从临床多组学整合调控网络探...关节炎的“效-毒”关联机制_王晓月.pdf

第 29 卷第 5 期2023年 3 月中国实验方剂学杂志Chinese Journal of Experimental Traditional Medical FormulaeVol.29，No.5Mar.，2023从临床多组学整合调控网络探究雷公藤多苷片治疗类风湿关节炎的“效-毒”关联机制王晓月1，张依1，陈文佳1，万磊2，刘健2，张彦琼1*，林娜1*（1.中国中医科学院 中药研究所，北京 100700；2.安徽中医药大学 第一附属医院，合肥 230031）摘要 目的：通过雷公藤多苷片治疗类风湿关节炎疗效应答和诱发药源性肝损伤相关分子互作网络挖掘，揭示雷公藤多苷片的“效-毒”关联机制。方法：利用前期已获得临床来源的雷公藤多苷片疗效应答相关基因表达谱和诱发药源性肝损伤相关蛋白质表达谱数据，根据生物分子间的相互作用信息，构建雷公藤多苷片“效-毒”关联网络，并通过计算网络节点的拓扑特征值，筛选关键网络靶标和挖掘其生物学功能。结果：经临床转录组学检测，获得雷公藤多苷片治疗类风湿关节炎疗效相关差异表达基因 119个。经临床蛋白质组学检测，获得雷公藤多苷片诱发药源性肝损伤相关差异表达蛋白质 49个。进一步的临床症状富集分析表明，雷公藤多苷片治疗类风湿关节炎的效应基因与痹症相关中医症状和医学临床表型紧密相关，雷公藤多苷片诱发药源性肝损伤相关的差异蛋白质最显著富集于消化系统异常的临床症状，可见，上述多组学数据能够表征雷公藤多苷片治疗类风湿关节炎和诱发药源性肝损伤的主要临床症状。进而，“效-毒”关联网络挖掘表明，雷公藤多苷片治疗类风湿关节炎的关键效应靶标和诱发药源性肝损伤核心致毒蛋白质均参与机体“免疫-炎症”失衡的调节，特别在中性粒细胞脱颗粒、补体级联反应、蛋白质翻译后修饰介导的免疫炎症反应等方面发挥着重要作用；二者还显著富集于肾素-血管紧张素系统（RAS）和丝裂原活化蛋白激酶（MAPK）信号通路等与细胞增殖和细胞周期调控密切相关的信号通路，以及参与糖原代谢和氧化还原反应等肝脏功能直接相关的生物学通路。结论：本研究采用临床表型组学、临床转录组学和蛋白质组学整合的研究策略，系统阐释了雷公藤多苷片“效-毒”的关联特征和分子机制，为探索雷公藤多苷片“增效减毒”机制奠定良好的数据基础。关键词 雷公藤多苷片；类风湿关节炎；药源性肝损伤；多组学整合；“效-毒”关联网络中图分类号 R2-0；R33；R593.22 文献标识码 A 文章编号 1005-9903（2023）05-0049-09doi 10.13422/ki.syfjx.20221545 增强出版附件 内容详见 http：/或 http：/网络出版地址 https:/ 2022-07-07 17:05:33Exploration on Efficacy-toxicity Association Mechanisms of Tripterygium wilfordii Polyglycoside Tablets Against Rheumatoid Arthritis Based on Multi-omics Integrated Regulatory NetworkWANG Xiaoyue1，ZHANG Yi1，CHEN Wenjia1，WAN Lei2，LIU Jian2，ZHANG Yanqiong1*，LIN Na1*（1.Institute of Chinese Materia Medica，China Academy of Chinese Medical Sciences，Beijing 100700，China；2.The First Affiliated Hospital of Anhui University of Chinese Medicine，Hefei 230031，China）Abstract Objective：To explore the efficacy-toxicity association mechanisms of Tripterygium wilfordii polyglycoside tablets（TWPT）by establishing and analyzing an interaction network associated with the clinical efficacy of TWPT in the treatment of rheumatoid arthritis（RA）and TWPT-induced liver injury.收稿日期 2022-04-24基金项目 国家自然科学基金面上项目（81974526）；中国中医科学院科技创新工程项目（CI2021A03807）；北京市自然科学基金面上项目（7212186）；中央级公益性科研院所基本科研业务费专项（ZXKT22019）第一作者 王晓月，在读博士，从事中药药理研究，E-mail：通信作者 *张彦琼，研究员，博士生导师，从事中医药系统生物学研究工作，E-mail：；*林娜，研究员，博士生导师，从事中药药理研究，E-mail：49第 29 卷第 5 期2023年 3 月中国实验方剂学杂志Chinese Journal of Experimental Traditional Medical FormulaeVol.29，No.5Mar.，2023Method：On the basis of the TWPT efficacy-related gene expression profile and TWPT-induced liver injury-related protein expression profile which were both obtained from our clinical cohorts，the efficacy-toxicity association network of TWPT was constructed，and the key network targets were identified by calculating the topological values of the nodes，including the degree，closeness and betweenness.After that，the biological functions and pathways of the key network targets were investigated by enrichment analysis.Result：A total of 119 differentially expressed genes（58 up-regulated and 61 down-regulated）between RA patients with TWPT well and weak response were identified as TWPT efficacy-related genes by clinical transcriptomics，and 49 differentially expressed proteins（36 up-regulated and 13 down-regulated）were demonstrated to be TWPT-induced liver injury-related proteins by clinical proteomics.In addition，the clinical symptom enrichment analysis indicated that the TWPT efficacy-related genes were significantly associated with various clinical symptoms of arthralgia in traditional Chinese medicine and clinical phenotypes of modern medicine，and most of the TWPT-induced liver injury-related proteins were involved in digestive system abnormalities.Therefore，the aforementioned multi-omics data represented the main clinical symptoms of TWPT treating RA and inducing liver injury.Mechanically，the efficacy-toxicity association network revealed that both TWPT efficacy-related genes and TWPT-induced liver injury-related core proteins were involved in the immune-inflammatory imbalance，especially playing an important role in neutrophil degranulation，complement cascade reaction，and immune-inflammatory response mediated by protein post-translational modification.Notably，the above genes and proteins were also enriched in various signaling pathways related to cell proliferation and cell cycle regulation，such as RAS and mitogen-activated protein kinase（MAPK）signaling pathway，and in several liver functional processes，such as glycogen metabolism and redox reaction.Conclusion：This study systematically explained the efficacy-toxicity association characteristics and molecular mechanisms of TWPT by applying a research strategy integrating clinical phenomics，transcriptomics and proteomics，laying a good data foundation for exploring the efficacy enhancing and toxicity-reducing mechanisms of TWPT.Keywords Tripterygium wilfordii polyglycoside tablets；rheumatoid arthritis；drug-induced liver injury；multi-omics integration；efficacy-toxicity association network中药雷公藤为卫矛科植物雷公藤 Tripterygium wilfordii的干燥根或根的木质部，首载于明朝 滇南本草，味辛、苦，性温，有毒，入肝脾二经，行十二经络，治筋骨疼痛，风湿寒痹，麻木不仁，瘫痪痿软，湿气流痰，暖筋，止腰疼1-2。中药雷公藤的代表制剂雷公藤多苷片不仅包含雷公藤的主要化学成分，且临床治疗类风湿关节炎（RA）效果确切。近期多项双盲随机对照临床试验证明，雷公藤多苷片