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目的:研究沙利度胺(thalidomide,THD)对类风湿关节炎(rheumatoid arthritis,RA)动物模型CIA大鼠血清炎症因子白介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、血管内皮细胞生长因子(vascular endo-thelial cell growth factor,VEGF)、基质金属蛋白酶-1,-2,-9(matrix metalloproteinase-1,-2,-9,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,探讨THD治疗RA的可能机制。方法:采用Ⅱ型胶原建立CIA大鼠模型,通过免疫组化观察大鼠关节滑膜血管新生(微血管密度,MVD)情况;THD治疗后用West-ern Blot法检测IL-1β、TNF-α、VEGF、MMP-1,-2,-9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法检测不同浓度的THD处理后的CIA大鼠FLS中ERK和p-ERK表达情况。结果:CIA大鼠造模成功,通过THD治疗能降低CIA大鼠血清炎症因子IL-1β、TNF-α的表达,较造模组分别降低了56%,48%;并能降低血管新生相关因子VEGF、MMP-1,-2,-9表达,较造模组分别降低了52%,52%,13%,33%。并减轻了CIA大鼠的关节炎症及关节滑膜的血管新生;在原代培养CIA大鼠滑膜细胞中THD对FLS中ERK蛋白表达无明显影响,不同浓度的THD作用于FLS,可使p-ERK表达降低(P<0.05),低、中、高浓度的THD组p-ERK分别降低了54%,70%,84%。结论:THD能够调节炎症因子和血管新生相关因子的表达,从而发挥抗炎及抗血管新生的作用,其机制可能与调节ERK信号转导通路有关。
Objective: To investigate the effect of thalidomide (THD) on the serum levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNFα) in CIA rat model of rheumatoid arthritis tumor necrosis factor-alpha, TNF-alpha, vascular endothelial growth factor (VEGF), matrix metalloproteinase-1, -2, -9 , MMP-1, -2, -9), and the effect on the ERK signal transduction pathway of fibroblast-like synoviocytes (FLS), and to explore the possible mechanism of THD in the treatment of RA. Methods: CIA rat model was established by type Ⅱ collagen, synovial neovascularization (MVD) was observed by immunohistochemistry. After THD treatment, the expression of IL-1β, TNF-α, Expressions of VEGF, MMP-1, -2, -9 were detected. Primary FLS of normal rats and CIA rats were isolated by collagenase digestion. The expression of ERK and IL-6 in FLS of THP-treated CIA rats was detected by Western Blot p-ERK expression. Results: The model of CIA rats was successfully established. THD treatment reduced the expression of serum inflammatory cytokines IL-1β and TNF-α in CIA rats by 56% and 48% respectively compared with the model rats, and decreased the number of angiogenesis-related factors The expression of VEGF, MMP-1, -2, -9 were decreased by 52%, 52%, 13% and 33% respectively compared with the model group. And alleviated the joint inflammation and synovial angiogenesis in CIA rats. THD had no significant effect on the expression of ERK protein in FLS in primary cultured CIA synovial cells. Different concentrations of THD on FLS induced a decrease of p- ERK expression decreased (P <0.05), and the p-ERK decreased by 54%, 70% and 84% in low, medium and high concentration THD group respectively. CONCLUSION: THD regulates the expression of inflammatory cytokines and angiogenesis-related factors and thus exerts anti-inflammatory and antiangiogenic effects, which may be related to the regulation of ERK signal transduction pathway.