β_2GPI/抗β_2GPI抗体复合物激活THP-1细胞内TRIF途径  

The activation of TRIF-dependent signaling pathway in THP-1 cells induced by β_2GPI/anti-β_2GPI antibodies complex

作　　者：解鸿翔[1] 周红[1] 王海波[1] 陈东东[1] 王婷[1] 张先梅[1] 夏龙飞[1] 穆原[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12期1280-1283,1287,共5页Chinese Journal of Cellular and Molecular Immunology

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

摘　　要：目的:观察β2GPI/抗β2GPI抗体复合物能否激活单核细胞株THP-1的TRIF途径,以探讨TRIF依赖途径在抗磷脂综合征(APS)发病机制中的作用。方法:采用一定剂量β2GPI/抗β2GPI抗体复合物刺激THP-1细胞一定时间,收集细胞总RNA及总蛋白,荧光定量PCR(RT-PCR)检测细胞TRIF mRNA水平,Western blot检测细胞TRIF蛋白表达情况;进一步观察TLR4途径抑制剂-TAK-242是否干预β2GPI/抗β2GPI抗体复合物对TRIF的诱导表达以及相关细胞因子IL-6、IL-8、TNF-α的表达。结果:β2GPI/抗β2GPI抗体复合物(100 mg/L)能够诱导THP-1细胞表达TRIF(mRNA及蛋白),并显示时间效应,分别于刺激1 h和2 h时TRIF mRNA及蛋白表达至高峰。TAK-242(5μmol/L)能够明显抑制β2GPI/抗β2GPI抗体复合物对THP-1细胞TRIF的诱导表达,同时抑制IL-6、IL-8、TNF-α等炎症因子的表达。结论:TRIF依赖的TLR4途径参与了β2GPI/抗β2GPI抗体复合物对THP-1细胞的激活,提示其在抗磷脂综合征的病理机制中发挥一定作用。AIM： To observe whether the TIR-domaincontaining adaptor inducing interferon-β （TRIF） is activated in THP-1 cells treated with β2GPI/anti-β2GPI complex and investigate the roles of TRIF-dependent signaling pathway of Toll-like receptor 4 （TLR4） in antiphospholipid syndrome （APS）. METHODS： The total RNA was extracted and the protein lysates were collected from THP-1 cells stimulated with β2GPI/anti-β2GPI complex. And the level of TRIF mRNA in THP-1 cells was detected by Real-time PCR （RT- PCR）, TRIF protein expression was investigated by western blotting, respectively. Furthermore, whether TLR4 inhibitor, TAK-242, could interrupt the expression of TRiF as well as some inflammatory cytokines such as IL-6, IL-8 and TNF-α in THP-1 cells stimulated with β2GPI/anti-β2GPI complex was also investigated. RESULTS： Both mRNA and protein levels of TRIF could be significantly increased in THP-1 cells with treatment of β2GPI/anti-β2GPI complex （100 mg/L）. The expression of TRIF was shown in a manner of time-dependence, with the maximal levels at 1 hour （mRNA） and 2 hour （protein） stimulation respectively. The β2GPI/anti- β2GPI complex-induced TRIF and inflammatory cytokines including IL-6, IL-8 and TNF-α expression in THP-1 cells could be inhibited by TAK-242 （5 μmol/L）. CONCLUSION： TRIF-dependent signaling pathway of Toll-like receptor 4 is involved in the activation of THP-1 cells induced by β2GPI/ anti-β2GPI complex, suggesting that TRIF may play an important role in the pathogenesis of antiphospholipid syndrome.

关 键 词：抗磷脂综合征 β2GPI/抗β2GPI抗体 TLR4 TRIF 炎症因子 

分 类 号：R593.2[医药卫生—内科学]