中医药导报2025，Vol.31Issue(6)：7-14,8.DOI:10.13862/j.cn43-1446/r.2025.06.002
基于网络药理学和实验验证探讨清营生脉汤对心肌缺血再灌注损伤的保护机制
The Protective Mechanism of Qingying Shengmai Decoction(清营生脉汤)on Myocardial Ischemia Reperfusion Injury Based on Network Pharmacology and Experimental Verification
摘要
Abstract
Objective:To explore the protective mechanism of Qingying Shengmai decoction on myocardial ischemia reperfusion injury(MIRI)based on network pharmacology and molecular docking methods,and to verify some key targets and signaling pathways through in vitro experiments.Methods:The active ingredients and targets of Qingying Shengmai decoction were obtained by traditional Chinese medicine pharmacology database and analysis platform(TCMSP)database and BATM-TCM database,and the disease targets of MIRI were screened by GeneCards database and OMIM database.The intersection of drug targets and disease targets was obtained through Venny platform.The intersection target PPI network of Qingying Shengmai decoction in the treatment of MIRI was constructed based on String database to obtain the core targets of drugs and diseases.The enrichment results of gene ontology(GO)enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were visualized using the bioinformatict platform.The"components-target-pathway"interaction network was established with Cytoscape v3.9.1.Molecular docking of core compo-nents was performed by AutoDockTools and PyMol software.In vitro,H9c2 cardiomyocytes were induced with H2O2 to construct oxidative stress model,and the optimal drug concentration was screened by CCK-8.The content of lactate dehydrogenase(LDH)and cardiac troponin T(c-TnT)was detected by enzyme-linked im-munosorbent assay(ELISA),and the content of reactive oxygen species(ROS)and mitochondria was detected by laser confocal microscopy.Western blotting was used to verify the predicted pathway,and real-time quantitative PCR was used to detect the mRNA levels of core targets.Results:There were 109 active ingredients in Qingying Shengmai decoction,corresponding to 1 002 targets,of which 122 targets were related to the treatment of MIRI.GO enrichment analysis obtained a total of 2 494 items.A total of 195 pathways were obtained by KEGG enrichment analysis,mainly involving TNF signaling pathway,HIF-1 signaling pathway,JAK-STAT signaling pathway,etc.The results of molecular docking showed that the binding forces of quercetin,luteolin,kaempferol with TNF,IL-6,HIF-1α,JAK2,STAT3 and other key target proteins were stable.Cell experiments showed that the serum containing Qingying Shengmai decoction drug-containing serum could reduce the levels of LDH and c-TnT(P<0.05),inhibit the generation of ROS,and increase the content of mitochondria(P<0.05).The results of Western blotting and PCR showed that the Qingying Shengmai decoction drug-containing serum could down-regulate the expression of HIF-1α and up-regulate the protein and mRNA expression of JAK2(P<0.05).Conclusion:Qingying Shengmai decoction has a multi-component,multi-target and multi-pathway therapeutic effect on myocardial ischemia reperfusion injury.The Qingying Shengmai decoction drug-containing serum may alleviate oxidative stress-induced myocardial injury by regulating HIF-1α and JAK2 signaling pathway.关键词
心肌缺血再灌注损伤/清营生脉汤/网络药理学/分子对接/氧化应激/H9c2细胞Key words
myocardial ischemia reperfusion injury/Qingying sheng mai decoction/network pharmacology/molecular docking/oxidative stress/H9c2 cells分类
医药卫生引用本文复制引用
武文硕,付殷,张璐艳,谢蝶,张洋..基于网络药理学和实验验证探讨清营生脉汤对心肌缺血再灌注损伤的保护机制[J].中医药导报,2025,31(6):7-14,8.基金项目
黑龙江省博士后资助项目(LBH-Q19058) （LBH-Q19058）
黑龙江省中医药经典普及化专项课题项目(ZYW2023-019) （ZYW2023-019）
