins; Morphinans; Reactive Oxygen Species; TOR Serine-Threonine Kinases

Surfactants generally have been used as edge activators of transfersomes. However, surfactants edge activated transfersomes frequently lead to cutaneous irritation, skin lipid loss and other side effects after dermal administration. In this study, mixed monoterpenes edge activated PEGylated transfersomes (MMPTs) were prepared by ethanol injection process with sinomenine hydrochloride as a model drug. The formulation of MMPTs was optimized by an orthogonal design. We investigated skin permeation/deposition characteristics and pharmacokinetics of sinomenine hydrochloride loaded in MMPTs by comparing with liposomes using in vitro skin tests and in vivo cutaneous microdialysis. In in vitro study, the accumulative skin permeated quantity (ASPQ) and skin permeation rate (SPR) of simonenine (SIN) in the optimized MMPTs were prominently higher than that in the other MMPTs. The optimized MMPTs had a SIN ASPQ of over three times of SIN ASPQ in the liposomes and much larger SPR of SIN compared with the latter. In contrast, the drug deposition of the optimized MMPTs in the stratum corneum was much less than that of the conventional liposomes. It was noteworthy that the drug deposition curve in the whole skin (stratum corneum-stripped skin, either) for the optimized MMPTs increased initially and then decreased with an obvious peak deposition amount at 12h, while, a relatively steady curve was observed for the liposomes. In in vivo cutaneous pharmacokinetic study, the steady state concentration (C

Topics: Administration, Cutaneous; Animals; Drug Compounding; Drug Delivery Systems; Male; Microdialysis; Monoterpenes; Morphinans; Polyethylene Glycols; Rabbits; Rats, Sprague-Dawley; Skin; Skin Absorption; Skin Irritancy Tests

Mast cells are unique immunocytes that function as sentinel cells in host defense reactions such as immediate hypersensitivity responses and anaphylactic responses. The mast cell specific receptor MRGPRX2 (Mas-related G protein-coupled receptor X2) triggers mast cell degranulation-a key process in anaphylactic reactions. We sought to better understand anaphylactic reaction induced by sinomenine hydrochloride (SH). MRGPRX2-related pseudo-allergic reactions induced by SH were investigated using the hindpaw swelling and extravasation assay in vivo and mast cell degranulation assays in vitro. MrgprB2 knockout mice exhibit a reduced SH-induced inflammation effect. Furthermore, MRGPRX2 (the orthologous gene of MrgprB2) related human mast cells are activated by SH in a dose-dependent manner; however, MRGPRX2 knockdown mast cells showed reduced degranulation. The results showed a kind of mechanism that SH-induced anaphylactoid reactions were mediated by MRGPRX2 via activating PLC molecular signaling pathways to provoke mast cells Ca

Topics: Anaphylaxis; Animals; Antirheumatic Agents; Calcium Signaling; Cell Line; Cells, Cultured; Gene Expression Regulation; Humans; Mast Cells; Mice; Mice, Knockout; Morphinans; Nerve Tissue Proteins; Peritoneum; Receptors, G-Protein-Coupled; Receptors, Neuropeptide

Evidence suggests that the dopamine receptor rate-limiting enzyme, tyrosine hydroxylase (TH), and the glutamate receptor, N-methyl-D-aspartate receptor 2B (NR2B), contribute to morphine dependence. Previous studies show that chronic exposure to morphine changes the expression of opioid receptors. In this study, we focus on the effects of sinomenine on morphine-dependent mice and its related neural mechanisms. Conditioned place preference (CPP) mouse model was established using morphine (9 mg/kg, s.c.), and their expression levels of TH and NR2B were observed by immunohistochemistry. Moreover, their mu opioid receptor (MOR) and delta opioid receptor (DOR) contents were assessed using quantitative reverse transcription polymerase chain reaction. Results showed that high sinomenine dose (80 mg/kg) effectively attenuated the behavior of CPP mice and reversed increased expression levels of TH and NR2B induced by morphine. Moreover, compared with the morphine group, sinomenine up-regulated the content of MOR to a normal level but did not significantly affect the DOR expression. In summary, these data indicate that sinomenine can inhibit morphine dependence by increasing the expression levels of TH, NR2B, and MOR in the mouse brain; however, DOR may not contribute to this effect.

Topics: Animals; Brain; Mice; Morphinans; Morphine; Morphine Dependence; Neurons; Receptors, N-Methyl-D-Aspartate; Receptors, Opioid; Tyrosine 3-Monooxygenase

New strategies for the treatment of clear-cell renal cell carcinoma (ccRCC) are urgently needed. Recently, accumulating evidence has demonstrated that sinomenine hydrochloride (SH), extracted from the plant sinomenium acutum, has potent anti-cancer activity in multiple human malignant solid tumors. However, the underlying molecular mechanism by which SH treatment exerts its antagonizing tumorigenic effects has not been clearly elucidated. In this study, we showed that SH treatment exerted profound effects on cell growth in ccRCC in a dose- and time-dependent manner in vitro. Furthermore, treatment with SH significantly suppressed the migration, invasiveness and angiogenesis of ccRCC cells in vitro. Mechanistically, we revealed that SH treatment blocked epithelial-mesenchymal transition (EMT) through the downregulation of the expression of the transcription factors Snail1 and Twist in ccRCC cells. Additionally, the depletion of p-Smad2 and p-Smad3 was required for SH treatment to inhibit EMT effectively. Taken together, these findings indicate an anti-cancer role for SH in ccRCC cells and reveal a potential molecular mechanism by which Smad / EMT axis functions in ccRCC, as its hyperactivation has been associated with cell proliferation, migration, invasiveness and angiogenesis in this cancer type. These observations suggest that SH can act as an efficacious adjuvant chemotherapeutic candidate that targets the Smad/EMT axis in patients with ccRCC.

Topics: Carcinoma, Renal Cell; Cell Movement; Cell Proliferation; Dose-Response Relationship, Drug; Drug Delivery Systems; Epithelial-Mesenchymal Transition; Human Umbilical Vein Endothelial Cells; Humans; Kidney Neoplasms; Morphinans; Neoplasm Invasiveness; Neovascularization, Pathologic; Smad Proteins

To investigate the effects of sinomenine on growth and apoptosis of MCL Jeko-1 cell line and its mechanism.. The proliferation rate of Jeko-1 cells treated by different doses of sinomenine was assayed by MTT method; and the cell apoptosis was detected by flow cytometry. The expressions of Cyclin D1, BCL-2, BAX, Caspase-3, Akt signaling pathway protein Akt, phosphorylated-Akt (p-Akt), and phosphorylated-mTOR (p-mTOR), phosphorylated- P70S6K(p-P70S6K) were determined by Western blot.. The growth of Jeko-1 cell line was inhibited by Sinomenine. The apoptosis rates of Jeko-1 cells treated by 0, 1, 2, and 4 mmol /L of Sinomenine for 24 hours were (2.21±1.05) %, (11. 29±2.42)%, (18.79±2.84) %, (31.05±3.52) % respectively, and with very statistically significant difference(P<0.01). The expressions of p-Akt, p-mTOR, p-P70S6K were down-regulated, but total Akt expression was not changed. The expressions of cyclin D1 and BCL-2 were down-regulated, but that of BAX, and Caspase-3 were up-regulated.. The sinomenine can inhibit Jeko-1 cell proliferation, which may be realized through down-regulating the phosphorylation level of p-Akt, p-mTOR, and p-P70S6K, thus inhibiting the Akt signaling pathway and promoting the cell apoptosis.

Topics: Apoptosis; Cell Line, Tumor; Cell Proliferation; Humans; Lymphoma, Mantle-Cell; Morphinans; Proto-Oncogene Proteins c-akt

Rheumatoid arthritis (RA) is a systemic autoimmune disorder mainly characterized by inflammation of the synovial tissue that can lead to destruction of bone and cartilage. Sinomenine is an alkaloid extracted from the stem of the Chinese medicinal plant Sinomenium acutum. It has been reported that sinomenine has immunosuppressive and anti-inflammatory properties. However, the molecular mechanism underlying the effect of sinominine on IL-1β-induced human RA fibroblast-like synoviocytes (RAFLS) is poorly understood. Therefore, in this study, we investigated the effect of sinomenine on the expression of inflammatory cytokines in IL-1β-treated human RAFLS in vitro and the underlying mechanism. RAFLS viability was evaluated using the MTS assay after sinomenine treatment. The levels of inflammatory cytokines were measured with ELISA, RT-PCR and western blot, respectively. The levels of TLR4 and its downstream signaling targets were determined by western blot analysis. We found that sinomenine suppressed not only NO and PGE2 production but also iNOS and COX-2 expression in IL-1β-induced RAFLS. It also inhibited the expression of TNF-α and IL-6 in IL-1β-stimulated RAFLS. Furthermore, sinomenine prevented IL-1β-induced TLR4, MyD88 and p-NF-κB p65 expression. Taken together, these results demonstrated that sinomenine prevented IL-1β-induced inflammation in human RAFLS at least in part by inhibiting the TLR4/MyD88/NF-κB signaling pathway, suggesting that sinomenine could be a potential agent in the treatment of RA.

Topics: Antirheumatic Agents; Arthritis, Rheumatoid; Blotting, Western; Cell Survival; Cells, Cultured; Cytokines; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Gene Expression Regulation; Humans; Inflammation; Morphinans; Myeloid Differentiation Factor 88; NF-kappa B; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Sinomenium; Synoviocytes; Toll-Like Receptor 4

Topics: Analgesics; Animals; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Chronic Pain; Freund's Adjuvant; Gyrus Cinguli; Inflammation; Mice; Morphinans; Phosphorylation; Receptors, N-Methyl-D-Aspartate; Signal Transduction; Up-Regulation

Diabetic cardiomyopathy is a severe complication of diabetes mellitus (DM). The goal of current work was to study the effects of sinomenine on streptozotocin-induced cardiomyopathy in rats. DM in rats was induced by intraperitoneal injection of streptozotocin. Cardiac function was evaluated by measuring left ventricle end-diastolic diameter, left ventricle end-systolic diameter and ejection fraction. Cardiac inflammation was evaluated by the degree of infiltration of T lymphocytes and the levels of pro-inflammatory cytokines. Sinomenine attenuated diabetic symptoms without affecting plasma glucose. Cardiac dysfunction in the sinomenine-treated diabetic rats was significantly improved, as reflected by decreased levels of left ventricle end-diastolic diameter, left ventricle end systolic diameter and an increased level of ejection fraction. Sinomenine observably reduced cardiomyocyte hypertrophy in DM rats. Moreover, sinomenine reduced infiltration of CD3+ and CD68+ positive cells and decreased the levels of tumor necrosis factor-α, interlukin-1 and interlukin-6. Finally, sinomenine-treated rats showed a reduced expression of NF-κB and an increased expression of IκB in the myocardium compared with the myocardium of untreated diabetic rats. Our results indicate sinomenine significantly improves cardiac function in diabetic rats, which may be attributed to the deactivation of NF-κB and the blockade of inflammatory cytokine-mediated immune reactions.

Topics: Animals; Cardiotonic Agents; Cytokines; Diabetic Cardiomyopathies; Dose-Response Relationship, Drug; Immunosuppressive Agents; Inflammation Mediators; Male; Morphinans; NF-kappa B; Rats; Rats, Wistar; Treatment Outcome

Type 2 diabetes mellitus (T2DM) accounts for more than 90% of all cases of diabetes mellitus (DM). Diabetic neuropathic pain (DNP) is a common complication of T2DM. Sinomenine is a natural bioactive component extracted from the Sinomenium acutum and has anti-inflammatory effects. The aim of our study was to investigate the effects of sinomenine on DNP mediated by the P2X

Topics: Animals; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Diabetic Neuropathies; Ganglia, Spinal; HEK293 Cells; Humans; Hyperalgesia; Male; Morphinans; Pain Threshold; Purinergic P2X Receptor Antagonists; Rats; Rats, Sprague-Dawley; Receptors, Purinergic P2X3

Sinomenine hydrochloride (SH) has been investigated for its anti-tumor growth effect. We have previously reported that SH inhibited breast cancer cell proliferation via MAPKs signaling. However, whether SH could inhibit tumor metastasis has not been fully explored. In this study, we found that SH suppressed the metastasis potential of breast cancer cells. The wound healing and transwell assays showed that SH inhibited the migration and invasion ability of both 4T1 and MDA-MB-231 breast cancer cells. The orthotopic mouse model of 4T1 and the experimental mouse model of MDA-MB-231-luc (MDA-MB-231 cell line expressing firefly luciferase) demonstrated that SH treatment inhibited breast cancer metastasis by inhibiting epithelial-mesenchymal transition (EMT) and cancer stem cell (CSC) properties without obvious hepatotoxicity and renal toxicity. We also found that SH decreased spleen volume and weight in both mouse models, especially in the 4T1 mouse model. IL-6, a strong inflammatory factor causing EMT, was remarkably reduced. Overall, this anti-metastasis effect of SH could be possibly caused by attenuating inflammatory reaction, which led to inhibition of EMT and CSC characteristics of breast cancer cells. This study, together with our previous one, provides more evidence of SH as a potential drug for breast cancer therapy.

Topics: Animals; Antirheumatic Agents; Cell Line, Tumor; Disease Models, Animal; Epithelial-Mesenchymal Transition; Female; Humans; Inflammatory Breast Neoplasms; Mammary Neoplasms, Experimental; Mice; Morphinans; Neoplasm Metastasis; Neoplastic Stem Cells; RAW 264.7 Cells

Sinomenine (SN), a purified alkaloid from Chinese herb Sinomenium acutum that was used preferentially in the treatment of rheumatoid diseases, has exerted neuroprotective effects and anti-inflammatory properties in many previous studies. Some studies have revealed that the antioxidant property of SN, acting mainly through inhibiting NADPH oxidase activation, was involved in the beneficial effects of SN. However, SN belongs to the family of dextrorotatory morphinan analogues, which may initiate elevation of reactive oxygen species (ROS) levels. Thus in the present report, we conducted studies to examine its impact and mechanism on the resistance of PC12 neuronal cells to oxidative stress. Precondition with SN (0.1-5 μM) for 12 h significantly decreased H

Topics: Animals; Animals, Newborn; Antioxidants; Apoptosis; Cell Survival; Cells, Cultured; Cytoprotection; Dose-Response Relationship, Drug; Hydrogen Peroxide; Morphinans; Neurons; Oxidative Stress; PC12 Cells; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Up-Regulation

The purpose of the present study was to prepare and optimize sinomenine (SIN) pluronic lecithin organogels system (PLO), and to evaluate the permeability of the optimized PLO in vitro and in vivo. Box-Behnken design was used to optimize the PLO and the optimized formulation was pluronic F127 of 19.61%, lecithin of 3.60% and SIN of 1.27%. The formulation was evaluated its skin permeation and drug deposition both in vitro and in vivo compared with gel. Permeation and deposition studies of PLO were carried out with Franz diffusion cells in vitro and with microdialysis in vivo. In vitro studies, permeation rate (Jss) of SIN from PLO was 146.55 ± 2.93 μg/cm(2)/h, significantly higher than that of gel (120.39 μg/cm(2)/h) and the amount of SIN deposited in skin from the PLO was 10.08 ± 0.86 μg/cm(2), significantly larger than that from gel (6.01 ± 0.04 μg/cm(2)). In vivo skin microdialysis studies showed that the maximum concentration (Cmax) of SIN from PLO in "permeation study" and "drug-deposition study" were 150.27 ± 20.85 μg/ml and 67.95 μg/ml, respectively, both significantly higher than that of SIN from gel (29.66 and 6.73 μg/ml). The results recommend that PLO can be used as an advantageous transdermal delivery vehicle to enhance the permeation and skin deposition of SIN.

Topics: Administration, Cutaneous; Animals; Antirheumatic Agents; Drug Delivery Systems; Drug Evaluation, Preclinical; Gels; Lecithins; Male; Morphinans; Organ Culture Techniques; Poloxamer; Rats; Rats, Sprague-Dawley; Skin Absorption

Sinomenine hydrochloride-loaded dissolving microneedles (SH-DM) were fabricated by maltose and poly-lactic-co-glycolic acid using a casting method. The mechanical strength of SH-DM was investigated by an insertion test. In vivo transdermal absorption experiment was performed to evaluate the percutaneous absorption of SH-DM, sinomenine hydrochloride gel (SH-G) was used as a control. The results demonstrated that prepared SH-DM was morphologically intact with sufficient mechanical strength after inserting into aluminum foil and rat skin. The value of area under curve obtained after administration of SH-DM through transdermal in rabbits showed a significantly rise and was 1.99 times higher than that of SH-G, and the relative bioavailability value was 199.21%. These results showed that SH-DM enhanced bioavailability and permeability of SH.

Topics: Administration, Cutaneous; Animals; Biological Availability; Drug Delivery Systems; Female; Male; Morphinans; Needles; Rabbits; Rats; Skin; Skin Absorption; Solubility

In this work, a new and facile method was introduced to prepare molecularly imprinted polymers (MIPs) based on nano clay hectorite (Hec) for sinomenine hydrochloride (SM) analysis. Hec was firstly dissolved in distilled water in order to swell adequately, followed by a common precipitation polymerization with SM as the template, methacrylic acid as monomer, ethylene glycol dimethacrylate as a crosslinker and 2,2-azobisisobutyronitrile as an initiator. Hec@SM-MIPs were characterized by Fourier transform infrared spectrometer, transmission electron microscopy, scanning electron microscopy, energy dispersive X-ray spectroscopy and X-ray diffraction. The maximum binding capacity of Hec@SM-MIPs, SM-MIPs and non-imprinted polymers (NIPs) (Hec@NIPs) was 57.4, 16.8 and 11.6 mg/g, respectively. The reason for this result may be that Hec@SM-MIPs have more binding sites and imprinted cavities for template molecule. Equilibrium data were described by the Langmuir and Freundlich isotherm models. The results showed that the Hec@SM-MIPs adsorption data correlated better with the Langmuir equation than the Freundlich equation under the studied concentration range. In vitro drug release experiment, Hec@SM-MIPs have a better ability to control SM release than SM-MIPs. Therefore, Hec@SM-MIPs were successfully applied to extraction of SM and used as the materials for drug delivery system.

Topics: Adsorption; Drug Carriers; Drug Liberation; Kinetics; Molecular Imprinting; Morphinans; Polymerization; Polymethacrylic Acids; Silicates

Mast cells are vital mediators of drug allergy and, therefore, studying the relationship between drug allergy and mast cells is essential. Sinomenine is the principal active component of Sinomenium acutum, which has anti-inflammatory and anti-immune effects, and is used to treat various rheumatoid diseases. However, allergic responses to sinomenine are frequently reported. Therefore, this study assessed the effects of sinomenine on mast cell activation to characterize its allergic effects and the underlying mechanisms. Enzyme-linked immunosorbent assay (ELISA), western blot analyses, and degranulation assays were performed to measure pro-inflammatory and allergic mediators in P815 cells. The allergenic effects of sinomenine were also determined in mice by using active general anaphylaxis (ASA). The results indicated that sinomenine induced inositol-1,4,5-trisphosphate (IP

Topics: Anaphylaxis; Animals; Anti-Allergic Agents; Anti-Inflammatory Agents; Calcium; Cell Degranulation; Cell Line, Tumor; Drug Hypersensitivity; Inflammation; Inositol 1,4,5-Trisphosphate Receptors; Interleukin-6; Male; Mast Cells; Mice; Mice, Inbred BALB C; Morphinans; Phospholipase C gamma; Phosphorylation; Signal Transduction; src-Family Kinases; Up-Regulation

Sinomenine is originally derived from medicinal herb and used preferentially in treatment of rheumatoid diseases in Far East regions. SIN has strong anti-inflammatory and immune-regulatory properties, acting mainly through inhibiting NF-kB signaling. Although the upstream target through which SIN affects NF-kB activity is unknown, evidence suggests that SIN might regulate inflammation through Nrf2 signaling. In this study we explored the role of Nrf2 in mediating SIN's anti-inflammation and kidney protection in a mouse model of obstructive nephropathy. We found that SIN is an activator of Nrf2 signaling. It markedly increased Nrf2 protein level, Nrf2 nuclear translocation, Nef2 transcription capacity, and the downstream protein expression. We further demonstrated that SIN activation of Nrf2 is likely due to its repression of the Nrf2 inhibitor Keap1 since it drastically reduced Keap1 protein through the PKC-sensitive ubiquitination-proteasomal degradation. SIN treatment of nephropathy mice effectively reduced the kidney damage and inflammatory responses, balanced renal oxidative stress, and improved the pathological protein expression in an Nrf2 dependent manner. In addition, SIN also Nrf2-dependently modulated macrophage M1/M2 polarization and inhibited the IkBα phosphorylation and NF-kB nuclear translocation, hence revealing an important upstream event that contributed to its anti-inflammation and tissue protection. Taken together our study has identified a novel pathway through which SIN exerts its anti-inflammation and renal protective functions, and provided a molecular basis for SIN potential applications in the treatment of kidney and other inflammatory disorders.

Topics: Acute Kidney Injury; Animals; Disease Models, Animal; Heme Oxygenase-1; Humans; Inflammation; Kidney; Macrophages; Mice; Morphinans; NF-E2-Related Factor 2; NF-kappa B; Oxidative Stress; Signal Transduction

Asthma is an inflammatory disease that involves airway inflammation and remodeling. Sinomenine (SIN) has been demonstrated to have immunosuppressive and anti‑inflammatory properties. The aim of the present study was to investigate the inhibitory effects of SIN on airway inflammation and remodeling in an asthma mouse model and observe the effects of SIN on the transforming growth factor‑β1 (TGF‑β1)/connective tissue growth factor (CTGF) pathway and oxidative stress. Female BALB/c mice were sensitized by repetitive ovalbumin (OVA) challenge for 6 weeks in order to develop a mouse model of asthma. OVA‑sensitized animals received SIN (25, 50 and 75 mg/kg) or dexamethasone (2 mg/kg). A blank control group received saline only. The area of smooth muscle and collagen, levels of mucus secretion and inflammatory cell infiltration were evaluated 24 h subsequent to the final OVA challenge. mRNA and protein levels of TGF‑β1 and CTGF were determined by reverse transcription‑quantitative polymerase chain reaction and immunohistology, respectively. The indicators of oxidative stress were detected by spectrophotometry. SIN significantly reduced allergen‑induced increases in smooth muscle thickness, mucous gland hypertrophy, goblet cell hyperplasia, collagen deposition and eosinophilic inflammation. The levels of TGF‑β1 and CTGF mRNA and protein were significantly reduced in the lungs of mice treated with SIN. Additionally, the total antioxidant capacity was increased in lungs following treatment with SIN. The malondialdehyde content and myeloperoxidase activities in the lungs from OVA‑sensitized mice were significantly inhibited by SIN. In conclusion, SIN may inhibit airway inflammation and remodeling in asthma mouse models, and may have therapeutic efficacy in the treatment of asthma.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Connective Tissue Growth Factor; Dexamethasone; Drug Evaluation, Preclinical; Female; Lung; Mice, Inbred BALB C; Morphinans; Oxidative Stress

This study examined the antinociceptive effects of sinomenine in a rat model of postoperative pain.. Male and female rats were subjected to a surgical incision in the right hind paw, and the von Frey filament test was used to measure mechanical hypersensitivity after drug or vehicle treatment (p.o. or i.p.). Rats were treated daily with sinomenine before or after the surgery and the AUCs of the antinociceptive effects measured during a 4 h period were calculated to determine the ED50 values of sinomenine. The anti-hyperalgesic effects of different doses of a combination of sinomenine and acetaminophen (paracetamol) were assessed in another group of rats. Dose combinations were determined by using a fixed ratio dose-addition analysis method.. Sinomenine (5-80 mg·kg(-1) ) produced dose-dependent antinociceptive effects in rats that had been subjected to surgery and this effect lasted for 4 h. The potency of sinomenine, given i.p. or p.o., did not differ between male and female rats. However, sinomenine was fourfold more potent when given i.p. than p.o. The GABAA receptor antagonist bicuculline blocked the antinociceptive effects of sinomenine. The antinociceptive effect of a daily treatment with sinomenine remained stable throughout the course of postoperative pain. Pretreatment with sinomenine did not alter the mechanical hypersensitivity post-surgery. The combination of sinomenine with acetaminophen produced an infra-additive interaction.. Sinomenine demonstrated significant antinociceptive activity against postoperative pain and may be a useful novel pharmacotherapy for the management of postoperative pain.

Topics: Acetaminophen; Analgesics; Animals; Dose-Response Relationship, Drug; Female; Male; Morphinans; Pain, Postoperative; Rats; Rats, Sprague-Dawley

Sinomenine, a pure alkaloid extracted from the Chinese medicinal plant Sinomenium acutum, and sinomenine hydrochloride (SN) has been successfully used for the therapy of rheumatoid arthritis (RA) and kidney diseases. Autophagy is a cytoprotective mechanism used by podocytes and other cells to alleviate the effects of oxidative stress, and angiotensin II (Ang II) significantly promotes podocyte autophagy. However, excessive autophagy may lead to cell death and podocyte depletion. The present study evaluated the effect of SN in podocytes induced by Ang II.. Podocytes were pretreated with graded concentrations (10(-8) M ∼ 10(-4) M) of SN and then stimulated with Ang II. The LC3B protein and the p47-phox membrane fraction were measured by Western blot. Autolysosomes were assessed by transmission electron microscopy. FACS was used to quantify the ROS produced by podocytes. The translocation of p47-phox to the membrane was investigated by immunofluorescence.. The 10(-8) M ∼ 10(-4) M of SN alone did not effect ROS generation or podocyte autophagy. The 10(-8) M and 10(-6) M SN attenuated Ang II-induced autophagy in podocytes. Furthermore, SN decreased the level of ROS generation in Ang II-induced podocytes via inhibition of NOX subunit p47-phox translocation to the membrane.. The appropriate concentration of SN attenuated Ang II-induced podocyte autophagy through ROS generation, at least in part, by regulating NOX subunit p47-phox translocation to the membrane.

Topics: Angiotensin II; Autophagy; Cell Line, Transformed; Exocytosis; Humans; Morphinans; NADPH Oxidases; Oxidative Stress; Podocytes; Reactive Oxygen Species

Sinomenine (SIN) is an anti-inflammatory and anti-arthritic alkaloid derived from Sinomenioum acutum. Effects of SIN on lipopolysaccharide (LPS)-induced osteolysis have not been reported. Here, we found that SIN reduced LPS-induced erosion of skull bones in C57BL/6 mice significantly. LPS can induce bone-absorbing osteoclast formation independent of RANKL in pre-osteoclastic RAW264.7 cells in vitro. Here, SIN suppressed LPS-induced osteoclast formation and osteoclast survival in RAW264.7 cells. Expression of osteoclastic-specific marker genes was also inhibited by SIN during osteoclast differentiation and osteoclast survival stimulated with LPS. SIN showed much stronger inhibitory effects on expression of Fra-1 and MMP-9 mRNA in osteoclast differentiation rather than osteoclast survival. SIN dramatically inhibited LPS-induced TNF-α production in vitro and in vivo. Further signaling studies revealed that SIN suppressed the activation and relative gene expression of three notable nuclear factors (NF-κB, AP-1, NFAT), reduced intracellular levels of Ca(2+), and down-regulated phosphorylation of MAPK p38 (but not JNK) in LPS-induced osteoclastogenesis. Focusing on upstream signals after LPS stimulation, SIN decreased expression of TLR4 and TRAF6 during osteoclast differentiation, and reduced expression of TLR4 (but not TRAF6) in osteoclast survival. These data suggest that SIN might be a potential agent for the treatment of osteolysis caused by Gram-negative bacteria infection or inflammation due to its inhibition of osteoclastogenesis through reduction of TLR4/TRAF6 expression and downstream signal transduction.

Topics: Animals; Calcium; Cell Survival; Lipopolysaccharides; MAP Kinase Signaling System; Mice; Morphinans; NF-kappa B; NFATC Transcription Factors; Osteoclasts; Osteogenesis; Osteolysis; RAW 264.7 Cells; TNF Receptor-Associated Factor 6; Toll-Like Receptor 4; Transcription Factor AP-1; Tumor Necrosis Factor-alpha

The objective of this study is to investigate the therapeutic effect of sinomenine (SIN) on rat cerebral ischemia-reperfusion (IR) injury and the molecular mechanism.. One hundred thirty-five rats were equally randomized into sham-operated group, middle cerebral artery occlusion (MCAO) group, and SIN group, and reversible rat MCAO model was made according to the Longa method for the MCAO and SIN groups. Then, 15 rats from each group were decapitated at 6, 12, and 24 hours after reperfusion to obtain brain tissue samples. Rats in the SIN group were injected with sinomenine by tail vein (90 mg/kg) 1 hour before ischemia; rats in the MCAO and sham-operated groups were administrated with the same volume of saline. Neurological severity score (NSS), infarction volume, ischemic brain water content, and blood-brain barrier (BBB) permeability were determined at corresponding time points. Acid-sensing ion channel (ASIC) 1a mRNA level was determined by quantitative real-time polymerase chain reaction; ischemic brain contents of lactic acid (LD), lactic dehydrogenase (LDH), ATPase, and inflammatory factors were determined by spectrophotometric method.. At 12 hours after reperfusion and since then, NSS in the SIN group decreased obviously; infarction volume, brain water content, and BBB permeability in the SIN group were lower than those in the MCAO group (P < .05). IR injury resulted in the upregulation of the contents of ASIC1a mRNA, LD, LDH, and inflammatory factors and the downregulation of the contents of ATPase, while SIN could reverse the upregulation/downregulation effect induced by IR injury (P < .05).. Through its anti-inflammation effect, which alleviates acidosis, improves energy metabolism, and inhibits ASIC1a level, SIN protects ischemic rat brain against cerebral IR injury.

Topics: Acid Sensing Ion Channels; Acidosis; Animals; Anti-Inflammatory Agents; Blood-Brain Barrier; Brain; Capillary Permeability; Cytoprotection; Disease Models, Animal; Energy Metabolism; Infarction, Middle Cerebral Artery; Inflammation Mediators; Male; Morphinans; Neuroprotective Agents; Rats, Sprague-Dawley; Reperfusion; Reperfusion Injury; Time Factors

Osteosarcoma is the most common primary malignant tumor of the bone. The long-term survivals continue to be unsatisfactory for patients with metastatic and recurrent disease. Metastasis is still a severe challenge in osteosarcoma treatment. Sinomenine, an alkaloid from traditional Chinese medicine, has been proved to possess potent antitumor and anti-invasion effect on various cancers. However, the effect of sinomenine on human osteosarcoma and the underlying mechanisms remains unknown. We report here that sinomenine inhibited proliferation by inducing S phase arrest and suppressing the clone formation. Significant inhibitory effects were found in invasion and metastasis in osteosarcoma, but little cytotoxicity was observed in tested concentrations. Exposure to sinomenine resulted in suppression of invasion and migration in osteosarcoma cells as well as tube formation ability in the human umbilical vein endothelial cells (HUVEC) and U2OS cells. Furthermore, it demonstrated that CXCR4 played a key role contributing to invasion in osteosarcoma which is considered to be a core target site in sinomenine treatment. Sinomenine inhibited invasion by suppressing CXCR4 and STAT3 phosphorylation then downregulating the expression of MMP-2, MMP-9, RANKL, VEGF downstream. In addition, then RANKL-mediated bone destruction stimulated by osteoclastogenesis and VEGF-related neovascularization were restrained. Importantly, in vivo, sinomenine suppressed proliferation, osteoclastogenesis and bone destruction. Through these various comprehensive means, sinomenine inhibits metastasis in osteosarcoma. Taken together, our results revealed that sinomenine caused S phase arrest, inhibited invasion and metastasis via suppressing the CXCR4-STAT3 pathway and then osteoclastogenesis-mediated bone destruction and neovascularization in osteosarcoma. Sinomenine is therefore a promising adjuvant agent for metastasis control in osteosarcoma.

Topics: Angiogenesis Inhibitors; Animals; Bone Neoplasms; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Gene Expression Regulation, Neoplastic; Human Umbilical Vein Endothelial Cells; Humans; Mice; Morphinans; Neoplasm Metastasis; Osteosarcoma; Receptors, CXCR4; Rho Guanine Nucleotide Exchange Factors; S Phase Cell Cycle Checkpoints; Signal Transduction; Xenograft Model Antitumor Assays

The investigation was undertaken to evaluate the effect of sinomenine (Sin) on experimental adjuvant arthritis rats stimulated by Freund's complete adjuvant and explore the corresponding potential molecular mechanism. The content of proinflammatory cytokines such as tumor necrosis factor-alpha, interleukin-1 beta and interleukin-6 were detected. Besides, canonical nuclear factor kappa B (NF-κB) pathway was also assessed to evaluate the antiarthritic potential of sinomenine. Pathological sections of rat paws showed sinomenine and diclofenac sodium significantly alleviated articular cartilage lesion, cellular infiltration, epithelial cell degeneration, synovial tissue vasodilation and congestion. The phosphorylations of inhibitor of kappaB alpha and NF-κB subunit p65 were downregulated with the treatment of sinomenine in dose dependent manners, as well as proinflammatory cytokines. Therefore, it was assumed that sinomenine might be a new therapeutic candidate to treat arthritis. © 2016 IUBMB Life, 68(6):429-435, 2016.

Topics: Adaptor Proteins, Signal Transducing; Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Experimental; Cytokines; Edema; Freund's Adjuvant; I-kappa B Proteins; Morphinans; Nuclear Proteins; Nuclear Receptor Interacting Protein 1; Phosphorylation; Rats, Sprague-Dawley; Transcription Factor RelA

Renal fibrosis is the common feature of chronic kidney disease and mainly mediated by TGFβ-associated pro-fibrogenic signaling, which causes excessive extracellular matrix accumulation and successive loss of kidney functions. Sinomenine (SIN), an alkaloid derived from medicinal herb extensively used in treatment of rheumatoid arthritis and various inflammatory disorders, displays renal protective properties in experimental animals; however its pharmacological potency against renal fibrosis is not explored. In this study we report that SIN possesses strong anti-renal fibrosis functions in kidney cell and in mouse fibrotic kidney. SIN beneficially modulated the pro-fibrogenic protein expression in TGFβ-treated kidney cells and attenuated the renal fibrotic pathogenesis incurred by unilateral ureteral obstruction (UUO), which correlated with its activation of Nrf2 signaling - the key defender against oxidative stress with anti-fibrotic potentials. Further investigation on its regulation of Nrf2 downstream events revealed that SIN significantly balanced oxidative stress via improving the expression and activity of anti-oxidant and detoxifying enzymes, and interrupted the pro-fibrogenic signaling of TGFβ/Smad and Wnt/β-catenin. Even more impressively SIN achieved its anti-fibrotic activities in an Nrf2-dependent manner, suggesting that SIN regulation of Nrf2-associated anti-fibrotic activities constitutes a critical component of SIN's renoprotective functions. Collectively our studies have demonstrated a novel anti-fibrotic property of SIN and its upstream events and provided a molecular basis for SIN's potential applications in treatment of renal fibrosis-associated kidney disorders.

Topics: Animals; Antirheumatic Agents; Catalase; Fibrosis; Glutathione Peroxidase; HEK293 Cells; Humans; Kidney Diseases; Mice; Morphinans; NF-E2-Related Factor 2; Oxidative Stress; Signal Transduction; Superoxide Dismutase; Transforming Growth Factor beta1

The degranulation of mast cells and basophils is often initiated by a number of pathophysiological responses, especially in allergic and inflammatory conditions. Efficient techniques and methods for determining the level of such degranulation are highly demanded for laboratory and clinical studies. In this work, a rapid and sensitive assay based on the particle analysis of granules in RBL-2H3 cells, a cell line widely used as a convenient model system to study the degranulation of mast cells and basophils, was developed to detect cell degranulation using a Nanosight NS300 in light scatter mode and dynamic light scattering (DLS) on a Malvern Zetasizer Nano-ZS instrument. Using this method, drug-induced mast cell degranulation and systemic anaphylaxis were efficiently determined both in cell culture medium and blood samples from animals in the current study. This promising method is expected to be widely used for screening anti-allergic and anti-inflammatory drugs both in vitro and in vivo models, as well as for determining the level of mast cell degranulation of the patients in the clinic.

Topics: Animals; Basophil Degranulation Test; Basophils; Biosensing Techniques; Calcium; Cell Degranulation; Cell Line, Tumor; Dynamic Light Scattering; Equipment Design; Exocytosis; Female; Ionomycin; Mast Cells; Morphinans; p-Methoxy-N-methylphenethylamine; Predictive Value of Tests; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Time Factors

As an early sign of diabetic cardiovascular disease, endothelial dysfunction may contribute to progressive diabetic nephropathy (DN). Endothelial hyperpermeability induced by hyperglycemia (HG) is a central pathogenesis for DN. Sinomenine (SIN) has strong anti-inflammatory and renal protective effects, following an unknown protective mechanism against HG-induced hyperpermeability. We herein explored the role of SIN in vitro in an HG-induced barrier dysfunction model in human renal glomerular endothelial cells (HRGECs). The cells were exposed to SIN and/or HG for 24 h, the permeability of which was significantly increased by HG. Moreover, junction protein occludin in the cell-cell junction area and its total expression in HRGECs were significantly decreased by HG. However, the dysfunction of tight junction and hyperpermeability of HRGECs were significantly reversed by SIN. Furthermore, SIN prevented HG-increased reactive oxygen species (ROS) by activating nuclear factor-E2-related factor 2 (Nrf2). Interestingly, activation of RhoA/ROCK induced by HG was reversed by SIN or ROCK inhibitor. HG-induced hyperpermeability was prevented by SIN. High ROS level, tight junction dysfunction and RhoA/ROCK activation were significantly attenuated with knockdown of Nrf2. Mediated by activation of Nrf2, SIN managed to significantly prevent HG-disrupted renal endothelial barrier function by suppressing the RhoA/ROCK signaling pathway through reducing ROS. We successfully identified a novel pathway via which SIN exerted antioxidative and renal protective functions, and provided a molecular basis for potential SIN applications in treating DN vascular disorders.

