Study The Effect Of SFlt-1 And VEGF In Pathogenic Mechanism Of Glomerular Endothelial Cell Dysfunction

Objective: To explore the role of soluble fins-like tyrosine kinase receptor 1 (sFlt-1) in serum acquired from normal pregnancy and preeclampsia, the relation between the level of sFlt-1 protein and monolayer glomerular endothelial cell dysfunction.Methords:1. Serum concentration of sFlt-1 protein in specimen of 12 normal pregnant women, 12 mild preeclampsia women and 12 severe preeclampsia women was detected by performing enzyme linked immunosorbent assay (ELISA).2. Immunohistochemical detection of VEGF, FactorⅧand CD31 which are located in the cytoplasm of GENC are performed.3. The culture of human glomerular endothelial cell lines was intervened with maternal serum samples. The effect of serum on human glomerular endothelial cell dysfuntion was detected by multifunction microplate reader: monolayer barrier function was estimated by transferrring fluorescently-labeled BSA across GENC monolayer.Result:1. The level of sFlt-1protein in serum of severe preeclampsia was observably higher than that of normal pregnancy, whereas the level of sFlt-1 in severe preeclampsia was significantly higher than that of mild preeclampsia.2. Permeability in severe preeclampsia group was significantly increased compared with normal group. By contrast with severe preeclampsia group, the permeability of monolayer GENC in mild preeclampsia group decreased significantly. Intervention of exogenous VEGF decreased significantly the permeability of GENC in preeclampsia groups.3. The relationship between level of sFlt-1 in serum of preeclampsia and glomerular endothelial cell dysfunction was positive.Conclusion: The results of our study suggest that sFlt-1 increased GENC permeability by damaging integrity of endothelial barrier function, which may lead to the pathogenesis of preeclampsia. Objective: To investigate the effects of VEGF against sFlt-1 over-expression induced glomerular endothelial cell dysfunction.Methords:1. sFlt-1/pcDNA3 with liposome transfected into trophoblast cells. The level of sFlt-1 protein and mRNA in transfected cell was detected by performing reverse transcriptase-polymerase chain reaction (RT-PCR), real-time polymerase chain reaction (Real-Time PCR) and enzyme-linked immunosorbent assay (ELISA).2. The effect of sFlt-1 over-expression on endothelial cell dysfunction was determined on the basis of following aspects: momolayer barrier function was evaluated by transferring fluorescently-labeled BSA acrosss GENC monolayer.Result:1. sFlt-1 was transfected into trophoblast cells for 72h, compared with the normal control group, the expression of sFlt-1 mRNA was significantly increased, while the medium of free sflt-1 protein levels were also significantly higher.2. Transfected with sFlt-1 in glomerular endothelial cell function was impaired: The monolayer endothelial cell of bovine serum albumin concentrations were significantly increased, the endothelial cell monolayer barrier function was impaired, and by adding external VEGF-derived endothelial cells across the monolayer of bovine serum albumin concentrations were significantly lower.Conclusion: VEGF plays an important role in maintaining the phsiological functions of glomerular endothelial cells. Over-expression of sFlt-1 in trophoblast cells can lead to glomerular endothelial dysfunction, which may be the resaon of clinical sysmptom in preeclampsia.