通窍活血汤治疗缺血性脑卒中作用机制研究  

Mechanism of Tongqiao Huoxue Decoction in the Treatment of Ischemic Stroke

作　　者：汪雅玲 王虎 刘爽 陈辉 刘海林 WANG Yaling;WANG Hu;LIU Shuang;CHEN Hui;LIU Hailin(Chongqing Medical Science and Technology University,Chongqing,China 400061;Chongqing Liangjiang New Area People′s Hospital,Chongqing,China 401121)

机构地区：[1]重庆市医药科技学校,重庆400061 [2]重庆两江新区人民医院,重庆401121

出　　处：《中国药业》2025年第5期35-41,共7页China Pharmaceuticals

基　　金：重庆市自然科学基金面上项目[CSTB2024NSCQ-MSX0085];重庆市技术创新与应用发展专项面上项目[cstc2019jscx-msxmX0096]。

摘　　要：目的 探讨通窍活血汤治疗缺血性脑卒中(IS)的作用机制。方法 通过TCMSP,OMIM,GeneCards等数据库获取IS与通窍活血汤的共有靶点;通过Cytoscape 3.7. 1软件对药物-成分-靶点网络进行拓扑分析,获取核心活性成分及核心靶点,并对其进行基因本体论(GO)功能和京都基因与基因组百科全书(KEGG)通络富集分析;利用分子对接技术验证核心活性成分与核心靶点的结合能力。将40只大鼠随机分为空白对照组(A组),模型对照组(B组,1 mL/100 g蒸馏水),通窍活血汤高、中、低剂量组(C1组、C2组、C3组,0.72,0.36,0.18 mL/100 g),各8只。采用中动脉栓塞(t-MCAO)法构建缺血再灌注损伤(I/R)模型大鼠,建模成功后,各组大鼠分别灌胃相应药物。采用改良Longa五步四分评分法评估神经功能,通过苏木精-伊红(HE)染色法观察皮层脑组织形态学,采用酶联免疫吸附试验(ELISA)法检测大鼠脑损伤部位炎性因子[肿瘤坏死因子-α(TNF-α)、白细胞介素1β(IL-1β)]的水平,采用免疫印迹(Western blot)法检测大鼠脑损伤部位TP53、c-Jun、胱天蛋白酶3(Casp3)的蛋白表达水平。结果 共获得通窍活血汤活性成分69个,药物-疾病共有靶点120个,核心活性成分包括槲皮素、山柰酚、桑色素等,核心靶点包括TP53,c-Jun,Casp3等,信号通路包括丝裂原活化蛋白激酶(MAPK)、白细胞介素17(IL-17)等。分子对接结果显示,核心活性成分与核心靶点的结合能均≤-6.0 kcal/mol。与B组比较,C1组、C2组、C3组大鼠的神经功能缺损评分,脑损伤部位中TNF-α,IL-1β表达水平,脑损伤部位中TP53,c-Jun,Casp3的蛋白表达水平,损伤分数均显著降低(P <0.05)。结论 通窍活血汤可通过多成分、多靶点、多通路减轻脑缺血再灌注损伤。Objective To investigate the mechanism of Tongqiao Huoxue Decoction in the treatment of ischemic stroke(IS).Methods The common targets of IS and Tongqiao Huoxue Decoction were obtained through databases such as TCMSP,OMIM,GeneCards,etc.Topological analysis of the drug-ingredient-target network was performed by the Cytoscape 3.7.1 software to obtain key active components and key targets.Gene Ontology(GO)functional and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis of common targets were performed.Molecular docking technology was used to verify the binding ability between key active components and key targets.Forty rats were randomly divided into the blank control group(group A),model control group(group B,1 mL/100 g distilled water),and Tongqiao Huoxue Decoction high-,medium-,and low-dose groups(groups C1,C2,C3;0.72,0.36,0.18 mL/100 g),with eight rats in each group.The ischemia-reperfusion(I/R)injury model rats were constructed by the transient middle cerebral artery occlusion(t-MCAO)method.After successful modeling,rats in each group were orally administered with the corresponding drugs.The modified Longa five-grade scoring standard was used to evaluate the neurological function,and the morphology of cortical brain tissue was observed by the hematoxylin-eosin(HE)staining.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of inflammatory factors[tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)]at the site of brain injury in rats,and Western blot was used to detect the expression level of TP53,c-Jun,caspase 3(Casp3)protein.Results A total of 69 active components and 120 drug-disease intersection targets were obtained for Tongqiao Huoxue Decoction.Key active components included quercetin,kaempferol and luteolin,etc.Key targets included TP53,c-Jun,Casp3,etc.Signal pathways included mitogen-activated protein kinase(MAPK),interleukin-17(IL-17),etc.The molecular docking results showed that the binding energy between the active components and the key targets was≤-6.0

关 键 词：通窍活血汤 缺血性脑卒中 神经保护 网络药理学 分子对接技术 动物实验 

分 类 号：R932[医药卫生—生药学] R285.5[医药卫生—药学]