Topics: Capillary Permeability; Cells, Cultured; Dose-Response Relationship, Drug; Endothelial Cells; Glucose; Humans; Kidney Glomerulus; Morphinans; Reactive Oxygen Species; rho-Associated Kinases; rhoA GTP-Binding Protein; Signal Transduction; Treatment Outcome

Phytantriol (PT), ethanol (ET) and water were used to prepare in situ cubic liquid crystal (ISV2). The pseudo-ternary phase diagram of PT-ET-water was constructed and isotropic solution formulations were chosen for further optimization. The physicochemical properties of isotropic solution formulations were evaluated to optimize the composition of ISV2. In situ hexagonal liquid crystals (ISH2) were prepared based on the composition of ISV2 with the addition of vitamin E acetate (VitEA) and the amount of VitEA was optimized by in vitro release behavior. The phase structures of liquid crystalline gels formed by ISV2 and ISH2 in excess water were confirmed by crossed polarized light microscopy and small angle X-ray scattering, respectively. Rheological properties of ISV2 and ISH2 were studied by a DHR-2 rheometer. In vitro drug release studies were conducted by using a dialysis membrane diffusion method. Pharmacokinetics was investigated by determination of sinomenine hydrochloride (SMH) concentration in synovial membrane after intra-articular injection of SMH-loaded ISH2 in adjuvant-induced arthritis rats. The optimal ISV2 (PT/ET/water, 64 : 16 : 20, w/w/w) loaded with 6 mg x g(-1) of SMH showed a suitable pH, injectable and formed a cubic liquid crystalline gel in situ with minimum water absorption in the shortest time. The optimal ISV2 was able to sustain the drug release for 144 h. The optimal ISH2 system was prepared by addition of 5% VitEA into PT in the optimal ISV2 system. This ISH2 (PT/VitEA/ET/water, 60.8 : 3.2 : 16 : 20, w/w/w/w) was an injectable isotropic solution with suitable pH. The new ISH2 was able to sustain the drug release for more than 240 h. Local pharmacokinetics study indicated that the retention time and AUC(0-∞) of ISH2 group were increased significantly compared with that of SMH solution group and the AUC(0-∞) of ISH2 group was 6.01 times higher than that of SMH solution group. The developed ISH2 was suitable for intra-articular injection that may apply to patients in the treatment of rheumatoid arthritis.

Topics: alpha-Tocopherol; Animals; Chemistry, Pharmaceutical; Diffusion; Ethanol; Fatty Alcohols; Gels; Injections, Intra-Articular; Liquid Crystals; Morphinans; Rats; Rheology; Water

Sinomenine (SIN), an anti-arthritis drug, has previously been proven to exert immunomodulatory activity in rats by inducing intestinal regulatory T-cells (Treg cells). Here, we assessed the effect of SIN on the generation and function of Treg cells in autoimmune arthritis, and the underlying mechanisms in view of aryl hydrocarbon receptor (AhR). The proportions of Treg cells and IL-17-producing T-cells (Th17 cells) differentiated from naive T-cells were analyzed by flow cytometric analysis. The AhR agonistic effect of SIN was tested by analyzing the activation of downstream signaling pathways and target genes. The dependence of intestinal Treg cell induction and arthritis alleviation by SIN on AhR activation was confirmed in a mouse collagen-induced arthritis (CIA) model. SIN promoted the differentiation and function of intestinal Treg cells in vitro. It induced the expression and activity of AhR target gene, promoted AhR/Hsp90 dissociation and AhR nuclear translocation, induced XRE reporter activity, and facilitated AhR/XRE binding in vitro, displaying the potential to be an agonist of AhR. In CIA mice, SIN induced the generation of intestinal Treg cells, and facilitated the immunosuppressive function of these Treg cells as shown by an adoptive transfer test. In addition, the induction of intestinal Treg cells and the anti-arthritic effect of SIN in CIA mice could be largely diminished by the AhR antagonist resveratrol. SIN attenuates arthritis by promoting the generation and function of Treg cells in an AhR-dependent manner.

Topics: Animals; Antirheumatic Agents; Arthritis; Cell Differentiation; Cytochrome P-450 CYP1A1; Lymphocyte Activation; Male; Mice, Inbred DBA; Morphinans; Receptors, Aryl Hydrocarbon; T-Lymphocytes, Regulatory

Many compound collections used in high-throughput screening are composed of members whose structural complexity is considerably lower than that of natural products. We previously reported a strategy for the synthesis of complex and diverse small molecules from natural products using ring-distortion reactions, called complexity-to-diversity (CtD), and herein, CtD is applied in the synthesis of 16 diverse scaffolds and 65 total compounds from the alkaloid natural product sinomenine. Chemoinformatic analysis shows that these compounds possess complex ring systems and marked three-dimensionality.

Topics: Chemistry Techniques, Synthetic; Drug Design; Molecular Structure; Morphinans; Sinomenium; Small Molecule Libraries; Stereoisomerism

Astrocyte-mediated neuroinflammation plays a critical role in ischemic stroke-induced secondary cerebral injury. Previous studies have suggested that the dopamine D2 receptor (DRD2) acts as a key target in regulating the neuroinflammatory response. However, the underlying molecular mechanisms are still unknown, and effective DRD2 agonists are lacking. In the present study, we examined the anti-inflammatory and neuroprotective effects of sinomenine (Sino), a monomeric compound with potential immunoregulatory properties in nervous system.. TTC staining, apoptosis assay, evaluation of brain edema, and neurological assessment were performed in the middle cerebral artery occlusion (MCAO) mouse model. Primary astrocytes exposed to oxygen glucose deprivation (OGD) were used in the in vitro experiments. Quantitative PCR was applied to assess the levels of inflammatory cytokines. Multi-labeling immunofluorescence, Western blot, co-immunoprecipitation, and electrophoretic mobility shift assay (EMSA) were also used to investigate the molecular mechanisms underlying the Sino-mediated anti-inflammatory effects in vivo and in vitro.. Sino remarkably attenuated the cerebral infarction and neuronal apoptosis, reduced the levels of inflammatory cytokines, and alleviated neurological deficiency in MCAO mice. Sino significantly inhibited astrocytic activation and STAT3 phosphorylation as well as increased DRD2 and αB-crystallin (CRYAB) expression after MCAO. In vitro, Sino blocked OGD-induced activation of STAT3 and generation of pro-inflammatory cytokines in primary astrocytes, and these effects were significantly abolished by either DRD2 or CRYAB knockdown. Additionally, Sino induced up-regulation and nuclear translocation of CRYAB in astrocytes and enhanced the interaction between CRYAB and STAT3, which further inhibited the activation and DNA-binding activity of STAT3.. Our study demonstrates that Sino activates astrocytic DRD2 and thereby suppresses neuroinflammation via the CRYAB/STAT3 pathway, which sheds some light on a promising therapeutic strategy for ischemic stroke.

Topics: alpha-Crystallin B Chain; Animals; Animals, Newborn; Antirheumatic Agents; Astrocytes; Brain Edema; Cells, Cultured; Cerebral Infarction; Disease Models, Animal; Encephalitis; Hypoxia; Infarction, Middle Cerebral Artery; Mice; Mice, Inbred C57BL; Morphinans; Nervous System Diseases; Receptors, Dopamine D2; RNA Interference; Signal Transduction; STAT3 Transcription Factor

Microglia polarization plays a vital role in brain inflammatory injury following intracerebral hemorrhage (ICH). Previous studies have shown that sinomenine possesses potential immunoregulatory capabilities. However, microglia polarization's exact mechanisms in ICH remain uncertain. Therefore, we examined the role of sinomenine on microglia polarization and brain inflammation following ICH. For the experiment, autologous blood models were constructed in C57/BL6 mice. Markers of classically activated (M1) and alternatively activated (M2) microglia were detected by real-time polymerase chain reaction, immunofluorescence, and flow cytometry. Microglial toxicity was assessed using MTT and FACS assays. In addition, the neurological deficit and cerebral water content of ICH mice were also observed. Sinomenine attenuated M1 markers while promoting M2 markers of microglia. Sinomenine also protected hippocampal neurons from indirect toxicity mediated by ICH-treated microglia. Additionally, administration of sinomenine inhibited matrix metalloproteinase (MMP) 3/9 expression, cerebral water content, and neurological deficit. Therefore, sinomenine protected brain function following ICH, perhaps via M2 microglia phenotype induction and MMP 3/9 inhibition. This result suggests that sinomenine is a promising therapeutical strategy in ICH.

Topics: Animals; Brain Injuries; Cell Polarity; Cell Survival; Cells, Cultured; Cerebral Hemorrhage; Encephalitis; Male; Mice; Mice, Inbred C57BL; Microglia; Morphinans

Neuroinflammation remains the primary cause of morbidity and mortality in stroke-induced secondary brain injury. The NOD-like receptor pyrin 3 (NLRP3) inflammasome is involved in diverse inflammatory diseases, including cerebral ischemia, and is thus considered an effective therapeutic target. In the present study, we investigated the neuroprotection of Sinomenine (SINO), a potent natural anti-apoptotic and anti-inflammatory molecule, against cerebral ischemia in a mouse model of middle cerebral artery occlusion (MCAO) in vivo and in an oxygen glucose deprivation (OGD)-treated astrocytes/microglia model in vitro. SINO administration intraperitoneally alleviated the cerebral infarction, brain edema, neuronal apoptosis, and neurological deficiency after MCAO induction. SINO also attenuated astrocytic and microglial activation in the ischemic hemisphere. NLRP3 inflammasome activation after MCAO and OGD induction, with the up-regulation of NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), cleaved caspase-1 and pro-inflammatory cytokines, was significantly inhibited by SINO treatment both in vivo and in vitro. In addition, SINO reversed the OGD-induced inhibition of AMPK phosphorylation in vitro. Further, the suppressive effect of SINO on NLRP3 inflammasomes was blocked by an AMPK inhibitor, Compound C. Our findings demonstrate that SINO exerts a neuroprotective effect in ischemic stroke by inhibiting NLRP3 inflammasomes via the AMPK pathway, which also provides evidence of a novel treatment for clinical stroke therapy.

Topics: AMP-Activated Protein Kinases; Animals; Anti-Inflammatory Agents; Astrocytes; Brain Ischemia; Cells, Cultured; Glucose; Humans; Inflammasomes; Male; Mice; Mice, Inbred C57BL; Morphinans; Neuroprotection; NLR Family, Pyrin Domain-Containing 3 Protein; Signal Transduction; Stroke

Sinomenine is extracted from Sinomenii caulis (a traditional Chinese medicine), and it is used as the active ingredient in rheumatic arthritis treatments. It has been used in clinical applications for decades. However, there are some disadvantages, including low activity in transdermal permeation and a high dosage being clinically required. To overcome these defects, sinomenine was used as a primer, and structural modification was performed. In our study, eight new compounds were screened out by transdermal permeation in vitro and anti-inflammatory response in vitro and in vivo. Compound 1a exhibited the most potent transdermal permeation and anti-inflammatory activity. Based on these results, further development of this compound may be warranted.

Topics: Administration, Topical; Animals; Anti-Inflammatory Agents; Cell Survival; Cytokines; Drug Evaluation, Preclinical; Gene Expression; Lipopolysaccharides; Male; Mice; Morphinans; Permeability; Rats, Sprague-Dawley; RAW 264.7 Cells; Skin

Topics: Analgesics; Arthritis, Rheumatoid; Humans; Hyperalgesia; Hypersensitivity; Morphinans; Pain; Rats, Sprague-Dawley

Background and aims We have previously reported that sinomenine, an alkaloid isolated from the root of the plant Sinomenium acutum, had antinociceptive effect in rodent models of acute inflammatory or neuropathic pain. As a traditional medicine, sinomenine is used in China to treat rheumatoid arthritis (RA). Methods In the present study, we evaluated the potential antinociceptive effect of sinomenine in a mouse model of RA, collagen type II antibody (CII Ab) induced arthritis (CAIA) after acute and chronic administration. Results As single administration, sinomenine at 40 or 80 mg/kg significantly reduced mechanical hypersensitivity both at the time of peak joint inflammation (days 11-19 after CII Ab injection) or during the post-inflammatory phase (days 35-54). No tolerance to the effect of 80 mg/kg sinomenine was observed during repeated injection twice a day for 5 days from day 11 to day 19 or from day 49 to day 53 after CII Ab injection in CAIA mice. Conclusions We have shown that sinomenine is effective in alleviating localized and spread hypersensitivities in CAIA mice both during acute inflammation and in post-inflammatory phase. Further, repeated sinomenine administration has elevated the baseline mechanical threshold without producing tolerance. Implications Sinomenine may be clinically useful to treat chronic pain in RA, including wide-spread pain which appears to be a difficult clinical problem despite the improvement in the acute treatment of RA by disease modifying agents.

Topics: Animals; Antirheumatic Agents; Arthritis, Rheumatoid; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Hyperalgesia; Mice, Inbred CBA; Morphinans; Time Factors; Touch

Sinomenine is an anti-rheumatoid arthritis (RA) drug derived from the Sinomenium acutum. The major site of RA treatment is within the synovial compartment. However, the pharmacokinetic and penetration into synovial fluid (SF) of sinomenine have not been reported. In our study, the pharmacokinetics and penetration into SF of systemic and electroporation administered sinomenine were investigated by microdialysis incorporated with HPLC-MS/MS. Sinomenine went into plasma and SF more rapidly with higher peak concentration (Cmax ) by intramuscular injection compared with oral administration. The area under the concentration-time graph (AUC0-∞ ) of intramuscularly injected sinomenine was 1,403,294.75 ± 125,534.567 ng min/mL in plasma and 456,116.37 ± 62,648.36 ng min/mL in SF, which were equivalent with those for an oral dose. These results indicated that equal amounts of sinomenine could penetrate into SF by the two administration routes, and the permeation ratios were approximately 1:3. The AUC0-∞ and Cmax were lower with electroporation compared with systemic administration, but the CSF /CPlasma (concentration of sinomenine in SF vs that of plasma) at 90, 120, 150, 180, 240 and 480 min by electroporation was 3- to 10-fold higher relative to systemic administration. This illustrated that sinomenine can be targeted into joints by electroporation, and electroporation is a potential technique for sinomenine's transdermal delivery.

Topics: Administration, Cutaneous; Animals; Electroporation; Injections, Intramuscular; Linear Models; Microdialysis; Morphinans; Rabbits; Reproducibility of Results; Sensitivity and Specificity; Synovial Fluid

Sepsis, a systemic inflammatory response to infection, is the major cause of death in intensive care units (ICUs). The mortality rate of sepsis remains high even though the treatment and understanding of sepsis both continue to improve. Sinomenine (SIN) is a natural alkaloid extracted from Chinese medicinal plant Sinomenium acutum, and its hydrochloride salt (Sinomenine hydrochloride, SIN-HCl) is widely used to treat rheumatoid arthritis (RA). However, its role in sepsis remains unclear. In the present study, we investigated the role of SIN-HCl in sepsis induced by cecal ligation and puncture (CLP) in BALB/c mice and the corresponding mechanism. SIN-HCl treatment improved the survival of BALB/c mice that were subjected to CLP and reduced multiple organ dysfunction and the release of systemic inflammatory mediators. Autophagy activities were examined using Western blotting. The results showed that CLP-induced autophagy was elevated, and SIN-HCl treatment further strengthened the autophagy activity. Autophagy blocker 3-methyladenine (3-MA) was used to investigate the mechanism of SIN-HCl in vitro. Autophagy activities were determined by examining the autophagosome formation, which was shown as microtubule-associated protein light chain 3 (LC3) puncta with green immunofluorescence. SIN-HCl reduced lipopolysaccharide (LPS)-induced inflammatory cytokine release and increased autophagy in peritoneal macrophages (PM). 3-MA significantly decreased autophagosome formation induced by LPS and SIN-HCl. The decrease of inflammatory cytokines caused by SIN-HCl was partially aggravated by 3-MA treatment. Taken together, our results indicated that SIN-HCl could improve survival, reduce organ damage, and attenuate the release of inflammatory cytokines induced by CLP, at least in part through regulating autophagy activities.

Topics: Adenine; Animals; Autophagy; Cecum; Cell Line; Cytokines; Disease Models, Animal; Kidney; Lipopolysaccharides; Liver; Lung; Macrophages; Male; Mice; Mice, Inbred BALB C; Microtubule-Associated Proteins; Morphinans; Sepsis; Survival Rate

Sinomenine (SIN) has long been used as a therapeutic agent of rheumatoid arthritis (RA) in China. However, the discrepancy between low oral bioavailability and higher minimal effective concentration made its action mode mysterious. The present study aimed to gain insight into the mechanisms by which SIN suppressed collagen-induced arthritis (CIA) in rats in view of Th17 and regulatory T (Treg) cell balance. SIN was orally administered, and the clinical symptoms of CIA rats were monitored; inflammatory cytokines levels in serum were measured by ELISA; pharmacokinetic studies were performed in normal and CIA rats; Th17 and Treg cell frequencies were analyzed by flow cytometry. The data showed that SIN treatment resulted in a dramatic decrease of arthritis scores and paw volume of CIA rats, which was accompanied by down-regulation of IL-17A and up-regulation of IL-10 in rat serum. The frequency of Treg cells was increased and the frequency of Th17 cells was decreased in the gut lymphoid tissues of SIN-treated rats. Immunohistochemistry assay demonstrated that more α4β7-positive cells were detained in joint tissues after SIN treatment. Moreover, the anti-arthritis efficacy of SIN disappeared when it was given by intraperitoneal injection, further confirming the action of SIN was gut-dependent. In conclusion, SIN exerts anti-RA action probably through modulating the frequencies of Treg cells and Th17 cells in intestinal lymph nodes and yielding a trafficking of lymphocytes (especially Treg cells) from gut to joint.

Topics: Animals; Anti-Inflammatory Agents; Arthritis, Experimental; Arthritis, Rheumatoid; Cell Differentiation; Collagen; Gastrointestinal Tract; Interleukin-10; Interleukin-17; Lymph Nodes; Morphinans; Rats; Rats, Wistar; T-Lymphocytes, Regulatory; Th17 Cells

Solid tumor vasculature is characterized by structural and functional abnormality and results in a hostile tumor microenvironment that mediates several deleterious aspects of tumor behavior. Sinomenine is an alkaloid extracted from the Chinese medicinal plant, Sinomenium acutum, which has been utilized to treat rheumatism in China for over 2000 years. Though sinomenine has been demonstrated to mediate a wide range of pharmacological actions, few studies have focused on its effect on tumor vasculature. We showed here that intraperitoneally administration of 100 mg/kg sinomenine hydrochloride (SH, the hydrochloride chemical form of sinomenine) in two orthotopic mouse breast cancer models for 14 days, delayed mammary tumor growth and decreased metastasis by inducing vascular maturity and enhancing tumor perfusion, while improving chemotherapy and tumor immunity. The effects of SH on tumor vessels were caused in part by its capability to restore the balance between pro-angiogenic factor (bFGF) and anti-angiogenic factor (PF4). However 200 mg/kg SH didn't exhibit the similar inhibitory effect on tumor progression due to the immunosuppressive microenvironment caused by excessive vessel pruning, G-CSF upregulation, and GM-CSF downregulation. Altogether, our findings suggest that SH induced vasculature normalization contributes to its anti-tumor and anti-metastasis effect on breast cancer at certain dosage.

Topics: Animals; Breast Neoplasms; Cell Line, Tumor; Cell Survival; Dose-Response Relationship, Drug; Female; Mice; Mice, Inbred BALB C; Morphinans; Neovascularization, Pathologic; Treatment Outcome

The effects of the water extract of Sinomeni Caulis et Rhizoma (SCR-WE) and its major constituents, sinomenine (SIN) and magnoflorine (MAG), on moderate hemolysis induced by lysophosphatidylcholine (LPC) were investigated in rat erythrocytes and compared with the anti-hemolytic effects of lidocaine (LID) and propranolol (PRO) as reference drugs. LPC caused hemolysis at concentrations above the critical micelle concentration (CMC), and the concentration of LPC producing moderate hemolysis (60 %) was approximately 10 μM. SCR-WE at 1 ng/mL-100 μg/mL significantly inhibited the hemolysis induced by LPC. SIN and MAG attenuated LPC-induced hemolysis in a concentration-dependent manner from very low to high concentrations (1 nM-100 μM and 10 nM-100 μM, respectively). In contrast, the inhibiting effects of LID and PRO on LPC-induced hemolysis were observed at higher concentrations (1-100 μM) but not at lower concentrations (1-100 nM). Neither SIN nor MAG affected micelle formation of LPC, nor, at concentrations of 1 nM-1 μM, did they attenuate the hemolysis induced by osmotic imbalance (hypotonic hemolysis). Similarly, SCR-WE also did not modify micelle formation or hypotonic hemolysis, except at the highest concentration. These results suggest that SIN and MAG potently protect the erythrocyte membrane from LPC-induced damage and contribute to the beneficial action of SCR-WE. The protective effects of SIN and MAG are mediated by some mechanism other than prevention of micelle formation or protection of the erythrocyte membrane against osmotic imbalance.

Topics: Animals; Aporphines; Cytoprotection; Drug Evaluation, Preclinical; Erythrocyte Membrane; Erythrocytes; Hemolysis; Lysophosphatidylcholines; Male; Micelles; Morphinans; Plant Extracts; Plant Stems; Rats; Rats, Sprague-Dawley; Rhizome; Sinomenium

A sensitive, precise and selective ultra-high performance liquid chromatography method coupled with triple-quadrupole mass spectrometry was developed and validated for the determination of trace amounts of sinomenine (ng/mL) in minute volumes of human plasma. Fifty microliter plasma samples were precipitated using methanol to extract sinomenine. Separation was carried out on a C18 column with a water and acetonitrile mobile phase gradient with formic acid as an additive. The mass spectrometry data were obtained in the positive ion mode, and the transition of multiple reactions was monitored at m/z 330.2→181.0 for sinomenine quantification. The working assay range for sinomenine was linear from 0.1173 to 15.02 ng/mL with the lower limit of quantification of 0.1173 ng/mL. The precision and accuracy of the method was less than 15% in intra-day and inter-day experiments with a matrix effect of less than 6.5%. After validation, the quantitative method was applied to analyze sinomenine levels in human plasma after transdermal delivery of the Zhengqing Fengtongning Injection. The results showed that some samples contained sinomenine within the concentration range 0.4131-4.407 ng/mL.

Topics: Administration, Cutaneous; Adult; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Female; Humans; Morphinans; Rheumatic Fever; Tandem Mass Spectrometry; Young Adult

Rheumatoid arthritis (RA) is a chronic, systemic inflammatory disorder that affects about 1% of the population worldwide. RA is mainly manifested by persistent synovitis and progressive joint destruction. The aim of the present study was to examine the anti-arthritis effects of SND-117, a sinomenine bivalent that is obtained from the structure modification of a clinically available anti-RA drug, sinomenine. The arthritis model (CIA) was established by immunizing DBA/1 mice with type II collagen, and the arthritis scores including inflammation, joint destruction and bone erosion were assessed after booster immunization for 3weeks. The levels of cytokines such as IL-1β, IL-6 and TNF-α were analyzed by quantitative PCR and ELISA. The TNF-α induced NF-κB activation in fibroblast-like synovial cells (FLSCs) was analyzed by Western blot. SND-117 significantly relieved the inflammatory symptoms of collagen-induced arthritis, reduced bone erosion and joint destruction in CIA mice. The serum levels of IL-1β, IL-6 and TNF-α of CIA mice were markedly decreased by SND-117. SND-117 also strongly inhibited the phosphorylation and nuclear translocation of NF-κB p65 in FLSCs upon TNF-α stimulation. These data demonstrated that SND-117 could effectively block the pathogenesis of collagen-induced arthritis in CIA mice via inhibition of NF-κB signaling, and might provide potential clinic benefits in rheumatoid arthritis management.

Topics: Animals; Antirheumatic Agents; Arthritis, Rheumatoid; Bone and Bones; Collagen Type II; Cytokines; Disease Models, Animal; Fibroblasts; Humans; Immunization, Secondary; Inflammation Mediators; Male; Mice; Mice, Inbred DBA; Morphinans; NF-kappa B; Sinomenium; Synovitis

Sinomenine (SIN), an alkaloid derived from the plant Sinomenium acutum, has anti-inflammatory and analgesic effects and has been used for rheumatoid arthritis treatment in China. This study aims to verify the hypothesis that SIN acts on α7 nicotinic acetylcholine receptor (α7nAChR) to inhibit the activation of macrophages stimulated by lipopolysaccharide. The prototypical α7nAChR antagonist α-bungarotoxin and mecamylamine attenuated the effect of SIN on tumor necrosis factor-α and interleukin-6 in RAW264.7 murine macrophage-like cells and primary peritoneal macrophages of mouse induced by lipopolysaccharide. With the knockdown of α7nAChR expression in RAW264.7 cells by small interfering RNA, the inhibitory effect of SIN on tumor necrosis factor-α and interleukin-6 was reversed. Sinomenine decreased p65 expression in nuclear and increased IκBα expression in cytoplasm, and these effects were reversed by the α7nAChR small interfering RNA as well. These results indicate that the anti-inflammatory effects of SIN on macrophages in vitro depend on α7nAChR.

Topics: alpha7 Nicotinic Acetylcholine Receptor; Animals; Anti-Inflammatory Agents; Bungarotoxins; Cytoplasm; I-kappa B Proteins; Interleukin-6; Lipopolysaccharides; Macrophages, Peritoneal; Male; Mecamylamine; Mice; Mice, Inbred BALB C; Morphinans; NF-KappaB Inhibitor alpha; RAW 264.7 Cells; RNA Interference; RNA, Small Interfering; Tumor Necrosis Factor-alpha

Sinomenine (SIN), an alkaloid derived from the Chinese medicinal plant Sinomenium acutum, is the major component of Zhengqing Fongtong Ning (ZQFTN), a pharmaceutical drug produced by Hunan Zhengqing Pharmaceutical Co. Ltd. in China for the treatment of rheumatoid arthritis and other autoimmune diseases. Some clinic reports indicate that ZQFTN may induce an anaphylactic reaction via potentiating the degranulation of immune cells. In the current study, we aimed to examine whether SIN is capable of inducing the degranulation of basophilic leukemia 2H3 (RBL-2H3) cells to elucidate how the anaphylactic reaction occurs. The results revealed that SIN could up-regulate β-hexosaminidase levels in RBL-2H3 cells without significant cytotoxicity, suggesting that SIN could induce the degranulation of RBL-2H3 cells. Furthermore, SIN increased the release of prostaglandin D2 (PGD2) and prostaglandin E2 (PGE2) in RBL-2H3 cells via promoting the expression of phosphorylated-extracellular signal-regulated kinase (P-ERK), the cleavage of Annexin A1 (ANXA1), and phosphorylated-cytosolic phospholipase A2 (P-cPLA2), as well as cyclooxygenase-2 (COX-2). The ERK inhibitor, PD98059, significantly attenuated the up-regulatory effect of SIN on cPLA2 phosphorylation. Interestingly, SIN did not significantly increase Ca(2+) influx in the cells. These findings not only explored the anaphylactic reaction and underlying mechanism of ZQFTN in RBL-2H3 cells, but may promote the development of relevant strategies for overcoming the adverse effects of the drug.

Topics: Anaphylaxis; Animals; Annexin A1; Arthritis, Rheumatoid; Basophils; Calcium; Cell Degranulation; Cell Line; Drug Hypersensitivity; Humans; MAP Kinase Signaling System; Morphinans; Phospholipases A2; Phosphorylation; Rats; Sinomenium

Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) is a vital transcription factor that regulates multiple important biological processes, including the epithelial-mesenchymal transition (EMT) and metastasis of breast cancer. Sinomenine is an isoquinoline well known for its remarkable curative effect on rheumatic and arthritic diseases and can induce apoptosis of several cancer cell types. Recently, sinomenine was reported as a tumor suppressor via inhibiting cell proliferation and inducing apoptosis. However, the role and mechanism of sinomenine in invasion and metastasis of breast cancer are largely unknown. Here, we report that sinomenine suppressed the invasion and migration of MDA-MB-231 and 4T1 breast cancer cells in a dose-dependent manner. We detected binding of NF-κB to the inhibitor of NF-κB (IκB) after the MDA-MB-231 cells were treated with 0.25, 0.5, and 1 mM sinomenine. Co-IP analysis revealed that sinomenine enhanced the binding of NF-κB and IκB in a dose-dependent manner, suggesting that sinomenine had an effect on inactivation of NF-κB. Western blotting and ELISA approaches indicated that the suppression effect was closely associated with the phosphorylation of IκB kinase (IKK) and its negative regulator CUEDC2. Sinomenine treatment decreased miR-324-5p expression, thus increased the level of its target gene CUEDC2, and then blocked the phosphorylation of IKK through altering the upstream axis. Finally, transfection of a miR-324-5p mimic inhibited the suppression of invasion and metastasis of MDA-MB-231 and 4T1 cell by sinomenine, providing evidence that sinomenine treatment suppressed breast cancer cell invasion and metastasis via regulation of the IL4/miR-324-5p/CUEDC2 axis. Our findings reveal a novel mechanism by which sinomenine suppresses cancer cell invasion and metastasis, i.e., blocking NF-κB activation.

Topics: Adaptor Proteins, Signal Transducing; Antineoplastic Agents, Phytogenic; Breast Neoplasms; Carrier Proteins; Cell Line, Tumor; Cell Movement; Cell Proliferation; Dose-Response Relationship, Drug; Female; Humans; I-kappa B Proteins; Interleukin-4; Membrane Proteins; MicroRNAs; Molecular Mimicry; Morphinans; Neoplasm Invasiveness; NF-kappa B; Phytotherapy

Topics: Aging; Calcium Channels; Electrophysiological Phenomena; Heart; Humans; Medicine, Kampo; Morphinans; Sinomenium; Vasodilation

Sinomenine is an isoquinoline-type alkaloid found in Sinomenium acutum (Thunb.) Rehd. et Wils and S. acutum (Thunb.) Rehd. et Wils var. cinereum Rehd. et Wils. When used as a medicine, this compound exhibits anti-inflammatory properties; however, sinomenine's use as a medication is limited by side effects, a short half-life, and low efficacy. Owing to these limits, attempts have been made to synthesize sinomenine derivatives with enhanced efficacy. In this study, the anti-inflammatory effects of novel sinomenine derivatives (S1a-S1f) were examined on the basis of lipopolysaccharide-induced inflammatory factor expression in Raw264.7 cells, dimethylbenzene-induced ear oedema, and Evan's blue leakage in mice, and carrageenan-induced paw oedema in rats. Compared with sinomenine, the derivatives significantly inhibited the expression of the inflammatory factors IL-1β and IL-6 at the transcriptional and translational levels. Topical application of 3.250mg/kg of the derivatives also alleviated ear oedema. Compared with the vehicle, the derivatives significantly inhibited carrageenan-induced rat paw oedema after 6h. Among the derivatives, S1a exhibited the most potent anti-inflammatory activity. S1a also significantly increased the sinomenine-induced inhibition of Evan's blue leakage. Thus, S1a may elicit the strongest anti-inflammatory effects of the tested compounds. Based on these results, further development of this compound may be warranted.

Topics: Animals; Anti-Inflammatory Agents; Capillary Permeability; Carrageenan; Coloring Agents; Edema; Evans Blue; Foot; Interleukin-1beta; Interleukin-6; Lipopolysaccharides; Male; Mice; Morphinans; Rats; Rats, Sprague-Dawley; Sinomenium; Tumor Necrosis Factor-alpha; Xylenes

A novel flow-injection chemiluminescence (FI-CL) method was described for the determination of sinomenine hydrochloride (SIN). The method was based on the inhibitory effect of SIN on the CL reaction of luminol and K3Fe(CN)6 in an alkaline solution, which was sensitized by CdTe/CdS quantum dots (QDs). Under the optimized conditions, the linear range for the determination of SIN was 1.0 × 10(-8) to 1.4 × 10(-6) mol/L. The detection limit was 7.5 × 10(-9) mol/L, and the relative standard deviation was 2.47% (n = 11). The current CL method was applied to determine SIN in pharmaceutical formulations and biological fluids with satisfactory results. The possible CL reaction mechanism was discussed briefly.

Topics: Cadmium Compounds; Ferricyanides; Flow Injection Analysis; Kinetics; Limit of Detection; Luminescent Measurements; Luminol; Morphinans; Quantum Dots; Sulfides; Tellurium

The effect of sinomenine (SIN) on the toll-like receptor (TLR) signal transduction pathway as well as the expression of myeloid differentiation factor 88 (MyD88) and tumor necrosis factor (TNF) receptor-associated factor-6 (TRAF6) was investigated. SIN inhibition of rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS) proliferation and RA cartilage and subchondral bone destruction was also investigated. RA-FLS were cultured in vitro and the intracellular alkaline phosphatase (ALP) activity was determined in order to obtain the optimal drug concentration. The rate of cell proliferation was determined. Fluorescence quantitative polymerase chain reaction (PCR) was applied to determine the MyD88 and TRAF-6 gene expression and western blot was used to detect the MyD88 and TRAF-6 protein expression. The ALP activity in the SIN groups was lower than that in the control group, among which the 0.5 mM SIN group had the lowest ALP activity (P < 0.01). The rate of RA-FLS proliferation detected by CCK-8 assay in the 0.5-mM SIN group was lower than that in the control group (P < 0.01) and was the highest 4 days after SIN induction. Gene and protein expression of MyD88 and TRAF-6 were downregulated significantly in the 0.5-mM SIN group compared to that in the control group (P < 0.01). SIN effectively inhibited MyD88 and TRAF-6 expression in RA-FLS, which may be one of the important molecular mechanisms involved in RA treatment and prevention of cartilage and subchondral bone destruction.

Topics: Alkaline Phosphatase; Analgesics; Anti-Inflammatory Agents; Arthritis, Rheumatoid; Arthroplasty; Cell Proliferation; Fibroblasts; Gene Expression Regulation; Humans; Joint Capsule; Morphinans; Myeloid Differentiation Factor 88; Primary Cell Culture; Signal Transduction; TNF Receptor-Associated Factor 6

To analyze the combination therapy of Sinomenine (SIN) and Methotrexate (MTX) in rheumatoid arthritis (RA), we herein demonstrated the combination effect of SIN and MTX on collagen-induced arthritis (CIA) in rats through their modulation on osteoclast-related cytokines.. CIA was induced by the immunization of type II collagen (CII) in SD rats. SIN and MTX were administrated alone or in combination after the onset of arthritis. Arthritis index and histological analysis were used to evaluate the effect of treatments. Effects of SIN and MTX on expression of receptor activator of NF-κB ligand (RANKL) and osteopontin (OPN) in synovial tissues were assayed by immunohistochemistry. RANKL, osteoprotegerin (OPG), IL-6, IL-17 and matrix metalloproteinases (MMPs) in rat serum were measured by ELISA. The expression of osteoclast-related cytokines in fibroblast-like synoviocytes (FLS) from RA patients was assayed by RT-PCR.. SIN and MTX combination additively reduced the inflammatory symptoms and joint damage in CIA. Combination of SIN and MTX significantly repressed synovial RANKL and OPN production. SIN and MTX exhibited complementary and synergistic effect upon down-regulating RANKL, IL-6, IL-17 and MMPs in rat serum. SIN and MTX also modulated the expression of RANKL and OPG in RA-FLS.. SIN and MTX have additive effects, decreasing inflammation and joint damage in CIA rats by modulating osteoclast-related cytokines. These results are indicative of the combined effect of SIN and MTX for anti-arthritic treatment in RA.

Topics: Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Cells, Cultured; Collagen Type II; Cytokines; Drug Synergism; Drug Therapy, Combination; Female; Humans; Joints; Methotrexate; Morphinans; Osteoclasts; Osteopontin; RANK Ligand; Rats; Rats, Sprague-Dawley

Sinomenine (SIN) is an alkaloid found in the roots and stems of Sinomenium acutum, which has been used to treat rheumatic arthritis in China and Japan. In this study we investigated the effects of SIN on osteoclast survival in vitro and the mechanisms of the actions.. Mature osteoclasts were differentiated from murine monocyte/macrophage cell line RAW264.7 through incubation in the presence of receptor activator of NF-κB ligand (RANKL, 100 ng/mL) for 4 d. The cell viability was detected using the CCK-8 method. The survival and actin ring construction of the osteoclasts were scored using TRACP staining and phalloidin-FITC staining, respectively. The apoptosis of the osteoclasts was detected by DNA fragmentation and Hoechst 33258 staining, and the cell necrosis was indicated by LDH activity. The activation of caspase-3 in osteoclasts was measured using Western blotting and the caspase-3 activity colorimetric method.. SIN (0.25-2 mmol/L) inhibited the viability of mature osteoclasts in dose-dependent and time-dependent manners, but did not affect that of RAW264.7 cells. Consistently, SIN dose-dependently suppressed the survival of mature osteoclasts. The formation of actin ring, a marker associated with actively resorbing osteoclasts, was also impaired by the alkaloid. SIN (0.5 mmol/L) induced the apoptosis of mature osteoclasts, which was significantly attenuated in the presence of the caspase-3 inhibitor Ac-DEVD-CHO. SIN increased the cleavage of caspase-3 in mature osteoclasts in dose-dependent and time-dependent manners. Furthermore, SIN dose-dependently enhanced caspase-3 activity, which was blocked in the presence of Ac-DEVD-CHO.. Sinomenine inhibits osteoclast survival in vitro through caspase-3-mediated apoptosis, thus it is a potential agent for treating excessive bone resorption diseases.

