矮地茶治疗骨质疏松症的网络药理学分析及实验研究  

Network pharmacological analysis and experimental study of Aidicha(Ardisiae Japonicae Herba)in treating osteoporosis

作　　者：谭雅文 原茵 吴紫璇 郭亚萍 王瑾源 彭雁飞 郑纺 TAN Yawen;YUAN Yin;WU Zixuan;GUO Yaping;WANG Jinyuan;PENG Yanfei;ZHENG Fang(School of Integrated Chinese and Western Medicine,Tianjin University of Chinese Medicine,Tianjin 301617,China)

机构地区：[1]天津中医药大学中西医结合学院,天津301617

出　　处：《湖南中医药大学学报》2023年第8期1468-1477,共10页Journal of Hunan University of Chinese Medicine

基　　金：国家自然科学基金青年项目(81703828);天津市自然科学基金面上项目(17JCYBJC26500);天津中医药大学中西医结合学院研究生创新基金(ZXYCXLX202117)。

摘　　要：目的基于网络药理学及体外实验探究矮地茶治疗骨质疏松症(osteoporosis,OP)的作用机制。方法使用TCMSP、GeneCards数据库筛选矮地茶治疗骨质疏松症的有效成分及关键靶点(疾病靶点与药物活性成分靶点的交集);使用Cytoscape 3.7.1软件构建“中药-活性成分-交集靶点-疾病”网络;使用STRING数据库构建关键靶点蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络;通过DAVID数据库对关键靶点进行GO分析和KEGG信号通路富集分析。体外实验使用矮地茶醇提物干预成骨诱导的ST-2细胞,通过碱性磷酸酶染色(alkaline phosphatase,ALP)、RT-qPCR、Western blot等检测其对成骨分化的影响并验证网络药理学信号通路预测结果。结果共获得槲皮苷、山柰酚、射干醇A、丹皮酚、肉桂醛等193个矮地茶主要活性成分;预测出82个矮地茶与OP的关键靶点,这些关键靶点主要参与调控雌激素信号通路、凋亡、Wnt/β-catenin信号通路等。体外实验显示,矮地茶能有效提高ST-2细胞成骨转录因子表达(P<0.05),同时上调Wnt/β-catenin信号通路关键蛋白β-catenin的表达(P<0.05),并下调通路抑制因子Sfrp1的mRNA水平(P<0.05)。结论矮地茶可通过多靶点、多通路治疗OP,主要与Wnt/β-catenin信号通路有关。Objective To investigate the mechanism of action of Aidicha(Ardisiae Japonicae Herba)in treating osteoporosis(OP)based on network pharmacology and in vitro experiments.Methods TCMSP and GeneCards databases were used to screen active ingredients and key targets(intersection of disease targets and drug active ingredient targets)for Aidicha(Ardisiae Japonicae Herba)treating OP;Cytoscape 3.7.1 software was applied to constructing"herb-active ingredient-intersection target-disease"network;STRING database was used to construct the key target protein-protein interaction(PPI)network.GO analysis and KEGG signaling pathway enrichment analysis of the key targets were performed by DAVID database.In vitro experiments were performed using Aidicha(Ardisiae Japonicae Herba)alcohol extracts to intervene in osteogenic-induced ST-2 cells.Alkaline phosphatase(ALP)staining,RT-qPCR,and Western blot were applied to detecting their effects on osteogenic differentiation and validating the prediction made by network pharmacological signaling pathway analysis.Results A total of 193 major active ingredients of Aidicha(Ardisiae Japonicae Herba),including quercetin,kaempferol,belamcandol A,paeonol,and cinnamaldehyde,were obtained;82 key targets(intersection targets)of Aidicha(Ardisiae Japonicae Herba)and OP were predicted,and they were mainly regulating estrogen signaling pathway,apoptosis,Wnt/β-catenin signaling pathway,etc.In vitro experiments showed that Aidicha(Ardisiae Japonicae Herba)effectively increased the expressions of osteogenic transcription factors in ST-2 cells(P<0.05).Meanwhile,it up-regulated the expression ofβ-catenin,a key protein of Wnt/β-catenin signaling pathway(P<0.05),but down-regulated the mRNA level of Sfrp1,a pathway suppressor(P<0.05).Conclusion Aidicha(Ardisiae Japonicae Herba)can treat OP through multiple targets and pathways,mainly related to Wnt/β-catenin signaling pathway.

关 键 词：骨质疏松症 矮地茶 成骨分化 网络药理学 WNT/Β-CATENIN信号通路 

分 类 号：R274.9[医药卫生—中西医结合]