目的 基于网络药理学、分子对接及实验验证探讨益气通脉散（人参、丹参、三七、水蛭、土鳖虫、大 黄）抗脑缺血再灌注损伤（CIRI）的作用机制。方法 （1）通过 TCMSP、TCMID、ETCM 数据库筛选益气通脉散各 中药活性成分及其作用靶点；通过 GeneCards、OMIM、TTD 疾病数据库筛选 CIRI 疾病相关靶点；通过 Venny 2.1 在线平台获得上述靶点的交集靶点，即益气通脉散治疗 CIRI 的潜在作用靶点。通过 Cytoscape 3.7.1 软件构 建“药物-活性成分-靶点”网络，并筛选出益气通脉散治疗 CIRI 的潜在活性成分。通过 STRING 11.0 数据库 进行潜在作用靶点的蛋白互作（PPI）分析，筛选核心靶点。通过 Metascape 数据库对潜在作用靶点进行 GO 功能 及 KEGG 通路富集分析。采用 AutoDockTools 1.5.7 软件对关键活性成分与核心靶点进行分子对接验证。（2）采用 大脑中动脉阻塞再灌注（MCAO/R）术建立 CIRI 大鼠模型。将 SD 大鼠随机分为：假手术组、模型组、益气通脉 散低剂量组（0.27 g·kg-1）、益气通脉散高剂量组（1.08 g·kg-1）、尼莫地平组（30 mg·kg-1），每组 10 只。造模前 3 d 开始预给药，每日 1 次，连续 7 d。采用改良神经功能缺损程度评分（mNSS）法对 MCAO/R 大鼠的神经功能缺 损进行评估；TTC 染色法检测脑梗死体积；尼氏染色法观察脑组织神经元受损情况；ELISA 法检测大鼠血清炎 症因子及氧化应激相关指标；TUNEL 染色法检测脑组织细胞凋亡情况；Western Blot 法检测脑组织中 Bax、 Bcl-2 蛋白表达水平。结果 （1）共得到益气通脉散的 46 个有效活性成分，178 个益气通脉散治疗 CIRI 的潜在 作用靶点。分析出槲皮素、毛地黄黄酮、山柰酚、缬氨酸、尿嘧啶等关键活性成分；TNF、IL-6、STAT3、 VEGFA、AKT1、IL-1β、CASP3、TP53、MAPK3、EGFR 等核心靶点；潜在作用靶点参与炎症反应、氧化应 激、细胞增殖分化、细胞凋亡等生物过程，涉及 cAMP、NF-κB、PI3K-Akt 等信号通路。主要活性成分槲皮 素、毛地黄黄酮、山柰酚、缬氨酸与 TNF、IL-6、STAT3、VEGFA 等靶蛋白有较好的结合活性。（2）与模型组 比较，各给药组大鼠的神经功能缺损评分明显降低（P＜0.05，P＜0.01），脑梗死面积均显著缩小（P＜0.01）， 脑组织缺血坏死区病理变化得到改善，脑组织缺血区神经元数量显著增加（P＜0.01），神经细胞凋亡率显著降 低（P＜0.01），血清 IL-1β、IL-6、TNF-α 水平及 MDA 含量显著降低（P＜0.05，P＜0.01），SOD、GSH-Px 活 性显著升高（P＜0.05，P＜0.01），脑组织中 Bax 蛋白表达显著下调（P＜0.01）， Bcl-2 蛋白表达显著上调（P＜ 0.01）。结论 益气通脉散可能通过通过减轻炎症及氧化应激反应，抑制细胞凋亡，发挥抗 CIRI 的作用。

Objective To investigate the mechanism of Yiqi Tongmai Powder （Ginseng Radix et Rhizoma，Salviae Miltiorrhizae Radix et Rhizoma，Notoginseng Radix et Rhizoma，Hirudo，Eupolyphaga Steleophaga，Rhei Radix et Rhizoma） against cerebral ischemia-reperfusion injury （CIRI）based on network pharmacology，molecular docking and experimental verification. Methods （1） The active components of Yiqi Tongmai Powder and their action targets were screened by TCMSP， TCMID and ETCM databases， the disease related targets of CIRI were screened by GeneCards， OMIM and TTD disease databases， and the intersection targets of the above targets were obtained through Venny 2.1 online platform，that is，the potential targets of Yiqi Tongmai Powder in the treatment of CIRI. The“drugs-active components-targets”network was constructed by Cytoscape 3.7.1 software， and the potential active components of Yiqi Tongmai Powder in the treatment of CIRI were screened. Protein-protein interaction （PPI） analysis of potential targets was carried out by STRING 11.0 database to screen core targets. The GO function and KEGG pathway enrichment of potential targets were analyzed by Metascape database. AutoDockTools 1.5.7 software was used to verify the molecular docking between the key active components and the core targets. （2） The rat model of CIRI was established by middle cerebral artery occlusion and reperfusion （MCAO/R）. SD rats were randomly divided into sham operation group，model group，Yiqi Tongmai Powder low-dose group （0.27 g·kg-1）， Yiqi Tongmai Powder high-dose group （1.08 g·kg-1） and Nimodipine group （30 mg·kg-1），with 10 rats in each group. Pre-administration began three days before the establishment of the model，once a day for seven days. The neurological deficit of MCAO/R rats was evaluated by modified neurological deficit score （mNSS）. The volume of cerebral infarction was measured by TTC staining. Nissl staining was used to observe the damage of neurons in brain tissue. ELISA method was used to detect serum inflammatory factors and oxidative stress related indexes. TUNEL staining was used to detect brain tissue apoptosis. Western Blot method was used to detect the protein expressions of Bax and Bcl-2 in brain tissue. Results （1） A total of 46 active components and 178 potential targets of Yiqi Tongmai Powder in the treatment of CIRI were obtained. The key active components such as quercetin， digitalis flavonoids， kaempferol， valine and uracil were analyzed， and the core targets such as TNF， IL-6， STAT3， VEGFA，AKT1，IL-1β，CASP3，TP53，MAPK3 and EGFR were analyzed. The potential targets are involved in inflammation， oxidative stress， cell proliferation and differentiation， apoptosis and other biological processes， including cAMP，NF- κB，PI3K-Akt and other signal pathways. The main active components quercetin，flavonoids of digitalis，kaempferol and valine have good binding activity to target proteins such as TNF，IL-6，STAT3 and VEGFA. （2） Compared with the model group，the neurological deficit score of rats in each treatment group was significantly decreased （P＜0.05，P＜0.01），the area of cerebral infarction was significantly decreased （P＜0.01）， and the pathological changes of ischemic necrotic area of brain tissue were improved. The number of neurons in ischemic area of brain tissue was significantly increased （P＜0.01）， and the rate of neuronal apoptosis was significantly decreased （P＜0.01）. The levels of serum IL-1β， IL-6， TNF- α and MDA were significantly decreased，while the activities of SOD and GSH-Px were significantly increased （P＜0.05）. The protein expression of Bax in brain tissue were significantly decreased and the protein expression of Bcl-2 significantly increased （P＜ 0.05）. Conclusion Yiqi Tongmai Powder may play an anti-CIRI effect by reducing inflammation and oxidative stress， inhibiting cell apoptosis.

R285.5；R857.3

国家自然科学基金项目（81573919，81673943）；国家中医药管理局中医药科学技术研究专项课题（GZY-KJS-2021-017）；河南省中医药 科学研究专项重点项目（20-21ZY2205，2022ZYZD07）；河南省科技攻关项目（212102311123）