ABSTRACT
Corydalis hendersonii(CH) is a Tibetan folk medicine with the functions of clearing heat, detoxifying, cooling blood, checking diarrhea, and lowering blood pressure. It is often used to treat high altitude polycythemia, vasculitis, peptic ulcer, and diarrhea. Nine compounds were separated from the ethanol extract of CH by silica gel, ODS, Sephadex LH-20 chromatography and semi-preparative HPLC. Their structures were identified as hendersine H(1),hendersine I(2), dehydrocheilanthifoline(3), protopine(4), izmirine(5), 6,7-methylenedioxy-1(2H)-isoquinolinone(6), icariside D_2(7), ethyl 4-(ß-D-glucopyranosyloxy)-3-methoxybenzoate(8), 3-hydroxy-4-methoxybenzoic acid(9), respectively, by the spectroscopic data analysis and comparison with those in the literature. Among them, compounds 1 and 2 are new isoquinoline alkaloids, and compounds 7-9 are reported the first time for Corydalis. The hypoglycemic model of H9c2 cardiomyocytes and the inflammatory model of H9c2 cardiomyocytes induced by conditional supernatant were employed to determine the activities of the above compounds. The results showed that 20 µmol·L~(-1) compound 1 had a protective effect on H9c2 cardiomyocytes and 10 µmol·L~(-1) compounds 4 and 5 inhibited H9c2 cardiomyocyte inflammation induced by conditional supernatant.
Subject(s)
Alkaloids , Corydalis , Humans , Corydalis/chemistry , Alkaloids/pharmacology , Alkaloids/chemistry , Inflammation , Spectrum Analysis , Isoquinolines/pharmacologyABSTRACT
Hepatic lipid deposition is one of the basic manifestations of obesity, and nowadays pharmacological treatment is the most important tool. Punicalagin(PU), a polyphenol derived from pomegranate peel, is a potential anti-obesity substance. In this study, 60 C57BL/6J mice were randomly divided into a normal group and a model group. After establishing a model of simple obesity with a high-fat diet for 12 weeks, the successfully established rat models of obesity were then regrouped into a model group, an orlistat group, a PU low-dose group, a PU medium-dose group, and a PU high-dose group. The normal group was kept on routine diet and other groups continued to feed the high-fat diet. The body weight and food intake were measured and recorded weekly. After 8 weeks, the levels of the four lipids in the serum of each group of mice were determined by an automatic biochemical instrument. Oral glucose tole-rance and intraperitoneal insulin sensitivity were tested. Hemoxylin-eosin(HE) staining was applied to observe the hepatic and adipose tissues. The mRNA expression levels of peroxisome proliferators-activated receptor γ(PPARγ) and C/EBPα were determined by real-time quantitative polymerase chain reaction(Q-PCR), and the mRNA and protein expression levels of adenosine 5'-monophosphate-activated protein kinase(AMPK), anterior cingulate cortex(ACC), and carnitine palmitoyltransferase 1A(CPT1A) were determined by Western blot. Finally, the body mass, Lee's index, serum total glyceride(TG), serum total cholesterol(TC), and low-density lipoprotein cholesterol(LDL-C) levels were significantly higher and high-density lipoprotein cholesterol(HDL-C) levels were significantly lower in the model group as compared with the normal group. The fat deposition in the liver was significantly increased. The mRNA expression levels of hepatic PPARγ and C/EBPα and the protein expression level of ACC were increased, while the mRNA and protein expression levels of CPT-1α(CPT1A) and AMPK were decreased. After PU treatment, the above indexes of obese mice were reversed. In conclusion, PU can decrease the body weight of obese mice and control their food intake. It also plays a role in the regulation of lipid metabolism and glycometabolism metabolism, which can significantly improve hepatic fat deposition. Mechanistically, PU may regulate liver lipid deposition in obese mice by down-regulating lipid synthesis and up-regulating lipolysis through activation of the AMPK/ACC pathway.
