抗β_2GPI/β_2GPI复合物激活THP-1细胞表达炎性相关因子  

Anti-β_2GPI/β_2GPI complex stimulates THP-1cells to express inflammatory factors

作　　者：王海波[1,2] 周红[1] 解鸿翔[1] 王婷[1] 穆原[1] 陈东东[1] 夏龙飞[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 [2]江苏大学附属人民医院儿科,江苏镇江212002

出　　处：《临床检验杂志》2012年第5期347-350,共4页Chinese Journal of Clinical Laboratory Science

基　　金：国家自然科学基金(30971301);江苏大学博士创新基金(09A079)

摘　　要：目的探讨抗β2糖蛋白I(抗β2GPI)自身抗体与β2糖蛋白I(β2GPI)复合物刺激单核细胞株THP-1表达炎性因子IL-6、IL-8、TNF-α及可能机制。方法用荧光定量PCR法检测抗β2GPI/β2GPI复合物刺激THP-1细胞不同时间(0.5、1、1.5、2、6 h)后IL-6、IL-8、TNF-αmRNA表达,用免疫荧光染色法及western blot观察THP-1细胞表达IL-6、IL-8、TNF-α蛋白的情况。荧光定量PCR及western blot观察Toll样受体4(TLR4)途径抑制物(TAK-242)干预抗β2GPI/β2GPI复合物对THP-1细胞的刺激作用。结果抗β2GPI/β2GPI复合物(100μg/mL)刺激THP-1细胞不同时间,可使细胞表达IL-6、IL-8及TNF-αmRNA水平逐渐上升,于刺激2 h时IL-6、IL-8及TNF-α达到峰值(P<0.01),免疫荧光染色及western blot证实细胞表达IL-6、IL-8及TNF-α蛋白水平也明显增加。TAK-242(5μmol/L)可显著抑制抗β2GPI/β2GPI复合物诱导的细胞IL-6、IL-8及TNF-α表达。结论抗β2GPI/β2GPI复合物通过TLR4途径,激活THP-1细胞表达炎性细胞因子IL-6、IL-8及TNF-α,参与抗磷脂综合征的病理机制。Objective To explore the possible mechanism of the anti-β2GPI/β2GPI complex stimulating monocyte cell line THP-1 to express inflammatory cytokines IL-6,IL-8 and TNF-α.Methods The expressions of IL-6,IL-8 and TNF-α mRNA in THP-1 cells following stimulating by anti-β2GPI/β2GPI complex at different points in time（0.5,1,1.5,2 and 6 h,respectively） were determined with real-time PCR,and their protein levels were measured with immunofluorescence and Western blot.The interference of inhibitor of TLR4 signal transduction pathway（TAK-242） for the effects of anti-β2GPI/β2GPI complex on THP-1 cells were investigated using real-time PCR and Western blot assays.Results The expression levels of IL-6,IL-8 and TNF-α mRNA in THP-1 cells stimulated with 100 μg/mL of anti-β2GPI/β2GPI complex were increased with the time of treatmenttime,and reached peak values at 2 hours after the stimulation（P〈0.01）.The protein levels of IL-6,IL-8 and TNF-α were also significantly increased as demonstrated with immunofluorescence and Western blot.However,the expressions of IL-6,IL-8 and TNF-α mRNA in THP-1 cells induced by anti-β2GPI/β2GPI complex were inhibited obviously by 5 μmol/L of TAK-242.Conclusion Anti-β2GPI/β2GPI complex may stimulate THP-1 cells to express inflammatory cytokines IL-6,IL-8 and TNF-α via TLR4 signal transduction pathway,which involves the pathogenesis of antiphospholipid syndrome.

关 键 词：抗磷脂综合征 抗β2GPI/β2GPI复合物 TOLL样受体4 炎性因子 THP-1细胞 

分 类 号：R593.2[医药卫生—内科学]