UPLC-Q-TOF-MS联合网络药理学和实验验证分析桃红四物汤治疗软组织损伤的作用机制  

Mechanism of Taohong Siwu Decoction in treating soft tissue injury based on UPLC-Q-TOF-MS,network pharmacology and experimental verification

作　　者：王伟军 段雪云[2,3] 汪伟[2,3] WANG Wei-jun;DUAN Xue-yun;WANG Wei(the First Clinical College^Hubei University of Chinese Medicine,Wuhan 430061,China;Hubei Provincial Hospital of Traditional Chinese Medicine,Wuhan 430061,China;Hubei Province Academy of Traditional Chinese Medicine,Wuhan 430074,China)

机构地区：[1]湖北中医药大学第一临床学院,湖北武汉430061 [2]湖北省中医院,湖北武汉430061 [3]湖北省中医药研究院,湖北武汉430074

出　　处：《中国中药杂志》2021年第12期3043-3051,共9页China Journal of Chinese Materia Medica

摘　　要：基于UPLC-Q-TOF-MS技术、网络药理学和实验验证方法探讨桃红四物汤(Taohong Siwu Decoction, THSWD)治疗软组织损伤(soft tissue injury, STI)的作用机制。采用UPLC-Q-TOF-MS技术鉴定THSWD化学成分;通过TCMSP数据库筛选THSWD活性成分及靶蛋白;Cytoscape软件构建“活性成分-靶点-通路”网络,STRING数据库进行蛋白互作分析;GeneCards和CTD数据库筛选STI相关靶点;DAVID数据库进行GO功能和KEGG通路富集分析;构建软组织损伤大鼠模型,通过蛋白免疫印迹法探讨THSWD对大鼠软组织损伤相关通路关键靶点的影响。筛选得到THSWD活性成分40种,潜在作用靶点141个,与STI相关靶点交集20个。活性成分作用靶点富集分析得到128条KEGG通路,主要富集在氨基酸合成与代谢、疾病信号通路、细胞凋亡、炎症等相关的通路。对“活性成分-靶点-通路”网络进行可视化分析,得到THSWD作用于STI的5种关键活性成分和6个关键蛋白。蛋白免疫印迹法实验结果表明,THSWD可以显著降低大鼠STI样本中细胞凋亡蛋白酶-3(CASP3)和B细胞κ轻肽基因增强子核因子1(NFKB1)的蛋白表达及磷酸化水平,降低前列腺素G/H合酶2(PTGS2)蛋白表达,增强细胞肿瘤抗原p53(TP53)蛋白表达。UPLC-Q-TOF-MS、网络药理学和实验验证结果表明,THSWD中活性成分可能通过调控NF-κB信号通路及细胞凋亡通路中的关键蛋白,发挥抗炎、抗氧化应激的作用,从而发挥治疗STI的作用。To explore the action mechanism of Taohong Siwu Decoction(THSWD) in the treatment of soft tissue injury(STI) based on UPLC-Q-TOF-MS technique, network pharmacology and experimental verification method. UPLC-Q-TOF-MS technique was used to identify the chemical constituents of THSWD. The active ingredients and predicted target proteins of THSWD were screened out through TCMSP database. Cytoscape software was used to construct the active component-target-pathway network, and STRING database was used for protein interaction analysis. GeneCards and CTD databases were used to screen out relevant targets of STI. GO function and KEGG pathway enrichment analysis were performed through DAVID database. The rat model of STI was constructed, and Western blot was used to verify the effect of THSWD on key targets of relevant pathways. The results showed 40 active ingredients in THSWD, and 141 potential targets and 20 targets of STI. Target enrichment analysis of the active components produced 128 KEGG pathways, which were mainly concentrated in amino acid synthesis and metabolism, disease signaling pathways, apoptosis, inflammation and other relevant pathways. Western blot showed that THSWD intervention could significantly decrease PTGS2, CASP3, NFKB1, p-CASP3 and p-NFKB1, while enhancing the expression of TP53 protein in the STI samples of rats. According to the results of UPLC-Q-TOF-MS, network pharmacology and experimental verification, active ingredients in THSWD may play anti-inflammatory and antioxidant effects in NF-κB signaling pathway and apoptotic pathway, thus playing a role in the treatment of STI.

关 键 词：桃红四物汤 网络药理学 软组织损伤 UPLC-Q-TOF-MS 信号通路 炎症 

分 类 号：R285.5[医药卫生—中药学]