oxLDL/β2GPI/抗β2GPI抗体复合物促进人脐静脉内皮细胞黏附分子表达的机制  

The mechanism of oxLDL/β2GPI/anti-β2GPI antibody complex promoting the expression of adhesion molecules in HUVECs

作　　者：张贵婷[1] 周红[1] 何超[1] 陈芋丹 欧阳航 张鹏[1] 王韧 

机构地区：[1]江苏大学医学院,江苏镇江212013

出　　处：《细胞与分子免疫学杂志》2017年第11期1472-1478,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81370614)

摘　　要：目的探讨氧化型低密度脂蛋白/β2糖蛋白I/抗β2糖蛋白I抗体(oxLDL/β2GPI/抗β2GPI抗体)复合物对人脐静脉内皮细胞(HUVEC)表达黏附分子的影响,以及Toll样受体4(TLR4)信号通路在其中发挥的作用。方法用单纯培养基、oxLDL、oxLDL/β2GPI复合物、oxLDL/抗β2GPI抗体复合物、oxLDL/β2GPI/抗β2GPI抗体复合物、LPS分组刺激HUVEC,收集总RNA和蛋白,利用实时定量PCR和Western blot法检测细胞间黏附分子1(ICAM-1)、血管黏附分子1(VCAM-1)和冯·维勒布兰德因子(v WF)的mRNA和蛋白水平;采用TLR4阻断剂TAK-242(5μmol/L)或p38丝裂原活化蛋白激酶(p38MAPK)阻断剂SB203580(10μmol/L)预处理的方式,探索阻断TLR4通路后,oxLDL/β2GPI/抗β2GPI抗体复合物对HUVEC黏附分子表达的影响,通过THP-1细胞黏附实验检测oxLDL/β2GPI/抗β2GPI抗体复合物对HUVEC黏附能力的影响。结果与培养基对照组相比,oxLDL/β2GPI/抗β2GPI抗体复合物能够显著提高HUVEC的ICAM-1、VCAM-1和v WF表达,并且能够增强HUVEC对THP-1细胞的黏附,TAK-242或SB203580处理能够抑制该过程。结论 oxLDL/β2GPI/抗β2GPI抗体复合物能够增强HUVEC的黏附分子表达和对THP-1细胞的黏附,TLR4和p38MAPK与该过程密切相关。Objective To investigate the effect of oxidative low-density lipoprotein/β2 glycoprotein I/anti-β2 glycoprotein I antibody（ oxLDL/β2GPI/anti-β2GPI-Ab） complex on the expressions of adhesion molecules in human umbilical vein endothelial cells（ HUVECs）,and the role of Toll-like receptor 4（ TLR4） signaling pathway during this process. Methods HUVECs were stimulated with simple medium, oxLDL, oxLDL/β2GPI complex, oxLDL/anti-β2GPI-Ab complex,oxLDL/β2GPI/anti-β2GPI-Ab complex and LPS separately,and then the total mRNA and protein were collected. The mRNA and protein expressions of intercellular adhesion molecule 1（ ICAM-1）,vascular cell adhesion molecule 1（ VCAM-1） and von Willebrand factor（ v WF） were detected by real-time quantitative PCR and Western blotting,respectively. Pre-treatment method was applied to explore the influence of TLR4 antagonist TAK-242（ 5 μmol/L） and p38 MAPK antagonist SB203580（10 μmol/L） on the expressions of adhesion molecules in HUVECs. THP-1 cell adhesion test was employed to evaluate the effect of oxLDL/β2GPI/anti-β2GPI-Ab complex on the ability of attracting monocytes of HUVECs. Results Compared with simple medium,oxLDL/β2GPI/anti-β2GPI-Ab complex obviously upregulated the expressions of ICAM-1, VCAM-1 and v WF in HUVECs and promoted the adhesion of THP-1 cells to HUVECs. In addition,these effects were inhibited by the pre-treatment of TAK-242 or SB203580. Conclusion Ox LDL/β2GPI/anti-β2GPI-Ab complex could up-regulate the expressions of adhesion molecules in HUVECs and promote the adhesion of THP-1 cells to HUVECs,in which TLR4 and p38 MAPK were closely involved.

关 键 词：oxLDL/β2GPI/抗β2GPI抗体复合物 Toll样受体4(TLR4) 人脐静脉内皮细胞(HUVEC) 黏附分子 

分 类 号：R392.5[医药卫生—免疫学]