MAPKs在anti-β_2 GPI/β_2 GPI刺激THP-1细胞表达TF过程中的作用探讨  

The roles of MAPKs in anti-β_2 GPI/β_2 GPI-induced tissue factor expression on THP-1 cells

作　　者：陈东东[1] 周红[1] 解鸿翔[1] 张先梅[1] 夏龙飞[1] 王婷[1] 王海波[1] 

机构地区：[1]江苏大学基础医学与医学技术学院,镇江212013

出　　处：《中国免疫学杂志》2012年第2期99-103,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金资助项目(No.30971301);江苏大学学生科研课题项目(No.10A108)资助

摘　　要：目的:探讨丝裂原活化蛋白激酶(Mitogen-activated protein kinases,MAPKs)在anti-β2 GPI/β2 GPI复合物诱导单核细胞株THP-1表达组织因子(TF)中的活化及其作用。方法:利用荧光定量PCR(Real-time PCR)、TF活性试剂盒等分别检测anti-β2 GPI/β2 GPI复合物诱导THP-1细胞表达TF mRNA及TF活性,Western blot检测细胞表达p38、磷酸化-p38(p-p38)、ERK1/2、磷酸化-ERK1/2(p-ERK1/2)、JNK、磷酸化-JNK(p-JNK)的情况。进一步采用p38、ERK1/2、JNK抑制剂(SB203580、U0126、SP600125)观察是否能阻断anti-β2 GPI/β2 GPI复合物诱导THP-1细胞表达TF。结果:Anti-β2 GPI/β2 GPI复合物(100μg/ml)能够显著增强THP-1细胞表达TF,并使p-p38、p-ERK1/2、p-JNK水平显著升高(P<0.05 vs control);其引发的MAPKs磷酸化具有时间效应性,均在刺激30分钟时达到高峰;对应的特异抑制剂SB203580(10μmol/L)、U0126(5μmol/L)、SP600125(90 nmol/L)单独或合并处理THP-1细胞后,anti-β2 GPI/β2 GPI复合物诱导细胞TF mRNA表达及TF活性的效应明显被阻断(P<0.01 vs control)。结论:Anti-β2 GPI/β2 GPI复合物诱导THP-1细胞表达TF过程中,MAPKs被激活进而发挥重要作用。Objective：To investigate whether mitogen-activated protein kinases（MAPKs） are involved in anti-β2 GPI/β2 GPI-induced tissue factor（TF） expression in THP-1 cells.Methods：TF expression in THP-1 cells was measured by Real-time PCR or TF activity after stimulating with anti-β2 GPI/β2 GPI complex.The activation of p38,ERK1/2,and JNK induced by anti-β2 GPI/β2 GPI complex were determined by Western blot using phosphor-specific antibodies.Inhibitors against p38,ERK1/2,JNK（SB203580,U0126,SP600125） were used to test whether the blockade of these pathway would interrupt the induction of TF expression in THP-1 cells by anti-β2 GPI/β2 GPI complex.Results：p38,ERK1/2,and JNK were active in a time-dependent manner in THP-1 cells upon treatment with anti-β2 GPI/β2 GPI complex（100 μg/ml）.Induction of TF mRNA expression and TF activity by anti-β2 GPI/β2 GPI complex（100 μg/ml） in THP-1 cells could be blocked by inhibitors against the MAPKs,SB203580（10 μmol/L）,U0126（5 μmol/L）,SP600125（90 nmol/L）.Conclusion：MAPKs can be activated by anti-β2 GPI/β2 GPI in THP-1 cells and is involved in the induction of TF expression.

关 键 词：抗磷脂综合征 Anti-β2 GPI/β2 GPI 丝裂原活化蛋白激酶 组织因子 

分 类 号：R593.2[医药卫生—内科学]