β2GPI/抗β2GPI抗体复合物对人绒毛膜细胞株JEG-3表达黏附分子及趋化因子的影响  

Influence of β2GPI/anti-β2GPI antibody complex on expression of adhesion molecules and chemokines in human chorionic cell line JEG-3

作　　者：王韧 周红[1] 王婷[1] 李娜 何超[1] 王婷[1] 张贵婷 陈芋丹[1] 张鹏 WANG Ren;ZHOU Hong;WANG Ting;LI Na;HE Chao;WANG Ting;ZHANG Gui-Ting;CHEN Yu-Dan;ZHANG Peng(School of Medicine,Jiangsu University,Zhenjiang 212013,China)

机构地区：[1]江苏大学医学院,镇江212013 [2]江苏省无锡市妇幼保健医院检验科,无锡214000

出　　处：《中国免疫学杂志》2019年第19期2310-2314,共5页Chinese Journal of Immunology

基　　金：江苏省自然科学基金青年基金资助项目(BK20150532);江苏省高校自然科学研究面上基金资助项目(15KJB310002)

摘　　要：目的:探讨β2糖蛋白I/抗β2糖蛋白I抗体(β2GPI/抗β2GPI抗体)复合物对人绒毛膜细胞株JEG-3表达黏附分子及趋化因子的影响,分析TLR4-MyD88信号通路在其中的作用。方法:将JEG-3细胞分组刺激:抗β2GPI抗体(100μg/ml)组,β2GPI(100μg/ml)/抗β2GPI抗体(100μg/ml)复合物组,同行对照抗体(RIgG,100μg/ml)组,无血清培养基(Media)组,β2GPI(100μg/ml)组,牛血清白蛋白组(BSA,100μg/ml)。应用RT-qPCR和Western blot及ELISA检测刺激后细胞间黏附分子VCAM-1和ICAM-1以及趋化因子IL-8在mRNA和蛋白水平上的表达;RT-qPCR和Western blot检测TLR4和MyD88在mRNA和蛋白水平上的表达;应用TLR4阻断剂TAK-242预处理的方式,进一步探讨TLR4在β2GPI/抗β2GPI抗体复合物对JEG-3黏附分子及趋化因子表达的影响。结果:β2GPI(100μg/ml)/抗β2GPI抗体(100μg/ml)复合物组相对于Media组能够显著提高JEG-3的VCAM-1、ICAM-1和IL-8表达,同时诱导TLR4-MyD88的表达,两组之间差异有统计学意义(P<0.01);TAK-242(5μmol/L)能够明显抑制该过程;抗β2GPI抗体(100μg/ml)单独组也能诱导TLR4-MyD88的表达。结论:β2GPI/抗β2GPI抗体复合物能够增强人绒毛膜细胞株JEG-3的黏附分子及趋化因子表达,TLR4/MyD88信号通路在其中发挥重要作用。Objective:To explore the effects ofβ2-glycoprotein I/anti-β2-glycoprotein I antibody(β2GPI/anti-β2GPI antibody)complex on the expression of adhesion molecules and chemokines in human chorionic cell line JEG-3,and the roles of TLR4-MyD88 signal pathway was evaluated.Methods:The JEG-3 cells were treated with different stimulus:anti-β2GPI antibody(100μg/ml),β2GPI(100μg/ml)/anti-β2GPI antibody(100μg/ml)complex,the homotype control antibody(RIgG,100μg/ml),serum-free medium,β2GPI(100μg/ml)and bovine serum albumin(BSA,100μg/ml)group respectively.The mRNAs and proteins expression of VCAM-1,ICAM-1 and IL-8 were evaluated by RT-PCR,Western blot and enzyme linked immunosorbent assay(ELISA),the mRNA and protein levels of TLR4 and MyD88 were also detected by RT-qPCR and Western blot,respectively.TLR4 antagonist TAK-242 was used to pre-treat the cells to further explore the influence of TLR4 on the expression of adhesion molecules and chemokines in JEG-3.Results:Compared with the Media group,theβ2GPI(100μg/ml)/anti-β2GPI antibody(100μg/ml)complex could significantly increase the expression of ICAM-1,VCAM-1 and IL-8 in JEG-3 cells(P<0.01).The expression of TLR4 and MyD88 was also enhanced byβ2GPI/anti-β2GPI antibody complex(P<0.01 vs media),TAK-242(5μmol/L)could markedly suppress the process.In addition,the expression of TLR4 and MyD88 could be enhanced by anti-β2GPI antibody alone.Conclusion:Theβ2GPI/anti-β2GPI antibody complex can enhance the expression of adhesion molecules and chemokines in human chorionic cell line JEG-3,and TLR4-MyD88 signal pathway plays an important roles in the process.

关 键 词：β2GPI/抗β2GPI抗体复合物 JEG-3 TLR4/MyD88信号通路 

分 类 号：R392[医药卫生—免疫学]