赵冯岩 杨浩澜 朱炎贞 林美妤 刘芳

〔摘要〕 目的 利用网络药理学方法探究补阳还五汤经由铁死亡途径调控缺血性脑卒中的作用机制。方法 通过TCMSP、PharmMapper数据库完成补阳还五汤处方中各药物的化学成分、靶点基因及其相应对接得分的收集，采用KEGG、GeneCards数据库获取疾病相关靶点;采用Cytoscape 3.6.1获取补阳还五汤调控缺血性脑卒中铁死亡途径的作用候选靶点网络图，并对该图进行模块分析和二次筛选;KOBAS 3.0对候选靶点与模块基因进行KEGG通路富集;采用Western blot技术检测PC12细胞氧化应激模型p-PI3K、PI3K、p-Akt、Akt、p-mTOR、mTOR蛋白表达。结果 共得到补阳还五汤主要活性成分10个，综合文献分析得缺血性脑卒中铁死亡途径关键靶点3个，主要KEGG通路7条。验证实验结果提示补阳还五汤通过PI3K-Akt-mTOR信号通路对氧化应激损伤的PC12细胞发挥保护作用。结论 补阳还五汤调控缺血性脑卒中铁死亡途径可能与HSP90AA1、EGFR、GRB2等靶点及MAPK、PI3K-Akt等信号通路有关，其中HSP90AA1靶点和PI3K-Akt信号通路可能是关键机制之一。

〔关键词〕 铁死亡;补阳还五汤;缺血性脑卒中;网络药理学

〔中图分类号〕R255.2       〔文献标志码〕A       〔文章编号〕doi：10.3969/j.issn.1674-070X.2021.07.016

〔Abstract〕 Objective To use network pharmacology to explore the mechanism of Buyang Huanwu Decoction regulating ischemic stroke through ferroptosis. Methods The chemical components， target genes of Buyang Huanwu Decoction and their corresponding docking scores were collected by TCMSP database or PharmMapper database， and the disease-related targets were obtained through KEGG and GeneCards database; the Cytoscape 3.6.1 software was used to get the network map of candidate targets of Buyang Huanwu Decoction effect on cerebral ischemic stroke by ferroptosis pathway， and the network map of candidate targets was screened twice and MCODE analysis subsquently; the candidate targets and module genes were enriched by KEGG pathway in KOBAS 3.0. Western blot were used to detect the expression of p-PI3K， PI3K， p-Akt， Akt， p-mTOR and mTOR protein in PC12 cells model with oxidative stress injury. Results There were 10 potential active ingredients in Buyang Huanwu Decoction， and 3 key targets and 7 main KEGG pathways in cerebral ischemic stroke by ferroptosis based on the literature analysis. And the validation experiment confirmed that Buyang Huanwu Decoction can protect oxidative stress induced PC12 cells injure via PI3K-Akt-mTOR signal pathway. Conclusion Buyang Huanwu Decoction in regulating cerebral ischemic stroke by ferroptosis may be related to HSP90AA1， EGFR， GRB2 and other signal pathways such as MAPK and PI3K-Akt. Among which the HSP90AA1 and PI3K-Akt signaling pathway may be one of the most crucial mechanisms.

〔Keywords〕 ferroptosis; Buyang Huanwu Decoction; cerebral ischemic stroke; network pharmacology

缺血性腦卒中是一类中枢神经系统疾病，指突发的脑组织局部供血不足或血流完全中断，使该局部脑组织崩解破坏，进而产生局部或全脑神经功能障碍的一种脑血管疾病。补阳还五汤出自《医林改错》[1]，以气虚血瘀立论，由黄芪、当归、赤芍、地龙、桃仁、川芎、红花组合成方，是治疗缺血性脑卒中的经典名方，现代临床应用也颇为广泛[2。现代药理研究[3-4]表明，补阳还五汤具有扩张血管、增加脑血流量、抗炎、抗缺氧、改善微循环以及抗血栓等作用，可通过增加血管内皮生长因子阳性细胞数量、抑制神经细胞凋亡、抗感染、抑制基质金属蛋白酶等多种途径来防治缺血性脑卒中。……