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目的:探讨肿瘤坏死因子受体相关因子6(TRAF6)在抗β2GPI/β2GPI复合物诱导单核细胞株THP-1表达组织因子(TF)中的作用。方法:采用一定剂量抗β2GPI/β2GPI复合物刺激THP-1细胞一定时间,收集细胞总RNA及总蛋白,实时定量PCR检测细胞TF mRNA水平,发色底物法检测细胞TF活性;RT-qPCR及Western blot分别检测细胞TRAF6mRNA和蛋白表达情况;进一步采用蛋白酶体抑制剂MG-132,观察是否能够干预抗β2GPI/β2GPI复合物对细胞的刺激效应。结果:抗β2GPI/β2GPI复合物(100 mg/L)能够刺激THP-1细胞表达TF mRNA及活性,与对照相比差异显著(P<0.05);使细胞TRAF6 mRNA和蛋白表达均增加,并显示时间相关性,分别在刺激15 min和30 min时表达至高峰;MG-132(5μmol/L)明显抑制抗β2GPI/β2GPI复合物(100 mg/L)对THP-1细胞TRAF6 mRNA和蛋白的刺激效应及TF的诱导表达。结论:抗β2GPI/β2GPI复合物诱导THP-1细胞表达TF过程中,TRAF6被激活并发挥重要作用。
AIM: To investigate the role of tumor necrosis factor receptor related factor 6 (TRAF6) in the expression of tissue factor (TF) induced by β2GPI / β2GPI complex in monocytic THP-1 cells. Methods: THP-1 cells were stimulated with a certain dose of anti-β2GPI / β2GPI complex for a certain period of time, total cellular RNA and total protein were collected. Real-time quantitative PCR was used to detect the level of TF mRNA. The expression of TRAF6mRNA and protein were detected by Western blot. The proteasome inhibitor MG-132 was further used to observe whether it could interfere with the stimulating effect of anti-β2GPI / β2GPI complex on cells. Results: The anti-β2GPI / β2GPI complex (100 mg / L) stimulated the expression of TF mRNA and the activity of TH mRNA in THP-1 cells, which was significantly different from the control The expression of TRAF6 mRNA and protein in THP-1 cells stimulated by anti-β2GPI / β2GPI complex (100 mg / L) was significantly inhibited by MG-132 (5μmol / L) Effect and induction of TF expression. CONCLUSION: TRAF6 is activated and plays an important role in the anti-β2GPI / β2GPI complex-induced THP-1 cell TF expression.