二妙散水提物调控胶原诱导性关节炎大鼠成纤维样滑膜细胞增殖、迁移和炎性因子的表达

Ermiao san aqueous extract regulates proliferation, migration, and inflammatory factor expression of fibroblast-like synovial cells in collagen-induced arthritis rats

背景:NF-κB信号转导通路的激活失调与类风湿关节炎成纤维样滑膜细胞的增殖及炎症密切相关,然而基于NF-κB信号转导通路的二妙散水提物干预胶原诱导性关节炎大鼠成纤维样滑膜细胞(collagen induced arthritis rat fibroblast-like synoviocytes,CIA-FLS)的作用机制未见相关报道。目的:探讨不同质量浓度二妙散水提物对CIA-FLS增殖、迁移和炎性因子表达的影响,并基于NF-κB信号通路探索其作用机制。方法:复苏CIA-FLS,传代培养后加入不同质量浓度二妙散水提物(200,400,800,1 600,3 200 mg/L)干预,MTT法、Transwell小室检测CIA-FLS增殖和迁移能力,qRT-PCR和Western blot检测CIA-FLS中NF-κB信号通路相关分子白细胞介素6、肿瘤坏死因子α、NF-κBp65 mRNA和蛋白表达水平。结果与结论:(1)不同质量浓度二妙散水提物作用48 h后,与空白对照组相比,二妙散水提物呈剂量依赖性抑制CIA-FLS增殖(P <0.05);(2)1 600 mg/L二妙散水提物极显著抑制CIA-FLS的增殖和迁移(P <0.01);同时,极显著抑制CIA-FLS中白细胞介素6、肿瘤坏死因子α、NF-κBp65m RNA和蛋白表达水平(P <0.01);(3)结果提示,二妙散水提物抑制CIA-FLS增殖、迁移和炎症因子表达的最佳质量浓度是1 600 mg/L。二妙散水提物可能通过抑制NF-κB信号通路的活化及炎症因子的生成,进而减轻CIA-FLS的增殖和炎症水平,为后期进一步研究二妙散治疗类风湿关节炎的作用机制提供实验依据。

BACKGROUND: The activation of nuclear factor-κB signaling pathway is closely related to the proliferation and inflammation of fibroblast-like synovial cells in rheumatoid arthritis. However, the mechanism by which ermiao San aqueous extract interferes with fibroblast-like synoviocytes of collagen-induced arthritis rats(CIA-FLS) via the NF-κB signaling pathway has not been reported. OBJECTIVE: To study the effects of different concentrations of ermiao San aqueous extracts on the proliferation, migration and inflammatory factors expression of CIA-FLS, and to explore its mechanism based on the nuclear factor-κB signaling pathway. METHODS: CIA-FLS cells were resuscitated and coped with different concentrations of ermiao San aqueous extracts(200, 400, 800, 1600, 3 200 mg/L) after subculture. The proliferation and migration ability of CIA-FLS were detected by MTT method and Transwell chamber, and real-time fluorescent quantitative PCR(q RT-PCR) and western blot assay were used to detect the NF-κB signaling pathway related molecules interleukin-6, tumor necrosis factor-α, nuclear factor-κBp65 m RNA and protein expression levels in CIA-FLS. RESULTS AND CONCLUSION: Different concentrations of ermiao San aqueous extracts acted on CIA-FLS for 48 hours. Compared with the blank control group, ermiao San aqueous extracts inhibited the proliferation of CIA-FLS in a dose-dependent manner(P < 0.05). 1 600 mg/L ermiao San aqueous extract significantly inhibited the proliferation and migration of CIA-FLS(P < 0.01), and at the same time, it significantly inhibited the expression of interleukin-6, tumor necrosis factor-α, nuclear factor-κBp65 mRNA and protein in CIA-FLS(P < 0.01). Therefore, the optimal concentration of ermiao San aqueous extract to inhibit the proliferation, migration and inflammatory factor expression of CIA-FLS is 1 600 mg/L. ermiao San aqueous extract may inhibit the activation of nuclear factor-κB signaling pathway and the production of inflammatory factors, thereby reducing the proliferation and inflammation of CIA-FLS. It provides certain research experimental basis for further research on the mechanism of ermiao San treatment of rheumatoid arthritis in the later period.