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.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
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)