Subject(s)
AMP-Activated Protein Kinases , PPAR gamma , Rats , Mice , Animals , Mice, Obese , AMP-Activated Protein Kinases/genetics , AMP-Activated Protein Kinases/metabolism , PPAR gamma/genetics , PPAR gamma/metabolism , Mice, Inbred C57BL , Liver/metabolism , Obesity/drug therapy , Obesity/genetics , Body Weight , Lipid Metabolism , Diet, High-Fat/adverse effects , Lipids , CholesterolABSTRACT
In this study, the compound search was completed through SciFinder and CNKI databases, and the drug-like properties were screened in FAFdrugs4 and SEA Search Server databases. In addition, based on the target sets related to acute myocardial ischemia(AMI) searched in disease target databases such as OMIM database, GeneCards database and DrugBank, a network diagram of chemical component-target-pathway-disease was established via Cytoscape to predict the potential active components of Corydalis Herba, a traditional Tibetan herbal medicine which derived from the aerial parts of Corydalis hendersonii and C. mucronifera against AMI. A protein-protein interaction(PPI) network was constructed through the STRING database and the core targets in the network were predicted. And the enrichment analyses of core targets were completed by DAVID database and R software. Furthermore, a molecular docking method was used to verify the binding of the components with core targets using softwares such as Autodock Vina. The present results showed that there were 60 compounds related to AMI in Corydalis Herba, involving 73 potential targets. The GO functional enrichment analysis obtained 282 biological processes(BP), 49 cell components(CC) and 78 molecular functions(MF). KEGG was enriched into 85 pathways, including alcoholism pathway, endocrine resistance pathway, calcium signaling pathway, cAMP signaling pathway, vascular endothelial growth factor signaling pathway and adrenergic signaling transduction pathway of myocardial cells. The results of network topology analysis showed that the key components of anti-AMI of Corydalis Herba might be tetrahydropalmatine, etrahydrocolumbamine, N-trans-feruloyloctopamine, N-cis-p-coumaroyloctopamine, N-trans-p-coumaroylnoradrenline and N-trans-p-coumaroyloctopamine, and their core targets might be CDH23, SCN4 B and NFASC. The results of molecular docking showed that the key components of Corydalis Herba had stable binding activity with the core targets. This study provides reference for further elucidation of the pharmacological effects of Corydalis Herba against AMI, subsequent clinical application, and development.
Subject(s)
Corydalis , Drugs, Chinese Herbal , Myocardial Ischemia , Drugs, Chinese Herbal/pharmacology , Medicine, Tibetan Traditional , Molecular Docking Simulation , Myocardial Ischemia/drug therapy , Vascular Endothelial Growth Factor AABSTRACT
Cardiovascular diseases seriously endanger human health and life. The accompanying myocardial injury has been a focus of attention in society. Chinese medicine,serving as a natural and precious reservoir for the research and development of new drugs,is advantageous in resisting myocardial injury due to its multi-component,multi-pathway,and multi-target characteristics. In recent years,with the extensive application of culture method for isolated cardiomyocytes,a cost-effective,controllable in vitro model of cardiomyocyte injury with uniform samples is becoming a key tool for mechanism research on cardiomyocyte injury and drug development.A good in vitro model can reduce experimental and manpower cost,and also accurately stimulate clinical changes to reveal the mechanism. Therefore,the selection and establishment of in vitro model are crucial for the in-depth research. This study summarized the modeling principles,evaluation indicators,and application of more than ten models reflecting different clinical conditions,such as injuries induced by hypoxia-reoxygenation,hypertrophy,oxidative stress,inflammation,internal environmental disturbance,and toxicity. Furthermore,we analyzed advantages and technical difficulties,aiming to provide a reference for in-depth research on myocardial injury mechanism and drug development.
