β-谷甾醇靶向调控PI3K3CG基因保护心肌肥厚作用研究  

β-sitosterol protect myocardial hypertrophy by regulating PI3K3CG

作　　者：李鹏宇 陈梦真 冉嘉欣 刘思宇 孙辉[2] LI Pengyu;CHEN Mengzhen;RAN Jiaxin;LIU Siyu;SUN Hui(Clinical College,Chengdu University of Traditional Chinese Medicine,Chengdu Sichuan 611137,China;Pharmaceutical Experiment Teaching Center,College of Pharmacy,Harbin Medical University,Harbin Heilongjiang 150081,China)

机构地区：[1]成都中医药大学临床学院,四川成都611137 [2]哈尔滨医科大学药学院药学实验教学中心,黑龙江哈尔滨150080

出　　处：《转化医学杂志》2022年第6期331-336,共6页Translational Medicine Journal

基　　金：黑龙江省大学生创新创业基金(S202110226059)。

摘　　要：目的 运用网络药理学方法建立通心络活性成分-作用靶点网络,明确靶点相应的生物功能,探讨β-谷甾醇与心肌肥厚之间的作用及途径。方法 通过中药系统药理学数据库和分析平台(TCMSP)收集通心络主要成分及药物潜在作用靶点,通过Cytoscape软件将通心络成分-作用靶点网络可视化,使用DAVID在线工具进行基因的基因本体论(GO)分析和KEGG通路富集分析。通过免疫荧光检测不同浓度的β-谷甾醇对于ISO诱导的H9C2细胞表面积的影响;使用qRT-PCR检测心肌肥厚相关因子的ANP,BNP及β-MHC的表达;使用Western blot检测PI3K3CG的变化;使用H&E染色及心脏超声检测β-谷甾醇对于心脏细胞结构以及心脏功能的改变。结果 通心络中主要有95个活性成分对应180个潜在靶点。通心络与心肌肥厚交集靶基因主要富集在钙离子信号通路、胆碱能突触信号通路及cAMP信号通路等。体内中β-谷甾醇逆转了由ISO诱导的心肌细胞表面积增加,同时,抑制ANP、BNP,PI3K3CG和β-MHC的表达。β-谷甾醇抑制ISO诱导的大鼠心肌细胞肥大作用,包括降低小鼠心肌细胞横截面积,降低IVSD和LVPWD,抑制肥大标志基因ANP、BNP、β-MHC以及PI3K3CG的表达。结论 β-谷甾醇通过抑制PI3K3CG水平,首先调控心肌肥厚相关因子的表达,进而影响心肌细胞肥厚,干预心肌肥厚发展。Objective Via establishing the component-target network of Tongxinluo to clarify the target’s biological function and explore the role and signaling pathway of β-sitosterol in cardiac hypertrophy.Methods TCMSP(Chinese Medicine System Pharmacology database and analysis platform) was used to collect the main components and potential targets of Tongxinluo. David was used to analysis enrichment of Gene Ontology(go) and KEGG pathway. Immunofluorescence was used to detect the effects of different concentrations of β-sitosterol on the surface area of H9C2 cells induced by ISO. QRT-PCR was used to detect the expression of ANP, BNP, and β-MHC, and Western blot was used to detect the changes of PI3K3CG. H&E staining and echocardiography were used to detect the changes in cardiac cell structure and cardiac function induced by β-sitosterol. Results 95 active components and 180 potential targets were found belong to Tongxinluo. The common related genes of Tongxinluo and myocardial hypertrophy were mainly involved in calcium signaling pathway, cholinergic synaptic signaling pathway, serotonin-containing signaling pathway,and cAMP signaling pathway. The calcium signaling pathway is the main pathway related to cardiac hypertrophy. β-sitosterol reversed the increase of myocardial cell surface area induced by ISO and inhibited the expression of ANP, BNP,PI3K3CG, and β-MHC. The effects of β-sitosterol on myocardial hypertrophy induced by ISO in rats include decreasing the cross-sectional area of cardiomyocytes, reducing IVSD and LVPWD, and inhibiting the expression of ANP, BNP, β-MHC, and PI3K3CG.Conclusion Tongxinluo treatment of myocardial hypertrophy is a multi-component, multi-target joint action. Among them, β-sitosterol can affect the expression of related factors of cardiac hypertrophy by inhibiting the level of PI3K3CG, affecting the hypertrophy of myocardial cells, and intervening in the development of cardiac hypertrophy.

关 键 词：网络药理学 通心络 心肌肥厚 Β-谷甾醇 

分 类 号：R542.2[医药卫生—心血管疾病]