复方雷公藤外敷对CIA大鼠成纤维样滑膜细胞中细胞因子和ERK通路表达的影响

Effects of compound tripterygium wilfordii external application on the expression of cytokines in fibroblast-like synoviocytes and ERK pathways of CIA rats

目的:通过观察复方雷公藤外敷对Ⅱ型胶原诱导的大鼠关节炎(CIA)模型大鼠膝关节成纤维样滑膜细胞中细胞因子表达以及ERK通路活化的影响,探讨复方雷公藤外敷治疗类风湿关节炎的抗炎、镇痛与减轻血管翳形成的作用机制。方法:采用CIA模型,各给药组分别用扶他林外敷、雷公藤多苷片口服和复方雷公藤外敷剂外敷。观察原代培养的成纤维样滑膜细胞中白介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、血管内皮生长因子(VEGF)和血管生成素1(Ang^(-1))表达差异及细胞外信号调节激酶(ERK)通路的活化情况。结果:与模型组比较,复方雷公藤外敷组显著降低成纤维样滑膜细胞原代培养上清中IL-1β、TNF-α、VEGF及Ang^(-1)含量水平(P<0.01);显著减少成纤维样滑膜细胞中IL-1β、TNF-α、VEGF和Ang^(-1) mRNA表达水平(P<0.05,P<0.01);显著抑制成纤维样滑膜细胞ERK通路蛋白的磷酸化。结论:复方雷公藤外敷剂通过降低CIA大鼠成纤维样滑膜细胞分泌炎性细胞因子水平减弱ERK通路活化,进而抑制滑膜组织增殖,减少滑膜中血管新生和血管翳形成。

Objective： Through the investigating the effect of compound tripterygium wilfordii external application on the expression of cytokines in fibroblast-like synoviocytes（FLS） and ERK pathways of CIA rats, to explore the mechanism of external application of compound tripterygium wilfordii for the treatment of anti-inflammatory, analgesia and the pannus reduced formation of rheumatoid arthritis of syndrome of stagnant dampness-heat. Methods： The type Ⅱ collagen induced arthritis（CIA） rat model was regarded as the study subject, and the experiment group was external application with votalin and compound tripterygium wilfordii, oral with triperygium wilfordii multiglucoside. Compared with the normal group and CIA model group, to observe the expression differences of interleukin 1β（IL-1β）, tumor necrosis factor α（TNF-α）, vascular endothelial growth factor（VEGF） and angiogenin-1（Ang-（-1）） and the activation of extracellular signal regulating kinase（ERK） pathway on the original cultured fibroid synovial cells. Results： Compared with the model group, the expression of IL-1β, TNF-α, VEGF and Ang-（-1） in the supernatant were significantly decreased of primary culture FLS compound with the tripterygium wilfordii external application group（P0.01）, and the mRNA levels of IL-1β, TNF-α, VEGF and Ang-（-1） in FLS were significantly reduced（P0.05, P0.01）, the protein phosphorylation of ERK pathway in FLS was significantly inhibited by compound tripterygium wilfordii external application. Conclusion： Compound tripterygium wilfordii can reduce the activation of ERK pathway by reducing inflammatory cytokine levels of FLS in CIA rat, so as to inhibit the proliferation of synovial tissues and reduce the angiogenesis and pannus formation.