基于网络药理学及实验验证探讨除风益损汤治疗角膜损伤机制  

Exploration on Machanism of Chufeng Yisun Decoction in the Treatment of Corneal Injury Based on Network Pharmacology and Experimental Validation

作　　者：李江伟 陈惠媚 张文清 欧晨 陈雄 姚小磊[1,2] 彭清华[2] LI Jiangwei;CHEN Huimei;ZHANG Wenqing;OU Chen;CHEN Xiong;YAO Xiaolei;PENG Qinghua(The First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007 [2]湖南中医药大学,湖南长沙410208

出　　处：《中国中医药信息杂志》2024年第8期23-28,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(82174443、81860870);国家中医药管理局人才支持项目-岐黄学者(2022年);国家中医药管理局中医眼科学重点学科建设项目(ZK1801YK015);湖南省自然科学基金(2021JJ30527);湖南省卫生健康高层次人才计划(2022年);湖南省教育厅科学研究项目(20A370);湖南省中医药科研计划项目(2021023);湖南省研究生科研创新项目(CX20220782);湖南中医药大学中医学一流学科开放基金(2021ZYX05)。

摘　　要：目的采用网络药理学结合实验验证方法研究除风益损汤治疗角膜损伤的机制。方法通过TCMSP、TCMID数据库获取除风益损汤组方药物活性成分及相关靶点,通过GeneCards、OMIM、TTD、NCBI-Gene数据库查找角膜损伤相关靶点。建立中药-活性成分-关键靶点网络,分析除风益损汤治疗角膜损伤的主要活性成分,并通过蛋白相互作用网络预测核心靶点。采用CCK-8法筛选除风益损汤含药血清促进细胞生长的最佳浓度,Westernblot检测自噬相关蛋白LC3、LAMP1及ERK2表达。结果除风益损汤治疗角膜损伤的主要活性成分为山柰酚、汉黄芩素、槲皮素及芍药苷,核心靶点为AKT1、TP53、MAPK1、JUN及TNF。除风益损汤含药血清干预人角膜成纤维细胞可升高LC3Ⅱ/LC3Ⅰ比值及LAMP1蛋白表达,降低ERK2蛋白表达,符合网络药理学预测。结论除风益损汤通过多成分、多靶点、多通路治疗角膜损伤,其促进自噬治疗角膜损伤的机制是通过下调ERK2表达、上调LC3和LAMP1表达实现的。Objective To study the mechanism of Chufeng Yisun Decoction in the treatment of corneal injury based on network pharmacology combined with experimental validation.Methods The active components and targets of Chufeng Yisun Decoction were obtained from TCMSP and TCMID databases.Related targets of corneal injury were searched through GeneCards,OMIM,TTD and NCBI-Gene databases.Chinese materia medica-active components-key target network was established.The main active components of Chufeng Yisun Decoction for the treatment of corneal injury were analyzed.The core targets were predicted through PPI network.CCK-8 method was used to screen the optimal concentration of serum containing Chufeng Yisun Decoction for promoting cell growth.Western blot was used to detect autophagy related protein expressions of LC3,LAMP1 and ERK2.Results The main active components of Chufeng Yisun Decoction in the treatment of corneal injury were kaempferol,wogonin,quercetin and paeoniflorin.The core targets were AKT1,TP53,MAPK1,JUN and TNF.The intervention of serum containing Chufeng Yisun Decoction on human corneal fibroblasts could increase the LC3I/LC3II ratio and LAMP1 protein expression,while decrease ERK2 protein expression,which was consistent with the prediction of network pharmacology.Conclusion Chufeng Yisun Decoction treats corneal injury through multiple components,targets and pathways.The mechanism of promoting autophagy therapy for corneal injury is achieved by down-regulating the expression of ERK2 and up-regulating the expression of LC3 and LAMP1.

关 键 词：除风益损汤 角膜损伤 自噬 炎症 网络药理学 

分 类 号：R285[医药卫生—中药学]