Anti-DR5 mAb诱导佐剂型关节炎大鼠滑膜细胞凋亡及机理初探

Induction of apoptosis by Anti-DR5 mAb in synovial cells of adjuvant arthritis rats and its possible mechanisms

目的:探讨Anti-DR5 mAb诱导佐剂型关节炎(Adjuvant Arthritis,AA)大鼠滑膜细胞凋亡作用及其可能机理。方法:注射弗氏完全佐剂建立AA大鼠模型,分离并体外培养大鼠滑膜细胞。MTT检测、DNA倍体分析及流式细胞术分析Anti-DR5 mAb对大鼠滑膜细胞的凋亡作用,Western blot检测大鼠滑膜细胞Caspase-3、Bcl-2蛋白的表达,Caspase抑制剂实验分析Anti-DR5 mAb对大鼠滑膜细胞凋亡作用的机制。结果:MTT实验表明Anti-DR5 mAb能抑制大鼠滑膜细胞的增殖,流式细胞术分析这种作用模式为细胞凋亡。Anti-DR5 mAb处理后的大鼠滑膜细胞中Caspase-3蛋白水平上升,Bcl-2蛋白水平下调。Cas-pase抑制剂实验证明Anti-DR5 mAb诱导大鼠滑膜细胞的凋亡是通过Caspase途径实现的。结论:Anti-DR5 mAb诱导大鼠滑膜细胞的凋亡,这种作用与滑膜细胞表面Caspase-3、Bcl-2蛋白表达相关。

Objective：To study the effects of Anti-DR5 mAb on inducing synovial cells of adjuvant arthritis （AA） rats and its mechanisms.Methods： AA was induced by CFA in rats.The synovial cells of rats were separated and cultured in vitro system. The apoptosis induced by anti-DR5 mAb on synovial cells was measured by MTT analysis, DNA fragmentation and flow cytometry. Caspase-3 and Bcl-2 expression in synovial cells were detected by Western blot. The involvement of the apoptotic pathway was further proved by a caspase inhibition assay. Resulls：MTT result showed that Anti-DR5 mAb could inhibit synovial cells growth. And flow cytometry suggested that the cell death mode was apoptosis. The protein level of caspase-3 in synovial cells treated with anti-DR5 mAb was raised, while Bcl-2 level declined. When the caspase inhibitor was added to the synovial cells treated with anti-DR5 mAb, it was showed in a dose-dependence inhibition effect, indicating that anti- DR5 mAb inducing apoptosis might be through the caspase pathway. Conclusion： Anti-DR5 mAb could induce synovial cell apoptosis through caspase pathway. And this effect may be related to the protein level of Caspase-3 and Bcl-2.