摘要
目的:采用网络药理学分析和实验验证相结合的方法,研究芪蛭通络胶囊对脑缺血再灌注损伤的治疗作用和作用机制。方法:利用网络药理学的方法获取芪蛭通络胶囊中的化学成分和潜在靶点并获得脑缺血再灌注病理过程相关的靶点,构建“中药-化合物-疾病网络图”,同时对核心靶点进行京都基因与基因组百科全书数据库(Kyoto Encyclopedia of Genes Genomes,KEGG)富集分析;采用线栓法制作大鼠右侧大脑中动脉缺血90 min再灌注模型,观察芪蛭通络胶囊0.8、1.6、3.2 g/kg和阳性药尼莫地平30 mg/kg给药7 d后的治疗作用;采用神经行为学评分和矿场试验检测大鼠神经功能和运动能力;TTC染色、苏木素伊红(hematoxylin-eosin,HE)染色、尼氏染色检测以及酶联免疫吸附法检测脑组织损伤和相关指标的变化;Western Blot检测Toll样受体4/髓样分化因子/核因子κB(toll-like receptor 4/myeloiddifferentiationfactor88/nuclear factor kappa-B,TLR4/MyD88/NF-κB)通路和NOD样受体蛋白3(nod-like receptor protein 3,NLRP3)炎症小体相关蛋白的变化。结果:网络药理学分析揭示,芪蛭通络胶囊治疗脑缺血再灌注的主要活性成分包括常春藤素、异欧前胡素、芒柄花黄素等19种化合物,其中涉及磷酯酰肌醇-3激酶/蛋白激酶B(phosphatidylinositol-3-kinase/protein serine threonine kinase,PI3K/AKT)信号通路、炎症信号通路、Toll样受体信号通路、NOD样受体信号通路等。实验研究表明,与模型对照组相比,芪蛭通络胶囊各组和尼莫地平30 mg/kg组均能明显改善脑缺血再灌注后大鼠神经功能和运动能力,减小缺血部位脑梗死体积(P<0.05或P<0.01),有效改善病理损伤,明显调控MyD88、NFκB、NLRP3、半胱氨酸天冬氨酸蛋白酶-1(cysteinyl aspartate specific proteinase-1,Caspase1)蛋白表达(P<0.05或P<0.01)。结论:网络药理学及实验验证表明芪蛭通络胶囊可能通过TLR4/MyD88/NFκB通路及NLRP3炎症小体治疗脑缺血再灌注损伤。
Objective:This article uses network pharmacological analysis and experimental verification to study the mechanism of Qizhi Tongluo Capsules in the treatment of cerebral ischemia reperfusion.Methods:The method of network pharmacology were used to obtain the chemical components and potential targets of Qizhi Tongluo capsules and the targets related to cerebral ischemia reperfusion.The Chinese medicine-compound-disease network diagram was constructed and the core targets were analyzed by Kyoto Encyclopedia of Genes Genomes(KEGG)enrichment.The right middle cerebral artery ischemia 90-minute reperfusion model was made by the suture method.Then therapeutic effect was obserbed by 7 days administration of Qizhi Tongluo Capsules 0.8、1.6、3.2 g/kg and nimodipine 30 mg/kg.The neurobehavioral scores and mine experiments were used to detect the neurological function and movement ability of rats.TTC staining,hematoxylin-eosin(HE)staining,nissl staining and enzyme-linked immunosorbent assay was performed to detect brain injury and changes of related indexes.Western blot was used to detect the expression levels of toll-like receptor 4(TLR4),myeloiddifferentiationfactor88(MyD88),nuclear factor kappa-B(NFκB)and nod-like receptor protein 3(NLRP3),inflammasome-related proteins.Results:Network pharmacological analysis revealed that the main active ingredients of Qizhitongluo Capsules in the treatment of cerebral ischemia and reperfusion includeD 19 compounds including hederin,isoperatorin,and formononetin,which involve phosphatidylinositol-3-kinase/protein serine threonine kinase(PI3 K-Akt)signaling pathway,Inflammation signaling pathway,Toll-like receptor signaling pathway,NOD-like receptor signaling pathway,etc.Experimental studies showed,compared with the model control group,all dozes of Qizhitongluo Capsules groups and the positive drug nimodipine 30 mg/kg group significantly improved the neurological function and exercise capacity of rats after cerebral ischemia and reperfusion,and reduced the volume of cerebral infarction(P<0.05 or P<0.01),it also effectively improved pathological damage,and significantly regulated the protein expression of MyD88,NFκB,NLRP3 and cysteinyl aspartate specific proteinase-1(Caspase1)(P<0.05 or P<0.01).Conclusion:Network pharmacology and experimental verification confirm that Qizhi Tongluo Capsules may treat cerebral ischemia reperfusion injury through TLR4/MyD88/NFκB pathway and NLRP3 inflammasome.
作者
刘珊
张雯
宋俊科
杨海光
姚静
李昆
王淑美
杜冠华
Liu Shan;Zhang Wen;Song Junke;Yang Haiguang;Yao Jing;Li Kun;Wang Shumei;Du Guanhua(Beijing Key Laboratory of Drug Target and Screening Research,Institute of Materia Medica,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050;Guangdong Pharmaceutical University,Guangzhou 510006;Beijing Zhendong Guangming Pharmaceutical Research Institute,Beijing 100085;Shanxi Zhendong Pharmaceutical Co.,Ltd.,Changzhi 046108)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2022年第1期155-162,共8页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家重点研发计划(编号:2020YFC2008302)
国家自然科学基金(编号:82004071)
关键词
芪蛭通络胶囊
脑缺血再灌注
炎症小体
网络药理学
Qizhi Tongluo Capsules
cerebral ischemia reperfusion
inflammasome
network pharmacology
作者简介
通信作者:杜冠华,博士,研究员,研究方向:神经药理,E-mail:dugh@imm.ac.cn;刘珊,硕士研究生,研究方向:神经药理学,E-mail:15841400281@163.com。
