基于转录组学与网络药理学探讨心阳片防治慢性心力衰竭的机制  

Study on the Mechanism of Xinyang Tablet on Prevention and Treatment of Chronic Heart Failure Based on Transcriptome and Network Pharmacology

作　　者：吴晗琴 程如熹 张璐[1,2] 何嘉琪[1,2] 陈鑫 王陵军[1,2,3] WU Hanqin;CHENG Ruxi;ZHANG Lu;HE Jiaqi;CHEN Xin;WANG Lingjun(Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China;The First Clinical College,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Lingnan Medical Research Center,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学 [2]广州中医药大学第一附属医院 [3]广州中医药大学岭南医学研究中心

出　　处：《中药新药与临床药理》2021年第12期1807-1816,共10页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81973776)。

摘　　要：目的基于转录组学与网络药理学探讨心阳片防治慢性心力衰竭(CHF)的关键机制。方法将32只小鼠随机分为假手术组、模型组、心阳片组(910 mg·kg^(-1))、培哚普利组(阳性对照组,0.607 mg·kg^(-1))。采用主动脉弓缩窄术(TAC)构建CHF小鼠模型。造模成功后,按照上述剂量灌胃给药(10 mL·kg^(-1)),每日1次,连续给药8周。给药结束后,对小鼠行心脏彩超检测,采集左室射血分数(LVEF)及左室短轴缩短率(LVFS)等心功能指标;采用Masson染色法观察心脏组织病理形态变化。取小鼠心脏组织提取RNA进行转录组测序,并对组间差异基因进行GO功能及KEGG通路富集分析。通过TCMSP平台以及文献检索筛选心阳片的活性成分,利用SwissTargetPrediction网站预测活性成分作用靶点。通过Venny平台对转录组测序得到的交集差异基因与活性成分作用靶点取交集,获得交集靶点(心阳片治疗CHF的关键靶点)。利用STRING平台构建交集靶点蛋白互作(PPI)网络;通过Cytoscape 3.8.0软件构建"中药-活性成分-关键靶点网络"网络,筛选出心阳片防治CHF的核心靶点,并对核心靶点进行KEGG通路富集分析;采用qPCR法检测核心靶点mRNA表达。结果与假手术组比较,模型组小鼠的LVEF、LVFS均显著降低(P<0.01),小鼠的心脏体积明显增大,心脏系数显著升高(P<0.01),心脏胶原容积分数明显升高(P<0.05)。与模型组比较,给药组小鼠的LVEF、LVFS均显著升高(P<0.01),小鼠的心脏体积明显缩小,心脏系数显著降低(P<0.01);心阳片组小鼠的心脏胶原容积分数明显下降(P<0.05),培哚普利组的变化无明显差异(P>0.05)。共得到心阳片有效成分83个;假手术组-模型组的差异基因与模型组-心阳片组差异基因的交集差异基因共有354个,进一步与心阳片主要活性成分的771个作用靶点取交集,共得到21个交集靶点,即心阳片治疗CHF的关键靶点。心阳片改善CHF的核心靶点包括CCNB1、CCNA2、CObjective To explore the pivotal mechanisms of Xinyang tablet on the prevention and treatment of chronic heart failure(CHF)based on transcriptome and network pharmacology.Methods The CHF models were induced by transverse aortic constriction(TAC)surgery,thirty-two mice were randomly divided into Sham operation group,model group,Xinyang tablet group(910 mg·kg^(-1))and positive control group(Perindopril 0.607 mg·kg^(-1)).After the model was successfully established,the above-mentioned doses of drugs were given by gavage(10 mL·kg^(-1))once a day for 8 consecutive weeks.After eight weeks,we collected left ventricular ejection fraction(LVEF)and left ventricular short axis shortening rate(LVFS) and other cardiac function data by echocardiographic testing.The pathological changes of heart tissue were evaluated by Masson staining.The mRNA was extracted from heart tissues and subjected to RNA-Seq for differentially expressed genes comparison,GO analysis and KEGG pathway enrichment analysis.Active components of Xinyang tablet were searched from TCMSP and literatures,the targets were predicted by SwissTargetPrediction platform.The common targets of transcriptomics and networkpharmacology(pivotal targets of Xinyang tablet in the treatment of CHF)were performed by Venny website.The common targets were used to construct protein-protein interaction(PPI) network through the STRING platform.The Drugsingredients-common targets network was constructed by Cytoscape 3.8.0 to screen core targets,the KEGG pathway enrichment analysis of the core targets was performed,and then,we tested the expression of core targets by Realtime fluorescence quantitative PCR.Results As compared with the Sham operation group,the LVEF,LVFS in model group were significantly decreased(P<0.01),the heart volume was increased and the cardiac coefficient was higher significantly(P<0.01),and the cardiac collagen volume fraction was significantly higher(P<0.05).As compared with the model group,the LVEF,LVFS in Xinyang tablet group were significantly increased(P<

关 键 词：心阳片 慢性心力衰竭 心功能 心脏纤维化 网络药理学 转录组学 小鼠 

分 类 号：R285.5[医药卫生—中药学] R857.3[医药卫生—中医学]