Topics: Animals; Apoptosis; Caspase 3; Cell Line; Macrophages; Mice; Morphinans; Osteoclasts

A collective synthesis of 8,14-dihydronorsalutaridine, 8,14-dihydrosalutaridine, norisosinomenine, and isosinomenine is reported. The strategy provides direct access to the correct oxidation level of the products. The combination of an organocatalyst guanidine superbase, a tertiary amine base, and a dehydrating agent was necessary for the successful Henry-Michael-dehydration cascade to form the phenanthrene motif. The required selective aliphatic nitro reduction could only be achieved under heterogeneous transfer-hydrogenation conditions.

Topics: Alkaloids; Amines; Hydrogenation; Molecular Structure; Morphinans; Oxidation-Reduction; Stereoisomerism

P-Glycoprotein-mediated multidrug resistance is a frequent event during chemotherapy and a key obstacle for bladder cancer therapy. Search for strategies to reverse multidrug resistance is a promising approach to improve the management of bladder cancer. In the present study, we reported a novel P-glycoprotein-mediated multidrug resistant cell model 253J/DOX, which was generated from human bladder cancer 253J cell line. Furthermore, we found that the multidrug resistant phenotype of 253J/DOX cells could be overcome by sinomenine, an alkaloid derived from the stem of Sinomenium acutum. Mechanistically, the chemosensitive effect by sinomenine was mediated by down-regulating P-glycoprotein expression, as well as triggering apoptotic pathways. The chemosensitive effect of sinomenine may make it a prime candidate agent to target bladder cancer.

Topics: Apoptosis; ATP Binding Cassette Transporter, Subfamily B, Member 1; Blotting, Western; Cell Line, Tumor; Coloring Agents; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Flow Cytometry; Fluorescent Antibody Technique; Fluorescent Dyes; Humans; Indicators and Reagents; Morphinans; Real-Time Polymerase Chain Reaction; Rhodamine 123; RNA, Small Interfering; Tetrazolium Salts; Thiazoles; Urinary Bladder Neoplasms

To synthesize the derivatives of sinomenine, 4-palmitoyl-sinomenine, and explore its therapeutic effect on lipopolysaccharide (LPS)-induced endotoxemia.. A highly efficient synthesis of sinomenine derivatives called 4-palmitoyl-sinomenine was made with a molecule of palmitic acid substitutions at C-16 position of ring A. One hour before endotoxemia induction by i.p. injection of 10 mg/kg LPS, high-dose treatment mice (n=5/group) received an i.p. injection of 5 mg/kg sinomenine or 4-palmitoyl-sinomenine while the low-dose treatment mice (n=5/group) received 2.5 mg/kg sinomenine or 4-palmitoyl-sinomenine. Untreated group and normal control group received normal saline. And their survival was monitored hourly for 24 hours. Examination of cytotoxicity of 4-palmitoyl-sinomenine on RAW264.7 cells was conducted at a concentration range of 1 to 125 μmol/L using MTT assay. RAW264.7 cells were exposed to 4-palmitoyl-sinomenine (0, 1, 2, 5, 10 μmol/L) for 24 hours, and then treated with LPS (1 μg/mL) for 6 hours. Then RAW264.7 cells were collected and the mRNA level of IL-6 was detected by real-time quantitative PCR(qRT-PCR).. Sinomenine derivatives were successfully synthesized to get 4-palmitoyl-sinomenine. The survival percentage of 4-palmitoyl-sinomenine treatment groups was higher than that of sinomenine treatment groups at the same treatment concentration. The 4-palmitoyl-sinomenine inhibited RAW264.7 cell proliferation and IL-6 gene transcription.. The 4-palmitoyl-sinomenine has an anti-inflammation probably through inhibiting the proliferation of RAW264.7 cells and decreasing the inflammatory gene expression and inflammatory cytokine release.

Topics: Animals; Anti-Inflammatory Agents; Cell Proliferation; Cells, Cultured; Interleukin-6; Lipopolysaccharides; Male; Mice; Mice, Inbred BALB C; Morphinans; RNA, Messenger

An accurate and reliable high-performance liquid chromatography-diode array detector (HPLC-DAD) method was developed and validated for determination of sinomenine (SI), paeoniflorin (PF) and paeonol (PA), which was further applied to assess the pharmacokinetics of SI, PF and PA in an anti-arthritic herbal product, Qingfu Guanjieshu (QFGJS) capsule, in rats. Successful separation was achieved with a C18 column and a mobile phase composed of acetonitrile and aqueous phase (containing 0.1% formic acid, adjusted with triethylamine to pH 3.5 ± 0.2). The method was validated with excellent precision, accuracy, recovery and stability in calibration ranges from 0.06 to 11.62 µg/mL for SI, from 0.09 to 35.70 µg/mL for PF, and from 0.15 to 4.53 µg/mL for PA (with r(2) > 0.999 for all three compounds). Our results showed that absorption of PF after administration of QFGJS was similar to that after oral administration of PF alone; the absorption of SI was decreased while the absorption of PA was increased after giving QFGJS orally compared with pure compounds. We may conclude that pharmacokinetic studies of complex herbal products are not only necessary but also feasible by using representative bioactive chemicals as indicators of establishing quality control standards and of determining pharmacokinetic behavior of herbal medicines.

Topics: Acetophenones; Administration, Oral; Animals; Chromatography, High Pressure Liquid; Drug Stability; Drugs, Chinese Herbal; Glucosides; Linear Models; Male; Monoterpenes; Morphinans; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Sensitivity and Specificity

The interaction between sinomenine and Lysozyme (Lys) in aqueous solution has been systemically investigated by fluorescence spectroscopic techniques at pH 7.4. The quenching rate constants and binding constants calculated indicated the static quenching mechanism and medium binding force. The effect of sinomenine on the conformation of Lys was analyzed using synchronous fluorescence and three-dimensional (3D) fluorescence. In addition, influence of pH on the binding of sinomenine to Lys was investigated and the binding ability of the drug to Lys deceased under other pH conditions (pH 9.0, 3.5, and 1.9) as compared with that at pH 7.4. As compared with the binding ability of sinomenine to native Lys, that of sinomenine to denatured Lys deceases dramatically. Furthermore, the effect of many metal ions on the binding constant of sinomenine with Lys was investigated.

Topics: Animals; Chemistry, Pharmaceutical; Chickens; Drug Design; Egg White; Hydrogen-Ion Concentration; Ions; Lysine; Metals; Morphinans; Muramidase; Protein Binding; Spectrometry, Fluorescence; Urea

Intracerebral hemorrhage (ICH) causes morbidity and mortality and commonly follows the reperfusion after an ischemic event. Microglial activation mediated cytokine and protease secretion contributes to brain injury in ICH. Previous studies have shown that sinomenine possesses potent immunoregulatory properties. However, little is known about its exact role in ICH. In the present study, to investigate the effect of sinomenine on microglial cells inflammation, we treated ICH-challenged BV2 microglial cells with sinomenine in vitro, and explored its neuroprotection role in intracerebral hemorrhage in vivo. Changes in inflammatory cytokines, such as TNF-α, IL-1β and IL-6, reactive oxygen species (ROS) and NF-κB activation NF-κB were observed. In addition, the neurological deficit and cerebral water content of ICH mice were studied. The results demonstrated that sinomenine could inhibit the release of these cytokines and attenuate ROS production in a dose-dependent manner, and reduce NF-κB activation. Furthermore, sinomenine markedly inhibited cerebral water content and neurological deficit. In conclusion, our findings suggest that sinomenine played the protective effects through inhibition of microglial inflammation, and the findings also provided a novel therapy to treat ICH induced brain injury.

Topics: Animals; Brain Injuries; Cell Line; Cell Movement; Cerebral Hemorrhage; Inflammation; Interleukin-1beta; Interleukin-6; Male; Mice; Mice, Inbred C57BL; Microglia; Morphinans; Neuroprotective Agents; NF-kappa B; Reactive Oxygen Species; Tumor Necrosis Factor-alpha

Chemoresistance in multidrug-resistant (MDR) cells over expressing P-glycoprotein (P-gp) encoded by the MDR1 gene, is a major obstacle to successful chemotherapy for colorectal cancer. Previous studies have indicated that sinomenine can enhance the absorption of various P-gp substrates. In the present study, we investigated the effect of sinomenine on the chemoresistance in colon cancer cells and explored the underlying mechanism. We developed multidrug-resistant Caco-2 (MDR-Caco-2) cells by exposure of Caco-2 cells to increasing concentrations of doxorubicin. We identified overexpression of COX-2 and MDR-1 genes as well as activation of the NF-κB signal pathway in MDR-Caco-2 cells. Importantly, we found that sinomenine enhances the sensitivity of MDR-Caco-2 cells towards doxorubicin by downregulating MDR-1 and COX-2 expression through inhibition of the NF-κB signaling pathway. These findings provide a new potential strategy for the reversal of P-gp-mediated anticancer drug resistance.

Topics: Antibiotics, Antineoplastic; ATP Binding Cassette Transporter, Subfamily B, Member 1; Caco-2 Cells; Celecoxib; Cell Line, Tumor; Cell Survival; Colonic Neoplasms; Cyclooxygenase 2; Dinoprostone; Doxorubicin; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Gene Expression Regulation, Neoplastic; Humans; Morphinans; NF-kappa B; Pyrazoles; Signal Transduction; Sulfonamides

Sinomenine, the main alkaloid extracted from the medicinal plant Sinomenium acutum, is known for its anti-inflammatory effects. Recent studies have suggested its anti-cancer effect in synovial sarcoma, lung cancer and hepatic cancer. However, the underlying molecular mechanism for its anti-cancer effect still remains unclear. This study investigated the anti-tumor activity of sinomenine hydrochloride (SH), a hydrochloride form of sinomenine, in human breast cancer cells in vitro and in vivo. We found that SH potently inhibited cell viability of a broad panel of breast cancer cell lines. Two representative breast cancer cell lines, namely ER(-)/PR(-) MDA-MB-231 and ER(+)/PR(+) MCF-7, were used for further investigation. The results showed that SH induced G1/S cell cycle arrest, caused apoptosis and induced ATM/Chk2- and ATR/Chk1-mediated DNA-damage response in MDA-MB-231 and MCF-7. The anti-cancer effect of SH was regulated by increased expression levels of p-ERK, p-JNK and p-38 MAPK. Further studies showed that SH resulted in an increase in reactive oxygen species (ROS) and inhibition of ROS by N-acetyl-L-cysteine (NAC) almost blocked SH-induced DNA damage but only mitigated SH-induced MAPK expression changes, suggesting that both ROS-dependent and -independent pathways were involved in MAPK-mediated SH-induced breast cancer cell death. The in vivo study demonstrated that SH effectively inhibited tumor growth without showing significant toxicity. In conclusion, SH induced breast cancer cell death through ROS-dependent and -independent pathways with an upregulation of MAPKs, indicating that SH may be a potential anti-tumor drug for breast cancer treatment.

Topics: Animals; Antineoplastic Agents; Apoptosis; Breast Neoplasms; Cell Line, Tumor; Female; Humans; MAP Kinase Signaling System; Mice, Inbred BALB C; Mitogen-Activated Protein Kinase Kinases; Morphinans; Reactive Oxygen Species

Various products containing sinomenine monomer and extracts of Sinomenium acutum have been widely applied in clinical treatments. The goal of the present study was to compare the pharmacokinetics of sinomenine in rats after oral administration of sinomenine monomer and Sinomenium acutum extract, and to attempt to explore potential component-component interactions between the constituents of this traditional Chinese herbal medicine. A reliable and specific reversed phase high performance liquid chromatography method was developed to analyze sinomenine in rat plasma. Pharmacokinetic parameters for sinomenine were processed by non-compartmental analysis. The results showed that the maximum concentration, the area under the concentration-time curve, clearance and the apparent volume of distribution of sinomenine in the Sinomenium acutum extract statistically differed from those of sinomenine monomer (p < 0.05); however, the mean residence time, time of peak concentration, and half-life did not show significant differences between the two groups. These findings suggested that some additional components in the Sinomenium acutum extract may decrease the absorption of sinomenine. The complex interactions between sinomenine and other components of the herbal extract could result in the altered pharmacokinetic behavior of sinomenine, which may subsequently cause different therapeutic and detoxification effects.

Topics: Administration, Oral; Animals; Chromatography, High Pressure Liquid; Humans; Inactivation, Metabolic; Morphinans; Plant Extracts; Rats; Sinomenium

It is reported that sinomenine (SIN) and 5-fluorouracil (5-FU) both are effective for colon cancer, but their cooperative suppressive effects and toxicity remain to be clarified in detail. This study aimed to determine suppressive effects and toxicity of sinomenine (SIN) plus 5-fluorouracil (5-FU) on LoVo colon carcinoma cells in vitro and in vivo. CCK-8, Hoechst 33258 staining and an annexin V-FITC/PI apoptosis kit were used to detect suppressive effects. Western blotting was applied to investigate the essential mechanism underlying SIN and 5-FU-induced apoptosis. SIN or 5-FU or both were injected into nude mice, and then suppressive effects and side effects were observed. SIN plus 5-FU apparently inhibited the proliferation of LoVo cells and induced apoptosis. Moreover the united effects were stronger than individually (p<0.05). The results of annexin V-FITC /PI staining and Hoechst 33258 staining showed that the percentage of apoptotic cells induced by SIN and 5-FU combined or alone was significantly higher than the control group (p<0.05). Expression of Bax and Bcl-2 was up-regulated and down-regulated respectively. SIN or 5-FU significantly inhibited effects on the volume of tumour xenografts and their combined suppressive effects were stronger (p<0.05). No obvious side effects were observed. It was apparent that the united effects of SIN and 5-FU on the growth of colorectal carcinoma LoVo cells in vitro and in vivo were superior to those using them individually, and it did not markedly increase the side effects of chemotherapy.

Topics: Animals; Antimetabolites, Antineoplastic; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; bcl-2-Associated X Protein; Cell Line, Tumor; Cell Proliferation; Colonic Neoplasms; Fluorouracil; Humans; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Morphinans; Proto-Oncogene Proteins c-bcl-2; Xenograft Model Antitumor Assays

Sinomenine is a principal ingredient of traditional Chinese medicine, Sinomenium Acutum, which has been reported to have various pharmacological effects including anti-rheumatism and immunomodulation. This study examined the effects of sinomenine in rats that received chronic constriction injury (CCI), a model of peripheral neuropathic pain. CCI injury on the right sciatic nerve led to long-lasting mechanical hyperalgesia. Acute sinomenine treatment (10-40 mg/kg, i.p.) significantly and dose-dependently reversed mechanical hyperalgesia. In addition, the antinociceptive effects of sinomenine remained stable during repeated daily treatment for up to 2 weeks. Although sinomenine did not alter the duration of immobility in the forced swimming test in healthy animals, it dose-dependently reversed the increased immobility time in rats receiving CCI, suggesting that sinomenine attenuated chronic pain-induced depressive-like behavior. The antinociceptive effects of sinomenine were blocked by the GABAa receptor antagonist bicuculine. The doses of sinomenine studied here did not significantly alter the spontaneous locomotor activity. Together, these results suggested that sinomenine exerts significant antinociceptive effects for neuropathic pain via GABAa-mediated mechanism, which suggests that sinomenine may be useful for the management of chronic painful conditions such as neuropathic pain.

Topics: Analgesics; Animals; Behavior, Animal; Dose-Response Relationship, Drug; Hyperalgesia; Male; Morphinans; Neuralgia; Pain Measurement; Rats; Rats, Sprague-Dawley; Touch; Treatment Outcome

Sinomenine (SIN) is a bioactive alkaloid extracted from the Chinese medicinal plant Sinomenium acutum, which is widely used in the clinical treatment of rheumatoid arthritis (RA). However, its role in acute lung injury (ALI) is unclear. In this study, we investigate the role of SIN in lipopolysaccharide (LPS)-induced ALI in mice. After ALI, lung water content and histological signs of pulmonary injury were attenuated, whereas the PaO2/FIO2 (P/F) ratios were elevated significantly in the mice pretreated with SIN. Additionally, SIN markedly inhibited inflammatory cytokine TNF-α and IL-1β expression levels as well as neutrophil infiltration in the lung tissues of the mice. Microarray analysis and real-time PCR showed that SIN treatment upregulated adenosine A(2A) receptor (A(2A)R) expression, and the protective effect of SIN was abolished in A(2A)R knockout mice. Further investigation in isolated mouse neutrophils confirmed the upregulation of A(2A)R by SIN and showed that A(2A)R-cAMP-PKA signaling was involved in the anti-inflammatory effect of SIN. Taken together, these findings demonstrate an A(2A)R-associated anti-inflammatory effect and the protective role of SIN in ALI, which suggests a potential novel approach to treat ALI.

Topics: Acute Lung Injury; Animals; Cells, Cultured; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Gene Expression Profiling; Gene Expression Regulation; Interleukin-1beta; Lipopolysaccharides; Mice; Mice, Knockout; Morphinans; Neutrophils; Receptor, Adenosine A2A; Signal Transduction; Sinomenium; Tumor Necrosis Factor-alpha

Sinomenine (SIN) is the active principle of the Chinese medical plant Sinomenium acutum which is widely used for the treatment of rheumatoid arthritis (RA) in China. Recently, several groups indicated that myeloid differentiation primary response protein 88 (MyD88) might be associated with disease progression of RA. Here, we observed the effect of SIN on MyD88 expression and showed its therapeutic role in RA. First, immunohistochemical staining in clinical specimens showed that MyD88 was mainly located in characteristic pathological structures of RA synovial tissues. Second, we found that MyD88 was overexpressed in the synovial tissues of the rats with adjuvant-induced arthritis (AIA). Treatment with SIN markedly decreased the expression of MyD88 in AIA rats. Finally, we provided evidences that SIN suppressed inflammation response and inflammation-induced joint destructive progression and arthritis symptoms in AIA rats. Therefore, SIN is an effective therapeutic agent for RA. Targeting MyD88 signaling may provide new methods for the treatment of RA.

Topics: Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Disease Progression; Drugs, Chinese Herbal; Inflammation; Joint Diseases; Male; Medicine, Chinese Traditional; Morphinans; Myeloid Differentiation Factor 88; Rats; Rats, Sprague-Dawley; Synovial Membrane; Toll-Like Receptor 2; Toll-Like Receptor 4

To explore the role of sinomenine in treatment of allergic rhinitis mice model and its possible mechanism.. We used ovalbumin (OVA) to make allergic rhinitis model of BALB/c mice. Saline was used in the control group. When we challenged the mice with OVA intranasally, the mice in sinomenine treatment group were feed by the food containing sinomenine. Mice were then killed 24 h after the last OVA challenge. The noses of mice from each group were removed en bloc and fixed, then each section was stained with hematoxylin and eosin. ELISA assay was used to measure the concentration of anti-OVA IgE, IL-4 and IFN-gamma. The proteins expressive level of T-bet and GATA3 were examined.. Nasal mucosa of the mice in sinomenine treatment group were not hyperplasia and without obvious infiltration of eosinophils. The concentration of anti-OVA IgE, IL-4 and IFN-gamma in the serum and T-bet and GATA3 expression levels of sinomenine treatment group were lower than those of allergic rhinitis group.. The sinomenine can be used to treat allergic rhinitis mice, and the mechanism may rely on the improvements of the Th1/Th2 imbalance.

Topics: Animals; Disease Models, Animal; Eosinophils; GATA3 Transcription Factor; Immunoglobulin E; Interferon-gamma; Interleukin-4; Male; Mice; Mice, Inbred BALB C; Morphinans; Nasal Mucosa; Phytotherapy; Rhinitis, Allergic; Rhinitis, Allergic, Perennial; T-Box Domain Proteins; Th1 Cells; Th2 Cells

To observe the analgesic effect of sinomenine on the neuropathic pain rat model induced by SSNI, and discuss its impact on monoamine neurotransmitters in striatal extracellular fluid.. Male SD rats were randomly divided into the sham operation group, the SSNI model group, the gabapentin group (100 mg x kg(-1)), the sinomenine high dose group (40 mg x kg(-1)) and the sinomenine low dose group (20 mg x kg(-1)). Mechanical hyperalgesia and cold pain sensitivity were evaluated by Von Frey hairs and cold spray. Striatum was sampled by microdialysis. High performance liquid chromatography-electrochemical detector (HPLC-ECD) were used to detect the content of such neurotransmitters as monoamine neurotransmitters noradrenaline (NE), dopamine (DA), 5-hydroxy tryptamine (5-HT) and their metabolites dihydroxyphenylacetic phenylacetic acid (DOPAC) and 5-hydroxyindoleacetic acid (5-HIAA).. SSNI model rats showed significant improvement in mechanical withdrawal threshold and cold pain sensitivity, significant decrease in intracerebral NE and notable increase in DA, 5-HT and their metabolites. Compared with the model group, the sinomenine high dose group showed significant increase in mechanical withdrawal threshold at 60, 90, 180 and 240 min after abdominal administration (P < 0.01), significant decrease in cold pain sensitivity score during 30-240 min (P < 0.05). Sinomenine can significantly up-regulated NE content in striatal extracellular fluid during 45-135 min (P < 0.05), remarkably reduce DA content and DOPAC at 45, 75 and 135 min (P < 0.05), 5-HT content during 45-135 min, DOPAC during 75-165 min (P < 0.05), and 5-HIAA during 45-135 min (P < 0.05).. Sinomenine has the intervention effect on neuropathic pain in SSNI model rats. Its mechanism may be related to disorder of monoamine neurotransmitters in striatal extracellular fluid.

Topics: Analgesics; Animals; Biogenic Monoamines; Disease Models, Animal; Extracellular Fluid; Male; Morphinans; Neostriatum; Neurotransmitter Agents; Rats; Rats, Sprague-Dawley; Sciatic Nerve

The aim of the present study was to develop and optimize sinomenine hydrochloride (SIN) in situ gel for uveitis treatment.. Carbopol 940 was used as the gelling agent in combination with hydroxypropyl methylcellulose (HPMC), which acts as a viscosity enhancer. The formulations were prepared using various concentrations of Carbopol 940 and HPMC. The prepared in situ gels were evaluated for gellation, drug release, ocular irritation, elimination time and pharmacokinetic studies. Furthermore, the effect of SIN on the development of experimental autoimmune anterior uveitis (EAAU) was assessed.. The optimum concentration of Carbopol was 0.1% (w/v), and that for HPMC was 0.4% (w/v). Which showed a significant enhancement in gel strength in the physiological condition while free flowing at non-physiological condition. Optimum formula F(2-3) consisting of 0.5% SIN was prepared and kept as gel group, and 0.5% SIN solution was prepared and kept as control group. Gel group provided sustained release of the drug over a period of 480 min. No evidence of overt toxicity and irritation was observed in any study. The elimination time of control group and gel group was completed within 10 min and 25 min, respectively. The area under the aqueous humor concentration vs. time curve (AUC(0-t)) and maximum concentration (C(max)) values of gel group was 2.70-fold and 1.79-fold higher than that of control group. Additionally, clinical examination showed that SIN suppressed inflammation in EAAU.. These results support the potential applications of SIN in situ gel for uveitis treatment.

Topics: Administration, Topical; Animals; Anti-Inflammatory Agents; Female; Gels; Molecular Structure; Morphinans; Rabbits; Rats; Rats, Inbred Lew; Specific Pathogen-Free Organisms; Uveitis

In this work, we assessed the effects of sinomenine (SN) on intestinal octreotide (OCT) absorption both in Caco-2 cell monolayers and in rats. We also investigated the molecular mechanisms of tight junction (TJ) disruption and recovery by SN-mediated changes in the claudin-1 and protein kinase C (PKC) signaling pathway. The data showed that exposure to SN resulted in a significant decrease in the expression of claudin-1, which represented TJ weakening and paracellular permeability enhancement. Then, the recovery of TJ after SN removal required an increase in claudin-1, which demonstrated the transient and reversible opening for TJ. Meanwhile, the SN-mediated translocation of PKC-α from the cytosol to the membrane was found to prove PKC activation. Finally, SN significantly improved the absolute OCT bioavailability in rats and the transport rate in Caco-2 cell monolayers. We conclude that SN has the ability to enhance intestinal OCT absorption and that these mechanisms are related at least in part to the important role of claudin-1 in SN-mediated, reversible TJ opening via PKC activation.

Topics: Administration, Intravenous; Administration, Oral; Animals; Biological Transport; Caco-2 Cells; Cell Death; Cell Membrane Permeability; Cell Polarity; Claudin-1; Electric Impedance; Fluorescent Antibody Technique; Humans; In Vitro Techniques; Intestinal Absorption; Male; Morphinans; Octreotide; Protein Kinase C-alpha; Rats, Sprague-Dawley; RNA, Messenger; Tight Junctions

The present study investigated the feasibility of ostrich oil utilizing as a promising vehicle for improved skin permeation of sinomenine with reference to vaseline matrix containing different content of chemical enhancers. The fatty acid composition of ostrich oil was analyzed by GC-MS. Penetration enhancing potential of ostrich oil on permeation of sinomenine across rat abdominal skin in vitro was studied using an automatic diffusion cell apparatus. The content of sinomenine percutaneous absorbed was determined by HPLC. Various parameters viz. steady-state skin flux (J(ss)), permeability coefficient (kP), cumulative amount of sinomenine (Q) and enhancement ratios (ER) were calculated from the permeation data. Fick's law of diffusion and Scheuplein kinetic were used to evaluate the transdermal absorbent enhancement of ostrich oil to sinomenine. Ostrich oil showed significant penetration effect on sinomenine compared with vaseline matrix containing different content of chemical enhancers, the density sequence as follow: 2% Azone > ostrich oil > 1% Azone plus 1% propylene glycol > 1% Azone > 3% Azone > 1% propylene glycol. The percutaneous endosmic rate constant (J(ss)) and permeability coefficient (k(P)) of sinomenine in ostrich oil through rat skin were 10.01 μg/cm²/h and 0.087, respectively. Ostrich oil produced stronger enhancement (ER = 24.31) with greater cumulative amount of drug permeated (255.53 μg/cm²) up to 24 h and caused no skin irritation. The drug release of sinomenine was coincided with Fick's equation. In summary, ostrich oil containing fatty acids is proposed as a promising adjuvant for use in cosmetics and pharmaceuticals for improved permeation of drug.

Topics: Administration, Cutaneous; Animals; Chromatography, High Pressure Liquid; Fatty Acids; Gas Chromatography-Mass Spectrometry; In Vitro Techniques; Male; Morphinans; Oils; Petrolatum; Pharmaceutical Vehicles; Rats; Rats, Sprague-Dawley; Skin; Skin Absorption; Stimulation, Chemical; Struthioniformes; Time Factors

Sinomenine (SIN) is a purified alkaloid from the Chinese herb Sinomenium acutum. Previous studies demonstrated that SIN possesses anti-inflammatory and anti-apoptotic properties. We thus in the present report conducted studies to examine its impact on ischemia reperfusion (IR) induced renal injury. Precondition of mice with 200 mg/kg of SIN provided significant protection for mice against IR-induced renal injury as manifested by the attenuated serum creatinine (Cre) and blood urea nitrogen (BUN) along with less severity for histological changes and tubular cell apoptosis. In line with these results, treatment of mice with SIN suppressed IR-induced inflammatory infiltration and the expression of chemokine CXCL-10, adhesion molecule ICAM-1, and cytokines TNF-а/IL-6. Mechanistic studies revealed that SIN inhibits NF-κB transcriptional activity to suppress IR-induced inflammatory response in the kidney, while it attenuates MAP kinase signaling to prevent tubular cells undergoing apoptosis after IR insult. Altogether, our data support that SIN could be a useful therapeutic agent for prevention and treatment of IR-induced renal injury in the clinical settings.

Topics: Animals; Anti-Inflammatory Agents; Apoptosis; Biomarkers; Blood Urea Nitrogen; Chemokine CXCL10; Creatinine; Cytoprotection; Disease Models, Animal; Inflammation Mediators; Intercellular Adhesion Molecule-1; Interleukin-6; Kidney Tubules; Male; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; Morphinans; Nephritis; NF-kappa B; Reperfusion Injury; Time Factors; Tumor Necrosis Factor-alpha

Sinomenine, a pure alkaloid isolated in Chinese medicine from the root of Sinomenium acutum, has been demonstrated to have anti-inflammatory and immunosuppressive effects. MicroRNAs (miRNAs) are gradually being recognized as critical mediators of disease pathogenesis via coordinated regulation of molecular effector pathways.. After colitis was induced in mice by instillation of 5% (w/v) 2,4,6-trinitrobenzenesulfonic acid (TNBS), sinomenine at a dose of 100 or 200 mg/kg was orally administered once daily for 7 days. We evaluated body weight, survival rate, diarrhea score, histological score and myeloperoxidase (MPO) activity. The mRNA and protein expression levels of miR-155, c-Maf, TNF-α and IFN-γ were determined by quantitative RT-PCR and immunohistochemistry, respectively. Sinomenine (100 or 200 mg/kg)-treated mice with TNBS-induced colitis were significantly improved in terms of body weight, survival rate, diarrhea score, histological score and MPO activity compared with untreated mice. Both dosages of sinomenine significantly decreased the mRNA and protein expression levels of c-Maf, TNF-α and IFN-γ, which elevated in TNBS-induced colitis. Furthermore, sinomenine at a dose of 200 mg/kg significantly decreased the level of miR-155 expression by 71% (p = 0.025) compared with untreated TNBS-induced colitis in mice.. Our study evaluated the effects and potential mechanisms of sinomenine in the anti-inflammatory response via miRNA-155 in mice with TNBS-induced colitis. Our findings suggest that sinomenine has anti-inflammatory effects on TNBS-induced colitis by down-regulating the levels of miR-155 and several related inflammatory cytokines.

Topics: Animals; Anti-Inflammatory Agents; Body Weight; Colitis; Interferon-gamma; Male; Mice; Mice, Inbred BALB C; MicroRNAs; Morphinans; Trinitrobenzenesulfonic Acid; Tumor Necrosis Factor-alpha

Receptor activator of NF-κB ligand (RANKL) is essential for osteoclastogenesis. Targeting RANKL signaling pathways has been an encouraging strategy for treating lytic bone diseases such as osteoporosis and rheumatoid arthritis (RA). Sinomenine (SIN), derived from Chinese medicinal plant Sinomenioumacutum, is an active compound to treat RA, but its effect on osteoclasts has been hitherto unknown. In the present study, SIN was found to ameliorate M. tuberculosis H37Ra (Mt)-induced bone loss in rats with a decreased serum level of TRACP5b and RANKL, and an increased level of osteoprotegerin (OPG). In vitro study also showed that SIN could inhibit RANKL-induced osteoclast formation and bone resorption. The osteoclastic specific marker genes induced by RANKL including c-Src, MMP-9, TRACP were inhibited by SIN in a dose dependent manner. Signal transduction studies showed that SIN could obviously reduce the expression of RANK adaptor molecule TRAF6 and down-regulate RANKL-induced NF-κB activation. It decreased the RANKL-induced p38, JNK posphorylation but not ERK1/2 posphorylation. SIN could also reduce RANKL-mediated calcium influx which is associated with TRAF6/c-Src complex. Finally, SIN suppressed RANKL induced AP-1 and NFAT transcription, as well as the gene expression of NFATc1 and AP-1 components (Fra-1, Fra-2, c-Fos). The protein expression of c-Fos and TRAF6 were also inhibited by SIN after RANKL stimulation. Taken together, SIN could attenuate osteoclast formation and Mt-induced bone loss by mediating RANKL signaling pathways.

Topics: Animals; Bone Resorption; Female; Mice; Mice, Inbred BALB C; Morphinans; Mycobacterium tuberculosis; Osteoclasts; RANK Ligand; Rats; Rats, Sprague-Dawley; Receptor Activator of Nuclear Factor-kappa B; Signal Transduction; TNF Receptor-Associated Factor 6

Ankylosing spondylitis (AS) is a common inflammatory rheumatic disease which lacks satisfactory treatment so far. Sinomenine (SIN) is an alkaloid and has recently been utilized in treating multiple rheumatic diseases including AS in China, but its exact mechanism remains to be explored. This study investigated the alteration of proteome in peripheral blood mononuclear cells (PBMCs) from AS patients.. Thirty AS patients were enrolled in this study. PBMCs from each AS patient were cultured in medium with or without SIN respectively. Then PBMCs proteins from both groups were separated by two-dimensional electrophoresis (2-DE) and analyzed by mass spectrometry (MS). Two differentially expressed proteins were then chosen to be verified using Western blotting.. Seven proteins, including a-synuclein (SNCA), calmodulin (CALM), acidic leucine-rich nuclear phosphoprotein 32 family member A (ANP32A), chloride intracellular channel protein 1 (CLIC1), guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1 (GNB1), gelsolin (GSN) and histone H2B type 1-M (HISTH2BM) were over-expressed, while coronin- 1A (CORO1A) was under-expressed in the SIN-treated PBMCs. Further bioinformatics search indicated that the changes of SNCA, ANP32A and CLIC1 pertained to apoptosis, while changes of GSN and CORO1A were associated with both apoptosis and inhibition of immunological function. Subsequently GSN and CORO1A were selected to validate by Western blotting and the results were consistent with those of 2-DE.. There were 8 differentially expressed proteins in the SIN-treated PBMCs, which might shed some light on the mechanism of SIN in the treatment of AS.

Topics: Adolescent; Adult; Blood Proteins; Blotting, Western; Cells, Cultured; Electrophoresis, Gel, Two-Dimensional; Female; Humans; Leukocytes, Mononuclear; Male; Middle Aged; Morphinans; Proteomics; Reproducibility of Results; Spondylitis, Ankylosing

Sinomenine is an alkaloid originally isolated from the root of the plant Sinomenium acutum. It is used in traditional medicine in China to treat rheumatic arthritis. In the present study, we evaluated the potential antinociceptive effects of sinomenine in rodents with nociceptive, inflammatory and neuropathic pain. In normal rats and mice, systemic sinomenine produced moderate antinociceptive effect in the hot plate and tail flick tests. Sinomenine also exerted analgesic effects on mechanical and heat hypersensitivity in mice after carrageenan induced inflammation. Finally, sinomenine effectively alleviated mechanical and cold allodynia in rats and mice after injury to peripheral nerve or spinal cord. The analgesic effect of sinomenine is not associated with side effects and is not reversed by the opioid receptor antagonist naloxone. Our results showed that sinomenine has a wide spectrum analgesic effect in rodent models of nociceptive, inflammatory and neuropathic pain.

Topics: Analgesics; Animals; Behavior, Animal; Carrageenan; Female; Hyperalgesia; Inflammation; Male; Mice; Morphinans; Neuralgia; Nociception; Rats; Sciatic Nerve; Spinal Cord Injuries

To investigate the inhibitory effects of sinomenine (SIN) combined with 5-fluorouracil (5-FU) on esophageal carcinoma in vitro and in vivo.. Esophageal carcinoma (Eca-109) cells were cultured in DMEM. The single or combined growth inhibition effects of SIN and 5-FU on the Eca-109 cells were examined by measuring the absorbance of CCK-8 dye in living cells. Hoechst 33258 staining and an Annexin V/PI apoptosis kit were used to detect the percentage of cells undergoing apoptosis. Western blotting was used to investigate the essential mechanism underlying SIN and 5-FU-induced apoptosis. SIN at 25 mg/kg and 5-FU at 12 mg/kg every 3 d, either combined or alone, was injected into nude mice and tumor growth inhibition and side effects of the drug treatment were observed.. SIN and 5-FU, both in combination and individually, significantly inhibited the proliferation of Eca-109 cells and induced obvious apoptosis. Furthermore, the combined effects were greater than those of the individual agents (P < 0.05). Annexin V/PI staining and Hoechst 33258 staining both indicated that the percentage of apoptotic cells induced by SIN and 5-FU combined or alone were significantly different from the control (P < 0.05). The up-regulation of Bax and down-regulation of Bcl-2 showed that the essential mechanism of apoptosis induced by SIN and 5-FU occurs via the mitochondrial pathway. SIN and 5-FU alone significantly inhibited the growth of tumor xenografts in vivo, and the combined inhibition rate was even higher (P < 0.05). During the course of chemotherapy, no obvious side effects were observed in the liver or kidneys.. The combined effects of SIN and 5-FU on esophageal carcinoma were superior to those of the individual compounds, and the drug combination did not increase the side effects of chemotherapy.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; bcl-2-Associated X Protein; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Dose-Response Relationship, Drug; Drug Synergism; Esophageal Neoplasms; Fluorouracil; Humans; Mice; Mice, Inbred BALB C; Mice, Nude; Morphinans; Proto-Oncogene Proteins c-bcl-2; Tumor Burden; Xenograft Model Antitumor Assays

Transferrin receptor (TfR) has been used as a target for the antibody-based therapy of cancer due to its higher expression in tumors relative to normal tissues. Great potential has been shown by anti-TfR antibodies combined with chemotherapeutic drugs as a possible cancer therapeutic strategy. In our study, we investigated the anti-tumor effects of anti-TfR monoclonal antibody (mAb) alone or in combination with sinomenine hydrochloride in vitro. Results suggested that anti-TfR mAb or sinomenine hydrochloride could induce apoptosis, inhibit proliferation, and affect the cell cycle. A synergistic effect was found in relation to tumor growth inhibition and the induction of apoptosis when anti-TfR mAb and sinomenine hydrochloride were used simultaneously. The expression of COX-2 and VEGF protein in HepG2 cells treated with anti-TfR mAb alone was increased in line with increasing dosage of the agent. In contrast, COX-2 expression was dramatically decreased in HepG2 cells treated with sinomenine hydrochloride alone. Furthermore, we demonstrated that the inhibitory effects of sinomenine hydrochloride and anti-TfR mAb administered in combination were more prominent than when the agents were administered singly. To sum up, these results showed that the combined use of sinomenine hydrochloride and anti-TfR mAb may exert synergistic inhibitory effects on human hepatoma HepG2 cells in a COX-2-dependent manner. This finding provides new insight into how tumor cells overcome the interference of iron intake to survive and forms the basis of a new therapeutic strategy involving the development of anti-TfR mAb combined with sinomenine hydrochloride for liver cancer.