Subject(s)
Apoptosis , Myocytes, Cardiac , Cell Hypoxia , Humans , Myocardium , Oxidative StressABSTRACT
Globally, the proliferation of shrubs within grasslands stimulates soil phosphorus (P) cycling and increases topsoil P storage beneath their canopies. However, little is known regarding the impact of shrub encroachment on subsoil P storage, and whether shrubs mediate changes in soil stoichiometry, like increasing P cycling. In grazed meadows on the Tibetan Plateau, soil and roots were sampled to 1 m depth in shrubby Hippophae rhamnoides ssp. sinensis groves and the surrounding grassy areas. Shrubs had higher P content than grasses, but lower C:P ratios in their leaves, litter, and roots. Similarly, shrubs had higher microbial P content than grasses, but lower microbial biomass C:P and N:P ratios in the soil. The larger microbial P stock in the 1 m of soil beneath shrubs responded to the larger root P stock there as well. Thus, both the plants and microbes acquired more P in shrubby areas than in grassy areas by accelerating P mineralization. The greater net production of available P in the topsoil and the synthesis of microbial P throughout the profile under shrubs increased the P solubility. Total P, inorganic P, and organic P stocks were lower under shrubs than under grasses in the top 1 m of soil. This decrease in soil P storage beneath shrubs is most likely attributable to P leaching due to higher P solubility, heavy rainfall, and larger soil gaps. Moreover, shrubs also had larger plant biomass P stock compared to grasses, and thus the depletion of P from the top 1 m of soil was further magnified via plant biomass removal. We concluded that shrubs increase P cycling to overcome the stoichiometric imbalance between their P requirement and the supply in the soil, and the fast P cycling under shrubs magnify P depletion within the rooted soil depth in alpine meadows.
Subject(s)
Phosphorus , Soil , Biomass , Carbon , Ecosystem , Grassland , TibetABSTRACT
Zerumbone(ZER), one of humulane-type sesquiterpenoids, showed its unique advantage against tumor activities. The main underlying mechanisms include inhibiting the growth and proliferation of cancer cells, inducing apoptosis of cancer cells and differentiation of cancer cells, regulating immune function, inhibiting invasion and metastasis of cancer cells, and reversing multidrug resistance of cancer cells. Studies on ZER focusing its cytotoxic or anti-tumor is one of hot topic. Currently, with the increasing studies on ZER, the clarified mechanisms are getting rich. The present paper describes a summary of its anti-tumor mechanism of action and helps to provide significant reference to more in-depth research.
Subject(s)
Antineoplastic Agents/pharmacology , Sesquiterpenes/pharmacology , Apoptosis/drug effects , Cell Line, TumorABSTRACT
Bawei Chenxiang Powder is a traditional Tibetan folk medicine formula, consisting of resinous wood of Aquilaria sinensis, kernel of Myristica fragrans, fruit of Choerospondias axillaris, travertine, resin of Boswellia carterii or B. bhaw-dajiana, stem of Aucklandia lappa, fruit of Terminalia chebula(roasted), and flower of Gossampinus malabarica. It has the function of clearing heart heat, nourishing heart, tranquilizing mind, and inducing resuscitation, which has been used for the treatment of coronary heart disease and angina pectoris. Modern research shows that the medicine materials of this formula mainly contain terpenoids like sesquiterpenes and triterpenes and polyphenols like flavonoids, lignans, and tannins, displaying some pharmacological activities such as anti-myocardial ischemia, anti-cerebral ischemia, and spatial learning and memory promotion. This review summaries the traditional uses, chemical constituents, and pharmacological activities research progress, hopefully to provide a reference for clarification of its pharmacological active ingredients.
Subject(s)
Drugs, Chinese Herbal , Terminalia , Flavonoids , Medicine, Tibetan Traditional , TibetABSTRACT
Saussurea lappa originates in India, and now mainly grow in Yunnan, Sichuan and other places in China. It is one of the commonly used traditional herbal medicines in Tibet and other minority regions, with effects in regulating qi to relieve pain and invigo-rating spleen to promote food. It has been used in clinic for gastrointestinal diseases, such as Qi stagnation syndrome of spleen and stomach, diarrhea and tenesmus. More than 200 compounds have been identified from S. lappa. Among them, sesquiterpenoids attracted much attention. In terms of the number of compounds, eudesmanetype is dominant, guaiane and germacranetypes have also been reported frequently. Pharmacological studies have involved extracts, volatile oils and monomeric components represented by dehydrocostus lactone. Anti-tumor, anti-inflammatory and anti-bacterial effects on digestive system have attracted great attention. However, due to the complex sources of S. lappa and widely used in clinical practice, there is few research progress on relevant chemical constituents and pharmacological activities. This paper systematically summarizes terpenes and the pharmacological effects of S. lappa, in order to provide basis for further studies and clinical applications.
