基于蛋白质组学研究驻景丸加减方治疗肝肾不足型年龄相关性黄斑变性的机制  

Mechanism of Modified Zhujing Pills(驻景丸)in Treatment of Age-related Macular Degeneration with Ganshenbuzu(肝肾不足)Based on Proteomics

作　　者：仲路[1] 张元钟[2] 惠沁怡 熊彩建 杨宁[3] 杭丽[1] 徐新荣[1] ZHONG Lu;ZHANG Yuanzhong;HUI Qinyi;XIONG Caijian;YANG Ning;HANG Li;XU Xinrong(Department of Ophthalmology,Jiangsu Province Hospital of Chinese Medicine,Nanjing 210029;Department of Ophthalmology,Nanjing Hospital of Chinese Medicine,Nanjing 210022;Nanjing University of Chinese Medicine,Nanjing 210023)

机构地区：[1]江苏省中医院眼科,南京210029 [2]南京市中医院眼科,南京210022 [3]南京中医药大学,南京210023

出　　处：《中药药理与临床》2024年第6期72-78,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金(编号:82074177);江苏省重点研发计划(编号:BE2018757)。

摘　　要：目的:基于血浆蛋白质组学探讨驻景丸加减方治疗肝肾不足型年龄相关性黄斑变性(Age-related macular degeneration,AMD)的机制。方法:采用TMT蛋白质组学技术,筛选正常人、肝肾不足AMD患者驻景丸加减方治疗前后血浆差异蛋白;对差异蛋白进行聚类分析、蛋白相互作用分析以及GO、Pathway功能注释和富集分析;选择蛋白相互作用分析度值最高的5个蛋白作为候选靶蛋白,ELISA法检测AMD患者血浆靶蛋白水平进行体内验证;选择Pathway富集分析主要信号通路,Western blot法检测驻景丸加减方含药血清干预ARPE-19细胞后相关通路蛋白表达水平进行体外验证。结果:筛选出主要候选靶蛋白5个,分别是CDH1、CDC42、PTPRK、CYCS和PTPRG,体内验证结果显示:与治疗前比较,治疗后血浆靶蛋白CDH1、PTPRK、CYCS升高(P<0.01),CDC42、PTPRG下降(P<0.01);Pathway富集分析获得主要信号通路是EMT和Wnt信号通路,体外验证结果显示驻景丸加减方能抑制EMT和Wnt信号通路。结论:驻景丸加减方治疗肝肾不足AMD的机制可能是调控EMT和Wnt信号通路。Objective:To investigate the mechanism of modified Zhujing Pills(驻景丸)in treating age-related macular degeneration(AMD)with Ganshenbuzu(肝肾不足)based on plasma proteomics.Methods:Using TMT proteomics technology,differential proteins in plasma before and after treatment with modified Zhujing Pills in normal individuals and AMD patients with Ganshenbuzu were screened.Differential proteins were subjected to clustering analysis,protein-protein interaction(PPI)analysis,GO,pathway functional annotation,and enrichment analysis.Five proteins with the highest degree of PPI analysis were selected as candidate target proteins.ELISA was used to detect the levels of plasma target proteins in AMD patients forin vivo validation.Pathway enrichment analysis was performed to identify the main signaling pathways.Western blot was used to detect the expression levels of pathway-related proteins in ARPE-19 cells after intervention with serum containing modified Zhujing Pills for in vitro validation.Results:Five main candidate target proteins were screened out,namely CDH1,CDC42,PTPRK,CYCS,and PTPRG.In vivo validation results showed that compared with the results before treatment,the plasma levels of target proteins CDH1,PTPRK,and CYCS increased after treatment(P<0.01),while CDC42 and PTPRG decreased(P<0.01).Pathway enrichment analysis revealed that the main signaling pathways were epithelial-mesenchymal transition(EMT)and Wnt signaling pathways.In vitro validation results showed that modified Zhujing Pills could inhibit EMT and Wnt signaling pathways.Conclusion:The mechanism of modified Zhujing Pills in treating AMD with Ganshenbuzu may involve the regulation of EMT and Wnt signaling pathways.

关 键 词：驻景丸加减方 年龄相关性黄斑变性 肝肾不足证 血浆 蛋白质组学 

分 类 号：R276.7[医药卫生—中医五官科学]