Topics: Antibodies, Monoclonal; Apoptosis; Carcinoma, Hepatocellular; Cell Division; Cyclooxygenase 2; Hep G2 Cells; Humans; Liver Neoplasms; Morphinans; Receptors, Transferrin; Vascular Endothelial Growth Factor A

Sinomenine, a pure alkaloid extract from Sinomenium acutum, has anti-inflammatory and immunoregulatory functions. This study investigated the efficiency and the signalling pathways involved in the effect of sinomenine on vascular smooth muscle cell (VSMC) dedifferentiation in response to platelet-derived growth factor (PDGF)-BB stimulation and vascular injury. VSMCs were isolated from rat aorta and preincubated with sinomenine before being stimulated with PDGF-BB. WST and BrdU incorporation assays were used to evaluate VSMC proliferation. Flow cytometric analysis was performed for testing the cell cycle progression. The cell migration of VSMCs were analysed using a Transwell system. The expression of VSMC specific genes and signalling proteins were tested by Western blot. For the animal study, C57/BL6 mice were fed either normal rodent chow diets or sinomenine chow diets that supplemented with 0.09 % sinomenine (w/w) in the normal chows for 14 days before carotid artery wire injury. PDGF-BB activated the dedifferentiation of VSMCs characterised by decreased expression of SMA, Smoothelin and SM22α. However, sinomenine treatment preserved the dedifferentiation in response to PDGF-BB. The activations of mitogen-activated protein kinase extracellular signal-regulated kinases, Akt, GSK3β and STAT3 induced by PDGF-BB were also inhibited in sinomenine-treated VSMCs. In vivo evidence with wire-injured mice exhibited a reduction in neointimal area and an increase in smooth muscle-specific gene expression in the sinomenine-treated group. In this study, we found that sinomenine-suppressed VSMC phenotype switching induced by PDGF-BB in vitro and neointimal formation in vivo. Therefore, sinomenine is a potential candidate to be used in the treatment of vascular proliferative disease.

Topics: Animals; Carotid Arteries; Cell Cycle; Cell Dedifferentiation; Cell Movement; Cell Proliferation; Cell Survival; Cells, Cultured; Gene Expression; Male; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; Morphinans; Muscle Proteins; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Neointima; Phenotype; Phosphorylation; Protein Processing, Post-Translational; Rats; Rats, Sprague-Dawley; Receptors, Platelet-Derived Growth Factor; STAT3 Transcription Factor

Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide. However, therapies against HCC to date have not been completely effective. Sinomenine hydrochloride (SH), an anti‑arthritis drug applied in clinical practice, has been reported to have in vitro anti‑neoplastic activity in various cancer cells. Whether SH inhibits HCC remains unknown. For this purpose, in this study, MTT assay was used to determine cell growth. Flow cytometry, Hoechst staining, DNA fragmentation, western blot analysis, immunohistochemisty and TUNEL staining were performed to investigate the mechanisms involved. The in vivo activity of SH was determined using a mouse xenograft model. SH inhibited the growth of various types of human HCC cells in vitro. We found that SH promoted cell cycle arrest in the G1 phase and sub‑G1 formation, associated with the increased p21/WAF1/Cip1 expression. Additionally, SH induced caspase‑dependent apoptosis, which involved the disruption of mitochondrial membrane potential, the increased release of cytochrome c and Omi/HtrA2 from the mitochondria into the cytoplasm, the downregulation of Bcl‑2 and the upregulation of Bax, the activation of a caspase cascade (caspase‑8, -10, -9 and -3) and PARP, as well as the decreased expression of survivin. The SH‑suppressed growth of human HCC xenografts in vivo occurred due to the decrease in proliferation and the induction of apoptosis, implicating the activation of caspase‑3, the upregulation of p21 and the downregulation of survivin. These findings suggest that SH exhibits anticancer efficacy in vitro and in vivo involving cell cycle and caspase‑dependent apoptosis and may serve as a potential drug candidate against HCC.

Topics: Animals; Apoptosis; bcl-2-Associated X Protein; Blotting, Western; Carcinoma, Hepatocellular; Caspases; Cell Cycle Checkpoints; Cell Proliferation; Cyclin-Dependent Kinase Inhibitor p21; Cytochromes c; Flow Cytometry; Humans; In Vitro Techniques; Liver Neoplasms; Male; Membrane Potential, Mitochondrial; Mice; Mice, Inbred BALB C; Mice, Nude; Morphinans; Poly(ADP-ribose) Polymerases; Proto-Oncogene Proteins c-bcl-2; Tumor Cells, Cultured

Sinomenine (1) is currently used for the treatment of rheumatoid arthritis (RA) in China and there is still room for the improvement of its efficacy. In present study, capillary based microfluidic system was effectively applied for the syntheses of two novel series of sinomenine derivatives. The Heck reactions in microreactor gave much higher conversions compared to the batch ones. The two-step synthesis of the isoxazoline in microreactor greatly shortened the reaction time without any isolation of intermediates. The inhibitory activity of synthesized compounds on the TNF-α-induced nuclear factor kappa B (NF-κB) activation was evaluated in vitro. Among the compounds, 3c and 3g showed the potent inhibitory activity. Furthermore, 3g exhibited the antiinflammatory effect in vivo.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Carrageenan; Cell Survival; Dose-Response Relationship, Drug; Edema; Mice; Mice, Inbred ICR; Microfluidic Analytical Techniques; Models, Molecular; Morphinans; NF-kappa B; NIH 3T3 Cells; Structure-Activity Relationship; Tumor Necrosis Factor-alpha

Sinomenine has been used to treat autoimmune diseases for centuries. However, little is known about its exact mechanisms of action. Whether sinomenine has an effect on programmed death‑1 (PD‑1) ligands (PD‑Ls) in vivo remains unclear. The present study aimed to determine the effect of sinomenine on the expression of PD‑L1 and PD‑L2 in peripheral blood mononuclear cells (PBMCs). A total of 25 patients with mesangial proliferative nephritis (MsPGN) were treated with sinomenine and followed up for 3 months. The expression of PD‑L1 and PD‑L2 was studied by using real‑time RT‑PCR and flow cytometric analysis, and recorded at months 0, 1 and 3 within the PBMCs. The intra‑renal expression of PD‑L1 and PD‑L2 was studied by immunohistochemistry. The results revealed that the PBMCs from the MsPGN patients expressed high levels of PD‑L1 at the mRNA and protein levels compared with the healthy donors. Immunohistochemistry revealed an increased PD‑L1 expression in the renal tissues from the MsPGN patients. Sinomenine was observed to have a significant effect in decreasing the PD‑L1 expression in the PBMCs. The present study therefore suggests a novel mechanism for the therapeutic effects of sinomenine on MsPGN in vivo.

Topics: Adult; B7-H1 Antigen; Female; Flow Cytometry; Gene Expression Regulation; Glomerular Mesangium; Humans; Leukocytes, Mononuclear; Male; Morphinans; Nephritis; Nephrotic Syndrome; Programmed Cell Death 1 Receptor; Sclerosis

The aim of the research was to investigate the antiendotoxin effects of Sinomenine, Fangchinoline, Stachydrine, Chuanxionggzine, Oxymartrine and Evodiamine. Endothelial cells were challenged with 1 μg/mL LPS for 3 h then treated respectively with six alkaloids at three concentrations (1, 5 and 10 μg/mL). The cells were incubated at 37°C in a cell incubator for 21 h. The supernatants were collected and analyzed the levels of interleukin-1α (IL-1α), thromboxane B(2) (TXB(2)), endothelin-1 (ET-1) and E-selectin by ELISA kits. The results revealed that Sinomenine, Oxymartrine and Evodiamine inhibited the production of IL-1α; Stachydrine, Chuanxionggzine and Evodiamine inhibited the secretion of TXB(2); Sinomenine and Oxymartrine down-regulated ET-1 expression; Fangchinoline and Evodiamine decreased the level of E-selectin. All these changes were significant. Taken together, the data suggested that six alkaloids may effectively reduce inflammatory response via these cytokines.

Topics: Alkaloids; Animals; Anti-Inflammatory Agents; Benzylisoquinolines; Cells, Cultured; Down-Regulation; Drugs, Chinese Herbal; E-Selectin; Endothelin-1; Endothelium, Vascular; Evodia; Humans; Interleukin-1alpha; Lipopolysaccharides; Morphinans; Proline; Pyrazines; Quinazolines; Quinolizines; Swine; Thromboxane B2

Sinomenine (1) is clinically available for the treatment of rheumatoid arthritis (RA), however, its efficacy is quite weak. In the present study, a library of novel sinomenine-based homodimers and monomers through variable-length linkers were designed and synthesized, and their bioactivities were evaluated using RAW264.7 cells and mice. Among the compounds, 2f and 3b possessed much more potent inhibitory effects on the production of nitric oxide (NO), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) than 1. Preliminary mechanism investigation revealed that 3b inhibited nuclear factor-κB (NF-κB) signaling pathway specifically, 2f suppressed both NF-κB and mitogen-activated protein kinase (MAPK) cascades. Moreover, 3b and 2f significantly alleviated the lipopolysaccharide (LPS)-induced mortality. These two compounds might serve as valuable candidates for anti-inflammatory drug discovery.

Topics: Animals; Anti-Inflammatory Agents; Blotting, Western; Cell Proliferation; Cells, Cultured; Enzyme-Linked Immunosorbent Assay; Interleukin-6; JNK Mitogen-Activated Protein Kinases; Lipopolysaccharides; Macrophages; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinases; Morphinans; NF-kappa B; Nitric Oxide; Nitric Oxide Synthase Type II; Shock, Septic; Signal Transduction; Tumor Necrosis Factor-alpha

A sensitive and selective liquid chromatographic tandem mass spectrometric method was developed and validated for the determination of sinomenine in human plasma. Plasma samples were precipitated using methanol with metronidazole as internal standard. Separation was carried out on an Inertsil ODS-3 column using a mixture of 0.2% ammonium acetate solution (A) and methanol (B) as the mobile phase with linear gradient elution as follows: 0 min (50%B)→1.5 min (80%B)→4.5 min (80%B)→4.6 min (50%B)→6.0 min (50%B). All mass data were obtained in the positive ion mode, and the fragmentation transitions for the selective multiple reaction monitoring were m/z 330→181 and 172→128 for sinomenine and metronidazole, respectively. The method was fully validated to be accurate and precise with a linear range of 0.5-500 ng/mL and applied to a single- and multiple-dose pharmacokinetics study of sustained-release capsules of sinomenine hydrochloride in 20 healthy Chinese volunteers. After oral administration of a single 60-mg dose, the T(max), C(max), AUC(0-96) and t(1/2) were 7.9±2.0h, 123±22 ng/mL, 3032±682 ng h/mL and 13.4±1.6 h, respectively. After oral administration of the 60 mg capsules twice-daily for 7 consecutive days, these parameters were 4.4±3.6 h, 279±69 ng/mL, 7333±2096 ng h/mL and 15.1±1.3 h, respectively. The AUC and C(max) values after multiple-dose treatment were significantly higher than those after a single-dose treatment (P<0.01), with an accumulation factor of 2.49±0.77.

Topics: Adult; Area Under Curve; China; Chromatography, Liquid; Delayed-Action Preparations; Drug Stability; Humans; Least-Squares Analysis; Male; Morphinans; Reproducibility of Results; Sensitivity and Specificity; Tandem Mass Spectrometry

Sinomenine (SIN) is a bioactive component derived from a Chinese medicinal plant. Our previous studies demonstrated that SIN has cytotoxic effects on human lung cancer cells. However, the antitumor molecular mechanisms of SIN have yet to be elucidated in detail. In the present study, we further explored the effects of SIN on NCI-H460 human lung cancer cell viability and apoptosis and investigated the regulation and function of PI3K/Akt and ERK signaling pathways during SIN-induced apoptosis in various lung cancer cell lines. NCI-H460 cells were incubated with 200 µg/ml SIN for the indicated times (0, 24, 48 or 72 h). Cell viability was assessed by MTT assay. Akt, p-Akt, ERK1/2 and p-ERK1/2 protein levels were detected by western blotting, respectively. Two different selective inhibitors (LY294002 for the PI3K pathway; PD98059 for the MEK/ERK pathway) were used to characterize the relative roles of PI3K/Akt and ERK in SIN-induced apoptosis. Apoptosis was determined by flow cytometry. SIN inhibited the proliferation of NCI-H460 cells in a time-dependent manner, which was accompanied with significant activation of pAkt and pERK. LY294002 and PD98059 both significantly increased SIN-induced apoptosis in NCI-H460, NCI-H226 and NCI-H522 cells. Our findings suggest that the activation of the PI3K/Akt and ERK signaling pathways antagonize SIN-induced lung cancer cell apoptosis and molecules that inhibit these pathways should potentiate the effects of SIN. This study represents a significant step forward in our understanding of the signal transduction pathways associated with the apoptosis elicited by SIN.

Topics: Apoptosis; Cell Line, Tumor; Cell Proliferation; Cell Survival; Enzyme Activation; Flavonoids; Humans; Lung Neoplasms; MAP Kinase Signaling System; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Morphinans; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Proto-Oncogene Proteins c-akt

Sinomenine (SN, 1) is a pure compound extracted from the Sinomenium acutum plant. We investigated the protective effects and mechanism of action of SN in a rat model of doxorubicin (DOX)-induced nephrosis. Nephrosis was induced by a single dose of 5 mg/kg DOX, and DOX-treated rats received a daily i.p. injection of 10 or 30 mg/kg SN, or saline (n = 6). Urine and serum biochemical parameters, serum TNF-α and IL-1β levels, nephrin, podocin, α-actinin-4, and peroxisome proliferator-activated receptor-α (PPAR-α) protein expression, and renal ultrastructure were examined at day 28. Compound 1 significantly attenuated the effect of DOX on urine and serum biochemical parameters. Electron microscopy demonstrated that 1 suppressed DOX-induced increases in foot process width. Compared with those in control rats, nephrin, podocin, and PPAR-α protein expressions decreased in the glomeruli of DOX-treated rats, and this effect was significantly attenuated by 1. However, no appreciable alterations were observed in the expression level of α-actinin-4. DOX significantly increased serum TNF-α and IL-1β compared with those in control rats, and 1 significantly reduced the serum levels of TNF-α and IL-1β. SN ameliorates DOX-induced nephrotic syndrome in rats, resulting in a modulation of renal nephrin, podocin expression, and thereby protecting podocytes from injury.

Topics: Animals; Doxorubicin; Interleukin-1beta; Intracellular Signaling Peptides and Proteins; Kidney Glomerulus; Male; Membrane Proteins; Models, Biological; Molecular Structure; Morphinans; Nephrosis; PPAR alpha; Rats; Tumor Necrosis Factor-alpha

To study the role of sinomenine (SIN) in prolonging high-risk corneal graft survival in rats.. All recipients were randomly assigned to SIN, cyclosporine A (CsA), SIN plus CsA and control groups. IL-2, IL-10, Fas-Fas ligand and CD4(+)CD25(+)FoxP3(+) T cells in peripheral blood were detected. In addition, rat corneal grafts' survival times were recorded.. Survival time was 15.88 ± 5.87 days in the SIN group, 17.67 ± 5.43 days in the CsA group and 20.75 ± 4.77 days in the drug combination group, which were longer survival times than those in the control group (p < 0.05). Compared with the SIN and CsA groups, levels of CD4(+)CD25(+)FoxP3(+) lymphocytes in the control group were decreased (p < 0.05) and were increased in the cotreated group (p < 0.05). IL-2 levels in the SIN-only and CsA-only groups were lower than those in the control group (p < 0.05) and higher than those in the cotreated group (p < 0.05); however, the results for IL-10 were different. The expressions of Fas and Fas ligand were least in the control group.. SIN could prolong allograft survival and might have potential clinical usage.

Topics: Animals; CD4 Antigens; Corneal Transplantation; Cyclosporine; Drug Therapy, Combination; Female; Forkhead Transcription Factors; Graft Rejection; Graft Survival; Humans; Interleukin-10; Interleukin-2; Interleukin-2 Receptor alpha Subunit; Models, Animal; Morphinans; Rats; Rats, Inbred Lew; T-Lymphocytes, Regulatory

While intra-articular injection of sinomenine hydrochloride has a therapeutic effect on osteoarthritis, it has a short half-life, and is thermolabile and photolabile. The aim of this research was to evaluate the sustained-release of sinomenine hydrochloride from an injectable sinomenine hydrochloride and sodium hyaluronate compound (CSSSI) and its therapeutic effect in a rabbit model of osteoarthritis following intra-articular injection.. An injectable compound consisting of 1% sodium hyaluronate and 2.5% sinomenine hydrochloride was prepared and kept as the experiment group, and 2.5% sinomenine hydrochloride was prepared and kept as the control group. The cumulative mass release was measured at different time points in each group in vitro. Sixty-five male Zelanian rabbits were randomly divided into five groups: 15 (30 knees) each for the control, sodium hyaluronate, sinomenine hydrochloride, and CSSSI groups respectively, and five (10 knees) for the modeling group. Papain was injected into both knees of each rabbit for model establishment. Subsequently, 0.2 ml of the corresponding drugs was injected into the articular cavities of the remaining experiment groups, while the control group was treated with 0.2 ml normal saline. All groups were treated once a week for 4 weeks. Seven days after the last treatment, knees were anatomized to perform pathological observations and Mankin's evaluation of the synovium. Four groups were compared using the SPSS 13.0 software package.. In the in vitro sustained-release experiments, 90% of the drug was released in the experiment group 360 minutes following the injection. Comparison of the Mankin's evaluations of the four groups illustrated statistical discrepancies (P < 0.05). In further paired comparisons of the CSSSI group vs. modeling control/sodium hyaluronate/sinomenine hydrochloride groups, statistical significance was uniformly obtained. Moreover, sodium hyaluronate and sinomenine hydrochloride treatments showed significant improvement over the modeling control (P < 0.05), whereas sodium hyaluronate vs. sinomenine hydrochloride comparison failed to reach significance (P > 0.05).. CSSSI has a sustained-release effect on sinomenine hydrochloride. Intra-articular injection of CSSSI was significantly better than the sole sodium hyaluronate or sinomenine hydrochloride for the treatment of osteoarthritis in a rabbit model.

Topics: Animals; Hyaluronic Acid; Injections, Intra-Articular; Male; Morphinans; Osteoarthritis; Rabbits; Random Allocation

In order to search for drugs with excellent anti-inflammatory activities, a series of novel sinomenine derivatives were designed, synthesized, and evaluated for their inhibition activities against NF-κB activation induced by lipopolysaccharide (LPS). Compared with the natural parent sinomenine, compounds 2a-w showed higher activity, while compounds 1a-o showed similar activity against NF-κB. Moreover, a molecular model for the binding between compound 2v and the active site of p50 was provided on the basis of the computational docking results.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Dose-Response Relationship, Drug; Drug Design; Lipopolysaccharides; Models, Molecular; Molecular Structure; Morphinans; NF-kappa B; Stereoisomerism; Structure-Activity Relationship

To evaluate the immunosuppressive efficacy of alkaloid sinomenine (SIN) and the synergistic effects in combination with cyclosporin A (CsA) in acute rejection after rat renal allograft.. Animals were treated with saline in group 1, SIN (30 mg/kg/d) in group 2, CsA (2.5 mg/kg/d) in group 3 and SIN (30 mg/kg/d) + CsA(2.5 mg/kg/d) in group 4. Another 12 syngeneic renal transplantation animals were treated with saline as control. Survival time is observed. The levels of serum creatinine (Scr) and blood urea nitrogen (Bun) were detected; the secretion of IFN-γ, IL-1β, TNF-α and IL-6 were detected by ELISA. The kidneys were fixed to perform histological staining.. The mean survival time was 8.00 ± 2.10 days in group 1, 10.67 ± 1.21 days in group 2, 11.00 ± 1.41 days in group 3 and 19.67 ± 2.80 days in group 4, while all the recipients survived more than 180 days in the control group. The 24-h urinary volume and urinary time of the other three groups were increased significantly compared with group 1. The levels of Scr and Bun, levels of IFN-γ, IL-1β, TNF-α and IL-6 were significantly higher in group 1 than that in the other three groups; there were significant differences between group 4 and group 2 or 3.. Our study showed that SIN had immunosupression effects in rat renal allograft models, it also had a synergistic effect in combination with CsA, which provided a new immunosuppressant for clinical application.

Topics: Animals; Blood Urea Nitrogen; Creatinine; Cyclosporine; Drug Synergism; Drug Therapy, Combination; Female; Graft Rejection; Humans; Immunosuppressive Agents; Interferon-gamma; Interleukin-1beta; Interleukin-6; Kidney; Kidney Transplantation; Male; Models, Animal; Morphinans; Rats; Rats, Inbred F344; Rats, Wistar; Transplantation, Homologous; Tumor Necrosis Factor-alpha

Sinomenine is an alkaloid compound and a prominent anti-inflammatory agent found in the root of the climbing plant Sinomenium acutum. However, its effects on the mechanism of human mast cell line (HMC)-1-mediated inflammation remained unknown.. To provide insight into the biological effects of sinomenine, we examined its influence on the pro-inflammatory cytokine production in HMC-1 cells stimulated by phorbol 12-myristate-13-acetate (PMA) plus A23187 by evaluating the stimulated cells in the presence or absence of sinomenine. In the present study, the pro-inflammatory cytokine production was measured using ELISA, Reverse Transcription-polymerase chain reaction (RT-PCR) and nuclear factor (NF)-kappaB, mitogen-activated protein kinases (MAPKs) pathway activation, as determined by Western blot analysis. Also, cyclooxygenase (COX)-2 expression was measured through Western blot and RT-PCR analysis.. Sinomenine inhibited the pro-inflammatory cytokine production induced by PMA plus A23187 in a dose-dependent manner. Furthermore, sinomenine inhibited the phosphorylations of extracellular signal-regulated kinase (ERK) and p38 MAPKs as well as the translocation of NF-kappaB p65 through reduced IkappaBalpha degradation. In addition, sinomenine suppressed COX-2 protein and mRNA expression dose-dependently.. Taken together, the results of this study indicate that the anti-inflammatory effects of sinomenine may occur via the inhibition of pro-inflammatory cytokine and COX-2 production through the inhibition of MAPKs and NF-kappaB pathway activation by PMA plus A23187 stimulation in HMC-1 cells.

Topics: Anti-Inflammatory Agents; Calcimycin; Cells, Cultured; Cyclooxygenase 2; Humans; Inflammation Mediators; Morphinans; NF-kappa B; Phosphorylation; Tetradecanoylphorbol Acetate

The binding of sinomenine to human serum albumin (HSA) in aqueous solution in the absence and presence of urea has been studied by fluorescence and the three-dimensional (3D) fluorescence at pH 7.40. Subdomain IIA binding site of human serum albumin (HSA) was characterized by examining the change in HSA fluorescence. The quenching rate constants and binding constants were calculated in the absence and presence of the denaturant. The results point to a static quenching mechanism operating in the complexes. However, the binding ability of sinomenine to denatured HSA is weaker than that of sinomenine to native HSA. Denaturation of HSA in the presence of urea is almost complete at [urea]≥ 8.0M. Upon unfolding, two fluorescence peaks were observed. One peak was assigned to the fluorescence of Trp-214 residue in a polar environment, and the other peak was assigned to the fluorescence of tyrosine residues. Compared to the free HSA, the HSA-sinomenine complex is more stable in the presence of urea.

Topics: Binding Sites; Humans; Models, Molecular; Morphinans; Protein Binding; Protein Structure, Tertiary; Protein Unfolding; Serum Albumin; Spectrometry, Fluorescence; Substrate Specificity; Urea

To investigate the effects of sinomenine (SIN) and methotrexate (MTX) on the proliferation and apoptosis of in vitro cultured fibroblast-like synoviocytes (FLS) in rheumatoid arthritis (RA) patients, as well as the expression of osteoclast differentiation factor in FLS.. FLS were isolated from the synovium of RA patients and cultured in vitro. FLS were incubated with different concentrations of SIN and MTX respectively or combined: 0.001, 0.010, 0.100, 1.000 mg/mL SIN; 0.001, 0.010, 0.100, 1.000 mg/mL MTX; 0.001 mg/mL SIN + 0.001 mg/mL MTX, 0.010 mg/mL SIN + 0.010 mg/mL MTX, 0.100 mg/mL SIN + 0.100 mg/mL MTX, 1.000 mg/mL SIN + 1.000 mg/mL MTX, namely SIN1, 2, 3, 4 groups; MTX1, 2, 3, 4 groups and the combination 1, 2, 3, 4 groups. The medium without drugs was used as a control group. There was a total of 13 groups, each group with 3 complex holes. MTT was applied to detect the growth of FLS. The flow cytometry was applied to detect the apoptosis of FLS. The expressions of FLS receptor activator of nuclear factor kappa B ligand (RANKL) mRNA and osteoprotegerin (OPG) mRNA were observed by semi-quantitative RT-PCR.. Compared with the control group, RA FLS proliferation OD values of all the drug groups were lower (P < 0.05). The RA FLS apoptosis OD value of the combination 3 group increased, the OPG mRNA expression increased, the expression of RANKL mRNA decreased with statistical difference (P < 0.05). The RA proliferation OD values of the SIN3 group and the MTX3 group increased when compared with the combination 3 group (P < 0.05).. SIN and MTX had synergistic effects in inhibiting FLS. This might be one of the mechanisms for inhibiting RA bone damage.

Topics: Apoptosis; Arthritis, Rheumatoid; Cell Proliferation; Cells, Cultured; Fibroblasts; Humans; Methotrexate; Morphinans; Synovial Membrane

CD147 expressed by monocytes, macrophages, and synoviocytes cells can stimulate the production of matrix metalloproteinases (MMPs) associated with the development of rheumatoid arthritis (RA). We investigated the effects of Sinomenine (SIN) on invasion and migration ability and gene expression of CD147, MMP-2, MMP-9 of fibroblast-like synoviocytes cells (FLS) co-cultured with activated human monocytic THP-1 cells (A-THP-1) in vitro. SIN is a pure alkaloid extracted from the Chinese medical plant Sinomenium acutum. FLS cells were co-cultured with THP-1 cells which were induced to differentiate into macrophages with phorbol 12-myristate 13-acetate (PMA). Cells were treated with different concentrations of SIN. Invasion and migration ability of cells was tested by transwell assays. Western blot analysis and zymographic analysis were adopted to detect the expression of CD147 and MMPs, respectively. RT-PCR was used to determine the expression of mRNA of CD147, MMP-2, and MMP-9. The invasion and migration ability of the co-cultured cells was significantly inhibited by SIN in a concentration-dependent fashion, and at the same time, the levels of CD147, MMP-2, MMP-9 were markedly down-regulated. This inhibitory effect was most notable at concentrations of 0.25 and 1.00 mM (P < 0.01). Our results point to a possible mechanism of SIN on treatment of RA is the inhibitory effect of SIN on cell invasion and migration ability, which strongly correlates with repressing the expression of CD147, MMP-2, and MMP-9.

Topics: Antirheumatic Agents; Basigin; Biomarkers; Cell Adhesion; Cell Movement; Cells, Cultured; Coculture Techniques; Down-Regulation; Fibroblasts; Gene Expression Regulation, Enzymologic; Humans; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Matrix Metalloproteinase Inhibitors; Matrix Metalloproteinases, Secreted; Monocytes; Morphinans; Synovial Membrane

Sinomenine is an alkaloid compound and a prominent anti-allergic agent found in the root of the climbing plant Sinomenium acutum. However, its effects on the bone marrow-derived mast cell (BMMC) mediated allergy and inflammation mechanism remain unknown. In this study, the biological effects of sinomenine were evaluated while focusing on its effects on the allergic mediator in PMA plus A23187-stimulated BMMCs. An investigation was also conducted to determine its effects on the production of several allergic mediators including interleukin-6 (IL-6), prostaglandin D(2) (PGD(2)), leukotriene C(4) (LTC(4)), β-Hexosaminidase (β-Hex), and cyclooxygenase-2 (COX-2) protein. The results revealed that sinomenine inhibited the PMA plus A23187-induced production of IL-6, PGD(2), LTC(4), β-Hex, and COX-2 protein. Taken together, these findings indicate that sinomenine has the potential for use in the treatment of allergy.

Topics: Animals; Anti-Allergic Agents; Bone Marrow Cells; Cell Survival; Cells, Cultured; Leukotriene C4; Male; Mast Cells; Mice; Mice, Inbred BALB C; Morphinans; Prostaglandin D2

Inhibition of the excessive NO production has been recognized as a potential means for the treatment of rheumatoid arthritis (RA). In order to discover more potent inhibitors and explore the preliminary structure activity relationship, a series of unique stereodimers of sinomenine analogues were designed and synthesized. Their inhibitory activity on NO production and cytotoxicity were evaluated using LPS-activated murine macrophages RAW264.7 assay and MTT method, respectively. Among these compounds, 1a, 2, 2a, 2b, and 4 showed potent inhibitory activity on NO production without obvious cytotoxicity. Furthermore, 2, 2a, and 2b significantly suppressed mRNA expression of iNOS. Interestingly, (S)-dimers displayed a better bioactivity than (R)-dimers. These compounds may sever as lead candidates in the development of novel therapeutic drugs for RA treatment.

Topics: Animals; Arthritis, Rheumatoid; Cell Line; Cell Survival; Dimerization; Gene Expression; Macrophages; Mice; Morphinans; Nitric Oxide; Nitric Oxide Synthase Type II; RNA, Messenger; Stereoisomerism

Sinomenine (SN), a bioactive alkaloid, has been utilized clinically to treat rheumatoid arthritis in China. Our preliminary experiments indicated that it could protect PC12 cells from oxygen-glucose deprivation-reperfusion (OGD-R), we thus investigated the possible effects of SN on cerebral ischaemia and the related mechanism.. Middle cerebral artery occlusion in rats was used as an animal model of ischaemic stroke in vivo. The mechanisms of the effects of SN were investigated in vitro using whole-cell patch-clamp recording, calcium imaging in PC12 cells and rat cortical neurons subjected to OGD-R.. Pretreatment with SN (10 and 30 mg·kg(-1) , i.p.) significantly decreased brain infarction and the overactivation of calcium-mediated events in rats subjected to 2 h ischaemia followed by 24 h reperfusion. Extracellular application of SN inhibited the currents mediated by acid-sensing ion channel 1a and L-type voltage-gated calcium channels, in the rat cultured neurons, in a concentration-dependent manner. These inhibitory effects contribute to the neuroprotection of SN against OGD-R and extracellular acidosis-induced cytotoxicity. More importantly, administration of SN (30 mg·kg(-1) , i.p.) at 1 and 2 h after cerebral ischaemia also decreased brain infarction and improved functional recovery.. SN exerts potent protective effects against ischaemic brain injury when administered before ischaemia or even after the injury. The inhibitory effects of SN on acid-sensing ion channel 1a and L-type calcium channels are involved in this neuroprotection.

Topics: Acid Sensing Ion Channels; Animals; Animals, Newborn; Blotting, Western; Brain Ischemia; Calcium Channels, L-Type; Cell Culture Techniques; CHO Cells; Cricetinae; Cricetulus; Cytoprotection; Disease Models, Animal; Glucose; Male; Morphinans; Nerve Tissue Proteins; Neurons; Neuroprotective Agents; Oxygen; Patch-Clamp Techniques; PC12 Cells; Rats; Rats, Sprague-Dawley; Sodium Channels

Two new morphinane alkaloids, 1-hydroxy-10-oxo-sinomenine (1) and 4,5-epoxy-14-hydroxy sinomenine N-oxide (2), have been isolated from the stems of Sinomenium acutum. Their structures were established by various spectral analyses, especially 2D NMR experiments. The structure of 2 was confirmed by single crystal X-ray diffraction. The absolute configurations of 1 and 2 were deduced by comparison of CD spectra with the known alkaloid sinomenine (3). Compound 1 was tested for DPPH inhibition and gave IC(50) of 27.9 μM. Compound 2 was tested for neuroprotective effect and showed significant activity against β-amyloid(25-35)-induced oxidative injury (*P < 0.05) at 10 μM in PC-12 cells.

Topics: Alkaloids; Animals; Biphenyl Compounds; Cyclic N-Oxides; Drugs, Chinese Herbal; Molecular Structure; Morphinans; Neuroprotective Agents; Nuclear Magnetic Resonance, Biomolecular; PC12 Cells; Picrates; Plant Stems; Rats; Sinomenium; X-Ray Diffraction

Sinomenine, an alkaloid isolated from the root of Sinomenium acutum, has been used to alleviate the symptoms of rheumatic diseases. Liang Miao San (LMS), composed of the herbs Rhizoma Atractylodis (Cangzhu) and Cotex Phellodendri (Huangbai), is another traditional Chinese medicine formula for rheumatoid arthritis (RA) treatment. Although numerous studies have demonstrated the potential anti-inflammatory activities of sinomenine and LMS, the underlying intracellular mechanisms regulating the anti-inflammatory activities of sinomenine and LMS on human primary fibroblast-like synoviocytes (FLS) from RA patients and normal control subjects have not been elucidated.. We investigated the in vitro anti-inflammatory activity of sinomenine and LMS on inflammatory cytokine tumor necrosis factor (TNF)-α-mediated activation of human normal and RA-FLS. The underlying intracellular signaling molecules were analyzed quantitatively using flow cytometry.. Sinomenine was found to significantly inhibit TNF-α induced cell surface expression of vascular cell adhesion molecule (VCAM)-1 and release of inflammatory cytokine and chemokine IL-6, CCL2 and CXCL8 from both normal and RA-FLS (all p<0.05). Moreover, the suppression of sinomenine on TNF-α induced VCAM-1 expression and IL-6 release of RA-FLS was significantly higher than that of normal FLS (p<0.05). LMS significantly inhibited TNF-α-induced inflammatory chemokines CXCL10 and CCL5 release from both normal and RA-FLS, with significantly higher suppression on CXCL10 secretion in RA-FLS than that of normal FLS (all p<0.05). Further investigations showed that sinomenine and LMS could significantly suppress TNF-α-induced phosphorylation of inhibitor κBα and extracellular signal-regulated protein kinase, the central signaling molecules mediating TNF-α-induced VCAM-1 expression and chemokine production.. Our results therefore provide a new insight into the differential anti-inflammatory activities of sinomenine and LMS through the suppression of TNF-α-activated FLS by modulating distinct intracellular signaling pathways in RA.

Topics: Anti-Inflammatory Agents; Antirheumatic Agents; Arthritis, Rheumatoid; Case-Control Studies; Cells, Cultured; Chemokine CCL2; Chemokine CCL5; Chemokine CXCL10; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Extracellular Signal-Regulated MAP Kinases; Fibroblasts; Flow Cytometry; Humans; I-kappa B Proteins; Inflammation Mediators; Interleukin-6; Interleukin-8; Medicine, Chinese Traditional; Morphinans; NF-kappa B; NF-KappaB Inhibitor alpha; Phosphorylation; Signal Transduction; Synovial Membrane; Tumor Necrosis Factor-alpha; Vascular Cell Adhesion Molecule-1

Neuroinflammation is an important contributor to the development of neurodegenerative diseases, including Alzheimer's disease. Thus, there is a keen interest in identifying compounds, especially from herbal sources, that can inhibit neuroinflammation. Amyloid-β (Aβ) is a major component of the amyloid plaques present in the brains of Alzheimer's disease patients. Here, we examined whether sinomenine, present in a Chinese medicinal plant, prevents oligomeric Aβ-induced microglial activation and confers protection against neurotoxicity.. Oligomeric amyloid-β was prepared from Aβ(1-42). Intracellular reactive oxygen species production was determined using the dye 2',7'-dichlorodihydrofluorescin diacetate. Nitric oxide level was assessed using the Griess reagent. Flow cytometry was used to examine the levels of inflammatory molecules. BV2-conditioned medium was used to treat hippocampal cell line (HT22) and primary hippocampal cells in indirect toxicity experiments. Toxicity was assessed using MTT reduction and TUNEL assays.. We found that sinomenine prevents the oligomeric Aβ-induced increase in levels of reactive oxygen species and nitric oxide in BV2 microglial cells. In addition, sinomenine reduces levels of Aβ-induced inflammatory molecules. Furthermore, sinomenine protects hippocampal HT22 cells as well as primary hippocampal cells from indirect toxicity mediated by Aβ-treated microglial cells, but has no effect on Aβ-induced direct toxicity to HT22 cells. Finally, we found that conditioned medium from Aβ-treated BV2 cells contains increased levels of nitric oxide and inflammatory molecules, but the levels of these molecules are reduced by sinomenine.. Sinomenine prevents oligomeric Aβ-induced microglial activation, and confers protection against indirect neurotoxicity to hippocampal cells. These results raise the possibility that sinomenine may have therapeutic potential for the treatment of Alzheimer's diseases as well as other diseases that involve neuroinflammation.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Cells, Cultured; Culture Media, Conditioned; Hippocampus; Humans; Microglia; Morphinans; Neuroprotective Agents; Nitric Oxide; Peptide Fragments; Reactive Oxygen Species

Herbal products can interfere with allopathic medicinal treatment through pharmacokinetic and pharmacodynamic interactions. Although pharmacokinetic interactions that alter drug absorption may cause variable and unsatisfactory drug bioavailability, a drug absorption enhancement effect of a herb may be used to ensure sufficient absorption of poorly absorbable drugs. The effect of the hydrochloride salt of sinomenine, an alkaloid obtained from the plant Sinomenium acutum, on the transepithelial electrical resistance and transport of different compounds (including cimetidine, vitamin C, rutin, luteolin and insulin) across Caco-2 epithelial cell monolayers was investigated in this study. Sinomenine HCl induced a concentration dependent lowering effect on the transepithelial electrical resistance of Caco-2 cell monolayers, which was completely reversible. Sinomenine HCl significantly increased the transport of all the test compounds in the apical-to-basolateral direction compared with the control group and decreased the transport of cimetidine, a P-glycoprotein substrate, in the basolateral-to-apical direction. From these results it can be concluded that sinomenine HCl increases drug absorption across the intestinal epithelium by means of one or more mechanisms including a transient opening of the tight junctions (as indicated by a reduction in transepithelial electrical resistance) to allow for paracellular transport and/or inhibition of active drug efflux transport (as indicated by inhibition of basolateral-to-apical transport of cimetidine).

