PBEF Activates P38MAPK/ERK Signaling Pathways To Increase Anoxia-reoxygenation-injured Permeability Of Endothelial Cells

Objective:According to the preliminary study,anoxia and reoxygenation of normal cultured human umbilical vein endothelial cells for a certain time to imitate pathophysiological changes of pulmonary endothelial cells during pulmonary ischemia-reperfusion in clinical cardiac surgery.Through RNA interference inhibiting PBEF gene to research the relationship : PBEF and TNF-α、 IL-1β、IL-6 inflammatory factors 、PBEF and cell-cell adhesion 、cell-matrix adhesion,to investigate the relationship between PBEF and P38 MAPK / ERK signaling pathway in the regulation of endothelial cell permeability induced by anoxia-reoxygenation.Methods:1.According to the previous study,HUVECs anoxia(20 h)/reoxygenation(9 h)model was established,synthesis of PBEF siRNA sequences.2.HUVECs were treated with differently,divided into anoxia-reoxygenated PBEFsiRNA transfection group,anoxia-reoxygenation siRNA negative control group,anoxia-reoxygenation group and normal control group.3.We used western blot to dectect the levels of PBEF in each group,ELISA to detect the contents of TNF-α,IL-1β,and IL-6.4.We used western blot to dectect the levels of MLC and its phosphorylation in each group.5.The expression of VE-cadherin、FAK and their phosphorylation and axillin、β-Catenin were detected by Western blot.6.Western blot was used to detect the expression of P38,ERK and their phosphorylation.Results:1.The expression of PBEF was significantly increased in HUVECs induced by anoxia-reoxygenatio,and PBEF increases the expression and release of TNF-α,IL-1β,IL-6 inflammatory mediators(P<0.01).2.PBEF can alter the cell contractility,cell-cell adhesions,cell-matrix adhesions by promoting the expression of MLC,VE-cadherin and FAK phosphorylation and β-Catenin,paxillin protein,then change the structure of endothelial cells(P<0.01).3.PBEF participates in changes in permeability of HUVECs via P38 MAPK and ERK signaling pathways(P<0.01).Conclusions:1.PBEF increases the expression of TNF-α,IL-1 β,IL-6 inflammatory mediators after HUVECs anoxia-reoxygenation,then involves the inflammation response.2.PBEF could regulate the shrinkage of endothelial cells 、 destroy cell-cell adhesion and cell-matrix adhesion through P38 MAPK / ERK signaling pathway,then destroy the barrier function of endothelial cells,then increase the permeability of pulmonary vascular endothelial cells induced by anoxia-reoxygenation...