雷公藤多甙对胶原诱导关节炎大鼠的治疗作用及其作用机制

目的探讨雷公藤多甙对类风湿关节炎(rheumatoid arthritis,RA)动物模型,胶原诱导关节炎(collagen induced arthritis,CIA)大鼠血清炎症因子白细胞介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子-α(tumornecrosis factor-α,TNF-α)、血管内皮生长因子(vascuoar endothelial cell growth factor,VEGF)、基质金属蛋白酶(matrix metalloproteinases,MMP)-1、2、9表达的影响,及对成纤维样滑膜细胞(fibroblast-like synoviocytes,FLS)ERK信号转导通路的影响,研究雷公藤多甙治疗RA的可能机制。方法采用Ⅱ型胶原建立CIA大鼠模型;观察雷公藤多甙治疗后模型大鼠关节肿胀情况,用免疫组化法观察大鼠关节炎症及滑膜微血管密度;用Western blot法检测CIA大鼠血清IL-1β、TNF-α、VEGF、MMP-1、2、9的表达,并用胶原酶消化法分离培养正常大鼠及CIA大鼠原代FLS;用Western blot法检测不同浓度雷公藤多甙治疗后CIA大鼠FLS中ERK和p-ERK的表达。结果 CIA大鼠造模成功,雷公藤多甙治疗能降低CIA大鼠血清炎症因子及血管新生相关因子表达(P0.05);免疫组化结果显示雷公藤多甙能抑制CIA大鼠关节滑膜血管新生(P0.05);不同浓度雷公藤多甙对FLS中ERK蛋白表达无明显影响,但可降低FLS的p-ERK表达(P0.05)。结论雷公藤多甙对CIA大鼠抗炎及抑制血管新生的作用可能与调节ERK信号转导通路有关。

Treatment Effect and Mechanism of Tripterygium in Collagen Induced Arthritis Rats

Objectives To investigate the possible treatment mechanism of Tripterygium in rheumatoid arthritis（RA）,we evaluate the effect of Tripterygium on the expressions of IL-1β,TNFα,VEGF,and MMP-1,2,9 in collagen induced arthritis（CIA） rats,and on the ERK signal transduction pathway of fibroblast-like synovial cells（FLS）.Methods CIA rat model that was induced with collagen type Ⅱ,treated with Tripterygium,and was scored with the degree of paw swelling.The microvessel density（MVD） of synovial membrane and expressions of IL-1β、 TNF-α、 VEGF、 MMP-1、 2、 9 in serum of CIA rats treated with Tripterygium were detected by Immunohistochemistry and Western blot respectively.The primary FLS of the knee joint from CIA rats were separated with collagenase digestion and cultured.The expressions of ERK and p-ERK in FLS were detected by Western blot on FLS incubated under different concentration of Tripterygium.Results The CIA rat model was successfully constructed.Western blot analysis showed that the Tripterygium can reduce the expressions of inflammatory factors（IL-1β and TNFα） and angiogenic relevant factors（VEGF,MMP-1,2,9） in serum of CIA rats（P 0.05）,and reduce the CIA in rats and synovial angiogenesis.Tripterygium significantly inhibited the expression of p-ERK（P 0.05） in a dose-dependent manner while the expression of ERK in FLS was not affected by Tripterygium.Conclusion Tripterygium can regulate the expression of inflammatory cytokines and angiogenesis-related factors,and play an anti-inflammatory and anti-angiogenesis effect,Tripterygium may regulate ERK signal transduction pathway.