Topics: Ascorbic Acid; Caco-2 Cells; Cell Membrane Permeability; Cimetidine; Epithelial Cells; Humans; Insulin; Intestinal Absorption; Intestinal Mucosa; Luteolin; Morphinans; Rutin

The purpose of this work was to investigate feasibility of a promising topical drug delivery system (TDDS) for sinomenine hydrochloride (SMH), extracted from the Chinese medicinal plant sinomenine acutum and currently used for the treatment of rheumatoid arthritis. It was found that SMH was a weak base (pK(a), 7.98 +/- 0.04) with pH-dependent solubility and partition coefficient. The result of in vitro permeation studies demonstrated that the permeation enhancer azone was the most effective. In contrast, spray had higher accumulative permeated amounts of SMH than patch, but permeated duration of spray was shorter than that of patch. The efficacy on Freund's complete adjuvant-induced arthritis suggested that there was near arthritis index for SMH spray with medium dose (i.e., 15 mg/rat) and oral solution at a dose of 12 mg/rat, indicating that topical SMH delivery system could achieve the similar anti-inflammatory efficacy with oral administration. Pharmacokinetic parameters including C(max) and AUC for both topical preparations were lower than those for oral preparation, which hinted that systemic side effect could be ignored. Therefore, the spray and patch were promising formulations for successful topical delivery of SMH through the skin instead of oral administration with side effects.

Topics: Administration, Topical; Animals; Antirheumatic Agents; Arthritis, Rheumatoid; Drug Delivery Systems; Drugs, Chinese Herbal; Humans; Male; Morphinans; Rabbits; Rats; Rats, Wistar; Sinomenium; Skin; Skin Irritancy Tests

Developing a sustained release drug to cure arthritis is needed. Sinomenine (SIN) is abstracted from sinomenium acutum and widely used in the treatment of various rheumatism and arrhythmia with few side effects. The primary aim of this study is to develop SIN microcapsules with polyelectrolyte multilayers for controlled drug release.. SIN microcrystals were encapsulated with chitosan, gelatin, and alginate by layer-by-layer technique, such as (gelatin/alginate)(4) and (chitosan/alginate)(6). The size distribution, zeta-potential, stability, and morphology of the microcapsules were characterized by a particle size analyzer, zetasizer, ultraviolet spectroscopy, and transmission electron microscope, respectively. The in vitro controlled release pattern of SIN was studied using a diffusion cell assembly at physiological pH of 6.8 or 1.4.. Light stability of these microcapsules was improved after microencapsulation. Compared with release rate of the SIN microcapsules coated by the poly(dimethyldiallyl ammonium chloride)/alginate and gelatin/alginate multilayers, release rate of the SIN microcapsules coated with chitosan/alginate multilayers was fast. Release rate progressively decreased with the increase of chitosan/alginate bilayer number and the decrease of pH value of release medium.. These novel SIN microcapsules may be developed into oral controlled drug delivery for rheumatism and arthritis.

Topics: Administration, Oral; Adsorption; Analgesics; Biological Availability; Capsules; Delayed-Action Preparations; Drug Delivery Systems; Drug Stability; Morphinans; Particle Size; Temperature

Sinomenine (SIN) is an alkaloid isolated from the Chinese medicinal plant Sinomenium acutum. It is widely used as an immunosuppressive drug for treating autoimmune diseases. Due to its poor efficiency, the large-dose treatment presents some side effects and limits its further applications. In this study, we used chemical modification to improve the therapeutic effect of SIN in vitro and in vivo. A new derivative of sinomenine, named 1032, demonstrates significantly improved immunosuppressive activity over that of its parent natural compound (SIN). In an experimental autoimmune encephalomyelitis (EAE) model, 1032 significantly reduced encephalitogenic T cell responses and induced amelioration of EAE, which outcome was related to its selective inhibitory effect on the production of IL-17. By contrast, SIN treatment only led to a moderate alleviation of EAE severity and the expression level of IL-17 was not significantly reduced. Furthermore, 1032 exhibited suppression of Th17, but not Treg, cell differentiation, a result probably related to its inhibitory effect on IkappaB-alpha degradation as well as on IL-6 and TNF-alpha secretion in BMDCs. We speculate that 1032 as a novel anti-inflammatory agent may target DC to block IL-6 production, which in turn would terminate Th17 cell development. Thus, SIN derivative 1032 presents considerable potential in new drug development for treating autoimmune and inflammatory disease.

Topics: Animals; Dendritic Cells; Encephalomyelitis, Autoimmune, Experimental; I-kappa B Proteins; Immunosuppressive Agents; Interleukin-17; Interleukin-6; Male; Mice; Mice, Inbred C57BL; Morphinans; T-Lymphocytes; T-Lymphocytes, Helper-Inducer

The effects of sinomenine (SIN, an alkaloid extracted from the Chinese medicinal plant Sinomenium acutum used for centuries to treat rheumatic disease, including rheumatoid arthritis) on apatitic nucleation and matrix vesicle (MV)-induced mineral formation were compared with those of cysteine, levamisole, and theophylline. We found that SIN was not an inhibitor of tissue non-specific alkaline phosphatase (TNAP), a marker of biological mineralization, but confirmed that cysteine, levamisole, and theophylline were. Further, none of these four molecules directly affected the nucleation of hydroxyapatite (HA) formation, in contrast to pyrophosphate (PP(i)) which did. Incubation of 0.25-1.0mM cysteine, theophylline, or levamisole with MVs in synthetic cartilage lymph (SCL) containing AMP and Ca(2+), but not inorganic phosphate (P(i)), prolonged the induction time of mineral formation, apparently by inhibiting TNAP activity. SIN at the same levels neither inhibited TNAP activity nor affected the induction time of MV mineral formation. However, SIN did markedly delay MV-induced mineral formation in SCL containing P(i) (instead of AMP) in a manner similar to theophylline, but to a lesser extent than levamisole. Cysteine did not delay, in fact it slightly accelerated MV-induced mineral formation in Pi-containing SCL. These findings suggest that levamisole, SIN and theophylline may directly affect Ca(2+) and/or P(i) accretion during mineral formation; however, TNAP was not directly involved. The possible roles of annexins and other ion transporters, such as proteins of the solute carrier family implicated in Ca(2+) and P(i) influx are discussed.

Topics: Adenosine Monophosphate; Animals; Antirheumatic Agents; Calcification, Physiologic; Cartilage; Chick Embryo; Cysteine; Durapatite; Extracellular Matrix; Humans; Levamisole; Minerals; Morphinans; Phosphates; Phosphodiesterase Inhibitors; Theophylline; Transport Vesicles

Boi and its original plant Sinomenium acutum from Japan were compared with Seifuto and its botanical origins from China in terms of their internal transcribed spacer (ITS) sequences and major chemical components. Boi, Seifuto, and their botanical origins overall showed seven variable sites in the ITS sequence and six genotypes. Japanese S. acutum and Boi had one nucleotide variation at position 593 to show two genotypes (J1 and J2) and their heterozygote (J3). Seifuto samples and their botanical origins, S. acutum and S. acutum var. cinereum from China, showed three genotypes (C1, C2, and C3), which did not agree with the botanical classification, indicating that they cannot be distinguished according to their ITS sequences. All Seifuto samples from Henan market showed the same ITS genotype (C1). The Japanese and Chinese genotypes differed in the nucleotide position 424, which can be used to distinguish the country of origin of these materials. In the HPLC analysis of six major components, sinomenine (1), magnoflorine (2), menisperine (3), 6-O-methyllaudanosoline glucoside (4), liriodendrin (5), and menisdaurin (6), all were detected in Boi, whereas five (all except for menisdaurin) were detected in Seifuto. The main component in the rhizome of Seifuto was sinomenine, whereas magnoflorine was the main component in the rhizome and the climbing stem of Boi. The content of sinomenine in Seifuto was almost twice that in Boi. Although the individual content of alkaloids 1-4 differed between Boi and Seifuto, the total contents of these alkaloids were comparable between them both in the climbing stem and rhizome.

Topics: Alkaloids; Aporphines; China; Chromatography, High Pressure Liquid; DNA, Ribosomal Spacer; Furans; Genotype; Glucosides; Glycosides; Japan; Morphinans; Sinomenium

Ischemia/reperfusion (I/R) injury can be characterized as an inflammatory response including recruitment of inflammatory cells to a post-ischemic organ or tissue and a cascade of mediators. Sinomenine (SIN), a pure alkaloid extracted from the Chinese medical plant Sinomenium acutum, has been used to treat various inflammatory diseases including rheumatism and arthritis. However, whether SIN can attenuate I/R injury has not previously been examined. Using a syngeneic orthotopic liver transplantation model in rats, we investigated the effect of SIN on hepatic I/R injury, in particular its effect on heme oxygenase-1 (HO-1) induction and its hepatocellular protective effect. To our knowledge, our results were the first to show that: (a) SIN pretreatment was able to induce HO-1 expression in donor livers in a dose dependent manner; (b) SIN pretreatment protected the liver graft from cold I/R injury; and (c) the protective effect of SIN was, at least in part, mediated by HO-1, as proved by the fact that inhibiting HO-1 activity with zinc protoporphyrin (ZnPP) reduced the protection. Thus, SIN deserves further exploration as a novel agent to attenuate I/R injury.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cold Ischemia; Heme Oxygenase (Decyclizing); Liver; Liver Transplantation; Male; Morphinans; Protoporphyrins; Rats; Rats, Sprague-Dawley; Reperfusion Injury

To prepare sinomenine hydrochloride delayed-onset sustained-release tablets.. The tablets containing sinomenine hydrochloride were prepared by dry-compression coating technique with the ratio of HPMC in core tablet and the ratio of HPMC in coating film as the influence factors and the lag-time and release rate as the evaluation parameters. Experiments were done on the central composite design, the data were simulated by using multi-linear equation and second-order polynomial equation. The possibly optimal formulation was predicted by response surface method. The dissolution date (lag-time and release rate) of the tablets prepared under the optimum condition were compared with the predicted. The drug released mechanism of the tablet were studied by Model-fitted of drug released within 6-15 h with zero-order, Higuchi and Peppas equation, respectively.. The lag-time and release rate were simulated using second-order polynomial equation, regression coefficients of the two parameters were 0.9901 and 0.9876, respectively. Bias between the observed and predicted values of lag-time and release rate were -3.15% and -0.34%, respectively. The lag-time of the tablet prepared under the optimum condition in vitro was about 6 h, then drug released from the tablet within 6-15 h was found to conform to zero-order kinetics and was controlled by bulk erosion mechanism.. Sinomenine hydrochloride delayed-onset sustained-release tablets release drug slowly after lag time. The models developed in this study are proved to be highly predictable.

Topics: Chemistry, Pharmaceutical; Delayed-Action Preparations; Drug Delivery Systems; Excipients; Morphinans; Pharmaceutical Preparations; Tablets; Technology, Pharmaceutical

Sinomenine (SIN), an alkaloid extracted from the stem of the Chinese medicinal plant sinomenium acutum, has been used for treating rheumatoid arthritis. But little is known whether SIN has a protective effect on osteoarthritis (OA). In this study, we investigated the protective effect of SIN on IL-1beta-induced proteoglycan degradation and apoptosis in rabbit articular cartilage and chondrocytes. Treatment with 10 ng/ml IL-1beta increased the level of glycosaminoglycan (GAG) released into the culture media, and up-regulated the activity and mRNA expression of matrix metalloproteinase 13 (MMP-13) and down-regulated the activity and mRNA expression of tissue inhibitor of metalloproteinase 1 (TIMP-1) in cartilage explants, as confirmed by the methods of GAG quantitation, MMP-13/TIMP-1 enzyme-linked immunosorbent assay (ELISA) and real-time quantitative RT-PCR. Treatment with 10 ng/ml IL-1beta resulted in marked apoptosis in chondrocytes, as demonstrated by decreased cell viability, occurrence of DNA laddering and increased caspase-3 activity and annexin V binding of phosphatidylserine. However, simultaneous treatment with SIN (10, 50 or 250 microM) inhibited the GAG release and the activity and mRNA expression of MMP-13, and enhanced the activity and mRNA expression of TIMP-1 in a dose-dependent manner in cartilage explants. Furthermore, DNA fragment, caspase-3 activity and apoptosis rate were down-regulated, and cell viability was up-regulated dose-dependently in chondrocytes. Thus, SIN has the protective capacity to antagonize cartilage degradation and chondrocyte apoptosis, which suggest that SIN may act as an agent for pharmacological intervention in the progress of OA.

Topics: Animals; Apoptosis; Cartilage, Articular; Caspase 3; Cell Survival; Cells, Cultured; Chondrocytes; Dose-Response Relationship, Drug; Glycosaminoglycans; Matrix Metalloproteinase 13; Morphinans; Osteoarthritis; Rabbits; Tissue Inhibitor of Metalloproteinase-1

To investigate the effect of sinomenine on the level of tumor necrosis factor-alpha (TNF-alpha) and nuclear factor-kappaB (NF-kappaB) and in the heterotopic tissue in rats with endometriosis.. The rats with endometriosis were divided into sinomenine lavage group, blank control group, model group and danazol group, and the levels of TNF-alpha and NF-kappaB in the heterotopic tissues of the rats were detected with immunohistochemistry.. Sinomenine lavage and danazol treatment both significantly decreased the levels of levels of TNF-alpha and NF-kappaB in the heterotopic tissues of the rats as compared with the model group (P<0.05), and lesions were significantly smaller in sinomenine lavage group than in danazol group.. Sinomenine can inhibit the production and activity of TNF-alpha and NF-kappaB to suppress the adhesion, implantation, infiltration and growth of the endometrial cells in the rat model of endometriosis.

Topics: Animals; Choristoma; Danazol; Endometriosis; Female; Morphinans; NF-kappa B; Rats; Rats, Wistar; Tumor Necrosis Factor-alpha

To improve and validate analytical methods based on HPLC and liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) for the quantitative measurement of sinomenine in rat plasma and brain tissue.. The separation of analytes and the internal standard (IS), chloramphenicol, was performed on an Agilent TC-C18 column (250×4.6 mm, 5 μm). Blood samples were measured with a Surveyor photodiode array (PDA) detector at a wavelength of 263 nm. The LCQ DECA XP(Plus) mass spectrometer was operated in the multiple reactions monitoring mode using positive electrospray ionization, and the transition from the precursor ion (m/z 279) to the product ion (m/z 224) for sinomenine was measured in brain tissue.. Measurements were linear over the concentration range of 0.1-100 μg/mL for sinomenine in plasma and over the range of 0.01-5.00 μg/g for sinomenine in brain tissue. The intra- and inter-day variabilities were less than 10% of the relative standard deviation (RSD), and the extraction and recovery of sinomenine was 72.48%-80.26% from plasma and 73.75%-80.26% from brain tissue. The limit of quantification (LOQ) was 0.1 μg/mL for plasma, and 0.01 μg/g for brain tissue. Identification of sinomenine was reproducible at 0.5, 5, and 50 μg/mL in the plasma and at 0.05, 0.50, and 2.00 μg/g in brain tissue. The concentration of sinomenine measured in brain tissue after a single ip dose had a neuroprotective effect on H₂O₂-induced injury in PC12 cells in vitro.. Our methods offered a sensitivity within a wide linear concentration range for sinomenine. These methods were successfully applied to evaluate sinomenine pharmacokinetics over time in rat brain tissue after a single ip dose of 30 mg/kg.

Topics: Animals; Brain; Calibration; Cell Culture Techniques; Cell Survival; Chromatography, High Pressure Liquid; Female; Hydrogen Peroxide; Male; Morphinans; Neuroprotective Agents; Oxidants; PC12 Cells; Rats; Rats, Wistar; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization; Tissue Distribution

To explore the pharmacokinetic of Sinomenine transdermal patch.. The plasma drug concentration of Beagle dogs was determined after administration with HPLC-UVD as analysis tools. The pharmacokinetics parameters were fitted with kinetica software package.. The exclusive analysis method was established with the following pharmacokinetics parameters: T (peak) = 8 h, Cmax = 366 ng/mL, MRT = 13 h. The pharmacokinetic characteristics was accordance with one-order rate and two-compartment model.. The method is preferable to be applied to the pharmacokinetics research and further applied pharmacological study which will play a reference role in clinical application.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Area Under Curve; Biological Availability; Chromatography, High Pressure Liquid; Dogs; Drugs, Chinese Herbal; Male; Models, Animal; Morphinans; Sinomenium; Skin Absorption; Spectrophotometry, Ultraviolet; Time Factors; Transdermal Patch

To assess feasibility of plasma protein-binding percentage by using microdialysis method.. Sinomenine hydrochloride was selected as model drug. The in vitro rat plasma protein-binding percentage of three kinds of concentration was determinated by using microdialysis sampling tool and HPLC-UV. At the same time the classical study method, such as ultrafiltration method, was also used to determinate the protein-binding percentage to compare the difference between the two methods.. The in vitro rat plasma protein-binding percentage of sinomenine hydrochloride using microdialysis method was about 26% and kept relative stable at the concentration range. There was no significant difference between the two methods.. The results shows that the binding percentage is relative low. Microdialysis method is a new method for the study of protein-binding degree.

Topics: Animals; Blood Proteins; Chromatography, High Pressure Liquid; Microdialysis; Morphinans; Plasma; Protein Binding; Rats; Ultrafiltration

To study the alkaloids in the stems and leaves of Stephania cepharantha Hayata.. The dried stems and leaves of Stephania cepharantha Hayata were percolated with 95% ethanol and the solvent was removed by rotary evaporation to give a concentrate, and the concentrate was extracted by petroleum ether and chloroform. Column chromatograghy on MCI CHP 20P, silica gel, Rp-18, Sephadex LH-20 and polyamide were applied for the isolation and purification of the chloroform fraction. The structures were elucidated by their physicochemical properties and spectral data.. Five alkaloids were obtained and identified as, Stephasunoline (I) Aknadinine (II), Discretamine (III), Acutumine (IV), Sinomenine (V).. Compounds I, III, IV are isolated from this plant for the first time, and compound IV is isolated from the genus for the first time.

Topics: Alkaloids; Berberine Alkaloids; Magnetic Resonance Spectroscopy; Morphinans; Plant Leaves; Plant Stems; Spiro Compounds; Stephania

Currently, there is a considerable interest in discovering G-quadruplex ligands. Plant-derived agents, because of their diversity in structure and bioactivity and low toxicity, may be a very diverse source of G-quadruplex ligands. However, up to now, the screening of G-quadruplex ligands from natural plant extract has not been reported. Herein, in order to develop a simple method for fast identifying G-quadruplex ligands from plant extract, we intended to substitute the spectral shift in the imino region (delta 10-12) in (1)H NMR spectra of G-quadruplex for in vitro bioassay to judge the existence/nonexistence of G-quadruplex ligand(s) in plant extract, and then couple G-quadruplex recognition with NMR based structure elucidation to identify the structure of the ligand(s) without the need of prior separation. In this paper, we successfully screened a G-quadruplex ligand from a simulated plant extract using this approach. This research work provides a promising tactic to find new leading compounds from nature plant extract.

Topics: Acetophenones; Ascorbic Acid; Berberine; Buffers; Circular Dichroism; Computer Simulation; G-Quadruplexes; Hydrogen-Ion Concentration; Ligands; Molecular Structure; Morphinans; Nuclear Magnetic Resonance, Biomolecular; Plant Extracts; Protons

New methods based on MEEKC coupling with field-amplified sample injection (FASI) induced by ACN were proposed for five isoquinoline alkaloids (berberine, palmatine, jatrorrhizine, sinomenine and homoharringtonine) in no salt and high salt sample solution (HS). For the separation of five isoquinoline alkaloids, a running buffer composed of 18 mM sodium cholate, 2.4% v/v butan-1-ol, 0.6% v/v ethyl acetate, 10% v/v (or 30% v/v) methanol and 87.0% v/v (or 67% v/v) 5 mM Na2B4O7~10 mM NaH2PO4 buffer (pH 7.5) was developed. In order to improve the sensitivity, FASI induced by ACN was applied to increase the detection sensitivity. The detection limit was found to be as low as 0.0002 microg/mL in no salt sample solution and 0.062 microg/mL in HS. The method has been applied for the analysis of human urine spiked with analytes, and the assay results were proved to be satisfactory, and also the determination of berberine in urine sample after oral administration berberine.

Topics: 1-Butanol; Acetates; Acetonitriles; Alkaloids; Berberine Alkaloids; Chromatography, Micellar Electrokinetic Capillary; Harringtonines; Homoharringtonine; Humans; Hydrogen-Ion Concentration; Methanol; Morphinans; Reproducibility of Results; Sensitivity and Specificity; Sodium Dodecyl Sulfate; Water

The aim of this study was to investigate the mechanism of the effects of Sinomenine (SIN) on the invasion and migration ability of activated human monocytic THP-1 cells (A-THP-1). Sinomenine is a pure alkaloid extracted from the Chinese medical plant Sinomenium acutum.. Human monocytic THP-1 cells were induced to differentiate into macrophages with phorbol 12-myristate 13-acetate (PMA). Cells were treated with different concentrations of SIN. The invasion and migration ability of cells was tested by in vitro transwell assays. The levels of CD147 and MMPs were evaluated by flow cytometric analysis and zymographic analysis, respectively. The mRNA expression of CD147, MMP-2, and MMP-9 was measured by RT-PCR.. The invasion and migration ability of A-THP-1 cells was significantly inhibited by SIN in a concentration-dependent fashion; at the same time, the levels of CD147, MMP-2, and MMP-9 were markedly down-regulated. This inhibitory effect was most notable at concentrations of 0.25 mmol/L and 1.00 mmol/L (P<0.01).. A possible mechanism of the inhibitory effect of SIN on cell invasion and migration ability is repression of the expression of MMP-2 and MMP-9, which strongly correlates with the inhibition of CD147 activity.

Topics: Arthritis, Rheumatoid; Basigin; Cell Differentiation; Cell Movement; Cell Survival; Cells, Cultured; Humans; Immunosuppressive Agents; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Matrix Metalloproteinase Inhibitors; Monocytes; Morphinans

Sinomenine has been used to treat autoimmune diseases for centuries. However, the mechanism underlying its therapeutic effects remains unknown. Increasing recognition of the importance of the Th1/Th2 imbalance in nephritis has raised the questions of whether there is a Th1/Th2 imbalance in patients with mesangial proliferative nephritis (MsPGN) and whether sinomenine can modulate the Th1/Th2 imbalance. In this study, 25 MsPGN patients were treated with sinomenine and followed for 3 months. The expression of T-bet and GATA-3 mRNA in peripheral blood mononuclear cells (PBMCs) and the serum levels of interferon-gamma (IFN-gamma), interleukin (IL)-4, and IL-10 were studied at month 0, month 1, and month 3. The intra-renal expression of T-bet and GATA-3 was studied via immunohistochemistry. Results reveal that PBMCs from MsPGN patients expressed high levels of T-bet mRNA and low levels of GATA-3 mRNA, and the T-bet/GATA-3 ratio in MsPGN patients was significantly higher than that in healthy donors. Meanwhile, MsPGN patients were found to have simultaneously elevated IFN-gamma values and decreased IL-10 values. Immunohistochemistry revealed increased T-bet and decreased GATA-3 expression in renal tissues from MsPGN patients. Moreover, sinomenine was found to cause a decrease in T-bet mRNA expression, resulting in a drop in the T-bet/GATA-3 ratio. Sinomenine was also found to elicit a decrease in the serum levels of IFN-gamma. These results suggest that a shift toward the Th1 pathway of Th cell activation occurs in MsPGN patients, and that sinomenine has the potential to counter this shift in the Th1/Th2 balance and thereby produce therapeutic effects.

Topics: Adult; Enzyme-Linked Immunosorbent Assay; Female; Follow-Up Studies; GATA3 Transcription Factor; Gene Expression Regulation; Glomerulonephritis, Membranoproliferative; Homeostasis; Humans; Immunohistochemistry; Interferon-gamma; Interleukin-10; Interleukin-4; Kidney; Male; Middle Aged; Morphinans; T-Box Domain Proteins; Th1 Cells; Th2 Cells

To investigate the effect of sinomenine on the expression of chemokines and chemokine receptors of dendritic cells in patients with rheumatoid arthritis in vitro.. The peripheral blood mononuclear cells isolated from 8 patients with rheumatoid arthritis were induced to differentiate into dendritic cells with GM-CSF and IL-4. The dendritic cells were exposed to sinomenine at high (5 mmol/L), moderate (2 mmol/L), and low (1 mmol/L) concentrations or treated with the control medium. The expression of CCR5 and CCR7 on the surface of the dendritic cells were measured by flow cytometry, and the CCR5 and CCR7 mRNA expressions were detected by semi-quantitative PCR. Enzyme-linked immunosorbent assay (ELISA) was used to measure the expressions of CXCL9 (MIG), CXCL10 (IP-10) and CXCL11 (ITAC).. Compared with the control cells, the dendritic cells treated with sinomenine, especially at high and moderate concentrations, showed significantly lowered mRNA and protein expressions of CCR5 and CCR7. Similar results were observed in the expressions of CXCL9 (MIG) and CXCL10 (IP-10), but not in CXCL11 (ITAC).. Sinomenine produces therapeutic effect on rheumatoid arthritis possibly by inhibiting the expression of chemokines and chemokine receptors in the dendritic cells to suppress the chemotactic migration of the dendritic cells.

Topics: Adult; Arthritis, Rheumatoid; Chemokines; Dendritic Cells; Gene Expression Regulation; Humans; Middle Aged; Morphinans; Receptors, Chemokine; RNA, Messenger

To optimize the formulation and preparation process of sinomenine liposomes.. Method of aether injection and mixture uniform design were adopted to determine the formulation of sinomenine liposomes is the proportion of phospholipids, cholesterol and Vitamin E with the index of entrapment efficiency. And the single-factor test was used to study the preparation process of the liposomes, including the volume of buffer solution, the preparation temperature and the ultrasonic time.. The optimized formulation was that the ratio of sinomenine : phospholipids : cholesterol : vitamin E mass ratio was 8.92 : 60.35 : 28.81 : 1.91. The volume of buffer solution was 50 mL x g(-1) membrane, the preparation temperature was 50 degrees C, and the ultrasonic time was 20 min.. Satisfactory shape and entrapment efficiency of the liposomes can be obtained by the optimized formulation and preparation process.

Topics: Chemistry, Pharmaceutical; Cholesterol; Dosage Forms; Drug Carriers; Drug Compounding; Drug Delivery Systems; Drug Stability; Liposomes; Morphinans; Particle Size; Phospholipids; Technology, Pharmaceutical

To study pharmacokinetic parameters and absolute bioavailability for oral use of sinomenine tablet in beagle dogs.. Applying to double cycle self crossover design, a single oral or intravenous dose of 10 mg x kg(-1) sinomenine was given to 10 beagle dogs. Drug concentrations in plasma were determined by HPLC. The pharmacokinetic parameters were calculated by 3P97 pharmacokinetic program.. The concentration-time curves of oral administration fitted to one compartment model in the beagle dogs. The T(max), C(max), t1/2 and AUC(0-T) was (82.5 +/- 13.9) min, (0.15 +/- 0.027) mg x L(-1), (87.6 +/- 28.3) min and (28.43 +/- 3.48) mg x min x L(-1), respectively. The concentration -time curves of i.v. fitted to two compartment model in the beagle dogs. The t1/2beta and AUC(0-T) was (106.7 +/- 120.2) min and (93.32 +/- 82.08) mg x min x L(-1). The absolute bioavailability for oral use was (30.46 +/- 4.24)%.. The absolute bioavailability of sinomenine is low, and the elimination of sinomenine tablet is fast.

Topics: Animals; Biological Availability; Dogs; Drugs, Chinese Herbal; Female; Injections, Intravenous; Male; Morphinans; Tissue Distribution

To prepare the gastric retenting and chronopharmacologic drug delivery tablets containing sinomenine hydrochloride as a model drug, evaluate the effects of the coating layers formulation and technics on drug release behavior, and to elucidate the mechanism of drug release based on obtained results.. The gastric retenting and chronopharmacologic drug delivery tablets were prepared by press-coated technics. The types of disintegrants were chosen according to the expanding rate and the lag-time. The effects of formulation and technics of coating layer on the release characteristic of the drug were investigated by dissolution testing. The mechanism of drug release was proved by erosion test.. The tablets had typical chronopharmacologic drug delivery properties with a lag time followed by a rapid release phase. CMS-Na was selected as the disintegrant. The lag-time was prolonged with the increase of the ratio of HPMC/carrrageenan and the amount of matrix material in coating layer. The compressing pressure and preparation method of coat material had minor influence on the lag-time of the tablets. Coating layer erosion and tablet core swelling were involved in the mechanism of drug release.. The tablets had typical chronopharmacologic drug delivery properties. A suitable lag-time can be achieved by adjusting formulation of coating layer to meet the requirement of chronotherapy.

Topics: Chemistry, Pharmaceutical; Drug Chronotherapy; Drug Delivery Systems; Humans; Morphinans; Stomach; Tablets, Enteric-Coated

To observe the effect of alcohol extracts from orientivne and its effective component sinomenine on withdrawal syndromes and neurotransmitter of morphine-abstinent mice and on intracellular calcium level in morphine-dependent neuronal-cells line. To study the detoxification of alcohol extracts from orientvine and sinomenine on morphine-dependent animal and explore the mechanism of its effect.. The effect of alcohol extracts from orientivne and sinomenineon on abstinent syndromes was observed by experiment study on morphine-dependent ex vivo ileum from guinea pigs and morphine-dependent mice. The morphine-dependent model mice were established by injection on dosage increasing by degrees. The hypothalamic monomine neurotransmitters such as NA, DA, 5-HT were tested by fluorospectrophotometry. Morphine-dependent cell line was established by administering morphine at different doses into the culture medium. The cells were stained with fluo-3 and the intracellular calcium level was measured by flow cytometry.. Alcohol extracts from orientvine and sinomenine could alleviate withdrawal contractile response of morphine-dependent ex vivo ileum from guinea pigs and withdrawal syndromes of morphine-dependent mice, decrease the concentration of the neurotransmitters, and elevate the intracellular calcium level and inhibit the decreasing of Ca2+ induced by naloxone.. Alcohol extracts from orientvine and sinomenine have some effects on withdrawal syndromes which may be related to inhibiting neurotransmitters and the regulation of intracellular calcium concentration.

Topics: Animals; Calcium; Disease Models, Animal; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Ethanol; Female; Guinea Pigs; Ileum; Male; Mice; Morphinans; Morphine; Morphine Dependence; Muscle Contraction; Neurons; Norepinephrine; Random Allocation; Rats; Rats, Sprague-Dawley; Sinomenium; Substance Withdrawal Syndrome

Topics: Animals; Aorta; Calcium; Cells, Cultured; Intracellular Space; Male; Mitogen-Activated Protein Kinases; Morphinans; Myocytes, Smooth Muscle; Protein Kinase C; Rats; Rats, Sprague-Dawley

To study the effects of sinomenine on apoptosis in cutured neonatal rat cardiomyocytes induced by H2O2 and its possible mechanism.. H2O2 was used to build an oxidative stress-induced injury model in neonatal rat cardiomyocytes after being treated with sinomenine (10, 30, 100 micromol L(-1)), the apoptosis rate, the content of malondialdehyde (MDA), the activity of superoxide dimutase (SOD), the activity of lactate dehydrogenase (LDH) and expression of NF-kappaB protein of the Cardiomyocytes were examined.. Compared with the model group, the apoptosis rate and the content of MDA, LDH decreased greatly (P < 0.01), and the activity of SOD increased distinctly (P < 0.01) after being treated by sinomenine (10, 30, 100 micromol x L(-1)).. Sinomenine can inhibit the apoptosis induced by H2O2 in neonatal rat cardiomyocytes. The protective mechanism could be related to its ability to reduce lipid pexosidation and to inhibit cardiomyocyte expression of NF-kappaB protein.

Topics: Animals; Apoptosis; Gene Expression Regulation; Hydrogen Peroxide; L-Lactate Dehydrogenase; Malondialdehyde; Morphinans; Myocytes, Cardiac; NF-kappa B; Rats; Rats, Wistar; Superoxide Dismutase

In a continuing study on discovery of more potent derivatives of sinomenine (1), a clinically available alkaloid for rheumatoid arthritis (RA) treatment, oxidation of sinomenine provided two unique stereoisomers, disinomenines 2 and 3. The structure of 3 was determined by MS, NMR, and X-ray analysis. The formation of 2 and 3 via oxidation of sinomenine by potassium permanganate (KMnO4) exhibited a pH-dependent stereoselectivity. The bioassay results using human synovial sarcoma cells (SW982) showed that 2 inhibited, while 3 stimulated, IL-6 production.

Topics: Cell Line, Tumor; Crystallography, X-Ray; Dimerization; Humans; Hydrogen-Ion Concentration; Interleukin-6; Morphinans; Oxidation-Reduction; Sarcoma, Synovial; Stereoisomerism

Sinomenine, one of the alkaloids extracted from roots or stems of Sinomenium acutum, is documented to show antinociceptive action but the action mechanism is still unclear. The present study was aimed to investigate the effect of sinomenine on opioid mu-receptor (OMR). In Chinese Hamster Ovary (CHO) cell transfected with OMR, the binding of [(3)H]naloxone was displaced by sinomenine in a concentration-dependent manner. This compound also raised the phosphorylation of OMR in these cells. In a tail-flick test, sinomenine produced dose-dependent antinociception in mice, which was dose-dependently inhibited by pretreatment of naloxonazine, a selective OMR antagonist. Long-term pretreatment with sinomenine may delay the analgesic tolerance of morphine. The obtained results suggest that sinomenine has an ability to activate OMR, implicating the potential of sinomenine to be applied in clinic.

Topics: Analgesics, Opioid; Animals; Binding, Competitive; CHO Cells; Cricetinae; Cricetulus; Dose-Response Relationship, Drug; Drug Interactions; Male; Mice; Mice, Inbred BALB C; Morphinans; Morphine; Naloxone; Narcotic Antagonists; Phosphorylation; Protein Binding; Receptors, Opioid, mu; Transfection

Rheumatoid arthritis (RA) is a chronic, inflammatory autoimmune disease that results in progressive joint destruction and substantial morbidity. The stem of the Chinese medicinal plant, Sinomenium acutum Rehder & Wilson (Family Menispermaceae), has been used to treat various rheumatic and arthritic diseases, of which the major bioactive component is sinomenine. We investigated the nature and molecular mechanisms of the anti-arthritic effect of sinomenine on collagen-induced arthritis in female Wistar rats. The results showed that sinomenine markedly suppressed the incidence and disease progression of established CIA, showing as dramatic reduction of paw swelling, ESR, and arthritic scores. Sinomenine suppressed the production of proinflammatory cytokines IL-1beta and IL-6 in serum, inhibited the protein expressions and activities of MMP-2 and MMP-9, and elevated the protein expressions and activities of TIMP-1 and TIMP-3 in rat paw tissues.

Topics: Animals; Antirheumatic Agents; Arthritis, Experimental; Arthritis, Rheumatoid; Female; Interleukin-1beta; Interleukin-6; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Matrix Metalloproteinase Inhibitors; Morphinans; Rats; Rats, Wistar; Tissue Inhibitor of Metalloproteinase-1; Tissue Inhibitor of Metalloproteinase-3

Sinomenine transdermal patch was prepared and its properties were studied. The patches were produced by salivation method. The releasing rate in vitro of the patch was determined by HPLC. Peel test was used to evaluate the adhesion. Acute skin irritation test was performed in comparison with formalin (0.8%) by using mouse model. The Sinomenine TDDS Patch was prepared. The releasing rate in vitro followed the Higuchi equation (r>0.99), the releasing amount was beyond 90% in 24h. The peel adhesion to steel (N/25 mm) is 10 or above. The skin irritation tests showed negligible erythema and edema. The Sinomenine transdermal patch was prepared successfully and it may be beneficial for topical use.

