TRAF6在抗β_2GPI/β_2GPI复合物诱导THP-1细胞表达组织因子时的活化  

Activation of tumor necrosis factor receptor-associated factor 6 in anti-β_2GPI/β_2GPI-induced tissue factor expression on THP-1 cells

作　　者：许国莹[1] 周红[1] 文海平[1] 郭东琳[1] 周芳[1] 陈东东[1] 解鸿翔[1] 

机构地区：[1]江苏大学基础医学与医学技术学院,江苏镇江212013

出　　处：《细胞与分子免疫学杂志》2011年第5期487-490,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30670907;30971301);江苏大学科研立项基金资助项目(09A079)

摘　　要：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 whether tumor necrosis factor receptor-associated factor 6（TRAF6） is involved in anti-β2GPI/β2GPI-induced tissue factor（TF） expression on THP-1 cells.METHODS： The total RNA was extracted and the protein lysates were collected from THP-1 cells stimulated with anti-β2GPI/β2GPI complex.And then the TF expression on THP-1 cells was detected by real-time quatitative PCR（RT-qPCR） and TF activity kit.TRAF6 mRNA and its protein expression were investigated by RT-qPCR and Western blotting,respectively.The proteasome inhibitor,MG-132,was used for inhibitory assays,in order to demonstrate the effect of anti-β2GPI/β2GPI complex on THP-1 cells.RESULTS： The TF expression（both mRNA and activity） on THP-1 cells was significantly up-regulated with the treatment of anti-β2GPI/β2GPI complex（100 mg/L）,compared with untreated cells（P0.05）.The TRAF6 mRNA and protein levels in THP-1 cells were also significantly increased with the treatment of anti-β2GPI/β2GPI complex.The expression of TRAF6 was shown in a time-dependent manner,with the maximal level at 15 minutes（mRNA） and 30 minutes（protein） respectively.All the stimulating effects of anti-β2GPI/β2GPI complex（100 mg/L） on THP-1 cells were inhibited by MG-132（5 μmol/L）.CONCLUSION： TRAF6 is up-regulated and contributs to TF expression on THP-1 cells induced with anti-β2GPI/β2GPI complex.

关 键 词：抗磷脂综合征 抗β2GPI/β2GPI tumor NECROSIS factor receptor-associated factor6 组织因子 

分 类 号：R593.2[医药卫生—内科学]