Study On The Effect Of RTEF-1 On Preventing Abnormal Increasing Of Endothelial Permeability

[Objective] To investigate the effect of Related transcriptional enhancer factor-1（RTEF-1）on endothelial permeability and the potential mechanism in human umbilical vein endothelial cells（HUVECss）.[Methods] We established RTEF-1 over-expressing HUVECss with PXJ40-RTEF-1 vectors containing RTEF-1 coding sequence（NM₀03213）, and RTEF-1 suppression HUVECss with small interfering RNA（siRNA）. The efficiency of RTEF-1 over-expression and RNA interfering transfection were identified by Real-time Quantitative PCR and western blot.HUVECss were divided into five groups: Control group: HUVECss transfected with blank vector PXJ40; RTEF-1 o/e Group: HUVECss transfected with PXJ40-RTEF-1 vectors; NC siRNA Group: HUVECss transfected With negative control siRNA; RTEF-1 siRNA1Group: HUVECss transfected with RTEF-1 siRNA1; RTEF-1 siRNA2 Group: HUVECss transfected with RTEF-1 siRNA2. Two-chamber transwell detection systems were used to measuring the monolayer paracellular permeability of HUVECss after incubation with lipopolysaccharide. Real-time Quantitative PCR and western blot were performed to detect RTEF-1- induced changes of gene expression of Edg-1, SPHK1, SPHK2 and junctional complexes in HUVECss.[Results] LPS increased the permeability of monolayer endothelium. RTEF-1over-expression could relief the abnormal increasing induced by LPS in HUVECs（P<0.05）.RTEF-1 over-expression in HUVECs upregulated Edg-1 expression. RTEF-1 interfering downregulated Edg-1, SPHK1, SPHK2 and CX43 expression, while upregulated claudin3,claudin4 expression（P<0.05）. No significant changes were deteced on the expression of ZO-1, ZO-2, occludin, VE-cadherin, CX37 and CX40 induced by RTEF-1.[Conclusions] RTEF-1 could prevent the abnormal increasing of endothelial permeability.Edg-1, SPHK1, SPHK2 and interendothelial Junctional molecular claudin3, claudin4 and CX43 were involved in RTEF-1 induced endothelial permeability protection.