Topics: Adhesiveness; Administration, Cutaneous; Animals; Antirheumatic Agents; Chemistry, Pharmaceutical; Dosage Forms; Edema; Erythema; Excipients; Kinetics; Male; Mice; Morphinans; Permeability; Skin; Skin Absorption; Skin Irritancy Tests; Solubility; Tissue Adhesives

To explore the feasibility and advantages of using microdialysis as sampling method for dynamic determination of sinomenine in topical skin.. In this study, sinomenine was administered to the rats by transdermal drug delivery system and celiac injection. With microdialysis technique for sampling, the concentration of sinomenine in dialysate was determined by high performance liquid chromatography (HPLC).. Under existing determination condition, topical drug concentration in the skin of rats was hard to be detected after sinomenine administered to the skin, but it could be detected after celiac injection.. Microdialysis sampling method can be used to determine topical drug concentration in the skin dynamically, and this method is better than traditional methods obviously.

Topics: Administration, Cutaneous; Animals; Chromatography, High Pressure Liquid; Feasibility Studies; Injections, Intraperitoneal; Male; Menispermaceae; Microdialysis; Morphinans; Rats; Rats, Sprague-Dawley; Skin

To investigate the effects of intra-articular injection of sinomenine on interleukin-1beta (IL-1beta) content in synovial fluid and serum in rabbits with osteoarthritis (OA).. Degenerative osteoarthritis in the knee joint of left posterior limb was induced in 36 rabbits by full extension using plaster cast for 6 weeks. Then the rabbits were randomly divided into untreated group, hyaluronic acid group and sinomenine group. Another 4 normal rabbits were selected as normal control group. Rabbits in the sinomenine group and the hyaluronic acid group received intraarticular injections of sinomenine and hyaluronic acid once a week for 5 weeks, respectively. The content of IL-1beta in synovial fluid and serum were measured before and after treatment by enzyme-linked immunosorbent assay. The pathological changes of cartilaginous tissue were analyzed by using Mankin's score.. Compared with those in the untreated group, synovial fluid and serum IL-1beta contents in the sinomenine group and the hyaluronic acid group were significantly decreased (P<0.01). And the synovial fluid and serum IL-1beta contents in the sinomenine group were lower than those in the hyaluronic acid group (P<0.05). The mean Mankin's score of cartilage in the sinomenine group was significantly lower than that in hyaluronic acid group (P<0.05).. Sinomenine may reduce the degree of articular degeneration in rabbit with OA through decreasing the content of IL-1beta in synovial fluid and serum.

Topics: Animals; Disease Models, Animal; Hyaluronic Acid; Interleukin-1beta; Male; Morphinans; Osteoarthritis, Knee; Rabbits; Serum; Synovial Fluid

To evaluate the possible role of alkaloid sinomenine (SIN) on chronic rejection in rat heart transplantation model.. After a brief course of cyclosporine A (CsA), DA recipients of PVG hearts were treated with placebo, SIN, CsA, or a combination of both drugs. Grafts were analyzed morphometrically and by immuno-histochemistry. Expressions of basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), and endothelin 1 (ET-1) were assessed by reverse transcription-polymerase chain reaction.. Cardiac grafts of SIN-treated rats showed a mild degree of vasculopathy compared with untreated rats or CsA-treated recipients. Degree of vasculopathy was significantly reduced in rats treated with combined SIN and CsA than rats receiving either drug alone. Treatment with SIN alone did not affect gene expressions of bFGF, VEGF, and ET-1 while expressions of bFGF, VEGF, and ET-1 were significantly reduced by combined treatment with SIN and CsA.. These results demonstrated a potential value of SIN, in combination with low-dose CsA to attenuate the vasculopathy in this rat model of chronic cardiac allograft rejection.

Topics: Animals; Disease Models, Animal; Graft Rejection; Graft Survival; Heart Transplantation; Male; Morphinans; Phytotherapy; Rats

To study bioequivalence of Sinomenine patch made by different preparation process, and to testify feasibility and superiority of microdialysis as a new method in topical bioequivalence study.. Normal gel patch and liposome gel patch of sinomenine were prepared by different preparation, nude mouse served as the experimental subjects sampling method of drug in the skin was tissue homogenization microdialysis, and drug concentration in dialysate was determined by HPLC.. Results of tissue homogenization showed that liposome gel patch leads more remainder drug in the skin of nude mouse than normal gel patch, and results of microdialysis showed that liposome gel patch led higher instantaneous drug concentration than normal gel patch. Concentration-time curve of sinomenine in the skin accorded with the results of most dermal delivery systems studies over the world.. Topical bioequivalence of liposome gel patch of sinomenine is higher than that of normal gel patch of sinomenine. Microdialysis can be used to study bioavailability and bioequivalence of different preparation.

Topics: Administration, Cutaneous; Animals; Chromatography, High Pressure Liquid; Male; Mice; Microdialysis; Morphinans; Skin; Therapeutic Equivalency

Two new morphinane alkaloid dimers, 2,2'-disinomenine (1) and 7',8'-dihydro-1,1'-disinomenine (2), and known 1, 1'-disinomenine (3), were isolated from ethanol extracts of stems of Sinomenium acutum. Their structures were elucidated on the basis of spectroscopic methods. The absolute configuration of alkaloids 1-3 was determined by direct comparison of their CD spectra with the known alkaloid sinomenine. The isolated alkaloids were tested for cytotoxicity against A549, P388, and HeLa cell lines, and 1 and 3 showed weak inhibition against A549 and Hela cells.

Topics: Alkaloids; Antineoplastic Agents, Phytogenic; Drug Screening Assays, Antitumor; Drugs, Chinese Herbal; HeLa Cells; Humans; Molecular Structure; Morphinans; Plant Stems; Plants, Medicinal; Sinomenium

The present study was designed to investigate the effect of sinomenine (SIN), an alkaloid extracted from sinomenium acutum, on the antigen-induced activation of RBL-2H3. For this investigation, the RBL-2H3 cells were sensitized with dinitrophenyl (DNP)-specific IgE overnight in 1.0 ml of Eagle's MEM (EMEM), and varying doses of SIN were added to the culture medium for 30 min and challenged with dinitrophenyl-human serum albumin (DNP-HSA) to induce mast cell degranulation before supernatants were collected. The effects of SIN on antigen-induced release of beta-hexosaminidase were measured by enzymatic assay, calcium influx by FACS, cytokines by ELISA, and signaling events by immunoblotting. The results showed that treatment with SIN was followed by a decrease in FcepsilonRI-mediated mast cell release of beta-hexosaminidase, production of IL-4 and TNF-alpha, phosphorylation of Gab2 (Scaffolding adapter Grb2-associated binder 2), Akt and p38 mitogen-activated protein kinase (MAPK). In addition, SIN had no effect on the phosphorylation of LAT and no significant difference on calcium mobilization was observed between control and SIN treated group. These results suggested that SIN might suppress the antigen-induced activation of RBL-2H3 cells via a Ca2+ independent pathway.

Topics: Adaptor Proteins, Signal Transducing; Animals; Antigens; beta-N-Acetylhexosaminidases; Calcium; Cells, Cultured; Interleukin-4; Mast Cells; Morphinans; p38 Mitogen-Activated Protein Kinases; Rats; Tumor Necrosis Factor-alpha

In search of more potent derivatives of sinomenine (1), a clinically available natural alkaloid for the treatment of rheumatoid arthritis (RA), biocatalyzed cross-coupling of sinomenine and guaiacol (2) by Antrodiella semisupina, provided two unique C-C coupled (3 and 4) and one C-O linked (5) novel metabolites. The structures of the metabolites were elucidated by means of MS, 2D NMR techniques and X-ray analysis. 4 exhibited more potent inhibitory activity on IL-6 production than 1 in human synovial sarcoma cell (SW982), and 5 stimulated IL-6 production.

Topics: Basidiomycota; Catalysis; Crystallography, X-Ray; Guaiacol; Magnetic Resonance Spectroscopy; Molecular Structure; Morphinans; Spectrometry, Mass, Electrospray Ionization

Topics: Animals; Brain; Male; Mice; Mice, Inbred Strains; Morphinans; Morphine Dependence; Nitric Oxide Synthase Type I

To explore the effects of sinomenine(Sin) on intracellular free calcium ([Ca2+]i) and the activity of PKC (protein kinase C) of the cultured aortic vascular smooth muscle cells (VSMC) during ischemia and hypoxia.. The effect of Sin on changes in [Ca2+]i were determined in cultured rabbit VSMC after exposure to high K+, norepinephrine (NE) and caffeine (Caf). Fluorescent Ca2+ -indicater fura-2/AM was used. The effects of Sin were compared with that of verapamil (Ver). The hypoxia model was made, then the activity of PKC was measured by y scintillation counting instrument.. Sin (10 x 10(-6) mol x L(-1), 3 x 10(-5) mol x L(-1) 10(-4) mol x L(-1)) inhibited the elevation of [Ca2+]i induced by high K+ -depolarization in a concentration dependent manner. In addition, Sin inhibited the elevation of [Ca2+]i induced by NE in the presence of extracellular Ca2+. In the absence of extracellular Ca2+, Sin (3 x 10(-5) mol.L(-1)) also had no blocking effect on the NE-induced [Ca2+]i increase. It was found that the activity of PKC treated with Sin in VSMC cytoplasm and cell membrane in normal condition increased, the activity of PKC in cytoplasm in ischemia and hypoxia situation increased, but the activity of PKC in cell membrane decreased. When VSMC was treated with Sin, the activity of PKC in cytoplasm decreased and that of cell membrane increased.. The results suggest that Sin might decrease the[Ca2+] i of VSMC by blocking both VDC and ROC, could regulate the PKC activities induced by ischemia and hypoxia.

Topics: Animals; Aorta; Calcium; Cell Hypoxia; Cells, Cultured; Cytoplasm; Morphinans; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Protein Kinase C; Rabbits

Sinomenine (7,8-didehydro-4-hydroxy-3,7-dimethoxy-17-methylmorphinan-6-one) is a pure alkaloid extracted from the Chinese medical plant. In this report a liquid chromatography-electrospray mass spectrometry (LC-ESI-MS) method with in vivo microdialysis for the pharmacokinetic study of free-form sinomenine in rat skin has been developed. A microdialysis probe was surgically implanted into the subcutaneous tissue of the rats and an isotonic phosphate buffer (PBS) was used as the perfusion medium. Samples were collected and then analyzed off-line by LC-ESI-MS. The chromatographic separation was achieved within 4.2 min by using a narrow-bore Xterra C(18) column (2.1 x 150 mm, 5 microm) with acetonitrile-(10 mmol/L ammonium acetate buffer, 0.1% acetic acid) (15:85, v/v). Ion signal m/z 330.1 for sinomenine was measured in the positive mode. Linearity was established for the range of concentrations of 2.0-10000.0 ng/mL with a coefficient of determination (r) of 0.9989. The intra- and inter-day reproducibility of the present method was better than 6%. The lower limit of quantification (LLOQ) was 1.0 ng/mL. The proposed method described provides more authentic information on pharmacokinetics and metabolism at the site of action by using the coupling of microdialysis to LC-ESI-MS technique than the traditional sampling methods.

Topics: Animals; Calibration; Chromatography, Liquid; Male; Microdialysis; Molecular Structure; Morphinans; Rats; Rats, Wistar; Sensitivity and Specificity; Skin; Spectrometry, Mass, Electrospray Ionization

Sinomenine, an herbal ingredient isolated from Sinomenium acutum, is used for the amelioration of arthritis. It has been found that this molecule could bind to human serum transferrin (Tf), the iron (III) transport protein in the blood, by using fluorescence, circular dichroism (CD) spectroscopy, and molecular modeling methods. The results provide possible usage of transferrin to transport sinomenine.

Topics: Algorithms; Circular Dichroism; Humans; Hydrogen-Ion Concentration; Models, Molecular; Morphinans; Protein Binding; Sinomenium; Spectrometry, Fluorescence; Transferrin

Cytochrome P450s superfamily expressed widely in organisms are known to play an important role in the biotransformation of many endogenous and exogenous substances. Inhibition or induction of cytochrome P450 isozymes is one of the major causes for clinical drug-drug interactions. Sinomenine can be metabolized to at least 2 metabolites in human, rat in vivo and in human liver microsomes. The major metabolite was identified to be N-demethylsinomenine. However, which CYP450 isozymes mediated by sinomenine in vivo and in vitro is not known.. In vitro study, 6 probe drugs were incubated with or without sinomenine respectively to study the effect of sinomenine on different cytochrome P450s activities in human microsomes. In vivo study, a 5-drug cocktail approach was used to study the inhibitive and inducing effect of sinomenine at normal clinical dose on cytochrome P450s activities.. Sinomenine (50 micromol/l) had no significant effects on the activities of CYP1A2, CYP3A4, CYP2C9, CYP2E1, and CYP2D6, but it decreased the activity of CYP2C19 by 69% (p=0.012) in human microsomes. In vivo, sinomenine showed almost no significant effects on the activities of CYP1A2, CYP3A4, CYP2E1, and CYP2D6, but enhanced the elimination of mephenytoin by 73% (p=0.032).. Sinomenine (50 micromol/l) inhibited the activity of CYP2C19 in human microsomes, but in vivo sinomenine at normal clinical dose enhanced the elimination of mephenytoin.

Topics: Anticonvulsants; Cytochrome P-450 Enzyme Inhibitors; Cytochrome P-450 Enzyme System; Humans; Mephenytoin; Microsomes, Liver; Morphinans

To observe the pathological characteristics of the rejection of rat cardiac allograft, study the effects and mechanisms of sinomenine on the rejection of rat cardiac allograft.. The rat model of heterotopic cardiac allograft was established. The rats were randomly divided into four groups: Group A(control group of healthy SD rats), Group B (cardiac allograft of SD to SD rats), Group C (cardiac allograft of SD to Wistar rats) and Group D (sinomenine treated group of cardiac allograft of SD to Wistar rats). ELISA and immunohistochemical staining were carried out to estimate the expression level of IL-2 and ICAM-1 in the grafts.. Little ICAM-1 and IL-2 was observed in both Group A and Group B and the infiltration of lymphocytes was not so obvious. In group C, ICAM-1 and IL-2 were increased significantly and the infiltration of lymphocytes was obviously. In group D, inflammatory reaction and infiltration of lymphocytes were much less than those in group C, and ICAM-1 and IL-2 is expressed weakly.. The expression level of ICAM-1 and IL-2 was relevant to the occurrence and development of graft rejection. Sinomenine could obviously reduce the expression of intercellular adhesion molecule and the infiltration of lymphocytes in the grafts and prolong grafts survival significantly.

Topics: Animals; Enzyme-Linked Immunosorbent Assay; Gene Expression Regulation; Heart Transplantation; Immunohistochemistry; Immunosuppressive Agents; Intercellular Adhesion Molecule-1; Interleukin-2; Male; Morphinans; Random Allocation; Rats; Rats, Sprague-Dawley; Rats, Wistar; Transplantation Tolerance

Sinomenine is an active component isolated from Sinomenium acutum and is widely used as an immunosuppressive drug for treating autoimmune diseases. CD4(+) T-cell population plays a key role in adaptive immune response and is related to some autoimmune diseases. In this study, we investigated the possible immunosuppressive effect of sinomenine on CD4(+) T cells and its underlying mechanism. Our data demonstrated that sinomenine remarkably suppressed the proliferation of CD4(+) T cells, blocked the cell cycle progression from G0/G1 phase to S plusG2/M phases. Finally, the immunosuppressive activity elicited by sinomenine in CD4(+) primary lymphocytes was found to be largely accounted for by caspase 3-dependent cells apoptosis. Sinomenine did not significantly alter the expression of bcl-2 in activated CD4(+) primary T cells, suggesting that bcl-2 might not be involved in sinomenine-induced T cells apoptosis. In sum, this study proposes a novel mechanism for the immunosuppressive function of sinomenine on primary mouse CD4(+) T cells.

Topics: Animals; Apoptosis; CD4-Positive T-Lymphocytes; Cell Proliferation; Female; Immunosuppressive Agents; Mice; Mice, Inbred C57BL; Morphinans

Sinomenine is a pure alkaloid extracted from the Chinese medical plant Sinomenium acutum. It was demonstrated that sinomenine had anti-inflammatory and immunosuppressive effects in the previous studies. The aim of the present study was to evaluate therapeutic effects of sinomenine on 2, 4, 6-trinitrobenzene sulfonic acid (TNBS) induced colitis in mice. Two hours following colonic instillation of TNBS, sinomenine with several doses (30, 100, 200 mg/kg) was given by gastric gavage once daily for 7 days. Comparing with the saline-treated mice with TNBS-induced colitis, sinomenine (100 mg/kg and 200 mg/kg)-treated mice with TNBS-induced colitis were shown improvements of weight loss, macroscopic score, histological score, and myeloperoxidase activity. Moreover, treatments with sinomenine (100 mg/kg and 200 mg/kg) decreased the up-regulated mRNA and protein levels of tumour necrosis factor-alpha(TNF-alpha) and interferon-gamma (IFN-gamma) caused by TNBS. Our findings suggest that sinomenine attenuates TNBS-induced colitis in mice and the therapeutic mechanism might be related to the reduction of up-regulated colonic TNF-alpha and IFN-gamma production caused by TNBS.

Topics: Animals; Anti-Inflammatory Agents; Body Weight; Colitis; Colon; Enzyme-Linked Immunosorbent Assay; Female; Interferon-gamma; Mice; Mice, Inbred BALB C; Morphinans; Peroxidase; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Trinitrobenzenesulfonic Acid; Tumor Necrosis Factor-alpha

Sinomenine, a purified alkaloid extracted from Sinomenium acutum, has been employed to treat arthritis for a long time. Previous studies have shown that sinomenine has favorable function of immunosuppression, but the exact mechanism has not been illustrated yet. In this study, we investigated the effect of sinomenine on human monocyte-derived dendritic cells (DCs). Lipopolysaccharide (LPS) induced DCs, exposed to sinomenine or only medium, were tested for their maturation parameters, cytokine expression, mixed lymphocyte reaction, activity and translocation of nuclear factor-kappa B (NF-kappaB) and phosphorylation of p38 stress-activated protein kinase (p38SAPK). Down-regulated membrane expression of CD40, CD80, CD83, CD86, and HLA-DR were found on DCs exposed to sinomenine. Similar result was noticed in their IL-12 production. Reduced activity and translocation of NF-kappaB was also detected in sinomenine treated DCs, except for the level of p38SAPK phosphorylation. Our results support the conclusion that sinomenine inhibits immune responses through suppressing the functions of antigen-presenting cells and NF-kappaB pathway is involved in their maturation cascade and T-cell activation. The findings indicate the potency for sinomenine to be generally used in DCs-mediated autoimmune diseases.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Blotting, Western; Cytokines; Dendritic Cells; Electrophoretic Mobility Shift Assay; Enzyme Activation; Flow Cytometry; Fluorescent Antibody Technique; Humans; Indicators and Reagents; Interleukin-12; Lymphocyte Culture Test, Mixed; Monocytes; Morphinans; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Phenotype; Reverse Transcriptase Polymerase Chain Reaction; Stimulation, Chemical; Translocation, Genetic

A Ru(bpy)(3)(2+)-based electrochemiluminescence (ECL) detection coupled with capillary electrophoresis (CE) has been established for the determination of sinomenine for the first time. Optimum separation was achieved with a fused-silica capillary column (50 cm x 25 microm i.d.) and a background electrolyte of 50 mM sodium phosphate (pH 5.0) at a separation voltage of 15 kV. The content of sinomenine was detected by ECL at the detection voltage of 1.15 V (versus Ag/AgCl) with 5 mM Ru(bpy)(3)(2+) in 75 mM phosphate solution (pH 8.0) when a chemically modified platinum electrode by europium(III)-doped prussian blue analogue (Eu-PB) was used as a working electrode. Under the optimized conditions, the ECL intensity was in proportion to sinomenine concentration in the range from 0.01 to 1.0 microg mL(-1) with a detection limit of 2.0 ng mL(-1) (3sigma). The relative standard derivations of migration time and ECL intensity were 0.93 and 1.11%, respectively. The level of sinomenine in Sinomenium acutum Rehd. et Wils was easily determined with recoveries between 98.6 and 102.7%.

Topics: Calibration; Electrochemistry; Electrodes; Electrolytes; Electrophoresis, Capillary; Ferrocyanides; Hydrogen-Ion Concentration; Luminescence; Medicine, Chinese Traditional; Morphinans; Phosphates; Plant Extracts; Platinum; Rubidium; Solvents

Dendritic cells (DC) excel at presenting antigen to T cells and thus make a key contribution to the induction of primary and secondary immune responses. Sinomenine has been used for centuries in the treatment of patients with autoimmune diseases as it possesses immunosuppressive and anti-inflammatory activities. However, the effect of sinomenine on the differentiation, maturation, and functionality of DC derived from monocytes has not been studied. We show here that DC differentiation is promoted when monocytes are treated with GM-CSF and IL-4 (IL-4) in the presence of sinomenine (200 microg/ml), as evidenced by the upregulation of CD1a while CD14 was decreased. In addition, incubation of immature DC with sinomenine significantly blunted lipopolysaccharide (LPS)-induced DC maturation, as shown by the reduction of expression of the maturation marker CD83 and co-stimulatory molecules, including CD86, B7-H1, and CD40. Moreover, sinomenine also prevented decreases in antigen (FITC-Dextran or Lucifer Yellow) uptake by LPS-treated DC. Mixed lymphocyte reactions (MLRs) revealed that sinomenine-treated DC impede the secretion of the cytokines IL-2 and IFN-gamma by co-cultured CD4(+) T cells. Therefore, modulation of DC differentiation, maturation, and functionality by sinomenine is of potential relevance to its immunomodulatory effects in controlling specific immune responses in autoimmune diseases, transplantation, and other immune-mediated conditions.

Topics: Antigens, CD; Cell Differentiation; Cells, Cultured; Coculture Techniques; Dendritic Cells; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Immunologic Factors; Immunophenotyping; Interferon-gamma; Interleukin-12; Interleukin-4; Lectins, C-Type; Lipopolysaccharides; Lymphocyte Culture Test, Mixed; Mannose Receptor; Mannose-Binding Lectins; Monocytes; Morphinans; Receptors, Cell Surface; Sinomenium; Time Factors; Up-Regulation

To explore the effect of sinomenine on the nitric oxide (NO)/neural nitric oxide synthase (nNOS) system in the cerebellum and spinal cord of morphine-dependent and morphine-withdrawal Kunming mice, mice were subjected to injection of morphine with an increasing dose for 5 d, and then were treated with sinomenine (40 mg/kg, i.p.) for another 5 d. Naloxone was used to develop acute withdrawal, and the withdrawal syndromes, including teeth chattering, twisting, straightening, sneezing and ptosis, were investigated. nNOS mRNA expressions in the cerebellum and spinal cord were determined by semi-quantitative RT-PCR. nNOS activity and NO level were determined by the chemistry-colorimetry and nitrate reductase-reduction, respectively. The results obtained were as follows: (1) Sinomenine restored the decrease in body weight and alleviated the signs of morphine-withdrawal in mice. (2) Sinomenine also reduced the increases in nNOS mRNA expression and nNOS activity resulting from morphine-dependence, and simultaneously attenuated the high level of NO in both tissues following morphine-withdrawal. (3) Administration of sinomenine alone did not develop physical dependence in mice. The results obtained indicate that sinomenine may attenuate morphine addiction and significantly alleviate morphine-withdrawal symptoms, and the mechanism may be associated with the effect of sinomenine on the NO/nNOS system in the cerebellum and spinal cord.

Topics: Animals; Body Weight; Cerebellum; Male; Mice; Morphinans; Nitric Oxide; Nitric Oxide Synthase Type I; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Spinal Cord; Substance Withdrawal Syndrome

For microdialysis studies in the skin, laboratory-made linear probes are often used. The application of the microdialysis technique to the investigation of pharmacokinetics and pharmacodynamics of drugs requires careful assessment of the linear probes' performance to ensure validity of the data obtained using this technique. The aim of this study was to establish and validate the microdialysis technique for investigation of the pharmacokinetics and pharmacodynamics of sinomenine.. Using different lengths of the dialysis membrance and different perfusion flow rates, a flow rate of 2 microL/min with 20-min sampling intervals was selected for the subsequent studies, based on the analysis of sinomenine from the microdialysis probe. In vitro recovery of sinomenine from the microdialysis probe was independent of concentration, stable over an 8-h period. Comparable in vitro recoveries were obtained by different established approaches including recovery estimation by gain, loss and the zero-net flux (ZNF) method. Recovery by loss was used to study the in vivo recovery of sinomenine from rat subcutaneous tissue.. The performance of the microdialysis system was stable over an 8-h study, resulting in a mean in vitro recovery of 51.91+/-1.29%. Recovery obtained using the ZNF plot was 52.43%. In vivo recovery of sinomenine was 34.46+/-0.76% and was stable over the 7-h study period.. The in vitro and in vivo performance of the microdialysis technique was established for the study of sinomenine. It would prove to be a useful and reliable tool to study the pharmacokinetics and pharmacodynamics of sinomenine. The data obtained in our study highlight the importance of a systematic examination of microdialysis linear probe validation.

Topics: Animals; Calibration; Drug Industry; In Vitro Techniques; Male; Microdialysis; Morphinans; Rats; Rats, Wistar; Reproducibility of Results; Skin

Preparative centrifugal partition chromatography (CPC) was successfully carried out for the separation of sinomenine from the methanolic extract of Sinomenium acutum stems and rhizomes. The optimum two-phase solvent system of CPC was composed of n-hexane/ethyl acetate/methanol/water at a volume ratio of 1:6:2:8 (v/v/v/v) with 0.1% triethylamine (TEA). Preparative CPC yielded 44.3 mg of sinomenine from 400 mg of MeOH extract with a purity of 96.9%.

Topics: Centrifugation; Chromatography; Molecular Structure; Morphinans; Plant Extracts; Sinomenium; Solvents

The therapeutic value of an antirheumatic alkaloid, sinomenine (SIN), was investigated in the acute experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis (MS). SIN is a bioactive alkaloid derived from the Chinese medicinal plant, Sinomenium acutum REHDER & E. H. WILSON (Family Menispermaceae). Chinese doctors have utilized this plant to treat rheumatic and arthritic diseases for over one thousand years. Experiments in which EAE-induced Lewis rats exhibit an acute monophasic episode of disease demonstrated that SIN is effective in preventing clinical signs of disease. The therapeutic effect on disease activity was observed at preonset administration times and at various doses tested. Consistent with disease activity in vivo, SIN-treated animals have reduced cellular infiltration within the spinal cord along with decreased TNF-alpha and IFN-gamma expression levels. SIN can significantly inhibit proliferation response of splenocytes induced by MBP(68-82). TNF-alpha and IFN-gamma, secreted by splenocytes induced by MBP(68-82) are inhibited by SIN by dose-dependence manner. The mRNA levels of CC chemokines, RANTES, MIP-1alpha and MCP-1, are inhibited in SIN-treated EAE rats. The data in this proof of concept study support the premise that SIN may be a promising new therapeutic intervention in MS.

Topics: Acute Disease; Animals; Anti-Inflammatory Agents, Non-Steroidal; Aquaporins; Cell Proliferation; Chemokine CCL5; Chemokines; Cytokines; Dose-Response Relationship, Drug; Encephalomyelitis, Autoimmune, Experimental; Eye Proteins; Female; Interferon-gamma; Membrane Glycoproteins; Morphinans; Rats; Rats, Inbred Lew; Receptors, CCR2; Receptors, Chemokine; Reverse Transcriptase Polymerase Chain Reaction; Spinal Cord; Tumor Necrosis Factor-alpha

1. Sinomenine is an alkaloid with a wide range of pharmacological actions. In the present study, we investigated the effect of sinomenine on blood pressure and its possible mechanisms of action. 2. Spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats were given intraperitoneal injections of sinomenine. At 30 min, 2.5-10 mg/kg sinomenine decreased systolic blood pressure (SBP) in a dose-dependent manner in SHR, but had no effect on the SBP in WKY rats. 3. The vascular effect of sinomenine was then examined in aortic rings isolated from Wistar rats. Sinomenine (0.1-10 micromol/L) produced concentration-dependent relaxation in aortic rings precontracted with phenylephrine (10 nmol/L) or KCl (40 mmol/L). Glibenclamide (1-100 micromol/L), a specific inhibitor of ATP-sensitive K(+) channels attenuated the sinomenine-induced relaxation, but this effect was not observed when inhibitors of other types of K(+) channels were used. 4. We further investigated the effects of sinomenine on changes in intracellular Ca(2+) concentrations ([Ca(2+)](i)) in cultured aortic smooth muscle (A7r5) cells by using the Ca(2+)-sensitive dye fura-2 as an indicator. Sinomenine, over the concentration range 0.1-10 micromol/L, decreased the increases in [Ca(2+)](i) elicited by phenylephrine (1 micromol/L) or KCl (40 mmol/L) in a concentration-dependent manner. Glibenclamide (1-100 micromol/L) abolished the effects of sinomenine. 5. In conclusion, sinomenine causes vascular relaxation by opening ATP-sensitive K(+) channels, thus decreasing [Ca(2+)](i).

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Aorta, Thoracic; Blood Pressure; Cell Line; Cells, Cultured; Dose-Response Relationship, Drug; Female; Fluorescent Dyes; Fura-2; Glyburide; Hypoglycemic Agents; KATP Channels; Male; Morphinans; Muscle Tonus; Muscle, Smooth, Vascular; Potassium Channel Blockers; Potassium Channels, Inwardly Rectifying; Rats; Rats, Inbred SHR; Rats, Wistar; Vasodilation

Sinomenine (SN), an alkaloid prepared from the root of Sinomenium acutum Rehd. Et wils, is used to alleviate the symptoms of rheumatism in Chinese medicine. In the present study, the potential inhibition of TNF-alpha-induced VCAM-1 expression on human umbilical vein endothelial cells (HUVECs) was evaluated in vitro. HUVECs were isolated from freshly collected umbilical cords. Positive controls were stimulated with TNF-alpha, omitting SN. Negative controls were cultured omitting TNF-alpha and SN. Experimental groups were co-cultured with TNF-alpha and SN at different concentrations (0.25, 0.5, and 1.0 mol/L), or TNF-alpha and Dexamethasone (Dex) at a concentration of 1.0 x 10(-6) mol/L. Cells were harvested after culturing with the above drugs for 12 h. VCAM-1 mRNA expression was detected by real-time quantitative PCR, and VCAM-1 expression was detected by flow cytometry. The experimental data indicated that VCAM-1 mRNA and VCAM-1 were induced by TNF-alpha. The relative VCAM-1 mRNA expression decreased in the experimental groups (p<0.05). Concentrations of SN at 0.5 and 1.0 mol/L inhibited expression of VCAM-1 (p<0.05). SN at concentration of 0.25 mol/L and Dex at concentration of 1.0 x 10(-6) mol/L did not show an inhibitory effect on VCAM-1 expression in TNF-alpha-induced HUVECs. Our preliminary data indicates that SN has an inhibitory effect in vitro on TNF-alpha-induced VCAM-1 expression at both mRNA level and protein level in HUVECs, and suggests that SN may be a novel method of immunotherapy for rheumatic carditis or rheumatic heart disease.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Cell Separation; Cell Survival; Cells, Cultured; Endothelial Cells; Endothelium, Vascular; Factor VIII; Flow Cytometry; Humans; Immunohistochemistry; Indicators and Reagents; Morphinans; NF-kappa B; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Stimulation, Chemical; Tumor Necrosis Factor-alpha; Umbilical Veins; Vascular Cell Adhesion Molecule-1

The mechanisms involved in the induction and regulation of inflammation resulting in dopaminergic (DA) neurotoxicity in Parkinson's disease (PD) are complex and incompletely understood. Microglia-mediated inflammation has recently been implicated as a critical mechanism responsible for progressive neurodegeneration.. Mesencephalic neuron-glia cultures and reconstituted cultures were used to investigate the molecular mechanisms of sinomenine (SN)-mediated anti-inflammatory and neuroprotective effects in both the lipopolysaccharide (LPS)- and the 1-methyl-4-phenylpyridinium (MPP+)-mediated models of PD.. SN showed equivalent efficacy in protecting against DA neuron death in rat midbrain neuron-glial cultures at both micro- and sub-picomolar concentrations, but no protection was seen at nanomolar concentrations. The neuroprotective effect of SN was attributed to inhibition of microglial activation, since SN significantly decreased tumor necrosis factor-alpha (TNF-alpha, prostaglandin E2 (PGE2) and reactive oxygen species (ROS) production by microglia. In addition, from the therapeutic point of view, we focused on sub-picomolar concentration of SN for further mechanistic studies. We found that 10(-14) M of SN failed to protect DA neurons against MPP+-induced toxicity in the absence of microglia. More importantly, SN failed to show a protective effect in neuron-glia cultures from mice lacking functional NADPH oxidase (PHOX), a key enzyme for extracellular superoxide production in immune cells. Furthermore, we demonstrated that SN reduced LPS-induced extracellular ROS production through the inhibition of the PHOX cytosolic subunit p47phoxtranslocation to the cell membrane.. Our findings strongly suggest that the protective effects of SN are most likely mediated through the inhibition of microglial PHOX activity. These findings suggest a novel therapy to treat inflammation-mediated neurodegenerative diseases.

Topics: Animals; Animals, Newborn; Anti-Inflammatory Agents; Cell Death; Cell Line; Coculture Techniques; Dopamine; Dose-Response Relationship, Drug; Encephalitis; Enzyme Inhibitors; Mice; Mice, Inbred C57BL; Mice, Knockout; Microglia; Morphinans; NADPH Oxidases; Neurons; Neuroprotective Agents; Parkinson Disease; Rats; Rats, Inbred F344; Tumor Necrosis Factor-alpha

Diabetic retinopathy involves an inflammatory response in the retina characterized by an increase in inflammatory cytokines and activation of microglia. The degree of microglia activation may influence the extent of retina injury following an inflammatory stimulus. Cytokines, released by activated microglia, regulate the influx of inflammatory cells to the damaged area. Thus, therapeutic strategy to reduce cytokine expression in microglia would be neuroprotective. Sinomenine, an alkaloid isolated from the stem and root of Sinomenium acutum, has long been recognized as an anti-inflammatory drug for rheumatoid arthritis and also inhibits macrophage activation. In this study, we activated retinal microglia in culture with advanced glycation end products (AGEs) treatment and attempted to determine whether sinomenine could reduce the production of cytokines from the activated microglia at both gene and protein levels. Changes in inflammatory cytokines, TNF alpha, IL-1 beta and IL-6, were measured by semi-quantitative RT-PCR and enzyme-linked immunosorbent assay (ELISA) both in the presence and absence of AGEs. The effect of sinomenine on levels of reactive oxygen species (ROS) and the nuclear translocation of NF-kB p65 were studied with a laser confocal scanning microscope. AGEs treatment induced a significant release of TNF alpha, IL-1 beta, and IL-6 from retinal microglia. Sinomenine could inhibit release of these cytokines. Sinomenine attenuated ROS production in a dose-dependent fashion and reduced the nuclear translocation of NF-kB p65 in AGEs-activated retinal microglia in culture.

Topics: Active Transport, Cell Nucleus; Animals; Animals, Newborn; Anti-Inflammatory Agents, Non-Steroidal; Blotting, Western; Cell Nucleus; Cells, Cultured; Gene Expression; Glycation End Products, Advanced; Interleukin-1beta; Interleukin-6; Microglia; Morphinans; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Retina; Reverse Transcriptase Polymerase Chain Reaction; Transcription Factor RelA; Tumor Necrosis Factor-alpha

The present study was designed to investigate the effect of sinomenine (SIN), an alkaloid extracted from sinomenium acutum on collagen-induced arthritis (CIA) in mice. For this investigation, mice were s.c. immunized with type II collagen (CII) emulsified with complete Freund's adjuvant (day 0). Varying doses of SIN were orally administered daily commencing on day 0 daily over a period of 55 days. The severity of arthritis was evaluated according to clinical score, the effect of SIN on immune responses were determined by measurement of proliferative responses of spleen cells, antibody levels in serum and cytokine assays. Anti-CII IgG2a and IFN-gamma were measured as indicators of Th1 immune responses and anti-CII IgG1, IgE and IL-5 as those of Th2 responses. IL-10 and TGF-beta were measured as indicators of T cell regulator responses. The results showed that treatment with SIN was followed by decreases in the incidence and severity of CIA, anti-CII IgG and the antigen-specific splenocyte proliferation. Production of all isotypes of antibodies including anti-CII IgG2a, IgG1 and IgE as well as secretion of cytokines such as IFN-gamma and IL-5 were suppressed by SIN. In addition, SIN enhanced the secretion of TGF-beta while it had no obvious effect on production of IL-10. These results suggest that the anti-arthritic effect of SIN may be related to the suppression of both Th1 and Th2 immune responses. TGF-beta may at least in part contribute to the suppression of Th1 as well as Th2 immune responses.

Topics: Administration, Oral; Alkaloids; Animals; Arthritis, Experimental; Autoantibodies; Cell Proliferation; Collagen Type II; Cytokines; Dose-Response Relationship, Immunologic; Female; Freund's Adjuvant; Mice; Morphinans; Spleen; Th1 Cells; Th2 Cells

Polycysteic acid based electrochemical oxidation of L-cysteine (CySH) and carbon nanotubes (CNTs) formed a composite thin film material at a glassy carbon electrode (GCE) that was used a novel modifier for electroanalytical determination of sinomenine which is used for rheumatoid arthritis treatment. The determination of sinomenine at the composite modified electrode was studied by differential pulse voltammetry (DPV). The peak current obtained at + 0.632 V (vs SCE) from DPV was linearly dependent on the sinomenine concentration in the range of 1.0 x 10(-7) to 6.0 x 10(-5) M in a B-R buffer solution (0.04 M, pH 1.81) with a correlation coefficient of 0.998. The detection limit (S/N = 3) was 5.0 x 10(-8) M. The electrochemical reaction mechanism of sinomenine was also discussed. This new method was then applied to the high-throughput determination of sinomenine in human serum samples with satisfactory results. This polycysteic acid/CNTs composite film may be considered to be a promising, low-cost, durable, and biocompatible material for the modification of sensors in applications to pharmaceutical and biomedical analysis.