Subject(s)
Saussurea , Sesquiterpenes , China , Plant Extracts , Terpenes , TibetABSTRACT
Usnic acid and its derivatives, a group of organic molecules with great importance, are characteristic to lichens, possessing pharmacological activities such as anti-virus, anti-bacteria, anti-humor, anti-inflammatory, analgesic, and anaesthetic effects. Many of them have been widely used as medicine, but also bring side effects such as dermatitis and liver damages. In the past decades, great efforts by isolation, organic synthesis, and structure modification methods were put on discovery of UA derivatives with higher biological activities or less side effects. This paper describes herein the most progress on natural sources, isolation and structure elucidation, structural characteristics, synthesis and modification results, pharmacological activities and toxicities of UA and its derivatives, hopefully to provide valuable reference for further research.
Subject(s)
Benzofurans/chemistry , Benzofurans/pharmacology , Lichens/chemistry , Biological ProductsABSTRACT
Meconopsis horridula is one of alpine plants belonging to family Papaveraceae, mainly distributed in Himalaya Range area. M. horridula is a rare alpine flower, and is a kind of traditional Tibetan medicine, which has been included in more than 40 compound formulae, having efficacies of clearing away heat and alleviating pain, activating blood circulation to remove stasis, traditionally used for the treatment of fractures, injuries, and chest and back pains. Modern research shows that the whole plant of M. horridula contains alkaloids, flavonoids, and terpenes, and its pharmacological activities including antitumor, antivirus and myocardial protection etc. However, due to various factors, the current research of M.horridula still faces many challenges. This paper summaries herein a progress of MH on its ecological resources, traditional uses, and studies on chemical constituents and pharmacological effects, hopefully to provide a useful reference for the ecological protection and applications.
Subject(s)
Alkaloids/pharmacology , Flavonoids/pharmacology , Medicine, Tibetan Traditional , Papaveraceae/chemistry , Phytochemicals/pharmacology , Terpenes/pharmacologyABSTRACT
Sixteen compounds were isolated from lichen Usnea longissima using of various chromatographic techniques including silica gel, Sephadex LH-20, ODS, and semi-preparative HPLC. By spectroscopic data analyses, their structures were identified by as useanol(1), lecanorin(2), 3-hydroxy-5-methylphenyl 2-hydroxy-4-methoxy-6-methylbenzoate(3), lecanorin E(4), 3'-methylevernic acid(5), evernic acid(6), barbatinic acid(7), 3,7-dihydroxy-1,9-dimethyldibenzofuran(8), orcinol(9), O-methylorcinol(10), methyl orsellinate(11), methyl everninate(12), 2,5-dimethyl-1,3-benzenediol(13), 2-hydroxy-4-methoxy-3,6-dimethyl benzoic acid(14), ethyl everninate(15), and ethyl 2,4-dihydroxy-6-methylbenzoate(16). Compound 1 was obtained as a natural product for the first time, and 3,4, 8,10,12, and 13 were isolated from Usneaceae family for the first time. Compound 1, 8, and 13 showed significant anti-inflammatory activity against NO production in RAW 267.4 cells with IC50 values of 6.8, 3.9 and 4.8 µmolâ¢L⻹, respectively, compared with the positive controls curcumin(IC50 15.3 µmolâ¢L⻹) and indomethacin(IC50 42.9 µmolâ¢L⻹).
Subject(s)
Phenols/isolation & purification , Usnea/chemistry , Animals , Chromatography, High Pressure Liquid , Mice , RAW 264.7 CellsABSTRACT
Litsea cubeba is one of aromatic medicinal plant belonging to family Lauraceae. The roots, stems and fruits of L. cubeba have been widely applied as folk medicines in some districts in China for relieving rheumatism and cold, regulating Qi (meridian) to alleviate pain. Previous studies revealed that this species contains major alkaloids, in specific aporphines, and minor flavonoids, lignans as well. Related pharmacological investigations demonstrated its activities and clinical applications on cardiovascular diseases, anti-cancer, against rheumatoid arthritis, relieving asthma and anti-allergic effects, as anti-oxidants, and so on. As an effort for further exploration of this bioactive ingredients and potential drug development, this paper summarizes most phytochemical and pharmacological results. Further, future prospects are also included.
