The Effects And Potential Mechanisms Of Osteoprotegerin On Leukocyte Adhesion To Person Umbilical Vein Endothelial Cells

PART 1 The effects and of ox-LDL and ATR on OPG expression in person umbilical vein endothelial cellObjectives to investigation the effects and primary mechanisms of ox-LDL and ATR on OPG expression in person umbilical vein endothelial cell.Methods Human umbilical vein endothelial cells(HUVECs)were cultured in vitro and divided into 3 groups:(1)ox-LDL stimulation group;(2) ox-LDL+ATR,PDTC and SB203580 group;(3)ATR stimulation group.the mRNA levels of OPG and NF-κBP65 genes were measured by real-time quantitative polymerase chain reaction.The OPG andp38 protein were analysed by western-blot,the OPG released from HUVECs were measured by ELISA.Results1)Expression of OPG was increased in HUVECs stimulated by ox-LDL for 24h compared with that of control group,the levels of NF-κBP65mRNA and P38 protein were also significantly increased;2)ATR,PDTC and SB203580 can suppress the expression of OPG induced by ox-LDL in HUVECs,the levels of NF-κBP65mRNA and P38 protein were also significantly decreased;3)Production of OPG was not changed in HUVECs stimulated by ATR for 24h compared with that of control group,the levels of NF-κBP65mRNA and P38 protein were also invariable.4)The results of correlation analysis showed that the Expression of OPG mRNA were positively correlated with NF-κBP65 mRNA.Conclusions:1)ox-LDL can increase the OPG expression of HUVECs in a dose-dependent manner2)The effect of ox-LDL on OPG production of HUVECs is take mainly through NF-κB signaling pathways,secondarily thoughs p38MAPK signaling pathways.3)ATR has not effect on OPG expression,but it can suppress the production of OPG induced by ox-LDL through NF-κB and the p38MAPK signaling pathways. PARTⅡThe effects of OPG on leukocyte adhesion to person umbilical vein endothelial cellsObjectives to investigation the effects and primary mechanisms of OPG on leukocyte adhesion to person umbilical vein endothelial cells(HUVECs).Methods HUVECs were cultured in vitro and divided into 2 groups:(1) OPG,OPG+PDTC and OPG+ SB203580 group;(2)ATR + OPG stimulation group.the mRNA levels of NF-κBP65,VCAM-1,ICAM-1,E-selectin,P-selectin and PSGL-1 mRNA genes were measured by real-time quantitative polymerase chain reaction.The p38 protein were analysed by western-blot.And the adhesiveness of THP-1 to HUVECs were measured by Fluorescence microscope.Results1)Expression of PSGL-1 was increased in HUVECs stimulated by OPG for 24h compared with that of control group,the levels of P38 protein were also significantly increased;OPG significantly promoted the adhesion of THP-1 to HUVECs.In addition,OPG had not effect on the expression of NF-κBP65,VCAM-1,ICAM-1,E-selectin and P-selectin mRNA.2)SB203580 can suppress the expression of PSGL-1induced by OPG in HUVECs,the levels of P38 protein were also significantly decreased.In addition,THP-1/ HUVECs adhesion induced by OPG was significantly decreased after HUVECs pre-incubated with SB203580.On the contrary, PDTC had not similar effect.3)ATR can suppress the expression of PSGL-1induced by OPG in HUVECs,the levels of P38 protein were also significantly decreased.In addition,THP-1/ HUVECs adhesion induced by OPG was significantly decreased after HUVECs pre-incubated with ATR.4)The results of correlation analysis showed that the Expression of PSGL-1 mRNA were positively correlated with ph-P38 protein(r=0.752, P＜0.01)。Conclusions1)OPG can increase the PSGL-1 expression of HUVECs and promote the adhesion of THP-1 to HUVECs in a dose-dependent manner.2)The effect of OPG on adhesion of HUVECs is take mainly through p38MAPK signaling pathways.3)ATR can suppress the production of PSGL-1 and THP-1/HUVECs adhesion induced by OPG through p38MAPK signaling pathways.