Topics: Carbon; Cysteic Acid; Electrochemistry; Electrodes; Humans; Hydrogen-Ion Concentration; Membranes, Artificial; Molecular Structure; Morphinans; Nanotubes, Carbon; Particle Size; Polymers; Reproducibility of Results; Sensitivity and Specificity

To screen the optimum formulation and prepare O/W sinomenine microemulsion and investigate its in vitro transdermal delivery ability.. The microemulsions were prepared with the formulation containing oleic acid-tween 80-dehydrated alcohol-water by the pseudo-ternary phase diagram. The permeation flux of sinomenine was determined in vitro by Franz diffusion cell fitted with rat skin. The sinomenine was determined by HPLC. The transdermal characteristics of sinomenine microemulsion were compared with that of sinomenine gels.. The steady state flux of sinomenine microemulsion was significantly higher than that of sinomenine gels. The average permeation rate of sinomenine microemulsion was 116. 44 microg x cm(-2) x h(-1) in vitro.. These results indicated that the studied microemulsion system with high permeation rate may be a potential vehicle for the transdermal delivery of sinomenine.

Topics: Administration, Cutaneous; Animals; Drug Compounding; Drug Delivery Systems; Emulsions; Ethanol; Male; Morphinans; Oleic Acid; Particle Size; Plants, Medicinal; Polysorbates; Rats; Sinomenium; Skin; Skin Absorption; Surface-Active Agents

To prepare Sinomenine hydrochloride-loaded bovine serum albumin microspheres (SM-BSA-MS).. SM-BSA-MS was prepared by spray drying technique. The morphology, drug-loading and release in vitro of SM-BSA-MS was studied.. The diameters of SM-BSA-MS were in the range of 1-3 m. The drug loading of microspheres, formulated with different drug/albumin ratios as 1, 2, 1:1, 2:1, were 31.6%, 47.7% and 67.9% , respectively. And the drug entrapment efficiencies of different drug/albumin ratios were higher than 94%. The results of in vitro release experiments showed that the drug loaded microspheres have the properties of sustained-release compared with the Sinomenine hydrochloride injection. Different release characteristics could be obtained by adjusting the prescription composition and the thermal denaturation condition.. Spray drying technique is a simple and feasible method for preparing SM-BSA-MS. The drug loaded microspheres had high drug-loading and sustained-release effect.

Topics: Delayed-Action Preparations; Drug Carriers; Drug Compounding; Microspheres; Morphinans; Particle Size; Plants, Medicinal; Serum Albumin, Bovine; Sinomenium

Paeoniflorin and sinomenine, derived from the root of Paeonia lactiflora Pall. (family Ranunculaceae) and the stem of Sinomenium acutum Rehder & Wilson (family Menispermaceae), respectively, have been, and are currently, widely used for treatment of rheumatic and arthritic diseases in China and Japan. Our previous studies demonstrated that sinomenine could significantly improve the bioavailability of paeoniflorin in rats, but the underlying mechanisms remain unknown. The present study aims to investigate the intestinal kinetic absorptive characteristics of paeoniflorin as well as the absorptive behavior influenced by co-administration of sinomenine using an in vitro everted rat gut sac model. The results showed a good linear correlation between the paeoniflorin absorption in sac contents and the incubation time from 0 to 90 min. However, the concentration dependence showed that a non-linear correlation exists between the paeoniflorin absorption and its concentrations from 10 to 160 microM, and the absorption was saturated at about 80 microM of the drug. Sinomenine at 16 and 136 microM concentrations could significantly enhance the absorption of paeoniflorin (20 microM) by 1.5- and 2.5-fold, respectively. Moreover, two well-known P-glycoprotein inhibitors, verapamil and quinidine, could significantly elevate the absorption of paeoniflorin by 2.1- and 1.5-fold, respectively. Furthermore, sinomenine in a pattern, which influenced paeoniflorin's absorption, manifested as similar to that of P-glycoprotein inhibitors. In conclusion, sinomenine significantly enhance the intestinal absorption of paeoniflorin, subsequently improve the bioavailability of paeoniflorin. The mechanism underlying the improvement of paeoniflorin's bioavailability was proposed that sinomenine could decrease the efflux transport of paeoniflorin by P-glycoprotein.

Topics: Animals; Anti-Inflammatory Agents; ATP Binding Cassette Transporter, Subfamily B, Member 1; Benzoates; Bridged-Ring Compounds; Digoxin; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Glucosides; In Vitro Techniques; Intestinal Absorption; Intestinal Mucosa; Intestines; Male; Models, Animal; Monoterpenes; Morphinans; Paeonia; Quinidine; Rats; Rats, Sprague-Dawley; Sinomenium; Time Factors; Verapamil

Sinomenine is an alkaloid with pharmacological effects of anti-inflammation, anti-angiogenesis, anti-arthritis and immunosuppression. This study aimed to investigate the effect of sinomenine on gene expression of human synovial sarcoma cells (Hs701.T) activated by IL-1 beta. The proliferative effect of sinomenine was examined in the presence or absence of IL-1 beta by the [3H]-thymidine incorporation and MTT assay, respectively. Using DNA microarray technology and RT-PCR, the activating action of IL-1 beta and modulatory effect of sinomenine on Hs701.T were simultaneously determined. Results showed that IL-1 beta could stimulate the proliferation and gene expression of Hs701.T cells. Sinomenine could significantly inhibit proliferation of IL-1 beta-activated Hs701.T cells and suppress expression of 17 genes including IL-6, PlGF, Daxx, and HSP27. These genes were found to be important in tumor progression through the mediation of inflammation, cell adhesion, proliferation, apoptosis and angiogenesis. In conclusion, our study provides supplementary information for the further studies on the pharmacological effects of sinomenine acting on synovial sarcoma.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Cell Division; Cell Line, Tumor; Cell Survival; DNA, Neoplasm; Gene Expression; Gene Expression Profiling; Humans; Interleukin-1; Morphinans; Oligonucleotide Array Sequence Analysis; Reverse Transcriptase Polymerase Chain Reaction; Sarcoma; Synovial Membrane

Vasodilating actions of sinomenine were examined using rat aorta ring strips. Sinomenine (0.1 to 100 microM) dilated norepinephrine (NE, 5 microM)-induced vasoconstriction in a concentration-dependent manner reaching 68.8+/-5.1% (n=6, P<0.01) at a concentration of 100 microM. Sinomenine (100 microM) also attenuated KCl (60 mM) and phorbol 12, 13-dibutyrate (PDB, a protein kinase C, PK-C, activator, 300 nM)-induced vasoconstriction by 86.9+/-8.5% (n=6, P<0.01) and 49.9+/-9.8% (n=6, P<0.01), respectively. Pretreatment with nicardipine (a Ca2+ channel antagonist), staurosporine (a PK-C inhibitor), NG-monomethyl-L-arginine acetate (L-NMMA, a nitric oxide, NO, synthesis inhibitor), and indomethacin (a cyclooxygenase inhibitor) were carried out. Nicardipine (0.1 microM) led to a significant decrease in the vasodilating potential of sinomenine (at 100 microM, 68.8+/-5.1% vs. 35.5+/-6.9%; n=5, P<0.001). Pretreatment with staurosporine (30 nM) reduced sinomenine-associated vasodilation from 68.8+/-5.1% to 49.5+/-7.7% (n=5, P<0.001), and L-NMMA (100 microM) and indomethacin (10 microM), to 25.3+/-2.3% (n=5, P<0.001) and to 37.1+/-9.3% (n=5, P<0.001), respectively. The responses were almost similar to the results without endothelium. Therefore, these results indicate that sinomenine causes the vasorelaxation by the mechanisms involved with the inhibitions of Ca2+ channel and PK-C activity, and also with the activations of NO and prostaglandin (PG) I2 syntheses in endothelium.

Topics: Animals; Aorta, Thoracic; Calcium Channel Blockers; Cyclooxygenase Inhibitors; Endothelial Cells; Enzyme Inhibitors; In Vitro Techniques; Male; Morphinans; Muscle, Smooth, Vascular; Myocardial Contraction; Myocytes, Cardiac; Nitric Oxide Synthase Type III; Protein Kinase C; Rats; Rats, Wistar; Vasoconstrictor Agents; Vasodilator Agents

To evaluate the effects of the alkaloid sinomenine (SIN), a COX-2 inhibitor, on the acute rejection in heart allografts.. Forty Wistar rats received the allograft of the hearts of 40 SD rats into the peritoneum. Then the recipients were randomly divided into 2 groups: SIN group, injected with SIN within 24 hours after the operation; and control group, injected with normal saline. The survival time was observed. The heartbeat was examined every day. The Wistar rats were killed 3 and 5 days after the operation respectively and the left ventricular tissues were taken to undergo pathological examination to detect the acute rejection and cell apoptosis. Immunochemistry and Western blotting were used to detect the COX-2 protein, and RT-RCR was used to detect the COX-2 mRNA. The mean numbers of apoptotic cardiomyocytes were determined with the terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL) technique.. The survival time of the SIN group was 12.5 +/- 2.6 days, significantly longer than that of the control group (6.8 +/- 0.5 days, P = 0.001). Examination 3 and 5 days after the treatment of SIN, the extents of inflammatory reaction, endovasculites, myocardial edema, and cardiomyocyte damage in the allografts of the SIN group were all significantly less, the mean numbers of apoptotic cardiomyocyte was significantly smaller compared with the control group (all P < 0.05). At day 5, the levels of COX-2 protein and mRNA of the SIN group were both significantly lower than those of the control group.. Inhibition of COX-2 prolongs the allograft survival and reduces the myocardial damage and inflammation during acute cardiac allograft rejection.

Topics: Animals; Apoptosis; Blotting, Western; Cyclooxygenase 2; Female; Gene Expression; Graft Rejection; Heart Transplantation; In Situ Nick-End Labeling; Male; Morphinans; Rats; Rats, Sprague-Dawley; Rats, Wistar; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transplantation, Homologous

To establish an HPLC method for the determination of entrapment efficiency of sinomenine liposomes.. The liposomes and dissociated drugs were separated by sephadex filtration, mini-column centrifugation and dialysis. The methodology study and the optimization of determining condition were carried out at the same time.. Sephadex filtration could effectively separate the sinomenine liposomes from dissociated sinomenine. The column recovery was 98.8%, the average entrapment efficiency of three tests was64.9%, RSD 2.67%.. The method was simple, exact, and had a good reappearance. It can be used to examine the entrapment efficiency of sinomenine liposomes.

Topics: Dextrans; Drug Carriers; Drug Delivery Systems; Filtration; Liposomes; Morphinans; Sinomenium; Technology, Pharmaceutical

Eighteen sinomenine derivatives on ring C were prepared, and their anti-inflammatory activities were also investigated. Most of these derivatives showed mild to moderate activities. Compounds 4a, 4c and 5b showed better anti-inflammatory activity. So further modification of the ring C in sinomenine should be worthwhile.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Female; Inflammation; Male; Mice; Molecular Structure; Morphinans

To determine the intestinal disposition mechanisms of paeoniflorin, a bioactive glucoside, and to investigate the mechanisms by which sinomenine increases paeoniflorin bioavailability.. A single-pass "four-site" rat intestinal perfusion model and a cultured Caco-2 cell model were employed.. In both model systems, paeoniflorin permeability was poor. In the perfusion model, maximal absorption and metabolism of paeoniflorin occurred in duodenum and jejunum, which were significantly decreased by a glucosidase inhibitor gluconolactone (20 mM). On the other hand, paeoniflorin absorption in terminal ileum increased significantly but its metabolism did not in the presence of sinomenine and cyclosporine A. In the Caco-2 cell model, paeoniflorin was transported 48-fold slower than its aglycone (paeoniflorigenin). Absorptive transport of paeoniflorin was significantly (p < 0.05) increased by sinomenine (38%), verapamil (27%), and cyclosporine A (41%), whereas its secretory transport was significantly (p < 0.01) decreased by sinomenine (50%), verapamil (35%) and cyclosporine A (37%). In contrast, MRP inhibitors MK-571 and leukotriene C4 did not affect transport of paeoniflorin. Lastly, sinomenine was also shown to significantly increase the absorptive transport of digoxin (a prototypical p-glycoprotein substrate) and to significantly decrease its secretory transport.. Poor permeation, p-gp-mediated efflux, and hydrolysis via a glucosidase contributed to the poor bioavailability of paeoniflorin. Sinomenine (an inhibitor of the p-gp-mediated digoxin efflux) increased paeoniflorin's bioavailability via the inhibition of p-gp-mediated paeoniflorin efflux in the intestine.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; ATP Binding Cassette Transporter, Subfamily B, Member 1; ATP-Binding Cassette Transporters; Benzoates; Biological Availability; Biological Transport; Bridged-Ring Compounds; Caco-2 Cells; Cells, Cultured; Chromatography, High Pressure Liquid; Digoxin; Drug Interactions; Glucosides; Humans; Intestinal Absorption; Male; Monoterpenes; Morphinans; Rats; Rats, Sprague-Dawley

The present study was designed to investigate the effect of sinomenine (SIN), an alkaloid extracted from Sinomenium acutum, on Th1 and Th2 immune responses in mice. For this investigation, mice were S. C. immunized with ovalbumin (OVA) emulsified with complete Freund's adjuvant (day 0). Varying doses of SIN were orally administered daily over a period of 21 days, commencing on day 0. On day 21, anti-OVA IgG and proliferative responses of spleen cells to the antigen were measured. Anti-OVA IgG2a and IFN-gamma were measured as indicators of Th1 immune responses and anti-OVA IgG1, IgE, and IL-5 as those of Th2 responses. TGF-beta was measured as an indicator of Th3 immune responses. The results showed that treatment with SIN was followed by decreases in anti-OVA IgG and the antigen-specific splenocyte proliferation. Production of all isotypes of antibodies including anti-OVA IgG2a, IgG1 and IgE as well as secretion of cytokines such as IFN-gamma and IL-5 was suppressed by SIN, although the suppression of anti-OVA IgG2a and IFN-gamma by the alkaloid appeared to be greater than that of anti-OVA IgG1, IgE, and IL-5. In addition, SIN enhanced the secretion of TGF-beta. These results suggest that SIN appears to have suppressive effects on both Th1 and Th2 immune responses. The results also suggest that Th1 responses may be more preferentially suppressed by the Sinomenium acutum-derived alkaloid compared to Th2 responses. TGF-beta may at least in part contribute to the suppression of Th1 as well as Th2 immune responses.

Topics: Administration, Oral; Alkaloids; Animals; Cell Proliferation; Female; Immune System; Immunologic Factors; Mice; Mice, Inbred DBA; Morphinans; Ovalbumin; Phytotherapy; Plant Extracts; Sinomenium; Spleen; Th1 Cells; Th2 Cells

To determine in vitro the rat plasma protein binding rate by using microdialysis method.. The binding rate was determined by using microdialysis probe as sampling tools and zero-net flux method as calibrating method. The regression equation was made by the difference of concentrations between the dialysis sample and the perfusate. The x-intercept of regression equation was the free drug concentration (Cf). The plasma protein binding rate was calculated by using the following equation: f = ( C0 - Cf)/C0.. The binding rate was kept relatively stable in the studied concentration range.. It is feasible that the plasma protein binding rate can be determined by using microdialysis method.

Topics: Animals; Blood Proteins; Chromatography, High Pressure Liquid; Male; Microdialysis; Morphinans; Plants, Medicinal; Protein Binding; Rats; Rats, Sprague-Dawley; Regression Analysis; Sinomenium

To evaluate the effects of Caulis Sinomenii and sinomenine on conditioned place preference (CPP) induced by morphine and brain histamine level in mice.. Sixty mice were randomized into 6 equal groups and morphine (Mor) was injected subcutaneously (9 mg/kg) for 6 consecutive days to induce CPP using a shuttle box. Since the 4th day of training, the mice in 5 of the groups were treated for 3 consecutive days with Caulis Sinomenii (10 g/kg), sinomenine (60 mg/kg), diphenhydramine (30 mg/kg), CP48/80 (5 mg/kg) and L-histidine (750 mg/kg) in addition to morphine (9 mg/kg) treatment, respectively, leaving the other group with exclusive morphine treatment. Another 10 mice received saline injection to serve as saline control group. The content of histamine (HA) in the mouse brain was measured by fluorospectrophotometry.. In morphine group, the mice showed significantly extended stay in morphine-paired compartment whose HA content in the brain was markedly increased (P<0.01). Treatment with Caulis Sinomenii and sinomenine resulted in significantly reduced time of stay in morphine-paired compartment and brain HA level (P<0.01).. CPP induced by morphine in mice is associated with increased HA level in the brain. Caulis Sinomenii and sinomenine can suppress the acquisition of place preference induced by morphine and modulate HA level in the central nervous system in morphine-dependent mice.

Topics: Animals; Arginine; Brain; Conditioning, Operant; Diphenhydramine; Histamine; Male; Mice; Morphinans; Morphine; Morphine Dependence; Motor Activity; Random Allocation; Sinomenium

Caulis Sinomenii is the dried plant stems of Sinomenium acutum and Sinomenium acutum var. cinereum and has been used in Chinese medicine for treating rheumatic diseases for over a thousand years. Previous studies have demonstrated that sinomenine is a major active constituent in both plants and can be utilized as an indicator of quality of the medicinal herb Caulis Sinomenii. Currently, S. acutum and S. acutum var. cinereum are growing over a wide geographical range in China, with equally wide variations in growing conditions. The objectives of this research were to determine whether there were difference between the species and varieties, and whether the different growing conditions could result in different quality by determining the content of sinomenine in different samples. A modified HPLC method using a diode array detector (DAD) has been developed for efficiently quantifying sinomenine in the plants. Using this method, fourteen samples of S. acutum var. cinereum and eleven samples of S. acutum from growing regions as well as eighteen herbal samples of Caulis Sinomenii from wholesale herbal markets were evaluated. The results showed that there was no marked difference in the content of sinomenine between the species and varieties collected from growing regions; however, a very large variation was found among the samples collected from different regions. Moreover, the content of sinomenine in the plants of large size (stem diameter>3 cm) was much higher than those of small size (stem diameter<1 cm). This implies that the growing region has greater impact on the quality of Caulis Sinomenii in terms of the content of sinomenine than the species and varieties. The results also showed that the content of sinomenine in commercial Caulis Sinomenii was markedly lower than that in the plants collected directly from growing regions. This suggests that to obtain the herb with higher content of sinomenine and thus ensure greater efficacy, both in clinical applications and in pharmacological investigations, the plant of Caulis Sinomenii with controlled stem size collected directly from growing regions is preferable.

Topics: China; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Morphinans; Plant Preparations

For exploring the mechanism of the anti-inflammatory and anti-rheumatic effect of sinomenine (SN), the actions of different dosage of SN were observed in vitro on the expression of cytokines, tumor necrosis factor (TNF-alpha) and interleukin-1beta (IL-1beta), as well as the activity of nuclear factor-kappaB (NF-kappaB) and the inhibitory kappaB-alpha (IkappaB-alpha) protein level of peritoneal macrophages (PMs) and synoviocytes in adjuvant arthritis (AA) rats. In this study, the experimental rat model of AA was used and PMs and synoviocytes were collected. The mRNAs of TNF-alpha and IL-1beta were detected with reverse transcription-polymerase chain reaction (RT-PCR) and NF-kappaB activity was measured by electrophoretic mobility shift assay (EMSA). The IkappaB-alpha protein level in the cytoplasma was detected by Western blot. Our results showed that expression of mRNAs of TNF-alpha and IL-1beta and NF-kappaB activity by PMs and synoviocytes were markedly increased compared to control group (P<0.05). In a definite concentration ranging from 30 to 120 microg/ml, SN showed inhibiting effect on the NF-kappaB activity and the expression of the mRNAs of TNF-alpha and IL-1beta in AA rats in a concentration-dependent manner (P<0.05). Positive correlations were found between changes of NF-kappaB activity and expression of TNF-alpha mRNA and IL-1beta mRNA (P<0.01). IkappaB-alpha protein level was increased by various dosages of SN in comparison with control group (P<0.01). In conclusion, SN decreases the mRNA expression of TNF-alpha and IL-1beta by inhibiting the NF-kappaB binding activity, which is mediated through up-regulating the IkappaB-alpha expression of PMs and synoviocytes in AA rats.

Topics: Animals; Arthritis, Experimental; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; I-kappa B Proteins; Interleukin-1; Macrophages, Peritoneal; Male; Morphinans; NF-kappa B; Plants, Medicinal; Rats; Rats, Wistar; RNA, Messenger; Synovial Membrane; Tumor Necrosis Factor-alpha; Up-Regulation

Sinomenine (SN), an immunnosuppressive compound derived from the Chinese medicinal plant Sinomenium acutum, has been used to treat autoimmune diseases effectively. Previous studies show SN can inhibit lymphocytes proliferation and macrophage production of pro-inflammatory factors. However, little is known about the mechanisms by which SN inhibits macrophage functions. In this study, we demonstrated that SN could inhibit the proliferation of murine macrophages RAW264.7 by inducing apoptosis in a dose- and time-dependent manner. We found activation of extracellular signal-regulated protein kinase (ERK) in SN-treated macrophages, and requirement for ERK activation in SN-induced apoptosis of macrophages. Contemporarily, the expression of p27/KIP1, proapoptotic factor Bax increased, and expression of Bcl-2 decreased, which might cooperate to induce apoptosis. Inhibiting ERK activation reduced the increased expression of p27 and Bax, but had no effect on the decreased expression of Bcl-2, suggesting the involvement of ERK activation in the SN-induced increased expression of p27 and Bax. These results demonstrated that SN could induce apoptosis of macrophages through activation of ERK, and ERK activation might partially involve in the increased expression of p27 and Bax in apoptotic macrophages. Therefore, induction of macrophage apoptosis through ERK activation may be one of mechanisms by which SN exhibits its immunosuppressive function.

Topics: Animals; Apoptosis; Cell Proliferation; Dose-Response Relationship, Drug; Extracellular Signal-Regulated MAP Kinases; Immunosuppressive Agents; Macrophages; Mice; Morphinans

Paeonia lactiflora Pall. (Ranunculaceae) root and Sinomenium acutum Rehder and Wilson (Menispermaceae) stem are two herbs widely used in Chinese medicine to treat rheumatoid arthritis. While, in theory, either herb could be used alone, in practice, Chinese medicine practitioners prescribe them together. Studies on pharmacokinetic interaction between the active constituents of these two herbs (paeoniflorin and sinomenine, respectively) provide empirical evidence to support their clinical practice. A single dose of paeoniflorin (150 mg/kg) alone and with sinomenine hydrochloride (90 mg/kg) was administered by gastric gavage to unrestrained conscious male Sprague-Dawley rats (n=5 or 6, 240-270 [corrected] g). Blood samples were collected periodically via a jugular vein before and after dosing from 10 min to 12 h. A high-performance liquid chromatographic (HPLC) assay was developed to determine the plasma concentrations of paeoniflorin. Non-compartmental pharmacokinetic profiles were constructed by using the software PK Solutions 2.0. The pharmacokinetic parameters were compared using unpaired Student t-test. After co-administration of sinomenine, the peak plasma concentration of paeoniflorin was elevated (P<0.01), the peak time was delayed (P<0.01), the AUC(0-t) was increased (P<0.001), the mean residence time (MRT) was prolonged (P<0.01), the C(L) was decreased (P<0.01) and the V(d) was reduced (P<0.05). These results indicate that sinomenine hydrochloride at 90 mg/kg significantly improved the bioavailability of paeoniflorin in rats.

Topics: Animals; Area Under Curve; Benzoates; Biological Availability; Bridged-Ring Compounds; Calibration; Chromatography, High Pressure Liquid; Glucosides; Half-Life; Male; Monoterpenes; Morphinans; Rats; Rats, Sprague-Dawley; Reproducibility of Results

To observe the effects of sinomenine on the immune functions and apoptosis of murine lymphocyte as well as on human synovial fibroblast proliferation.. Both in vivo and in vitro tests were adopted. The lymphocyte proliferation induced by mitogens was assayed by MTT method. Spleen T lymphocyte subtypes were tested with flow cytometry. Spleen lymphocyte apoptosis was analyzed by flow cytometry and DNA ladder methods. In vitro test was adopted to observe the effects of sinomenine on the proliferation of human fibroblast of rheumatoid arthritis.. Sinomenine can inhibit the proliferation of mouse lymphocytes induced by ConA, LPS and anti-CD3 mAb but not PMA in vitro, and inhibit the proliferation induced by LPS and PMA in vivo. Sinomenine can reduce up-regulated CD4+/CD8+ ratio of T lymphocyte subtype in adjuvant arthritis rat. At the same concentration increased apoptosis ratio. As to human synovial fibroblast, sinomenine can significantly inhibit proliferation of human fibroblast.. Sinomenine can inhibit the immunological function and correct imbalance of CD4+/CD8+ ratio of T lymphocyte subtype. It can also increase apoptosis ratio of spleen lymphocyte. This may be the mechanism of its immunological inhibitory effect.

Topics: Animals; Apoptosis; Arthritis, Experimental; Arthritis, Rheumatoid; CD4-CD8 Ratio; Cell Proliferation; Humans; Lymphocytes; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Morphinans; Plants, Medicinal; Rats; Rats, Sprague-Dawley; Sinomenium; Spleen; Synovial Membrane

Sinomenine is a pure alkaloid with a variety of pharmacological actions including anti-inflammation, immuno-suppression, analgesia, antihypertension, and antiarrhythmia. Methods have been developed to measure sinomenine in rats and rabbits through high-performance liquid chromatography (HPLC). But only one report described a method for determination of sinomenine in humans.. We developed a method to quantitate sinomenine in human plasma based on liquid chromatography-ion trap mass spectrometry coupled with electrospray ionization (HPLC/ESI/ion trap mass spectrum).. The calibration curve (r2=0.9993) of sinomenine was established with standard solutions. The mean inter-day and intra-day CVs<15%. The limit of detection for sinomenine was 0.5 ng/ml. The relative recovery of high, middle and low concentration was 99%, 103%, and 118%, respectively.. The method is simple, rapid, sensitive, specific, and accurate enough for determination of sinomenine in human plasma and may be used to investigate the metabolism and pharmacokinetics of sinomenine.

Topics: Chromatography, High Pressure Liquid; Humans; Morphinans; Spectrometry, Mass, Electrospray Ionization

Previous studies have shown that sinomenine possesses potent immunoregulatory properties. However, little is known about its exact mechanisms of action. Increasing recognition of the importance of renal tubular epithelial cells (TECs) in renal diseases raises the question whether sinomenine can regulate TEC activity. In this study, cultured human TECs were exposed to proinflammatory factors interferon-gamma (IFN-gamma) and tumor necrotic factor-alpha (TNF-alpha) in presence or absence of sinomenine for 72 h, followed by analysis of surface expression of costimulatory molecules. Flow cytometric analysis revealed that various costimulatory molecules were expressed on TECs and that they were significantly up-regulated by the simulation of IFN-gamma and TNF-alpha. However, sinomenine especially down-regulated B7-H1 and B7-DC expression on TECs at both mRNA and protein levels. Moreover, the significant damping effect of sinomenine on B7-H1 and B7-DC signals could promote IL-2 and IFN-gamma production by co-cultured CD4(+) T cell. Our results indicated that sinomenine could regulate TECs activity via B7-H1 and B7-DC, in addition to previously reported its effects on some pro-inflammatory factors production by macrophages and peripheral blood mononuclear cells.

Topics: Antigens, CD; B7-1 Antigen; B7-H1 Antigen; CD4-Positive T-Lymphocytes; Cell Line; Cells, Cultured; Cyclosporine; Epithelial Cells; Humans; Immunologic Factors; Interferon-gamma; Interleukin-2; Kidney Tubules; Membrane Glycoproteins; Morphinans; Peptides; Programmed Cell Death 1 Ligand 2 Protein; Tumor Necrosis Factor-alpha

Supercritical carbon dioxide, with and without a methanol modifier, was used to extract sinomenine from Sinomenium acutum (Thumb) Rehd et Wils. Sinomenine determinations were carried out using high-performance liquid chromatography (HPLC). The results show that the yield obtained after 2.5 h extraction with methanol-modified supercritical carbon dioxide was the highest (7.47 mg/g), while that obtained with only supercritical carbon dioxide was the lowest (0.17 mg/g). The recovery obtained with supercritical carbon dioxide, with and without a methanol modifier, could not be increased greatly by the method of the alkalinization of sample. Higher recoveries were obtained than extraction using methanol in Soxhlet extractor.

Topics: Chromatography, High Pressure Liquid; Chromatography, Supercritical Fluid; Morphinans; Sinomenium

Sinomenine, an alkaloid derived from the Chinese medical plant Sinomenium acutum, was studied with regard to its pharmacokinetics and tissue distribution in rats, and to its protein binding ability in the plasma of rats and rabbits and in the solutions of albumin and alpha-1-acid-glycoprotein. An HPLC analytical method was developed for determining sinomenine. The results demonstrated that oral administration with a single dosage at a rate of 90 mg sinomenine/kg in rats achieved about 80% bioavailability, while most of the other pharmacokinetic parameters were similar to the data from the animals treated intravenously. This indicates that oral administration of sinomenine would be appropriate in clinics. In rats, at 45 min after oral dosage, the drug was found to distribute widely in the internal organs, with tissue concentrations (from highest to lowest) in the order of kidneys, liver, lungs, spleen and heart, brain and testicles. At 90 min after dosing, the tissue concentrations in the organs were markedly decreased. The liver and kidneys manifested as the dominant organs with high tissue concentrations that might be responsible for metabolism and elimination of sinomenine. Examination of the protein binding ability showed that sinomenine with 4 microg/ml concentration in the plasma of rats and rabbits or in the albumin solution achieved a protein binding rate of more than 60%, while in the solution of alpha-1-acid-glycoprotein the rate was only about 33%. This result suggests that sinomenine might have much more potent binding ability with albumin than with alpha-1-acid-glycoprotein, resulting from its acidic property.

Topics: Administration, Oral; Albumins; Animals; Chromatography, High Pressure Liquid; Injections, Intravenous; Male; Morphinans; Orosomucoid; Protein Binding; Rabbits; Rats; Rats, Sprague-Dawley; Tissue Distribution

To investigate the effect of release behavior of sustained-release dosage forms of sinomenine hydrochloride (SM.HCl) on its pharmacokinetics in beagle dogs.. The in vitro release behavior of two SM.HCl dosage forms, including commercial 12-h sustained-release tablets and 24-h sustained-release pellets prepared in our laboratory, was examined. The two dosage forms were orally administrated to beagle dogs, and then the in vivo SM.HCl pharmacokinetics was investigated and compared.. The optimal SM.HCl sustained-release formulation was achieved by mixing slow- and rapid-release pellets (9:1, w/w). The SM.HCl release profiles of the sustained-release pellets were scarcely influenced by the pH of the dissolution medium. Release from the 12-h sustained-release tablets was markedly quicker than that from the 24-h sustained-release pellets, the cumulative release up to 12-h was 99.9% vs 68.7%. From a pharmacokinetic standpoint, the 24-h SM.HCl sustained-release pellets had longer tmax and lower Cmax compared to the 12-h sustained-release tablets, the tmax being 2.67+/-0.52 h vs 9.83+/-0.98 h and the Cmax being 1 334.45+/-368.76 ng/mL vs 893.12+/-292.55 ng/mL, respectively. However, the AUC(0-tn) of two SM.HCl dosage forms was comparable and both preparations were statistically bioequivalent. Furthermore, the two preparations had good correlations between SM.HCl percentage absorption in vivo and the cumulative percentage release in vitro.. The in vitro release properties of the dosage forms strongly affect their pharmacokinetic behavior in vivo. Therefore, managing the in vitro release behavior of dosage forms is a promising strategy for obtaining the optimal in vivo pharmacokinetic characteristics and safe therapeutic drug concentration-time curves.

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents; Delayed-Action Preparations; Dogs; Drug Design; Male; Morphinans

The present study was designed to investigate the putative anxiolytic-like effect of sinomenine in three experimental models of anxiety in male rats and mice. Use of the elevated plus-maze test revealed that sinomenine (20 and 40 mg/kg, p.o.) increased the percentage of open arm entries and diazepam (2 mg/kg, p.o.) increased the percentage of open arm entries, the percentage of time spent on open arms and total arm entries in mice. In the light/dark transition test, sinomenine (20 and 40 mg/kg, p.o.) increased time spent in the light area and diazepam (2 mg/kg, p.o.) increased time spent in the light area and the overall movements in mice. In the social interaction test, the sinomenine-treated animals significantly increased social interaction time in low light unfamiliar (7 mg/kg, p.o.) and high light unfamiliar conditions (7 and 14 mg/kg, p.o.) as well as diazepam (3 mg/kg, p.o.). Sinomenine (28 mg/kg, p.o.) can also decrease squares entered in rats in social interaction test under low light unfamiliar condition. In the open-field test, sinomenine (160 mg/kg) decreased squares entered in mice. Thus, these findings indicated that sinomenine exhibited anxiolytic-like effect.

Topics: Animals; Anti-Anxiety Agents; Anxiety; Behavior, Animal; Darkness; Disease Models, Animal; Dose-Response Relationship, Drug; Light; Male; Maze Learning; Mice; Morphinans; Rats; Rats, Wistar; Social Behavior; Species Specificity

Sinomenine is an alkaloid extracted from the Chinese medicinal plant, Sinomenium acutum, which has been utilized to treat rheumatoid arthritis (RA) in China for over 2000 years. Sinomenine has been shown to mediate a wide range of pharmacological actions which includes anti-inflammatory and anti-rheumatic effects. RA has been classified as a chronic immune-mediated disease that exhibits overlapping manifestation of inflammatory, abnormal cellular and hormonal immune responses with synovial hyperplasia. Since, angiogenesis is recognized to play a critical role in the development of RA and anti-angiogenic therapy has been proposed as a new therapeutic strategy for treatment of RA, we would like to see if sinomenine possesses anti-angiogenic property. In this study, sinomenine inhibited bFGF-induced proliferation of human umbilical vein endothelial cells (HUVEC) and arrested its cell cycle in G1 phase. Sinomenine disrupted tube formation of HUVEC on Matrigel and suppressed the chemotaxis of HUVEC. In addition, sinomenine reduced neovascularization in Matrigel plug assay as well as microvascular outgrowth in rat aorta ring sprouting assay. These results suggest that sinomenine inhibited bFGF-induced angiogenesis in vitro and in vivo. As the leukocytes-endothelial adhesive interactions also play an important role in inflammation, we found that sinomenine reduced the transmigration of granulocytic differentiated HL60 cells across IL-1beta activated HUVEC monolayer. Therefore, the inhibition of leukocytes migration across blood vessel walls and the anti-angiogenic effect of sinomenine may contribute towards its therapeutic mechanisms in alleviating the pathogenesis of RA.

Topics: Angiogenesis Inhibitors; Animals; Arthritis, Rheumatoid; Endothelial Cells; Female; Fibroblast Growth Factors; HL-60 Cells; Humans; In Vitro Techniques; Mice; Mice, Inbred BALB C; Morphinans; Neovascularization, Pathologic; Phytotherapy; Plant Extracts; Rats; Rats, Sprague-Dawley; Umbilical Veins

To explore the effects of long-term morphine exposure on cAMP and cGMP levels in primary cultured tuberomammillary nucleus (TM) neurons of neonatal rats and the effects of sinomenine on morphine-dependent TM cells.. TM neurons after a 7-day primary culture were further cultured in the medium containing 100 micromol/L morphine for 48 h to prepare the cell model of morphine dependence. Serial doses of histamine or sinomenine were administered 30 min naloxone treatment, the cAMP and cGMP levels of the TM cells were determined by enzyme immunoassay. cAMP and cGMP levels were also determined in normal TM cells treated by histamine or sinomenine.. After treatment with 100 micromol/L morphine for 48 h, cAMP and cGMP levels in the TM neurons were increased markedly. Treatment with 100 micromol/L naloxone added in the culture media caused an overshoot of cellular cAMP and a marked declination of cGMP, resulting in significantly increased cAMP/cGMP ratio. Sinomenine at 30 and 100 micromol/L and histamine at 40 micromol/L failed to obviously affect cAMP and cGMP levels in normal TM neurons, but sinomenine at 300 micromol/L and histamine at 80 micromol/L significantly increased the intracellular cAMP level. After pre-treatment with sinomenine at the above 3 doses or histamine at 40 micromol/L, the TM neurons with morphine dependence exhibited significant reduction in intracellular cAMP level but increment in cGMP level after naloxone treatment, with significantly reduced cAMP/cGMP ratio.. Long-term morphine (100 micromol/L) exposure for 48 h can induce marked changes of cAMP and cGMP levels in the TM neurons. The central histaminergic nervous system may be responsible for the development of morphine dependence and withdrawal. Sinomenine can significantly reduce the cAMP level and enhance cGMP level of morphine-dependent TM neurons precipitated by naloxone, which results in a near-normal ratio of cAMP and cGMP.

