Heat shock protein A12B decreases lipopolysaccharide-induced permeability of human umbilical vein endothelial cells

Abstract

Objective To explore the lipopolysaccharide(LPS)-induced changes of heat shock protein A12B(HSPA12B)expression and its effect on the permeability of human umbilical vein endothelial cells(HUVECs).Methods The HUVECs cultured in vitro were divided into four groups: control group(without any treatment); LPS group(1 μg / m L LPS); LPS + p IRES2-EGFPHSPA12B-3Flag group(The HSPA12 B gene overexpression plasmid was transiently transfected into HUVECs and then LPS of 1 μg / m L was added); and LPS + p IRES2-EGFP-3Flag group(The negative control plasmid was transiently transfected into HUVECs and then LPS of 1 μg / m L was added).The transendothelial electrical resistance(TEER) of HUVECs was measured by MERSST × 01 Electrode.The expression of VE-cadherin was studied by flow cytometry.RT-PCR and Western blotting analysis were used to examine the expression changes of HSPA12 B mRNA and protein in HUVECs stimulated with LPS at various time points.Results It was showed that after stimulated with LPS,HSPA12 B mRNA and protein were gradually upregulated,and peaked at 12 h.HSPA12 B significantly increased the TEER value of HUVECs(P 0.001),and it could completely offset the decline of TEER value induced by LPS(P 0.001).Besides,HSPA12 B could cause upregulation of VE-cadherin expression.Conclusion HSPA12 B can reduce the permeability of vascular endothelial cells by up-regulating the expression of VE-cadherin,thus protecting the vascular endothelial barrier function of endothelial cells.