基于网络药理学和体外实验探讨驻景丸治疗老年性黄斑变性的作用机制  

Based on Network Pharmacology and in vitro Experiments to explore the mechanism of Zhujing Pills in Treating Age-related Macular Degeneration

作　　者：焦肖宁 刘洺希 胥孜杭 朱杨壮壮 苏琳 侯怡飞 张宁[2] 王冰 邹纯朴[1] JIAO Xiaoning;LIU Mingxi;XU Zihang;ZHU Yangzhuangzhuang;SU Lin;HOU Yifei;ZHANG Ning;WANG Bing;ZOU Chunpu(School of Basic Medical Science,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Science and Technology Experimental Center,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Center for Pharmaceuticss Research,Shanghai Institute of Materia Medica Chinese,Academy of Sciences,Shanghai 201203,China)

机构地区：[1]上海中医药大学基础医学院,上海201203 [2]上海中医药大学科技实验中心,上海201203 [3]中国科学院上海药物研究所药物制剂研究中心,上海201203

出　　处：《中药新药与临床药理》2021年第8期1164-1172,共9页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：上海市进一步加快中医药事业三年行动计划(2018-2020)资助项目[ZY(2018-2020)-CCCX-2001-01]。

摘　　要：目的运用网络药理学和体外实验探讨驻景丸治疗老年性黄斑变性(AMD)的作用机制。方法检索Genecards、OMIM数据库与AMD相关的作用靶点,使用Jvenn在线工具获取驻景丸活性成分与AMD的交集靶点,构建“药物-成分-靶点”网络以及蛋白互作(PPI)网络。采用DAVID数据库对交集靶点的GO功能和KEGG通路富集分析,将网络中度值较高的成分与靶点进行分子对接。通过体外MTT实验和Western Blot实验,使用驻景丸有效成分干预人视网膜色素上皮细胞(ARPE19),进行核心靶点和通路的初步验证。结果网络药理学结果显示,共筛选出驻景丸中38个活性成分、150个交集靶点。蛋白互作网络显示关键靶点有MAPK1、Akt1、VEGFA等。GO富集分析涉及到247个生物过程、36个细胞组分和64个分子功能。KEGG通路分析得到HIF-1信号通路、PI3K-Akt信号通路、前列腺癌等85条相关通路。分子对接结果显示驻景丸治疗AMD的主要活性成分可以与MAPK1、Akt1、STAT3和MAPK3有较稳定的结合,其中Akt1与异鼠李素结合性最强。MTT结果显示驻景丸活性成分异鼠李素、槲皮素和山柰酚均可以促进碘酸钠损伤的ARPE19细胞增殖(P<0.05,P<0.01,P<0.001)。Western Blot结果证实碘酸钠通过激活STAT3信号通路诱导细胞凋亡相关蛋白Caspase-3、PARP、BAD表达(P<0.01,P<0.001),异鼠李素、槲皮素和山柰酚则可以通过抑制STAT3信号通路进而抑制Caspase-3、PARP、BAD凋亡蛋白表达(P<0.05,P<0.01,P<0.001)。结论本研究从网络药理学角度预测了驻景丸治疗AMD的药效物质基础及分子机制,并通过体外实验进行核心靶点和通路的初步验证,为驻景丸等中药复方研究提供了新的思路。Objective To explore the mechanism of Zhujing pills in treating age-related macular degeneration(AMD)by using network phamacology and in vitro experiments.Methods The targets related to AMD in Genecards and OMIM database were searched,and the intersection targets of active components of Zhujing pills and AMD were obtained by using Jvenn online tool,and the drug-component-target network and PPI protein interaction network were constructed.The DAVID database was used to analyze the GO function and KEGG pathway enrichment of the intersection targets,and finally molecular docking was carried out between the components with high moderate value of the network and the targets.Through MTT experiment in vitro and Western Blot experiment,the active ingredients of Zhujing pills were used to intervene in human retinal pigment epithelial cells(ARPE19)to conduct preliminary verification of core targets and pathways.Results A total of 38 active components and 150 intersection targets were screened out.The protein interaction network showed that the key targets were MAPK1,Akt1 and VEGFA.The GO enrichment analysis involved 247 biological processes,36 cell components and 64 molecular functions.The HIF-1 signaling pathway,PI3K-Akt signaling pathway,prostate cancer and other 85 related pathways were also given by KEGG pathway analysis.The results of molecular docking showed that the main active ingredients of Zhujing pills in the treatment of AMD can bind stably to MAPK1,Akt1,STAT3 and MAPK3,and Akt1 has the strongest binding to isorhamnetin.MTT results showed that the active ingredients of Zhujing pills such as isorhamnetin,quercetin and kaempferol can promote the proliferation of ARPE19 cells damaged by sodium iodate(P<0.05,P<0.01,P<0.001).Western Blot results confirmed that sodium iodate induces the expression of apoptosis-related proteins Caspase-3,PARP,BAD(P<0.01,P<0.001).Isorhamnetin,quercetin and kaempferol can inhibit the expression of Caspase-3,PARP and BAD apoptotic proteins by inhibiting the STAT3 signaling pathway(P<0.05

关 键 词：驻景丸 老年性黄斑变性 网络药理学 实验验证 分子对接 作用机制 

分 类 号：R285.5[医药卫生—中药学]