Li Changhong, Tang Shiyi, Hu Tianqi, Zhou Chenkang, Chen Yuxin, Hu Zhaoting, Pan Jingjing, Chen Jie, Wang Yumin
Department of Laboratory Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325015, China.
Cixi Biomedical Research Institute, Wenzhou Medical University, Zhejiang, China.
Comput Biol Chem. 2025 Apr;115:108314. doi: 10.1016/j.compbiolchem.2024.108314. Epub 2025 Jan 2.
As a widely recognized traditional Chinese medicine (TCM) decoction prescription in China, numerous studies have shown that Wutou-Guizhi decoction (WTGZD) exhibits significant therapeutic efficacy for Rheumatoid arthritis (RA). Nevertheless, the underlying molecular mechanisms have yet to be fully elucidated. This study aims to establish a database of active ingredients for WTGZD and identify RA-related target genes. The WTGZD-RA-Potential target gene network and protein-protein interaction network were constructed, followed by gene ontology (GO) analysis and functional enrichment analysis utilizing the Kyoto Encyclopedia of Genes and Genomes (KEGG). Cell proliferation was confirmed through CCK8 assay. Target gene identification was performed via real-time PCR using quantitative methods, and western blot analysis was conducted. In the course of this investigation, 95 active components of drugs and 34 targets associated with rheumatoid arthritis were identified. Through the utilization of network pharmacology analysis, we were able to identify a total of 17 essential active components of WTGZD and pinpoint 12 significant targets linked to rheumatoid arthritis (RA). Our findings suggest a consistent interaction between the key components of WTGZD and the critical targets associated with RA. Subsequent qPCR analysis revealed that stigmasterol, a principal constituent of WTGZD, exhibited inhibitory effects on the expression of various RA-related factors, such as TNF-α, IL-1β, MAPK8, MMP1, MMP3, and MMP9. Moreover, WTGZD effectively mitigated the increased protein expression of MMP-1 and MAPK8 induced by LPS stimulation, both of which are integral components of the IL-17 signaling pathway. These results suggest that WTGZD may play a significant role in the therapeutic intervention of rheumatoid arthritis by suppressing inflammatory immune responses.
作为中国广泛认可的中药汤剂处方，大量研究表明，乌头桂枝汤（WTGZD）对类风湿关节炎（RA）具有显著的治疗效果。然而，其潜在的分子机制尚未完全阐明。本研究旨在建立乌头桂枝汤的活性成分数据库，并鉴定与RA相关的靶基因。构建了WTGZD-RA-潜在靶基因网络和蛋白质-蛋白质相互作用网络，随后利用京都基因与基因组百科全书（KEGG）进行基因本体（GO）分析和功能富集分析。通过CCK8法确认细胞增殖。采用定量实时PCR进行靶基因鉴定，并进行蛋白质印迹分析。在本研究过程中，鉴定出95种药物活性成分和34个与类风湿关节炎相关的靶点。通过网络药理学分析，共鉴定出乌头桂枝汤的17种关键活性成分，并确定了12个与类风湿关节炎（RA）相关的重要靶点。我们的研究结果表明，乌头桂枝汤的关键成分与RA相关的关键靶点之间存在一致的相互作用。随后的qPCR分析表明，乌头桂枝汤的主要成分豆甾醇对多种RA相关因子如TNF-α、IL-1β、MAPK8、MMP1、MMP3和MMP9的表达具有抑制作用。此外，乌头桂枝汤有效减轻了LPS刺激诱导的MMP-1和MAPK8蛋白表达增加，这两者都是IL-17信号通路的组成部分。这些结果表明，乌头桂枝汤可能通过抑制炎症免疫反应在类风湿关节炎的治疗干预中发挥重要作用。
