The Influence Of β2GPI/anti-β2GPI Antibody Complex On Expression Of Adhesion Molecules And Chemokines In Human Chorionic Cell Line JEG-3

Objective: The persistence of antiphospholipid antibodies(a PLs)is one of the main factors of antiphospholipid syndrome(APS),which is closely related to the high pathogenicity of APS.The main clinical manifestations of APS include arteriovenous thrombosis and abortion,which in turn can be called autoimmune recurrent miscarriage.Antiphospholipid antibodies are heterogeneous populations of autoantibodies,of which anti-β2GPI antibodies(antiβ2-glycoprotein I antibodies)are the most pathological autoantibodies in obstetric APS and play an important role in autoimmune recurrent miscarriage.The anti-β2GPI antibody targets the placenta as a target antigen by binding to β2 glycoprotein I(β2-glycoprotein I,β2GPI)expressed by trophoblast cells.When placental trophoblast cells are activated by anti-β2GPI antibodies,their function changes and cytokine levels also change.Our previous study found that toll-like receptor 4(TLR4)plays an important role in the activation of endothelial cells and inflammation in the β2GPI/anti-β2GPI antibody.Our previous research did not address the effect of β2GPI/anti-β2GPI antibody complexes on placental trophoblast cells.Through pre-experiment and review of relevant literature,this experiment focused on the effect of β2GPI/anti-β2GPI antibody complex on the expression of abortion-associated adhesion molecules and chemokines in human chorionic cell line JEG-3,and whether TLR4/My D88 signaling pathway plays a role in it.Methods:(1)The protein and m RNA levels of VCAM-1,ICAM-1,IL-8 after anti-β2GPI antibody,β2GPI/anti-β2GPI complex,non-specific control antibody,media,β2GPI and BSA treatments were measured by Westernblot,Enzyme-linked immunosorbent assay(ELISA)and Real-time quantitative PCR(RT-q PCR).(2)The protein and m RNA levels of TLR4 and My D88 after anti-β2GPI antibody,β2GPI/anti-β2GPI complex,non-specific control antibody,media,β2GPI and BSA treatments were measured by Westernblot and Real-time quantitative PCR(RT-q PCR).(3)The expression of VCAM-1 and IL-8 induced by TLR4 in TAK-242 pretreatment and untreated JEG-3 were measured by RT-q PCR,Westernblot and ELISA.Results:(1)After treatment with β2GPI/anti-β2GPI antibody complex,the expression of VCAM-1,ICAM-1,IL-8 gene and protein levels in JEG-3 cells increased significantly compared with serum-free medium group.(p < 0.05).(2)The expression of TLR4 and My D88 in JEG-3 cells was significantly increased after treatment with β2GPI/anti-β2GPI antibody complex,and the difference was statistically significant(p < 0.05).Conclusions:(1)The anti-β2GPI antibody and the β2GPI/anti-β2GPI antibody complex can effectively promote the expression of adhesion molecules and chemokines VCAM-1,ICAM-1,IL-8 in human chorionic cell line JEG-3,and accelerate the adhesion and inflammatory response.(2)The anti-β2GPI antibody and the β2GPI/anti-β2GPI antibody complex can induce high expression of TLR4 and My D88 in JEG-3 cells;TLR4 pathway inhibitor TAK-242 can reduce the expression of VCAM-1 and IL-8 in the stimulation group,indicating that TLR4 signaling pathway play an important role in it.