诱骗受体DcR3对佐剂型关节炎大鼠模型的作用分析

Analysis of DcR3 in rat model of adjuvant arthritis

目的研究诱骗受体DcR3对佐剂型关节炎(AA)大鼠模型的作用及其机理。方法注射弗式完全佐剂建立大鼠佐剂型关节炎(AA)模型,尾静脉注射DcR3蛋白,观察大鼠关节肿胀度、间接ELISA检测血清和滑膜液中细胞因子IL-1β、TNF-α、IFN-γ的变化。RT-PCR检测滑膜和淋巴细胞中DcR3、Fas、FasL mRNA的表达以及脾脏中TGF-β、IFN-γ、TNF-α、IL-4、IL-10 mRNA的表达。Western blot分析滑膜细胞中Caspase-8、Caspase-3、Caspase-9、Bcl-2蛋白的表达。结果DcR3治疗AA大鼠后,足肿胀度降低;血液和滑膜液的IL-1β、TNF-α、IFN-γ水平下降;脾脏中TGF-β、IFN-γ、TNF-αmRNA表达下调,IL-4、IL-10 mRNA表达上调;滑膜细胞中Caspase-8、Caspase-3蛋白表达上调,Bcl-2蛋白表达下调。结论DcR3可以用于实验性大鼠AA的治疗,其治疗机制与调节滑膜细胞FasL、Fas mRNA的表达和血液淋巴细胞中Fas mRNA的表达,促进滑膜细胞和自身反应性淋巴细胞的凋亡;调节脾脏细胞Th1/Th2细胞因子平衡相关。本研究为进一步阐明RA的发病机理奠定了重要基础,为有效治疗RA提供了新思路。

Objective To study the immunoregulatory mechanisms of DcR3 on rats adjuvant arthritis （AA）. Methods AA was induced by complete freund＇s adjuvant （CFA） in rats, and the rats were injected subcutaneously with DcR3. The perimeters of rat hind soles were measured before and after the injection of CFA. The pathological changes of inflammatory knee joints were observed under optical microscope. The changes of IL-1β, TNF-α, and IFN-γ in serum and synovial liquid were detected by indirect ELISA, respectively. The DcR3, Fas, and FasL mRNA in lymphocytes and synovial cells, as well as the TGF-β, IFN-γ, TNF-α, IL-4, and IL-10 mRNA in synovial cells were detected by RT-PCR. Caspase-8, Caspase-9, Caspase-3, and Bcl-2 expression were analyzed by Western blotting, respectively. Results DcR3 could ameliorate arthritic symptoms and decrease the levels of IL-1β, TNF-α, and IFN-γ in serum and synovial liquid. DcR3 also could stimu- latesynovial cells and lymphocytes apoptosis, and decrease the expression of TGF-β, IFN-γ, TNF-α mRNA. DcR3 could increase the expressions of mRNA of IL-4 and IL-10, as well as increase the expression of Caspase-8 and Caspase-3 in synovial cells, but decrease the expression of Bcl-2. Conclusion DcR3 exerts anti-inflammatory role by suppressing the inflammatory cytokine production, stimulating the synovial cells and T cells apoptosis, and regulating the expressions of FasL and Fas mRNA in synovial cells, Fas mRNA in lymphocytes, and Th1/Th2 cell balance. The research provides valuable theoretical and experimental evidence for treating complicated inflammatory diseases.