Subject(s)
Litsea/chemistry , Plant Extracts/chemistry , Plant Extracts/pharmacology , Plants, Medicinal/chemistry , Animals , Drug Therapy , Humans , Molecular StructureABSTRACT
Persicae Semen (PS), a traditional Chinese medicine, has been widely used for the syndrome of blood stasis in China since the Eastern Han Dynasty. In the present study, we developed an HPLC-UV fingerprint analysis method for the quality control of PS. The HPLC fingerprint was performed on Shimadzu Inertsil C18 column (4.6 mm x 250 mm, 5 microm) at 35 degrees C. The mobile phases were composed of acetonitrile and water using a gradient elution. The flow rate was 1.0 mL x min(-1), and the detection wavelength was set at 210 nm. The fingerprint method was validated according to the Guidelines for Traditional Chinese Medicine Injection Fingerprint, and applied to determine 41 batches representative herbs collected from Xinjiang of China. The chromatographic peaks were characterized by UPLC-Q-TOF-MS, and nine of them were identified by comparison with the literature and/or reference standards. In order to classify and assess the samples, hierarchical clustering analysis and partial least square discriminant analysis were performed based on the common chromatographic peaks, and the samples were geographically classified into two classes, with six chemical compounds as classification markers which were significantly different between the two classes (P < 0.05).
Subject(s)
Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/chemistry , Prunus/chemistry , Seeds/chemistry , China , Chromatography, High Pressure Liquid/instrumentation , Drugs, Chinese Herbal/isolation & purification , Quality ControlABSTRACT
It was estimated that about 428 species of genus Corydalis are distributed all worldwide, with about 298, especially 10 groups and 219 species being uniquely spread in China. The genus Corydalis have been widely employed as folk medicines in China, especially as traditional Tibetan medicines, for treatment of fever, hepatitis, edema, gastritis, cholecystitis, hypertension and other diseases. The phytochemical studies revealed that isoquinoline alkaloids are its major bioactive ingredients. The extensive biological researches suggested its pharmacological activities and clinic applications against cardiovascular diseases and central nervous system, antibacterial activities, analgesic effects, anti-inflammatory, anti-oxidation and anti-injury for hepatocyte, and so on. As an effort in promoting the research of pharmacodynamic ingredients, this article presents an overview focusing on the distribution, phytochemical and pharmacological results of Corydalis species that have been applied in traditional Tibetan medicinal, hopefully to provide a reference for the new Tibetan medicine development from Corydalis plant resource.
Subject(s)
Anti-Infective Agents/chemistry , Anti-Infective Agents/pharmacology , Corydalis/chemistry , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacology , Alkaloids/chemistry , Alkaloids/pharmacology , Animals , Corydalis/classification , Humans , Mice , Molecular Structure , PhytotherapyABSTRACT
A phytochemical investigation on the roots and stems of Litsea cubeba led to the isolation of seven isoquinolone alkaloids. By spectroscopic analysis and comparison of their 1H and 13C-NMR data with those in literatures, these alkaloids were identified as (+)-norboldine (1), (+)-boldine (2), (+)-reticuline (3), (+)-laurotetanine (4), (+)-isoboldine (5), (+)-N-methyl-laurotetanine (6), and berberine (7), respectively. Among them, 7 was isolated from the genus for the first time. The evaluation of these compounds showed weak anti-inflammatory activity against NO production in RAW 267.4 and BV-2 cells.
