Shi Lu, Zhao Yiying, Feng Chenran, Miao Feng, Dong Linlin, Wang Tianquan, Stalin Antony, Zhang Jingyuan, Tu Jingru, Liu Kexin, Sun Wenyan, Wu Jiarui
Department of Pharmacology of Chinese Materia Medica, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, China.
Institute of Fundamental and Frontier Sciences, University of Electronic Science and Technology of China, Chengdu, China.
Front Pharmacol. 2022 Aug 17;13:967164. doi: 10.3389/fphar.2022.967164. eCollection 2022.
Shaogan Fuzi Decoction (SGFD), one of the classical prescriptions of Chinese Medicine, has a long history in the treatment of rheumatoid arthritis (RA), but definitive studies on its efficacy and mechanism of action are lacking. This study aims to elucidate the pharmacodynamic role of SGFD against RA and the potential mechanisms based on a combination of network pharmacology and experimental verification. The RA model in rats was induced by intradermal injection of bovine type Ⅱ collagen and incomplete Freund's adjuvant at the tail root. SGFD was administered once a day by oral gavage for 4 weeks. After SGFD administration, rat's arthritis index (AI) score and paw swelling decreased to some extent, and synovial inflammation, vascular hyperplasia, and cartilage destruction of the ankle joint were improved. Simultaneously, thymus and spleen index and serum levels of C-reactive protein (CRP) were lowered. Network pharmacology revealed that quercetin, kaempferol, naringenin, formononetin isorhamnetin and licochalcone A were the potentialiy active components, and IL6, TP53, TNF, PTGS2, MAPK3 and IL-1β were potential key targets for SGFD in the treatment of RA. Ingredients-targets molecular docking showed that the components had the high binding activity to these target proteins. The mechanism of SGFD for RA involves various biological functions and is closely correlated with TNF signaling pathway, Osteoclast differentiation, T cell receptor signaling pathway, mitogen-activated protein kinase (MAPK) signaling pathway, NF-κB signaling pathway, toll-like receptor signaling pathway, and so on. Western blot and ELISA showed that the expression of toll-like receptor 4 (TLR4), nuclear factor kappa-B (NF-κB) p65, phosphorylated c-Jun N-terminal kinase (p-JNK), p-p38, phosphorylated extracellular regulated kinase (p-ERK) and TNF-α was significantly upregulated in the synovium of RA rats, and the levels of serum inflammatory factors were significantly increased. SGFD inhibits the activation of the TLR4/NF-κB/MAPK pathway and the expression/production of pro-inflammatory cytokines. In summary, SGFD could improve the symptoms and inflammatory response in collagen-induced arthritis (CIA) rat model. The mechanism might be related to the regulation of TLR4/MAPKs/NF-κB signaling pathway and the reduction of inflammatory factor release, which partially confirms the results predicted by network pharmacology.
芍甘附子汤（SGFD）是中医经典方剂之一，在类风湿关节炎（RA）的治疗方面有着悠久的历史，但关于其疗效和作用机制的确定性研究尚显不足。本研究旨在结合网络药理学和实验验证，阐明芍甘附子汤对类风湿关节炎的药效作用及潜在机制。通过在大鼠尾根皮内注射牛Ⅱ型胶原蛋白和不完全弗氏佐剂诱导建立类风湿关节炎模型。芍甘附子汤通过灌胃给药，每天1次，连续给药4周。给药后，大鼠的关节炎指数（AI）评分和 paw肿胀有一定程度降低，踝关节的滑膜炎症、血管增生和软骨破坏得到改善。同时，胸腺和脾脏指数以及血清C反应蛋白（CRP）水平降低。网络药理学研究显示，槲皮素、山奈酚、柚皮素、芒柄花素、异鼠李素和甘草查尔酮A为潜在活性成分，白细胞介素6（IL6）、肿瘤蛋白p53（TP53）、肿瘤坏死因子（TNF）、前列腺素内过氧化物合酶2（PTGS2）、丝裂原活化蛋白激酶3（MAPK3）和白细胞介素1β（IL-1β）是芍甘附子汤治疗类风湿关节炎的潜在关键靶点。成分-靶点分子对接显示，这些成分与这些靶蛋白具有较高的结合活性。芍甘附子汤治疗类风湿关节炎的机制涉及多种生物学功能，与TNF信号通路、破骨细胞分化、T细胞受体信号通路、丝裂原活化蛋白激酶（MAPK）信号通路、核因子κB（NF-κB）信号通路、Toll样受体信号通路等密切相关。蛋白质免疫印迹法（Western blot）和酶联免疫吸附测定（ELISA）显示，类风湿关节炎大鼠滑膜中Toll样受体4（TLR4）、核因子κB（NF-κB）p65、磷酸化c-Jun氨基末端激酶（p-JNK）、磷酸化p38、磷酸化细胞外调节蛋白激酶（p-ERK）和肿瘤坏死因子-α（TNF-α）的表达显著上调，血清炎症因子水平显著升高。芍甘附子汤抑制TLR4/NF-κB/MAPK通路的激活以及促炎细胞因子的表达/产生。综上所述，芍甘附子汤可改善胶原诱导性关节炎（CIA）大鼠模型的症状和炎症反应。其机制可能与调节TLR4/MAPKs/NF-κB信号通路以及减少炎症因子释放有关，这部分证实了网络药理学预测的结果。
