基于网络药理学探讨复方苁蓉益智胶囊治疗轻度认知障碍的作用机制  

Mechanism of Fufang Congrong Yizhi Capsules in treatment of mild cognitive impairment based on network pharmacology

作　　者：韩沁 许宵钰 耿艺菲 刘静静 孙晓波[1] 罗云[1] HAN Qin;XU Xiao-yu;GENG Yi-fei;LIU Jing-jing;SUN Xiao-bo;LUO Yun(Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences,Beijing 100193,China;Lei Yun Shang Pharmaceutical Group Corporation Limited,Bozhou Anhui 136200,China)

机构地区：[1]中国医学科学院药用植物研究所,北京100193 [2]安徽雷允上药业有限公司,安徽亳州136200

出　　处：《中国药理学通报》2024年第2期334-343,共10页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金委-贵州省人民政府联合基金项目(No U1812403-5-3);中国医学科学院医学与健康科技创新工程项目(No 2022-I2M-2-002)。

摘　　要：目的基于网络药理学对复方苁蓉益智胶囊(fufang congrong yizhi capsule,FCYC)治疗轻度认知障碍(mild cognitive impairment,MCI)的作用机制进行预测和细胞实验验证。方法利用TCMSP、GeneCards、OMIM和TTD数据库、《中国药典》以及文献筛选FCYC的活性成分及治疗MCI的作用靶点,构建中药-化合物-靶点-疾病网络图以及交集靶点的蛋白相互作用图,通过易汉博生物信息平台进行GO和KEGG分析。利用Aβ25-35诱导神经细胞PC12损伤建立MCI细胞模型,FCYC预处理探究药效及机制:CCK-8法和LDH试剂盒测定细胞活力,JC-1检测细胞线粒体膜电位,ELISA法检测IL-1β、IL-6和TNF-α表达,免疫荧光法及Western blot法检测Bcl2、Bax和PI3K/AKT信号通路相关蛋白的表达。结果FCYC共有57个活性成分可能通过66个潜在靶点治疗MCI,GO富集分析结果显示这些靶点涉及339个生物过程、39个细胞组成以及65个分子功能;KEGG通路主要涉及PI3K-AKT、TNF信号通路等。实验结果表明,FCYC可明显提高Aβ25-35诱导损伤的PC-12细胞活力,降低PC-12细胞中IL-1β、IL-6和TNF-α的分泌,上调Bcl2/Bax的比例,并增加p-PI3K/PI3K和p-AKT/AKT的表达。结论FCYC通过多成分-多靶点-多通路治疗MCI,其可抑制神经炎症和神经细胞凋亡,其可能机制为通过调控PI3K/AKT信号治疗MCI。Aim To predict the mechanism of Fufang Congrong Yizhi Capsules(FCYC)in the treatment of mild cognitive impairment(MCI)by network pharmacology method,and further validate it in combination with cellular experiments.Methods TCMSP,GeneCards,OMIM and TTD databases,Chinese Pharmacopoeia and related literature were used to screen the active ingredients of FCYC and the targets of MCI treatment.The TCM-compound-target-disease network and PPI of intersection targets were constructed,and the GO and KEGG analysis were performed by the Ehamb bioinformation platform.GO and KEGG analysis were performed through Yihanbo biological information platform.Cell model of MCI was established by PC-12 injury induced by Aβ25-35,and its efficacy and mechanism were explored by pretreatment of compound FCYC.Cell viability was detected by CCK-8 and LDH kit.The mitochondrial membrane potential was detected by JC-1.The expressions of IL-1β,IL-6 and TNF-αwere detected by ELISA.The expressions of Bcl2,Bax and PI3K/AKT signaling pathways were detected by immunofluorescence and WB.Results A total of 57 active components of FCYC could be used to treat MCI through 66 potential targets.GO enrichment analysis showed that these targets were involved in 339 biological processes(BP),39 cell compositions(CC)and 65 molecular functions(MF).KEGG enrichment pathways were mainly involved in PI3K-AKT signaling pathway and TNF signaling pathway.The results of experiments showed that FCYC could significantly increase the activity of injured PC-12 cells induced by Aβ25-35,reduce the secretion of IL-1β,IL-6 and TNF-αin PC-12 cells,up-regulate the ratio of Bcl2/Bax,and increase the expression of p-PI3K/PI3K and p-AKT/AKT.Conclusions This study preliminarily shows that FCYC can treat MCI through multi-component,multi-target,and multi-pathway.In vitro experiments demonstrate that FCYC can significantly inhibit neuro-inflammation and neuronal apoptosis,and the possible mechanism may be the regulation of PI3K/AKT signaling for MCI.

关 键 词：轻度认知障碍 复方苁蓉益智胶囊 网络药理学 AΒ25-35 Bcl2/Bax PI3K/AKT信号通路 

分 类 号：R-332[医药卫生] R282.71R329.24R319R741