基于网络药理学和实验验证探讨补肾活血汤治疗中枢性老年性聋的机制  

Exploring Mechanisms of Bushen Huoxue Decoction in the Treatment of Central Presbycusis Based on Network Pharmacology and Experimental Verification

作　　者：周颖东 张梦娴 王青玲 康浩然 郭向东[2] ZHOU Yingdong;ZHANG Mengxian;WANG Qingling;KANG Haoran;GUO Xiangdong(Henan University of Chinese Medicine,Zhengzhou 450046 Henan,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000 Henan,China)

机构地区：[1]河南中医药大学,河南郑州450046 [2]河南中医药大学第一附属医院,河南郑州450000

出　　处：《中药新药与临床药理》2023年第10期1398-1408,共11页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81403439);河南省科技攻关计划项目(222102310604);河南省中医药科学研究专项重点课题(20-21ZY1045);河南省高等学校重点科研项目(23A360028)。

摘　　要：目的结合网络药理学与实验研究探究补肾活血汤(Bushen Huoxue Decoction,BSHXD)治疗中枢性老年性聋的潜在分子机制。方法通过TCMSP、ETCM、BATMAN-TCM数据库检索和筛选BSHXD的活性成分及作用靶点;通过Genecards、DisGeNET数据库检索老年性聋相关靶点;通过在线工具Venny 2.1获得BSHXD与老年性聋的共同靶点;通过Cytoscape软件构建BSHXD治疗中枢性老年性聋的“药物-成分-靶点-疾病”网络;使用DAVID数据库进行GO功能和KEGG通路富集分析;STRING数据库进行蛋白互作分析。采用自然衰老的C57BL/6J小鼠构建中枢性老年性聋的动物模型,以BSHXD进行灌胃干预。HE染色观察各组听皮层的病理改变;免疫印迹法检测听皮层组织PI3K、AKT、mTOR、p-PI3K、p-AKT、p-mTOR、Bax和Caspase-3的蛋白表达;免疫荧光染色观察听皮层Caspase-3的表达;TUNEL染色观察听皮层细胞的凋亡。结果通过网络药理学预测得BSHXD活性成分219种,BSHXD与老年性聋共同靶点1118个。GO与KEGG富集发现在BSHXD可能通过调控PI3K/AKT/mTOR信号通路与细胞凋亡途径治疗中枢性老年性聋。与对照组比,自然衰老小鼠听皮层p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR的比值升高(P<0.01,P<0.001),Bax和Caspase-3的表达明显上调(P<0.001),听皮层细胞凋亡明显增加(P<0.001),病理改变明显。BSHXD可明显降低p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR比值(P<0.01,P<0.001),并下调Bax与Caspase-3的表达(P<0.05,P<0.01,P<0.001),减少细胞凋亡并改善听皮层病理改变。结论BSHXD可以通过调控PI3K/AKT/mTOR信号通路与细胞凋亡途径延缓听皮层的衰老。Objective To explore the underlying mechanisms of Bushen Huoxue Decoction(BSHXD)in the treatment of central presbycusis based on integrating network pharmacology and experimental verification.Methods The active compounds and targets of BSHXD were searched and screened by TCMSP,ETCM and BATMAN-TCM databases.The age-related hearing loss(ARHL)-related targets were searched by Genecards and DisGeNET databases.The common targets of BSHXD and ARHL was obtained by Venny 2.1.The“drugs-components-targetsdiseases”network of BSHXD in the treatment of central presbycusis was constructed by Cytoscape software.The Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were performed by DAVID database.The protein-protein interaction analysis was performed by STRING database.The natural aging C57BL/6J mice were used as animal models of central presbycusis.HE staining was used to observe the pathological changes of auditory cortex,then Western Blot was used to detect the relative protein levels of PI3K,AKT,mTOR,p-PI3K,p-AKT,p-mTOR,Bax and Caspase-3,and immunofluorescence was used to observe the expressions of Caspase-3 in auditory cortex.Finally,TUNEL staining was used to investigate the apoptosis in auditory cortex.Results A total of 219 active compounds of BSHXD and 1118 common targets between BSHXD and ARHL were acquired.The results of GO and KEGG enrichment analysis suggested that BSHXD may treat central presbycusis by regulating PI3K/AKT/mTOR signal pathway and apoptosis pathway.Compared with the control group,the ratios of p-PI3K/PI3K,p-Akt/AKT and p-mTOR/mTOR in the auditory cortex of naturally aged mice were increased(P<0.01,P<0.001),the expressions of Bax and Caspase-3 were significantly up-regulated(P<0.001),apoptosis was significantly increased in the auditory cortex(P<0.001),and the pathological changes were obvious.BSHXD significantly decreased the ratios of p-PI3K/PI3K,p-Akt/AKT and p-mTOR/mTOR(P<0.01,P<0.001),down-regulated the expressions of Bax and Caspase-3(P<0.05,P<0.01,P<0.00

关 键 词：补肾活血汤 中枢性老年性聋 网络药理学 听皮层 衰老 PI3K/AKT/mTOR信号通路 细胞凋亡 小鼠 

分 类 号：R285.5[医药卫生—中药学]