基于TNF信号通路探讨三痹颗粒治疗类风湿关节炎模型大鼠的机制*

作者：尹玲桃1，谢湘雄1，禹勇军1，陈 达1，杨 蕙2

单位：1.娄底市中医医院，湖南 娄底 417000； 2.湖南中医药大学第一附属医院，湖南 长沙 410007

引用：引用：尹玲桃，谢湘雄，禹勇军，陈达，杨蕙.基于TNF信号通路探讨三痹颗粒治疗类风湿关节炎模型大鼠的机制[J].中医药导报,2025,31(9):55-60.

DOI：10.13862/j.cn43-1446/r.2025.09.009

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摘要：

目的：探究三痹颗粒对类风湿关节炎（RA）模型大鼠的影响及可能的作用机制。方法：采用大鼠尾根部真皮下层注射法注射完全弗氏佐剂+卡介苗油乳剂以构建RA模型，随机分为模型对照组、甲氨蝶呤组、三痹颗粒高剂量组、三痹颗粒中剂量组及三痹颗粒低剂量组，另设空白对照组，每组10只。三痹颗粒高、中、低剂量组予三痹颗粒灌胃，剂量分别为5.40 g/kg、2.70 g/kg、1.35 g/kg，3次/d。甲氨蝶呤组给予甲氨蝶呤（0.90 mg/kg）灌胃，2次/周。空白对照组和模型对照组给予蒸馏水（10 mL/kg）灌胃，3次/d。连续给药4周，检测大鼠踝关节肿胀度并进行HE染色观察，酶联免疫吸附试验法（ELISA）试剂盒检测血清肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6、IL-17和C反应蛋白（CRP），蛋白印迹法（Western Blotting）检测足趾组织TNF-α、IL-6、IL-17和CRP蛋白相对表达水平，实时定量聚合酶链式反应（PCR）检测TNF-α mRNA、IL-6 mRNA、IL-17 mRNA和CRP mRNA相对表达水平。结果：与模型组比较，三痹颗粒各剂量组和甲氨蝶呤组踝关节肿胀度降低（P<0.01），炎症细胞浸润减少。同时，三痹颗粒高剂量组和甲氨蝶呤组TNF-α蛋白及mRNA相对表达水平降低（P<0.05），表明TNF信号通路被抑制，其下游炎症因子IL-6、IL-17和CRP的蛋白及mRNA相对表达水平亦明显降低（P<0.05）。此外，三痹颗粒高剂量组踝关节肿胀度与甲氨蝶呤组比较，差异无统计学意义（P>0.05），TNF-α、IL-17和IL-6的蛋白及mRNA相对表达水平差异均无统计学意义（P>0.05）。结论：三痹颗粒能通过降低TNF-α表达水平以抑制TNF信号通路，并进一步下调下游炎症因子表达水平，缓解RA发展过程中出现的炎症细胞浸润和踝关节肿胀现象，从而减轻炎症反应。

关键词：类风湿关节炎；三痹颗粒；TNF信号通路；炎症反应；炎症因子

Abstract：

Objective: To explore the effect and possible mechanisms of Sanbi granules on rheumatoid arthritis (RA) model rats. Method: RA model was established by injecting complete Freund's adjuvant and BCG oil emulsion into the subcutaneous layer of rat tail root using injection method. The rats were randomly divided into model control group, methotrexate group, high dose group of Sanbi granules, medium dose group of Sanbi granules, and low dose group of Sanbi granules, with 10 rats in each group. A blank control group was also established. The high, medium, and low dose groups of Sanbi granules were given 5.40 g/kg, 2.70 g/kg, and 1.35 g/kg of Sanbi granules, respectively, 3 times a day. The methotrexate group was given methotrexate (0.90 mg/kg), twice a week. The blank control group and model control group were given distilled water at a dose of 10 mL/kg, 3 times a day. After continuous administration for 4 weeks, the swelling degree of the ankle joint in rats was detected and HE staining was implemented. The enzyme-linked immunosorbent assay (ELISA) kit was used to measure serum tumor necrosis factor (TNF)-α, interleukin-6 (IL-6), interleukin-17 (IL-17), and C-reactive protein (CRP). Western blotting analysis was used to measure the protein relative expression levels of TNF-α, IL-6, IL-17, and CRP in toe tissue. Real-time quantitative polymerase chain reaction (PCR) was used to measure the relative expression levels of TNF-α mRNA, IL-6 mRNA, IL-17 mRNA, and CRP mRNA. Results: Compared with the model control group, the ankle joint swelling degree was reduced in high, medium, and low dose groups of Sanbi granules and methotrexate group (P<0.01), and the inflammatory cell infiltration was reduced. At the same time, the TNF-α protein and TNF-α mRNA relative expression levels reduced in the high-dose group of Sanbi granules and  methotrexate group (P<0.05), indicating that the TNF signaling pathway was inhibited. The protein and mRNA relative expression levels of downstream inflammatory factors IL-6, IL-17, and CRP were also significantly reduced (P<0.05). In addition, compared with the methotrexate group, there was no statistically significant difference in ankle joint swelling and protein and mRNA relative expression levels of TNF-α, IL-17 and IL-6 in high-dose group of Sanbi granules (P>0.05). Conclusion: Sanbi granules can inhibit the TNF signaling pathway by reducing the expression level of TNF-α, and further down-regulate the expression level of downstream inflammatory factors, alleviate the inflammatory cell infiltration and ankle joint swelling that occur during the development of RA, and reduce the inflammatory response.

Key words：rheumatoid arthritis; Sanbi granules; TNF signaling pathway; inflammatory response; inflammatory factors

发布时间：2026-01-08

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