Topics: Animals; Animals, Newborn; Cells, Cultured; Cyclic AMP; Cyclic GMP; Female; Histamine; Hypothalamus; Morphinans; Morphine; Morphine Dependence; Neurons; Rats; Rats, Sprague-Dawley

The optimal extraction technology of Fengshitongbi capsule was investigated in this study.. Orthogonal tests were employed for optimum of extraction technology, the determination of the contents of Chlorogenic acid and Sinomenine were gained by HPLC and TLC-scanning respectively.. Six volume of water,extracting 3 times with half an hour each time, and seven volume of 70% ethanol, extracting two times with one and a half hour each time were considered the optimum extraction technology of Flos Lonicerae and Caulis Sinomenii as well as Rhizoma Smilacis Glabrae respectively.. The considerable extraction rate of active components in the drugs is achieved by applying the selected technology.

Topics: Capsules; Chlorogenic Acid; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Drug Combinations; Drugs, Chinese Herbal; Ethanol; Liliaceae; Lonicera; Morphinans; Plant Stems; Plants, Medicinal; Rhizome; Sinomenium; Technology, Pharmaceutical; Water

To study the effect of penetration enhancers on the transdermal penetration of sinomenine liposome patch.. We firstly established the experimental method of transdermal penetration, and the methodology study was carried out at the same time. Furthermore, we added 5% Azone, 10% Oleic acid, 1% PG + 4% Azone, 2% PG + 8% Oleic acid into the patch. The cumulative penetration amounts were determined during the test of transdermal penetration and the part lagged in the skin was determined after the test.. The cumulative penetration amounts released from the patch increased, while the part lagged in the skin decreased at the same time.. The results indicate that penetration enhancers can facilitate drug through corneum into dermis, but can't enhance the drug' s accumulation in the corneum. It is concluded that penetration enhancers can destroy the two-molecule structure of liposomes and cause drug leakage, so penetration enhancers should not be added into liposome patch of part functions.

Topics: Administration, Cutaneous; Animals; Azepines; In Vitro Techniques; Liposomes; Male; Menispermaceae; Morphinans; Oleic Acid; Propylene Glycol; Rats; Rats, Nude; Skin; Skin Absorption

To discuss the anti-inflammatory mechanism of sinomenine on inflammatory media in joint of adjuvant arthritis rats.. Rats were randomly divided into the normal group and the model group, the prednisone group, the small, medium, large of sinomenine group (30, 60, 120 mg x kg(-1)). Except for the rats in the normal group, animals were modeled to adjuvant arthritiswith freund's complete adjuvant. The arthritis index (AI) and the swelling degree of paws were recorded, and the activity of IL-1, TNF and the levels of NO, PGE2 in joint fluids of secondary arthritis were determined.. Compared with the normal group, the activity of IL-1, TNF and the levels of NO, PGE2 in joint fluids of secondary arthritis were increased significantly in the model group (P < 0.05). Compared with the model group, it was shown to exert a dramatic inhibitory effect on secondary reaction of freund's adjuvant arthritis of rats, and the activity of IL-1, TNF and the levels of NO, PGE2 in joint fluids of secondary arthritis were significantly decreased in the sinomenine group (P < 0.05).. Sinomenine has a remarkable treatment effect on RA. It is via NO to inhibit the activity of cytokines and decrease the level of inflammation mediators, which may be one of its curing RA mechanism.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Experimental; Dinoprostone; Drugs, Chinese Herbal; Interleukin-1; Male; Morphinans; Nitric Oxide; Phytotherapy; Random Allocation; Rats; Rats, Wistar; Sinomenium; Synovial Fluid

The root of Paeonia lactiflora and the stem of Sinomenium acutum are two herbs widely used in Chinese herbal medicine for the treatment of inflammatory and arthritic diseases. Studies on the interaction of the active constituents of these herbs, i.e., paeoniflorin and sinomenine, in pharmacokinetic parameters, tissues distribution, and protein binding ability could provide empirical data to support their clinical application. Following oral administration to rats, the pharmacokinetic alterations were compared. The results showed that the pharmacokinetic parameters (Cmax, Tmax, AUC, MRT, C(L), and Vd) of paeoniflorin were markedly enhanced when co-administrated with sinomenine. At 45 min after oral administration, the concentrations of paeoniflorin in the main internal organs were significantly increased when co-administrated with sinomenine. These phenomena were not ascribable to the alteration of the protein binding ability of paeoniflorin by sinomenine because obvious interactions of paeoniflorin and sinomenine in protein binding abilities in vitro to rat and rabbit plasma, human albumin, and alpha-1-acid-glycoprotein were not observed. However, with respect to the in vivo influence of paeoniflorin on sinomenine, the results showed that co-administration of paeoniflorin did not affect the pharmacokinetic parameters and tissue distribution of sinomenine.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Area Under Curve; Benzoates; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Dialysis; Drug Interactions; Glucosides; Male; Monoterpenes; Morphinans; Protein Binding; Rats; Rats, Sprague-Dawley; Reference Standards; Reproducibility of Results; Tissue Distribution

Cardiac actions of the constituents of Sinomeni Caulis et Rhizome (SCR) and Mokuboi-to, a traditional herbal drug, were investigated. In voltage-clamp experiments, sinomenine (1 mM) and tetrandrine (100 microM) inhibited the ionic currents concentration-dependently. The constituents affected the action potential configurations. In multicellular preparations, SCR (1 mg/ml) decreased the maximum rate of depolarization. Tetrandrine (30 microM) and sinomenine (300 microM) also had similar effects, but magnoflorine (1 mM) had less or no effect. Dysrhythmias were abolished under Ca(2+) overload conditions by sinomenine. These results indicate that at even acute administrations, these drugs exert the active electropharmacological actions and cardioprotections.

Topics: Action Potentials; Alkaloids; Animals; Aporphines; Benzylisoquinolines; Calcium Channel Blockers; Drugs, Chinese Herbal; Guinea Pigs; Heart; Ion Channels; Morphinans; Papillary Muscles

(1) To study the effect of concentration and flow rate on relative recovery (RR); (2) To study the difference Of relative recovery between probes; (3) To study the relationship between relative recovery and relative loss (RL).. HPLC.. (1) The sinomenine concentration has no obvious effect on RR in the same flow rate. (2) RR is reduced by exponent law as the flow rate increases. (3) There is significent difference between probes.. The RR isn't affected by sinomenine concentration as the flow rate is same. The flow rate has notable effect on RR as the sinomenine concentration is same. The RR of every probe should be assayed. We can take RL as RR to do in-vivo study.

Topics: Chromatography, High Pressure Liquid; Dialysis Solutions; In Vitro Techniques; Microdialysis; Morphinans; Pharmacokinetics; Reference Standards

To provide basic data for the synthesis of new sinomenine derivatives.. The C ring in sinomenine was modified.. Seven compounds were prepared and screened for anti-inflammatory and analgesic activities. Compounds 2 and 5 showed better activities.. Modification of the C ring in sinomenine should be worthy to be studied further.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Edema; Mice; Molecular Conformation; Molecular Structure; Morphinans; Pain Measurement; Plants, Medicinal; Rats; Sinomenium

(13)C DNMR studies of codeine and sinomenine (derivatives of N-Me morphinan) indicated that N-inversion-C-N rotation (NIR) is unusually fast for these substituted piperidines when compared with other N-Me piperidines. Since only broadening, but no signal splitting, was reached at low temperatures and the difference of chemical shifts (Delta delta) for individual conformers with the equatorially and axially oriented N-Me substituent was unavailable, the limits of the NIR barrier for these amines were determined by line shape analysis using Delta delta values provided by ab initio calculations. On the basis of the comparison of experimentally determined (13)C NMR chemical shifts for tropane conformers with the ones calculated at different theory levels for this N-Me piperidine, the B3LYP/6-31G(p)/GIAO level was chosen as a sufficiently accurate method for calculations of Delta delta. By this new "semiempirical" procedure of line shape analysis the NIR barrier for the studied morphinans lies within a 25-27 kJ mol(-1) (6.0-6.5 kcal mol(-1)) range. A low NIR barrier for morphine alkaloids is supposed to be an important factor in the activation of morphine receptor.

Topics: Alkaloids; Codeine; Crystallography, X-Ray; Kinetics; Molecular Conformation; Molecular Structure; Monte Carlo Method; Morphinans; Morphine; Nuclear Magnetic Resonance, Biomolecular; Structure-Activity Relationship; Thermodynamics

To study the effects of sinomenine (Sin) on cell proliferation, intracellular expression of cyclooxygenase-2 (COX-2), and production of PGE2 in lipopolysaccharide-induced PC-12 cells, To explore the Sin's mechanism on nerve cell.. PC-12 cells were cultured with nerve growing factors (NGF), and pretreated with Sin at various concentrations (0, 3 x 10(-6), 30 x 10(-6), 150 x 10(-6) mol x L(-1)) for 2 hours, then with or without stimulation of lipopolysaccharide (LPS). The proliferation activity of PC-12 cells was determined by 3H-TdR incorporation, and the production of PGE2 in culture supernatants of PC-12 cells was detected with competitive ELISA. Expression of COX-2 mRNA in PC-12 cells was analyzed by semi-quantitative RT-PCR, and expression of COX-2 protein was estimated by Western blot method and cellular enzyme immunoassay. Nuclear factor-kappa B (NF-kappaB) activity in whole-cell extract of PC-12 cells was also measured by an ELISA-based method.. The data showed that Sin down-regulated the expression of COX-2 mRNA and protein, and reduced the production of PGE2 in the LPS-stimulated PC-12 cells which correlated with Sin's concentrations positively. In addition, NF-kappaB activity in LPS-stimulated cells was suppressed significantly by Sin. No inhibition of proliferation of PC-12 cells due to Sin treatment was observed.. Sin mediates the down-regulation of expression of COX-2 and production of induced PGE2 in PC-12 cells by suppressing the activity of NF-kappaB.

Topics: Animals; Cell Proliferation; Cyclooxygenase 2; Dinoprostone; Down-Regulation; Lipopolysaccharides; Morphinans; NF-kappa B; PC12 Cells; Plants, Medicinal; Prostaglandin-Endoperoxide Synthases; Rats; RNA, Messenger; Sinomenium

Sinomenine, an herbal ingredient isolated from Sinomenium acutum, is used for the amelioration of arthritis. Using microdialysis and a specially constructed hepato-duodenal shunt probe, the present study investigated the pharmacokinetics of sinomenine in rat blood and bile and the effects of P-glycoprotein modulation and cytochrome P450 inhibition. The results indicated that the pharmacokinetics of sinomenine in rat blood appeared to be dose dependent in the 3 to 30 mg/kg range. The disposition of sinomenine in the bile exhibited a slow elimination phase, reaching a peak concentration in 20-40 min following intravenous administration. The area under the concentration versus time curves (AUC's) for sinomenine in the bile were significantly greater than those in the blood at dosages of 3, 10, and 30 mg/kg with the blood-to-bile distribution ratios (k = AUC(bile) / AUC(blood)) being 3.85 +/- 0.29 and 3.52 +/- 0.28 at 10 and 30 mg/kg, respectively, indicating active hepatobiliary excretion. Coadministration with 20 mg/kg of cyclosporin A 10 min prior to sinomenine administration resulted in a significant reduction of the bile AUC's for the dosages of 10 and 30 mg/kg., resulting in the bile/blood distribution ratio being significantly reduced to 0.47 +/- 0.05 and 0.49 +/- 0.05, respectively. On the other hand, proadifen treatment increased both the blood and bile AUC's, resulting in insignificant effects on the blood-to-bile distribution ratios. In conclusion, our results indicated that sinomenine underwent active hepatobiliary elimination which may be regulated by the P-glycoprotein and that P-450 was likely involved in its metabolism.

Topics: Animals; Area Under Curve; ATP Binding Cassette Transporter, Subfamily B, Member 1; Cyclosporine; Cytochrome P-450 Enzyme Inhibitors; Dose-Response Relationship, Drug; Enzyme Inhibitors; Male; Microdialysis; Morphinans; Proadifen; Rats; Rats, Sprague-Dawley; Sinomenium; Specific Pathogen-Free Organisms

To investigate the effects of sinomenine on morphine withdrawal response and acetylchline (Ach)-induced contracture in isolated guinea pig ileum.. The withdrawal contracture was elicited by subjecting isolated ileum incubated with morphine (3 micromol/L) at 37.5 degrees Celsius for 4 h to naloxone (1 micromol/L) treatment. Sinomenine (10, 50, 250 micromol/L) and nimodipine (Nim, 0.1 micromol/L) were administered 1 min before and after naloxone in morphine-dependent ilea bathed in Krebs solution containing morphine, to observe the changes in the withdrawal contracture of the ileum. The effect of sinomenine (10, 50, 250 micromol/L) on the contracture of untreated ileum in Krebs solution elicited by acetylcholine was also observed.. Naloxone-induced withdrawal contracture or acetylcholine-induced contracture of the ileum was significantly decreased in a dose-dependent manner, indicating that sinomenine can inhibit morphine withdrawal symptoms in guinea pigs.

Topics: Animals; Guinea Pigs; Ileum; Morphinans; Morphine; Morphine Dependence; Muscle Contraction; Naloxone; Nimodipine; Substance Withdrawal Syndrome

Chronic rejection (CR) with graft vasculopathy is recognized as a major cause of graft loss over time. Sinomenine (SN) has anti-inflammatory, antirheumatic, and immunomodulatory effects. Previously, we demonstrated antimacrophage and anti-T cell effects of SN in acute rejection. In the current study, we investigated the effect of SN in a rat cardiac allograft model of CR.. After a brief course of cyclosporine A (CsA), Lewis recipients of F344 hearts were treated with SN alone, CsA alone, or a combination of both drugs. Grafts were analyzed morphometrically and by immunohistochemistry. Expression of basic fibroblast growth factor (bFGF), vascular endothelial growth factor, and endothelin 1 was assessed by reverse transcription-polymerase chain reaction. Antidonor IgM formation was investigated by FACS.. Cardiac grafts from SN-treated rats showed less pronounced vasculopathy in comparison with untreated rats or CsA-treated recipients. After treatment with a combination of both drugs, rats had significantly less graft vasculopathy than rats receiving either drug alone. Treatment with CsA alone led to a decrease in bFGF expression, whereas SN alone did not affect gene expression. SN in combination with CsA, however, markedly reduced expression of bFGF, vascular endothelial growth factor, and endothelin 1. SN alone did not inhibit antidonor antibody formation.. These studies demonstrate for the first time the therapeutic value of SN in a model of chronic cardiac allograft rejection. SN in combination with low-dose T cell-targeted immunosuppression is effective in controlling tissue remodeling in the context of CR and is associated with inhibition of intragraft expression of mediators involved in angiogenesis, vascular tone, and tissue remodeling.

Topics: Adjuvants, Immunologic; Animals; Antibody Formation; Chronic Disease; Cyclosporine; Cytokines; Drug Therapy, Combination; Graft Rejection; Growth Substances; Heart; Heart Transplantation; Immunosuppressive Agents; Male; Morphinans; Myocardium; Rats; Rats, Inbred F344; Transplantation, Homologous; Ventricular Remodeling

To observe in vitro the effect of Sinomenine, a pure alkaloid extracted from the chinese medical plant Sinomenium acutum on the activity of cyclooxygenase (COX-1 and COX-2) and the expression of COX-1 and COX-2 mRNA.. Mononuclear leukocytes were obtained from healthy adults. Isolated mononuclear leucocytes from human peripheral blood (PBMC) were incubated (1 x 10(6).mL-1) with or without sinomenine (or indomethacin), after incubated for 24 hours at 37 degrees C with 5% CO2; the media were assayed for the PGE2 by radioimmunoassay (RIA). LPS was used to stimulate the monocytes at a concentration of 5 micrograms.mL-1. And by RT-PCR, both COX-1 and COX-2 mRNAs were detected in Mononuclear leukocytes after incubation for different hours with drug (sinomenine or indomethacin) or not.. LPS (stimulated) induced the production of PGE2 in PBMC increasing with high expression of COX-2 mRNA; sinomenine reduced PGE2 production in LPS stimulated human monocytes more than in non-stimulated human monocytes. In comparative experiments, indomethacin, a non selective COX inhibitor, reduced the production of PGE2 equally in both states. Meanwhile, neither sinomenine(0.1-1 mmol.L-1) nor indomethacin(0.5-10 mumol.L-1) inhibited the expression of both COX-1 and COX-2 mRNAs by RT-PCR with beta-actin as reference.. In contrast with indomethacin, Sinomenine shows a preferential inhibitory effect on COX-2 over COX-1, These results suggest that Sinomenine is a selective COX-2 inhibitor, which may be directly related to suppressing cyclooxygenase activity.

Topics: Adult; Cyclooxygenase 1; Cyclooxygenase 2; Dinoprostone; Humans; Isoenzymes; Leukocytes, Mononuclear; Membrane Proteins; Morphinans; Plants, Medicinal; Prostaglandin-Endoperoxide Synthases; RNA, Messenger; Sinomenium

To study on the detoxification effect of sinamine in morphine-dependent rats.. Morphine-dependent rats were induced by injecting morphine on dosage increasing by degrees, then treated with medication. The withdrawal symptoms, body weight and NE, DA, 5-HT in the brain were tested.. Sinamine could alleviate withdrawal symptom, reablement body weight, inhibit neurotransmitter in the brain.. Sinamine have effects on morphine-dependent rats which may relate to modulating neurotransmitter.

Topics: Animals; Biogenic Monoamines; Body Weight; Brain; Dopamine; Male; Morphinans; Morphine Dependence; Norepinephrine; Rats; Serotonin; Substance Withdrawal Syndrome

To study the permeance and diffusion trend of 125I-sinomenine via the transdermal delivery way in rats.. Assay the permeance depth and diffusion area of sinomenine labeled by isotopic 125I in subcutaneous of rats certain time after sinomenine was administrated on the cervical skin of rats.. The diffuse of sionmenine can reach cervical spine in longitudinal direction and form an area of 2 cm2 in transverse direction. The concentration gradient of both directions are significant.. 125I-sinomenine can not only be absorbed into blood via the transdermal delivery way to have whole effect, but also accumulate large amounts in the skin to have a local effect.

Topics: Administration, Cutaneous; Animals; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Iodine Radioisotopes; Morphinans; Plants, Medicinal; Rats; Rats, Sprague-Dawley; Sinomenium; Skin Absorption

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cyclosporine; Drug Therapy, Combination; Graft Rejection; Heart Transplantation; Immunosuppressive Agents; Morphinans; Rats; Transplantation, Homologous

Topics: Adjuvants, Immunologic; Animals; Cyclosporine; Graft Survival; Heart Transplantation; Morphinans; Rats; Rats, Inbred ACI; Rats, Inbred Lew; Time Factors; Transplantation, Homologous

The effect of Sinomenine on IL-8, IL-6, IL-2 and mIL-2R produced by peripheral blood mononuclear cells was investigated by using cell culture, radioimmunoassay and flow cytometry. It was showed that production of IL-8 and mIL-2R was inhibited, but the levels of IL-6 were enhanced by Sinomenine. Our results also demonstrated that Sinomenine did not have any effect on the production of IL-2. The study demonstrated that Sinomenine was able to regulate the production of cytokines. This may be one of the mechanisms by which Sinomenine works on rheumatoid arthritis.

Topics: Antirheumatic Agents; Arthritis, Rheumatoid; Humans; Interleukin-2; Interleukin-6; Interleukin-8; Leukocytes, Mononuclear; Morphinans

The alkaloid sinomenine extracted from the medicinal plant Sinomenium acutum is used in China for the treatment of various rheumatic diseases. It has immunomodulatory properties in a cardiac allograft transplantation model. Its antiproliferative effect on human mononuclear cells in combination with different immunosuppressive drugs was further analysed in vitro. Sinomenine dose-dependently attenuated thymidine incorporation, interleukin-2 synthesis, and cell cycle progression of activated T-lymphocytes. Cell proliferation was synergistically decreased by addition of sinomenine together with suboptimal concentrations of the established immunosuppressive drugs tacrolimus or mycophenolic acid, respectively.

Topics: Animals; Drug Synergism; Drugs, Chinese Herbal; Humans; Immunosuppressive Agents; Monocytes; Morphinans; Mycophenolic Acid; Plants, Medicinal; Rats; Tacrolimus

To study the effect of different animal skin on the transdermal speed constant of sinomenine.. Improved franz-cell was used for the transdermal study, pH6.8 phosphate buffer was used as the receptor and HPLC method was used for the determination of sinomenine through different animal skin.. The transdermal speed constant of the skin of hareless mice was much close to which of person. The higuchi equation of sinomenine through skin of hareless mice stored under -20 degrees C for 0d, 3d and 7d were Q = 40.34t1/2 + 196, Q = 38.01t1/2 + 25.3, Q = 41.55t1/2 + 24.7 respectively.. The results indicate that skin of hareless mice is suitable for the transdermal study of sinomenine, and the effect of storing time (0-7d) under -40 degrees C is not obvious.

Topics: Administration, Cutaneous; Animals; Chromatography, High Pressure Liquid; Mice; Mice, Nude; Morphinans; Permeability; Skin; Skin Absorption; Time Factors; Tissue Preservation

To observe the clinical effect and experimental result of long-acting analgesic in the treatment of anorectal post-operational pain.. Analysing the pathological features of anorectal post-operational pain, and select the Radix Stephana Tetrandra, which could disperse pathogenic Wind and dampness, relieve spasm and pain, as the major component. The authors extracted Sinomenine from the Stephana and by mixing with methylene blue and bupivacaine hydrochloride, an injection was prepared. Three groups were in the clinical observation: Long-acting analgesic group (102 cases), routine treatment group (80 cases) and the control group treated by Tramal (30 cases). The curative effect was observed and animal experiment was conducted.. The total analgesic rate of the long-acting analgesia was 56.86%. The clinical effect was better than using Bupivacaini hydrochloride or Tramal alone (P < 0.05). It was shown in experiment that long-acting analgesic had obvious effects of topical analgesic and muscle relaxant.. Long-acting analgesic has significant effect in relieving the anorectal post-operational pain.

Topics: Adult; Aged; Analgesics, Non-Narcotic; Anti-Inflammatory Agents, Non-Steroidal; Drugs, Chinese Herbal; Female; Fissure in Ano; Hemorrhoids; Humans; Male; Middle Aged; Morphinans; Pain, Postoperative

A RP-HPLC method was developed to determine the concentrations of sinomenine HCl in serum and urine and its pharmacokinetics was studied in healthy volunteers. C18H37 column was eluted with the mobile phase of acetonitrile--0.01 mol.L-1 sodium phosphate monobasic--N, N, N', N'-tetramethylenediamine (46:54:0.22 v/v, pH 6.9) and the ultraviolet absorbance was monitored at 263 nm. Triazolan was used as internal standard. The calibration curves were linear in the range of 6-480 ng.ml-1 in serum and 0.06-3 micrograms.ml-1 in urine, with mean recoveries of 75.46% and 91.38% respectively. The lowest detectable limits were 4 ng.ml-1 in serum and 40 ng.ml-1 in urine and the RSD for the intra-day and inter-day were less than 5%. A single oral dose of 80 mg sinomenine HCl tablet was given to 8 healthy male volunteers. The concentrations of sinomenine HCl in serum and urine were determined. The serum concentration--time curve was found to fit a two-compartment open model with first order elimination. The pharmacokinetic parameters were: T1/2 alpha 0.791 +/- 0.491 h, T1/2 beta 9.397 +/- 2.425 h, Tmax 1.040 +/- 0.274 h, Cmax 246.604 +/- 71.165 ng.ml-1, AUC 2651.158 +/- 1039.050 ng.h.ml-1, CL 0.033 +/- 0.010 ng.ml-1.

Topics: Adult; Anti-Inflammatory Agents, Non-Steroidal; Chromatography, High Pressure Liquid; Humans; Male; Morphinans

Extracts of the plant Sinomenium acutum have been used safely since ancient times in Chinese medicine for treatment of rheumatic diseases, and the purified alkaloid, sinomenine, was recently shown to have anti-inflammatory and antirheumatic effects. This study describes the effects of sinomenine in the high responder ACI-->Lewis cardiac transplant model in which allograft rejection occurred at 5 days posttransplant. Treatment with sinomenine (15-30 mg/kg/day i.p.) or a subtherapeutic dose of cyclosporine (CsA, 1.5 mg/kg/day, i.m.) prolonged allograft survival only marginally (mean survival of 5.4 and 7.8 days, respectively). In contrast, the combination of sinomenine and CsA had a statistically significant synergistic effect, with a mean survival of 42.2 days (P < 0.001). Allografts harvested at day 5 from recipients treated with either sinomenine or CsA showed dense mononuclear cell infiltrates with widespread subepicardial infarcts, edema, and microvascular platelet and fibrin deposition. Immunohistologic analysis showed that intragraft leukocytes consisted of >75% macrophages with approximately 10-20% T cells and <5% B or NK cells. Mononuclear cell activation was shown by expression of IL-2R (CD25, 10-20%) and labeling for IL-2 (approximately 10%), and IFN-gamma (10-20%), as well as TNF-alpha (>50%) and iNOS (>50%), but only low levels of IL-4 or IL-10 (<5%). Intragraft endothelial cells were activated, as shown by upregulation of MHC class II antigen and ICAM-1 (CD54) compared with only basal levels in normal donors hearts. Combined sinomenine/CsA therapy significantly enhanced graft morphology, resulting in only mild mononuclear cell infiltration, and an absence of infarcts, platelets, or fibrin deposition. Though residual intragraft mononuclear cells at day 5, as in control grafts, consisted primarily of macrophages plus small numbers of IL-2R+ T cells, these cells lacked expression of IL-2, had only low levels of IFN-gamma, but showed dense labeling for IL-4 and IL-10. In addition, TNF-alpha and iNOS were reduced to basal levels and no endothelial cell activation was observed, despite high titers of endothelium-bound IgM, IgG, and C3. Mitogen-induced in vitro proliferation of rat thymocytes was also more effectively decreased by the sinomenine/CsA combination than by either agent alone. These studies demonstrate the therapeutic value of sinomenine in transplantation, and indicate that this agent has novel and interesting antimacrophage, T cell, and

Topics: Adjuvants, Immunologic; Animals; Cell Division; Cyclosporine; Drug Therapy, Combination; Graft Rejection; Graft Survival; Heart Transplantation; Immunosuppressive Agents; Male; Morphinans; Rats; Rats, Inbred ACI; Rats, Inbred Lew; T-Lymphocytes

The effects of treatment with sinomenine, a pure alkaloid extracted from the chinese medical plant Sinomenium acutum, were investigated in rat adjuvant arthritis (AA) and antigen-induced arthritis (AIA). In AA, long-term, intraperitoneal (i.p.) treatment induced significant improvement of arthritic score, hind paw swelling, body weight and erythrocyte sedimentation rate (ESR) beginning past the clinical peak of the disease. In-acute AIA, short and middle-term treatment with sinomenine around and following induction of arthritis induced a dose-dependent decrease of both joint swelling and ESR, starting after the peak of arthritis, and a significant reduction of joint destruction on day 3. There was no rebound of the arthritic signs following suspension of treatment. Long-term treatment of chronic AIA partially ameliorated clinical parameters and significantly counteracted joint destruction. Maximal plasma concentrations of 22.5 micrograms/ml, fast wash out (half-life 4.24 +/- 0.99 h; mean +/- S.E.M.) and no evidence of accumulation of sinomenine were observed following single or repeated i.p. injection of 150 mg/kg. In vitro, sinomenine markedly inhibited proliferation of synovial fibroblasts from AIA or normal rats, both at rest and following activation with either transforming growth factor beta 2 (TGF-beta 2) or interleukin-1 beta (IL-1 beta). The effect was dose-dependent and half-maximal inhibition of proliferation occurred at 20.6 micrograms/ml, that is, within the in vivo therapeutic range of the drug. Late therapeutic effects of sinomenine in rat arthritic models despite early start of treatment may be related to its antiproliferative effects on synovial fibroblasts in addition to its previously reported anti-inflammatory properties.

Topics: Adjuvants, Immunologic; Animals; Anti-Inflammatory Agents, Non-Steroidal; Antigens; Arthritis, Experimental; Cell Division; Cells, Cultured; Cytokines; Disease Models, Animal; Female; Fibroblasts; Morphinans; Rats; Rats, Inbred Lew; Stimulation, Chemical; Synovial Membrane

We report a case of drug eruption caused by the crude drug Boi. A 41-year-old female patient had been diagnosed with chronic rheumatoid arthritis in the department of internal medicine. After ingestion of a decoction of the crude drug Boi for the alleviation of arthralgia, a slight fever developed, which was followed by systemic edematous erythema with itching. HPLC showed that the main components of the crude drug Boi are sinomenine and magnoflorine. The results of patch tests were negative for all oral drugs that the patient had been taking. Oral ingestion tests showed that the patient showed positive reactions to the as-is Boi boiling-water decoction and 1/10-volume sinomenine. Based on this, the drug eruption was judged to be caused by sinomenine. It is considered the first time that the causative component of a drug eruption was confirmed by oral ingestion tests with components of a crude drug of Kampo medicine (Sino-Japanese traditional medicine).

Topics: Adult; Anti-Inflammatory Agents, Non-Steroidal; Aporphines; Arthritis, Rheumatoid; Chromatography, High Pressure Liquid; Drug Eruptions; Drugs, Chinese Herbal; Erythema; Female; Humans; Morphinans; Plant Roots; Plant Shoots; Pruritus

The effects of sinomenine (7,8-didehydro-4-hydroxy-3,7-dimethoxy-17-methyl- 9 alpha,13 alpha,14 alpha-morphinan-6-one), a pure alkaloid extracted from the Chinese medical plant Sinomenium acutum, on different macrophage capacities were investigated in vitro using resident mouse peritoneal macrophages and the macrophage-like cell line RAW 264.7. Sinomenine markedly decreased prostaglandin E2 and leukotriene C4 synthesis of macrophages stimulated by zymosan or calcium ionophore and also significantly inhibited the nitric oxide production of RAW 264.7 cells activated by interferon-gamma/lipopolysaccharide. It can be considered that these effects are part of the analgesic, anti-inflammatory, and antirheumatic mechanisms of sinomenine.

Topics: Alkaloids; Animals; Cell Line; Depression, Chemical; Dinoprostone; Eicosanoids; Female; In Vitro Techniques; Leukotriene C4; Macrophages; Mice; Mice, Inbred DBA; Morphinans; Nitric Oxide; Spectrophotometry, Ultraviolet

The effect on lymphocyte proliferation of sinomenine, a pure alkaloid extracted from the Chinese medical plant Sinomenium acutum was investigated in vitro using mouse spleen cells and human peripheral blood mononuclear cells. It could be demonstrated that sinomenine markedly inhibited [3H]thymidine incorporation in mouse spleen cells activated with concanavalin A (IC50 = 400 microM) or by two-way mixed lymphocyte culture (IC50 = 60 microM) and also in human peripheral blood mononuclear cells activated with phytohemagglutinin, 12-O-tetradecanoylphorbol-13-acetate plus ionomycin, or mixed lymphocyte culture (IC50 ranging from 34 to 129 microM). Time kinetic experiments revealed that sinomenine was effective only when added within the first 48 h after the onset of mixed lymphocyte culture, which lasted for 5 days. Inhibition of lymphocyte proliferation by sinomenine was reversible. Accordingly, the drug showed no direct cytotoxicity in our cellular systems and had no inhibitory effect on the proliferation of the cytokine-independent growth of the human leukaemic T-cell lymphoblast cell line Jurkat. It can be considered that these anti-proliferative effects are part of the anti-inflammatory and anti-arthritic mechanisms of sinomenine obvious in clinical trials.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Division; Cell Survival; Concanavalin A; Depression, Chemical; Female; Humans; Ionomycin; Leukocytes, Mononuclear; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Mice; Mice, Inbred C57BL; Mice, Inbred DBA; Morphinans; Phytohemagglutinins; Spleen; Tetradecanoylphorbol Acetate

Sinomenine, an epimorphinan alkaloid, was tested for protecting hepatitis induced by lipopolysaccharide (LPS) in galactosamine (GalN)-sensitized mice. Sinomenine protected against the hepatic injuries in the dose range of 10-100 mg/kg in a dose-dependent manner and suppressed the production of tumor necrosis factor (TNF), which appeared in serum earlier than aminotransferases in GalN/LPS-treated mice. Sinomenine significantly suppressed the in vitro production of superoxide anion and hydrogen peroxide in the macrophage cultures stimulated with phorbol 12-myristate acetate. It is discussed that sinomenine prevents GalN/LPS-treated hepatic failure by suppressing TNF production and/or reactive oxygen generation.

Topics: Analgesics; Animals; Anti-Inflammatory Agents, Non-Steroidal; Free Radical Scavengers; Galactosamine; Hepatitis, Animal; Lipopolysaccharides; Macrophage Activation; Male; Mice; Mice, Inbred ICR; Morphinans; Reactive Oxygen Species; Tumor Necrosis Factor-alpha

Acute myocardial ischemia and reperfusion injury of rats were produced by ligating the left coronary artery for 15 min and reopening. The myocardial calcium contents were increased from 3.53 +/- 0.58 mumol/g dry wt in sham operation group to 6.02 +/- 1.19 mumol/g dry wt with reducing SOD/MDA ratio and showing ventricular extrasystole (VE), ventricular tachycardia (VT), and ventricular fibrillation (VF). Sinomenine (Sin) and verapamil (Ver) infusion 15 min before ischemia attenuated the elevated calcium contents to the level of the sham operation group, increased SOD/MDA ratio, and produced antagonistic effects on VE, VT, VF. These improvements indicated that Sin, similar to Ver, prevented myocardial injury by lowering intracellular Ca2+ accumulation.

Topics: Animals; Calcium; Free Radicals; Heart Rate; In Vitro Techniques; Morphinans; Myocardial Reperfusion Injury; Myocardium; Rats; Rats, Wistar

Pharmacokinetics of sinomenine in rabbit was studied by reverse-phase HPLC method. After i.v. injection of 20 mg/kg of sinomenine to rabbits, the pharmacokinetic characteristics were found to fit a two-compartment open model.

Topics: Animals; Chromatography, High Pressure Liquid; Female; Injections, Intravenous; Male; Morphinans; Rabbits

Topics: Animals; Carbon; Cyclic AMP; Cyclic GMP; Immunosuppressive Agents; Macrophages; Metabolic Clearance Rate; Morphinans; Phagocytosis; Rabbits

Topics: Acetylcholine; Action Potentials; Animals; Depression, Chemical; Female; Guinea Pigs; In Vitro Techniques; Male; Morphinans; Myocardial Contraction; Tetrodotoxin

Topics: Action Potentials; Animals; Barium; Guinea Pigs; In Vitro Techniques; Magnesium; Morphinans; Papillary Muscles

Topics: Anti-Inflammatory Agents, Non-Steroidal; Chromatography, Thin Layer; Densitometry; Medicine, Chinese Traditional; Morphinans; Plants, Medicinal; Seasons

Topics: Animals; Anti-Arrhythmia Agents; Female; Guinea Pigs; Heart Rate; In Vitro Techniques; Male; Mice; Morphinans; Rabbits; Rats

Sinomenine, an epimorphinan alkaloid, was tested for the immunosuppressive effect in mice. This compound produced a decrease of plaque-forming cells (PFC) to a T cell-dependent antigen, sheep red blood cells, in vivo. The depression of the PFC response induced with sinomenine was dose and time dependent. On the other hand, it failed to suppress the PFC response to a T cell-independent antigen, lipopolysaccharide. The immunosuppressive dose of sinomenine did not alter the cellularity of spleen, thymus, bone marrow and peripheral blood leucocytes, the DNA synthesis activity of bone marrow cells nor the proliferative responses of spleen cells induced by T cell and B cell mitogens in unprimed mice. These data suggest a selective effect of sinomenine on lymphoid cells. This compound has a potential for use in studies of immuno-deficiencies or clarifying some aspect of immunity.

Topics: Animals; Antibody Formation; DNA; Immunosuppressive Agents; In Vitro Techniques; Lymphoid Tissue; Male; Mice; Mice, Inbred Strains; Morphinans; T-Lymphocytes

Topics: Animals; Carbon Radioisotopes; Male; Mice; Morphinans; Muscle Relaxants, Central; Tissue Distribution

Topics: Antihypertensive Agents; Central Nervous System; Mice; Morphinans; Nervous System; Pharmacology; Plants, Medicinal; Research

Topics: Blood Pressure; Blood Pressure Determination; Dogs; Morphinans; Pharmacology; Plants, Medicinal; Rabbits; Rats; Research

Topics: Alkaloids; Conditioning, Classical; Morphinans; Pharmacology; Reflex; Toxicology

Topics: Alkaloids; Analgesia; Biological Assay; Mice; Morphinans; Pain; Pharmacology; Promethazine; Research; Toxicology

Topics: Alkaloids; Animals; Anura; Apnea; Appetite; Blood Cells; Blood Pressure; Dogs; Dyspnea; Gastrointestinal Tract; Haplorhini; Heart; Hypnotics and Sedatives; Ileum; Mice; Morphinans; Muscle, Smooth; Muscles; Muscular Diseases; Pharmacology; Rabbits; Rats; Research; Seizures; Tachycardia

Topics: Alcohols; Alkaloids; Morphinans

Topics: Alcohols; Alkaloids; Hydrolysis; Morphinans

Topics: Alcohols; Alkaloids; Morphinans

Topics: Alkaloids; Morphinans

Topics: Alkaloids; Morphinans

Topics: Alkaloids; Animals; Body Fluids; Dogs; Heparin; Histamine; Humans; Morphinans; Thiopental

Topics: Alkaloids; Animals; Histamine; Morphinans; Rats; Trypsin; Trypsin Inhibitors

Topics: Alkaloids; Animals; Histamine; Mast Cells; Methylamines; Morphinans; Rats

Topics: Alkaloids; Histamine; Histamine Release; Humans; Morphinans

Topics: Alkaloids; Histamine; Humans; Morphinans; Patient Discharge