Subject(s)
Drugs, Chinese Herbal/chemistry , Litsea/chemistry , Drugs, Chinese Herbal/isolation & purification , Molecular Structure , Plant Roots/chemistry , Plant Stems/chemistry , Spectrometry, Mass, Electrospray IonizationABSTRACT
OBJECTIVE: To investigate the role of anti-inflammatory effect and it's mechansim of Qufeng Zhitong capsule on the collagen-induced arthritis rat through the study of arthritis, joint inflammation and joint erosions. METHODS: Fifty male SD rat were assigned to 5 groups: normal control group, arthritis model group, and Qufeng Zhitong capsule treatment group (including low, medium and high doses), each group contains 10 rats. Arthritis were induced by immunization with type II collagen (CII) in complete Freund's adjuvant (CFA). After administered Qufeng Zhitong capsule 2 weeks, the volume of back foot of arthritis rat were detected, and the histology changes of the ankle joints of the arthritis rat were studied. The serum levels of IL-10 and TNF-alpha were measured by ELISA. RESULTS: After treated with Qufeng Zhitong capsule, the swelling of the paw of the rats were significantly decreased compared with the controls. Histology study demonstrated that the Qufeng Zhitong capsule treatment could reduce the synovium hyperplasia and inflammatory cell infiltration, the inhibitory effects were in accordance with the doses of the drug. The serum levels of pro-inflammatory cytokine of TNF-alpha in the arthritis rat were not changed significantly, while the serum levels of anti-inflammatory cytokine of IL-10 were elevated significantly after treated with Qufeng Zhitong capsule. CONCLUSION: Qufeng Zhitong capsule can inhibit the synovium hyperplasia and cartilage erosion by stimulating the serum levels of anti-inflammatory cytokine of IL-10 and suggest it can be a candidate of treatment of rheumatoid arthritis.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Arthritis, Experimental/drug therapy , Drugs, Chinese Herbal/pharmacology , Animals , Ankle Joint/drug effects , Ankle Joint/pathology , Anti-Inflammatory Agents/therapeutic use , Arthritis, Experimental/blood , Arthritis, Experimental/pathology , Capsules , Cell Proliferation/drug effects , Drugs, Chinese Herbal/therapeutic use , Interleukin-10/blood , Male , Rats , Rats, Sprague-Dawley , Synovial Membrane/drug effects , Synovial Membrane/pathology , Tumor Necrosis Factor-alpha/bloodABSTRACT
OBJECTIVE: To study the effects of sinomenine on apoptosis in cutured neonatal rat cardiomyocytes induced by H2O2 and its possible mechanism. METHOD: 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. RESULT: 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)). CONCLUSION: 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.
Subject(s)
Apoptosis/drug effects , Hydrogen Peroxide/toxicity , Morphinans/pharmacology , Myocytes, Cardiac/cytology , Myocytes, Cardiac/drug effects , Animals , Gene Expression Regulation/drug effects , L-Lactate Dehydrogenase/metabolism , Malondialdehyde/metabolism , Myocytes, Cardiac/metabolism , NF-kappa B/metabolism , Rats , Rats, Wistar , Superoxide Dismutase/metabolismABSTRACT
OBJECTIVE: To study the effect of total flaveos of Gymostemma pentaphyllum on the protein expression of apoptosis-associated Fas/FasL gene and tumor necrosis factor-alpha (TNF-alpha) concentration in cultured neonatal rat cardiomyocytes with hypoxia-reoxygenation (H/R). METHOD: A cultured primary neonatal rat cardiomyocytes model with H/R was erected, experiments were divided into six groups, (1)control group, (2)H/R group, (3)15 mg x L(-1) TFG plus H/R group, (4)45 mg x L(-1) TFG plus H/R group, (5) 105 mg x L(-1) TFG plus H/R group, (6)105 mg x L(-1) TFG group. TNF-aconcentration in cultured cardiomyocytes with H/R, was determined by ELISA method, the protein expression of Fas/FasL genes were estimated by immunohisto-chemistry. RESULT: After cardiomyocytes were made with H/R, Compared with control group, the positive expression index (PEI) of Fas/FasL proteins in cardiomyocytes increased significantly, Compared with H/R groups, the PEI of Fas/FasL proteins were lower significantly in groups with different dosages of TFG (P < 0.05). TFG inhibited the secretion of TNF-alpha from myocardial cells and increased the survival rate of myocardial cells. CONCLUSION: The protein expression of apoptosis-associated Fas/FasL genes increased during H/R. The TFG can protect myocardium against H/R injury by decreasing the production of TNF-alpha, downregulating the protein expression of Fas/FasL genes, and then inhibiting myocyte apoptosis.