基于网络药理学和大鼠体内验证的藏药红景天改善脑微循环障碍作用机制研究  

Study on mechanism of Tibetan medicine Rhodiola crenulata in treatment of cerebral microcirculatory disorders based on network pharmacology and experimental validation in rats

作　　者：马四清[1] 时宇静[2] 李园白[3] 杨阳[3] 李萌[3] 杜昱 李逸豪 刘方舟[3] MA Si-qing;SHI Yu-jing;LI Yuan-bai;YANG Yang;LI Meng;DU Yu;LI Yi-hao;LIU Fang-zhou(Qinghai Provincial People's Hospital,Xining 810007,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beiing 100700,China;Insitute of Information on Tradiional Chinese Medicine,China Academy of Chinese Medical Sciences,Beiing 100700,China)

机构地区：[1]青海省人民医院,青海西宁810007 [2]中国中医科学院中药研究所,北京100700 [3]中国中医科学院中医药信息研究所,北京100700

出　　处：《中国药理学通报》2024年第9期1781-1791,共11页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金青年项目(No 82204780);中国中医科学院科技创新工程重大攻关项目(No C12021A05311);青海科技计划应用基础研究(No 2020-ZJ-754)。

摘　　要：目的基于文献研究、网络药理学、分子对接与实验验证的方法,探究藏药红景天改善脑微循环障碍的核心靶点、关键成分和作用机制。方法通过文献和数据库收集红景天化学成分,利用反向药效团匹配预测红景天的潜在靶点;获取脑微循环障碍靶点,并与红景天靶点映射,构建交集靶点蛋白互作网络,获取核心靶点;构建“中药-成分-核心靶点-疾病”调控网络并获取关键成分;进行GO和KEGG富集分析,构建“核心靶点-信号通路-生物过程”网络;进行分子对接验证;采用RT-qPCR和Western blot进行动物实验验证,进一步证实网络药理学分析结果。结果从红景天中筛选出76个活性成分和285个靶点,获取脑微循环障碍靶点1074个,交集靶点97个,核心靶点6个,关键成分6个。分子对接结果表明,有3个关键成分与核心靶点的结合性大于核心靶点蛋白与其原始配体结合性。RT-qPCR结果显示红景天能下调核心靶点CASP3、AKT1 mRNA水平,Western blot检测结果显示藏药红景天能降低CASP3、AKT1蛋白表达(P<0.05)。结论藏药红景天可通过多成分、多靶点、多通路协同改善脑微循环障碍,该研究为临床应用藏药红景天治疗脑微循环障碍提供实验依据。Aim To explore the core target,key components and mechanism of Tibetan medicine Rhodiola crenulata in improving cerebral microcirculation based on literature research,network pharmacology,molecular docking and experimental verification.Methods The chemical components of Rhodiola were collected through literature and database,and the potential targets of Rhodiola crenulata were predicted by reverse pharmacophore matching.The related targets of cerebral microcirculation disorder were obtained and targets were mapping with Rhodiola crenulata.PPI network was constructed and the core targets were screened.The regulatory network of“herb-component-target-disease”was constructed and key components were screened.GO and KEGG enrichment analysis were conducted,and a“Core target-Pathway-Biological Process”network was constructed.Finally,molecular docking validation was carried out,and RT-qPCR and Western blot were used for animal experiments to further confirm the results of network pharmacology analysis.Results A total of 76 active components of Rhodiola crenulata were obtained and corresponding to 285 targets.Altogether 1074 related targets related to cerebral microcirculation disorder were obtained.Among them,there were 97 common targets and the main core targets were 6.The key components were 6.The results of molecular docking showed that the binding activity of three key components to the core target was greater than that of the core target protein and its original ligand.The result of RT-qPCR and Western blot demonstrated that Tibetan medicine Rhodiola crenulata could significantly reduce the expression of core target CASP3 and AKT1(P<0.01).Conclusions Tibetan medicine Rhodiola crenulata can improve the cerebral microcirculation disorder through multi components,multi targets and multi pathways.This study provides an experimental basis for clinical application of Tibetan medicine Rhodiola crenulata to treat cerebral microcirculation disorder.

关 键 词：脑微循环障碍 藏药 红景天 网络药理学 反向药效团匹配 分子对接 RT-QPCR Western blot 作用机制 

分 类 号：R-332[医药卫生] R282.71R319R331.37R743.31