膜联蛋白A_2介导β_2糖蛋白Ⅰ/抗β_2糖蛋白Ⅰ复合物刺激单核细胞上调表达组织因子  

Annexin A_2 mediated β_2GPⅠ/anti-β_2GPⅠstimulates up-regulation of tissue factor expression in monocytes

作　　者：俞颖[1] 谭潘丽[1] 钱丽丽[1] 

机构地区：[1]浙江中医药大学附属第一医院临床实验诊断中心,杭州310006

出　　处：《中国药物与临床》2010年第12期1337-1340,共4页Chinese Remedies & Clinics

基　　金：浙江省自然科学基金(Y206830);浙江省教育厅资助项目(20060746)

摘　　要：目的探讨膜联蛋白A2是否介导β2糖蛋白Ⅰ(β2GPⅠ)/抗β2糖蛋白Ⅰ结合于单核细胞表面,从而诱导单核细胞表达组织因子。方法利用纯化的抗磷脂综合征(APS)患者血清IgG、单克隆抗β2糖蛋白Ⅰ抗体、抗膜联蛋白A2抗体等刺激血液单核细胞及THP-1单核细胞株,通过凝血因子Ⅹa的生成量,判断细胞组织因子活性的表达;把带有膜联蛋白A2cDNA的载体转染入HEK293T细胞,然后用β2糖蛋白Ⅰ/抗β2糖蛋白Ⅰ复合物刺激,观察转染前后HEK293T细胞的组织因子表达情况;采用RNA干扰技术封闭单核细胞株THP-1膜表面的膜联蛋白A2,观察组织因子的表达情况。结果 APS患者血清IgG和抗β2糖蛋白Ⅰ抗体显著增强单核细胞组织因子的表达;HEK293T细胞转染膜联蛋白A2cDNA后组织因子的mRNA和蛋白的表达量较转染前增高;采用RNA干扰降低膜联蛋白A2表达后单核细胞株THP-1的组织因子表达显著降低。结论细胞表面膜联蛋白A2介导β2糖蛋白Ⅰ/抗β2糖蛋白Ⅰ诱导单核细胞表达组织因子活性,是APS血栓形成的机制之一。Objective To explore whether Annexin A2 mediated β2GPⅠ/ anti-β2GPⅠstimulates up-regulation of tissue factor （TF） expression in monocytes. Methods Purified IgG, anti-β2GPⅠand anti-Annexin A2 in sera from patients with antiphospholipid syndrome （APS） were used to stimulate monocytes and monocytic cell line THP-1. TF activity in monocytes was estimated with production of factor Ⅹa. The expression of TF in HEK293T cells before and after transfection with Annexin A2 cDNA and stimulation with β2GPⅠ/anti-β2GPⅠ was examined. RNA interference was used to block Annexin A2 on THP-1 membrane, and the expression of TF was again determined. Results Serum IgG and anti-β2GPⅠfrom APS patients were shown to significantly increase the TF expression in monocytes. The complex of Annexin A2 on monocyte membrane and β2GPⅠ/anti-β2GPⅠ also induced up-regulation of TF expression. Annexin A2 cDNA transfection of HEK293T cells resulted in enhanced expression of TF mRNA and protein. The TF expression was dramatically decreased in THP-1 cells with less Annexin A2 expression as a result of RNA interference. Conclusion Annexin A2 mediated β2GPⅠ/anti-β2GPⅠmay induce TF activity in monocytes, which may be one of underlying mechanisms for thrombosis in APS.

关 键 词：β2糖蛋白Ⅰ 抗体 抗独特型 单核细胞 凝血致活酶 膜联蛋白A2 RNA干扰 

分 类 号：R363[医药卫生—病理学]