基于网络药理学与分子对接探讨当归丹参药对治疗急性肝损伤的作用机制

A Study on Mechanism of ASM in the Treatment of ALI Based on Network Pharmacology and Molecular Docking

基于网络药理学与分子对接技术及试验验证探讨当归丹参药对(ASM)治疗急性肝损伤(ALI)的作用机制。本研究采用网络药理学方法筛选ASM治疗ALI的核心作用靶点,构建核心靶点进行蛋白互作网络和分子对接分析,通过构建大鼠ALI模型检测相关血清生化指标和肝脏蛋白的表达模式并验证分子对接结果。结果显示,从ASM中获得了67个活性成分,其中75个靶点与ALI相互作用。蛋白质–蛋白质相互作用(PPI)网络分析显示,TP53、CASP3、JUN、STAT3、AKT1、VEGFA、TNF、IL-6、MMP9和PTGS2可能是ASM治疗ALI的关键作用靶点。基因本体(GO)分析和京都基因和基因组百科全书(KEGG)信号通路分析表明,靶点主要富集于乙肝信号通路、脂质与动脉粥样硬化、糖基化终产物受体(RAGE)和细胞凋亡等信号通路。分子对接结果显示,核心靶点(TP53和CASP3)与β-谷甾醇、黄芩苷和隐丹参酮有良好的结合位点。体内试验结果表明,ASM能够有效改善四氯化碳(CCl4)诱导的大鼠肝损伤,降低大鼠血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)及丙二醛(MDA)的表达水平,提高肝脏超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性及谷胱甘肽(GSH)水平,上调肝脏组织中BCL2蛋白的表达水平,下调BAX、CASP3和TP53蛋白的表达水平。本研究表明,ASM改善ALI的机制可能是通过β-谷甾醇、黄芩苷和隐丹参酮等主要化学成分作用于TP53和CASP3等关键靶点,通过抑制氧化应激及调控细胞凋亡等信号通路来实现。本研究为临床上ASM治疗ALI提供理论与试验依据,为进一步研究和发展传统中医药提供新的思路和方法。

Based on network pharmacology and molecular docking technology,the mechanism of radix angelicae sinensis-radix salviae miltiorrhizae drug pair(ASM)in the treatment of acute liver injury(ALI)was studied and verified by experiment.In this study,network pharmacology and molecular docking methods were used to screen the core targets of ASM in the treatment of ALI,and the core targets were enriched by analysis,and protein interaction network was constructed.The core target was molecular docked with the active components of ASM,and the molecular docking was verified by constructing ALI rat model and detecting the expression of relevant serum biochemical indexes and liver proteins.The results showed that 67 active ingredients were obtained from ASM,of which 75 targets interacted with acute liver injury.Protein-protein interactions(PPI)network analysis showed that TP53,CASP3,JUN,STAT3,AKT1,VEGFA,TNF,IL-6,MMP9 and PTGS2 may be the key targets of ASM in the treatment of ALI.The core targets of GO and KEGG signaling pathway enrichment analysis were mainly concentrated in hepatitis B signaling pathway,lipid and atherosclerosis,AGE-RAGE,and apoptosis signaling pathways.Molecular docking results showed that the core targets(TP53 and CASP3)had good binding sites withβ-sitosterol,baicalin and cryptotanshinone.The results of in vivo experiments showed that ASM could reduce the expression levels of ALT,AST and MDA in serum of CCl4-induced model rats,and increase the activity of SOD,CAT and GSH in liver,thereby alleviating the CCl4-induced oxidative stress.The ASM could up-regulate BCL2 and down-regulate the expression levels of BAX,CASP3 and TP53.The results of this study indicate that ASM can effectively ameliorate ALI,which may be achieved throughβ-sitosterol,baicalin and cryptotanshinone and other chemical components,acting on key targets such as TP53 and CASP3,and through multiple signaling pathways such as p53 signaling pathway and apoptosis pathway.This study provides theoretical and experimental basis for clinical ASM treatment of ALI,and provides new ideas and methods for further research and development of traditional Chinese medicine.