Wu-Tou Decoction in Rheumatoid Arthritis: Integrating Network Pharmacology and In Vivo Pharmacological Evaluation.
Wu-Tou Decoction in Rheumatoid Arthritis: Integrating Network Pharmacology and In Vivo Pharmacological Evaluation.
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DOI：
10.3389/fphar.2017.00230
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发表时间：
2017
影响因子：
5.6
通讯作者：
Lu A
中科院分区：
文献类型：
--
作者：
Guo Q;Zheng K;Fan D;Zhao Y;Li L;Bian Y;Qiu X;Liu X;Zhang G;Ma C;He X;Lu A
目的：本研究旨在通过网络药理预测和实验验证，探讨五苓汤治疗类风湿关节炎的作用机制。研究方法：WTD的化合物和人类靶蛋白以及RA相关的人类基因分别来自TCM Database @ Taiwan、PubChem和GenBank。随后，分子网络和典型的途径，大概参与了治疗WTD对RA的独创性途径分析（IPA）软件。并以MIP-1β诱导的U937细胞模型和胶原诱导的关节炎（CIA）大鼠模型进行实验验证。结果如下：巨噬细胞中的CCR 5信号通路被证明是与细胞免疫应答和细胞因子信号传导相关的最大共享信号通路。此外，该通路中的蛋白激酶C（PKC）δ和p38可作为WTD治疗RA的靶蛋白。体外实验表明，WTD抑制MIP-1β诱导的U937细胞TNF-α、MIP-1α和RANTES的产生以及CCR 5、PKC δ和p38的磷酸化。在MIP-1β诱导的U937细胞中，WTD处理维持了对CCR 5敲低后TNF-α和RANTES产生的抑制作用。体内实验表明，温通汤能改善CIA大鼠的症状，降低CIA大鼠血清中IL-1β、IL-2、IL-6、TNF-α、MIP-1α、MIP-2、RANTES和IP-10的水平，降低CIA大鼠踝关节中MIP-1α、MIP-2、RANTES和IP-10的mRNA水平。WTD还能降低CIA大鼠踝关节及踝关节巨噬细胞中CCR 5、PKC δ和p38的磷酸化水平。结论：本研究表明，温通汤对类风湿关节炎炎症反应的抑制作用与其对巨噬细胞CCR 5信号通路的调节密切相关。
Purpose: This study aimed to explore underlying action mechanism of Wu-Tou decoction (WTD) in rheumatoid arthritis (RA) through network pharmacology prediction and experimental verification. Methods: Chemical compounds and human target proteins of WTD as well as RA-related human genes were obtained from TCM Database @ Taiwan, PubChem and GenBank, respectively. Subsequently, molecular networks and canonical pathways presumably involved in the treatment of WTD on RA were generated by ingenuity pathway analysis (IPA) software. Furthermore, experimental validation was carried out with MIP-1β-induced U937 cell model and collagen induced arthritis (CIA) rat model. Results: CCR5 signaling pathway in macrophages was shown to be the top one shared signaling pathway associated with both cell immune response and cytokine signaling. In addition, protein kinase C (PKC) δ and p38 in this pathway were treated as target proteins of WTD in RA. In vitro experiments indicated that WTD inhibited MIP-1β-induced production of TNF-α, MIP-1α, and RANTES as well as phosphorylation of CCR5, PKC δ, and p38 in U937 cells. WTD treatment maintained the inhibitory effects on production of TNF-α and RANTES in MIP-1β-induced U937 cells after CCR5 knockdown. In vivo experiments demonstrated that WTD ameliorated symptoms in CIA rats, decreased the levels of IL-1β, IL-2, IL-6, TNF-α, MIP-1α, MIP-2, RANTES, and IP-10 in serum of CIA rats, as well as mRNA levels of MIP-1α, MIP-2, RANTES, and IP-10 in ankle joints of CIA rats. Furthermore, WTD also lowered the phosphorylation levels of CCR5, PKC δ and p38 in both ankle joints and macrophages in ankle joints from CIA rats. Conclusion: It was demonstrated in this research that WTD played a role in inhibiting inflammatory response in RA which was closely connected with the modulation effect of WTD on CCR5 signaling pathway in macrophages.