基于网络药理学及实验验证探讨黑沙蒿有效物质群治疗类风湿关节炎的作用机制*

作者：呼格吉乐1，王青虎1，张小峰1，康秀荣2，高明霞1

单位：1.内蒙古民族大学蒙医药学院，内蒙古 通辽 028000； 2.通辽职业学院，内蒙古 通辽 028000

引用：引用：呼格吉乐，王青虎，张小峰，康秀荣，高明霞.基于网络药理学及实验验证探讨黑沙蒿有效物质群治疗类风湿关节炎的作用机制[J].中医药导报,2025,31(8):31-40,52.

DOI：10.13862/j.cn43-1446/r.2025.08.006

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摘要：

目的：从网络药理学、分子对接及细胞验证实验探讨黑沙蒿有效物质群（HSH）治疗类风湿关节炎（RA）作用机制。方法：利用Pubchem（化合物数据库）、BATMAN-TCM（中药分子机制生物信息学分析工具）、DisGeNet（疾病基因关联网络数据库）、GeneCards（基因卡片数据库）、NCBI（美国国家生物技术信息中心）、OMIM（在线人类孟德尔遗传数据库）、PharmGKB（药理基因组知识库）、GEO（基因表达综合数据库）等数据库及R语言获得HSH核心成分和成分靶点、疾病靶点及HSH治疗RA的潜在靶点。建立蛋白-蛋白相互作用（PPI）网络，进行基因本体（GO）和京都基因和基因组百科全书（KEGG）富集分析，选取核心成分和核心靶点进行分子对接。采用细胞计数试剂盒-8（CCK-8）和CFSE标记法检测HSH的无毒浓度和作用浓度。采用流式细胞技术、流式液相多重蛋白定量技术（CBA）、逆转录PCR（RT-PCR）等实验技术检测白细胞介素（IL）-17/T细胞受体信号通路相关细胞、关键性炎症因子及蛋白的变化。结果：对羟基肉桂酸、邻羟基肉桂酸、o-hydroxycapillene等3个核心成分与RA的关联度最高，其成分靶点有90个；经疾病相关靶点及RA差异表达基因交集获得875个RA疾病靶点。将RA疾病靶点与成分靶点相交，得到36个RA的潜在靶点；经PPI构建获取5个核心靶点，核心靶点与核心成分之间具有一定的结合活性；GO和KEGG富集分析显示IL-17/T细胞受体信号通路与HSH抗RA密切相关，HSH参与炎症反应及细胞因子产生和调节等多种生物过程，进而调控机体炎症和免疫反应发挥抗RA的作用。HSH的作用浓度为16 μg/mL，HSH能下调辅助性T细胞1型（Th1）水平，上调调节性T细胞（Treg）水平，降低Th17/Treg的比值，也能显著降低IL-21、IL-6、IL-2、γ干扰素（IFN-γ）、转化生长因子β1（TGF-β1）水平。RT-PCR实验结果显示，HSH显著下调维甲酸相关孤儿受体γt（RORγt）mRNA的表达，而叉头翼状螺旋转录因子（Foxp3）mRNA的表达升高无显著性差异。结论：HSH对RA具有良好的治疗作用，其作用机制可能与降低Th1，升高Treg，调节Th17/Treg的平衡和降低IL-21、IL-6、IL-2、IFN-γ、TGF-β1及RORγt 表达水平有关。

关键词：类风湿关节炎；黑沙蒿；有效物质群；网络药理学；分子对接；细胞验证实验

Abstract：

Objective: To explore the mechanism of the effective substance groups from Artemisia ordosica Krasch in the treatment of rheumatoid arthritis (RA) by network pharmacology, molecular docking and cell validation experiments. Methods: The databases such as Pubchem, BATMAN-TCM, DisGeNet, GeneCards, NCBI, OMIM, PharmGKB, GEO, and R language were used to obtain HSH core components and component targets, disease targets, and potential targets for HSH treatment of RA. The protein-protein interaction (PPI) network was established and the gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis were conducted to select core components and core targets for molecular docking. The CCK-8 and CFSE labeling method were used to detect the non-toxic concentration and effective concentration of HSH. The experimental techniques such as flow cytometry, CBA, and RT-PCR were used to detect changes in cells, key inflammatory factors, and proteins related to the interleukin (IL) -17/T cell receptor signaling pathway. Results: The three core components, namely hydroxycinnamic acid, o-hydroxycinnamic acid, and o-hydroxycapillene, had the highest correlation with RA, with 90 target points. Totally 875 RA disease targets were obtained by intersecting disease-related targets and differentiable expressed genes in RA. Intersection of RA disease targets with component targets resulted in 36 potential targets for RA. A total of 5 core targets were obtained through PPI construction, and there was a certain binding activity between the core targets and the core components. GO and KEGG enrichment analysis results showed that the IL-17/T cell receptor signaling pathway was closely related to the anti-RA effect of HSH, which also participated in regulating various biological processes such as inflammatory response, cytokine production, and regulation, thereby regulating the body's inflammation and immune response to exert anti-RA effects. The effective concentration of HSH was 16 μg/mL. HSH could down-regulate Th1 levels, up-regulate Treg levels, reduced Th17/Treg ratios, and significantly reduced the levels of IL-21, IL-6, IL-2, interferon-γ (IFN-γ), and transforming growth factor-β1 (TGF-β1). The RT-PCR experiment results showed that HSH significantly down-regulated the expression of RORγt mRNA, while there was no significant difference in the expression of Foxp3 mRNA. Conclusions: HSH has good therapeutic effect on RA, and its mechanism may be related to reducing Th1, increasing Treg, regulating Th17/Treg balance, and reducing the expression levels of IL-21, IL-6, IL-2, IFN-γ, TGF-β1, and RORγt.

Key words：rheumatoid arthritis; Artemisia ordosica Krasch; effective substance groups; network pharmacology; molecular docking; cell validation experiment

发布时间：2026-01-06

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