目的:探讨丝裂原活化蛋白激酶(Mitogen-activated protein kinases,MAPKs)在anti-β2GPI/β2GPI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-β2GPI/β2GPI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-β2GPI/β2GPI复合物诱导THP-1细胞表达TF.结果:Anti-β2GPI/β2GPI复合物(100 μg/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(10 μmol/L)、U0126(5 μmol/L)、SP600125(90 nmol/L)单独或合并处理THP-1细胞后,anti-β2GPI/β2GPI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-β2GPI/β2GPI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-β2GPI/β2GPI-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-β2GPI/β2GPI 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-β2GPI/β2GPI complex. Results:p38,ERKl/2,and JNK were active in a time-dependent manner in THP-1 cells upon treatment with anti-β2GPI/β2GPI complex ( 100 μg/ml). Induction of TF mRNA expression and TF activity by anti-β2GPI/β2GPI 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-β2GPI/β2GPI in THP-1 cells and is involved in the induction of TF expression.
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