基于网络药理学探讨生脉散降低钙泄漏保护糖尿病大鼠心肌收缩功能的机制研究  

The mechanism of ShengMaiSan reducing calcium leak and protecting myocardial contractile function in diabetic rats based on network pharmacology

作　　者：黄聪 孙明杰[1] 崔海峰[1] 孙丽华[1] 武乾[1] 翟取 石晓路[1] HUANG Cong;SUN Ming-jie;CUI Hai-feng;SUN Li-hua;WU QIAN;ZHAI Qu;SHI Xiao-lu(Key Laboratory of TCM Basic Research on Prevention and Treatment of Major Disease,Experimental Research Center,China Academy of Chinese Medical Sciences,Beijing 100700,China;National Medical Products Administration Institute of Executive Development,Beijing 100073,China)

机构地区：[1]中国中医科学院医学实验中心,北京市中医药防治重大疾病基础研究重点实验室,北京100700 [2]国家药品监督管理局高级研修学院,北京100073

出　　处：《药学学报》2022年第10期3115-3123,共9页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(82004244);北京市自然科学基金青年基金项目(7194301);中央级公益性科研院所基本科研业务费专项资金资助项目(ZZ2019011)。

摘　　要：本研究结合网络药理学和动物实验,探讨生脉散(ShengMaiSan,SMS)降低钙泄漏保护糖尿病大鼠心肌收缩功能的作用机制。通过TCMSP和BATMAN数据库获取SMS活性成分,利用Swiss Target Prediction和GeneCards分别预测SMS活性成分和糖尿病心肌病的潜在作用靶点。获取了1288个SMS潜在作用靶点,1066个糖尿病心肌病疾病靶点,得到180个交集靶点。使用String数据库构建蛋白互作关系网络,Cytoscape软件进行拓扑学分析获得核心靶点,Metascape平台进行GO(gene ontology)和KEGG(Kyoto encyclopedia of genes and genomes)富集分析。筛选出39个核心作用靶点,KEGG通路分析涉及钙信号通路等159条通路。结合文献报道筛选与糖尿病心肌病收缩功能相关的靶点,最终聚焦于钙信号调节通路。腹腔注射链脲佐菌素诱导1型糖尿病大鼠模型。将大鼠分为对照组、模型组、SMS组和曲美他嗪(trimetazidine,TMZ)组,检测左心室血流动力学,急性分离大鼠心肌细胞后进行心肌细胞收缩和钙瞬变同步检测,同时检测肌浆网(sarcoplasmic reticulum,SR)钙库容量和钙泄漏水平及钙泄漏相关蛋白。结果显示,与正常组相比,模型组大鼠左室和心肌细胞收缩功能降低,钙转运紊乱;与模型组相比,SMS组和TMZ组都可使左心室最大收缩压增高,最大收缩舒张速率增高,心肌细胞收缩幅度增高,降低舒张期Ca2+浓度,降低SR钙泄漏水平,使雷诺丁受体2(ryanodine receptor 2,RyR2)磷酸化水平降低,两给药组无显著性差异。SMS可能通过钙信号通路靶点,下调RyR2磷酸化水平、降低SR钙泄漏,增强心肌钙敏感性,增加心肌细胞收缩幅度,增加左室收缩压、左室最大收缩舒张速率,保护糖尿病大鼠心肌收缩功能。本研究动物福利和实验过程均遵循中国中医科学院医学实验中心实验动物伦理委员会的规定。In this study,network pharmacology research and animal experiments were used to predict and validate the potential targets of ShengMaiSan(SMS)in the treatment of diabetic cardiomyopathy.The active components of SMS were obtained through TCMSP and BATMAN databases.The potential targets of the active components and diabetic cardiomyopathy were predicted by Swiss Target Prediction and GeneCards databases,respectively.The protein-protein interaction(PPI)network was constructed by String database.Cytoscape software was adopted to perform topological analysis to select the core action target.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analyses were performed using Metascape platform.To validate the potential targets,a type I diabetic rat model which induced by intraperitoneal injection of streptozotocin(STZ)was prepared.Rats were divided into sham group,model group,SMS group and trimetazidine(TMZ)group.Left ventricular hemodynamics was detected after 4 weeks administrated of SMS or TMZ.Myocardial contraction and calcium transients were detected synchronously in cardiomyocytes,as well as sarcoplasmic reticulum calcium content,calcium leak level and ryanodine receptor 2(RyR2)expression were detected.Based on network pharmacology,1288 targets of SMS,1066 targets of diabetic cardiomyopathy,and 180 overlapped targets were obtained.The 39 core targets were screened,and 159 pathways including calcium signaling pathway were screened by KEGG pathway analysis.According to the previous studies and focusing on the contractility of diabetic cardiomyopathy,this study was involved calcium signaling regulation pathway in the SMS protection mechanism.The results showed that,compared with sham group,the systolic function of left ventricular and myocardial cells were decreased,and the calcium transport was in disorder in model group;compared with model group,both SMS group and TMZ group increased the maximum systolic pressure of left ventricle and the maximum systolic rate of left ventricular cont

关 键 词：生脉散 糖尿病心肌病 钙转运 钙泄漏 网络药理学 

分 类 号：R966[医药卫生—药理学]