摘要
通过网络药理学、分子对接及动物实验探究麝香通心滴丸(Shexiang Tongxin Dropping Pills,STDP)防治糖尿病心肌病(diabetic cardiomyopathy,DCM)的作用机制。借助BATMAN、TCMSP、GeneCards等数据库筛选STDP防治DCM的活性成分及作用靶点,利用STRING数据库及Cytoscape软件构建STDP防治DCM作用靶点的蛋白-蛋白相互作用(protein-protein inte-raction,PPI)网络及“药物-活性成分-靶点”网络,通过DAVID数据库对作用靶点进行GO和KEGG富集分析,采用AutoDock Vina软件对关键信号通路核心受体蛋白与其对应活性成分进行分子对接验证。采用高脂喂养联合腹腔注射链脲佐菌素的方法建立DCM大鼠模型,分为对照组、模型组、STDP低剂量组(20 mg·kg^(-1))、STDP高剂量组(40 mg·kg^(-1))、二甲双胍组(200 mg·kg^(-1));连续给药8周后超声心动图检测各组大鼠心功能,苏木精-伊红(hematoxylin-eosin,HE)染色观察各组大鼠心肌病理改变,天狼猩红染色检测各组大鼠心肌胶原纤维沉积,WGA染色检测心肌肥大程度,蛋白免疫印迹法检测各组大鼠心肌组织中p38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38)、磷酸化p38(phosphorylation-p38,p-p38)、c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)、磷酸化JNK(phosphorylation-JNK,p-JNK)、caspase-3及C-caspase-3蛋白的表达。网络药理学筛选得到STDP活性成分199种,对应靶点1655个,与463个DCM疾病靶点取交集,得到134个STDP防治DCM的潜在作用靶点,并筛选出糖尿病并发症中的AGE-RAGE信号通路;分子对接结果显示miltirone、dehydromiltirone和tryptanthrin与RAGE均有较好的结合力。动物实验结果证实,STDP可有效保护DCM大鼠心功能,与模型组比较,STDP低、高剂量组p-p38、p-JNK、C-caspase-3蛋白表达水平均显著降低。综上,SDTP可能通过影响AGE-RAGE信号通路发挥对DCM大鼠心功能的保护作用。
This study aimed to explore the mechanism of Shexiang Tongxin Dropping Pills(STDP)in treating diabetic cardiomyopathy(DCM)based on network pharmacology,molecular docking,and animal experiments.BATMAN,TCMSP,and GeneCards were searched for the active ingredients and targets of STDP against DCM.STRING and Cytoscape were used to build the protein-protein interaction(PPI)network and"drug-active ingredient-target"network.Gene Ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis of the targets were carried out based on DAVID.The molecular docking of key receptor proteins with corresponding active ingredients was performed using AutoDock Vina.The rat model of DCM was established by a high-fat diet combined with intraperitoneal injection of streptozotocin.Rats were assigned into control,model,low-(20 mg·kg^(-1))and high-dose(40 mg·kg^(-1))STDP,and metformin(200 mg·kg^(-1))groups.After 8 weeks of continuous administration,the cardiac function,myocardial pathological changes,and myocardial collagen fiber deposition of rats in each group were detected by echocardiography,hematoxylin-eosin(HE)staining,and Sirius red staining,respectively.The myocardial hypertrophy was detected by WGA staining.The expression levels of p38 mitogen-activated protein kinase(p38),phosphorylation-p38(p-p38),c-Jun N-terminal kinase(JNK),phosphorylation-JNK(p-JNK),caspase-3,and C-caspase-3 in the myocardial tissue of rats in each group were measured by Western blot.The network pharmacology predicted 199 active ingredients and 1655 targets of STDP and 463 targets of DCM.One hundred and thirty-four potential targets of STDP for treating DCM were obtained,and the AGE-RAGE signaling pathway in diabetic complications was screened out.Molecular docking results showed that miltirone,dehydromiltirone,and tryptanthrin had strong binding affinity with RAGE.The results of animal experiments confirmed that STDP effectively protected the cardiac function of DCM rats.Compared with the DCM model group,the STDP gr
作者
白亚利
崔鑫钰
袁悦莹
张雨婷
王伟
刘天华
吴晏
BAI Ya-li;CUI Xin-yu;YUAN Yue-ying;ZHANG Yu-ting;WANG Wei;LIU Tian-hua;WU Yan(School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China;Key Laboratory of Basic Research on Syndromes and Prescriptions,Ministry of Education,Beijing University of Chinese Medicine,Beijing 100029,China;Beijing Key Laboratory of Basic Research on Symptoms and Prescriptions,Beijing 100029,China;Research Institute of Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2024年第7期1905-1914,共10页
China Journal of Chinese Materia Medica
基金
国家自然科学基金重点项目(81930113)
北京中医药大学重点攻关项目(2020-JYB-ZDGG-013)。
关键词
麝香通心滴丸
糖尿病心肌病
网络药理学
分子对接
动物实验
AGE-RAGE信号通路
心功能
Shexiang Tongxin Dropping Pills
diabetic cardiomyopathy
network pharmacology
molecular docking
animal experiments
AGE-RAGE signaling pathway
cardiac function
