Effects of maternal serum on permeability of glomerular endothelial cell membrane

Abstract

The mechanism of injury on the human glomerular endothelial cells (ciGENC) induced by preeclampsia serum was investigated. Concentration of maternal serum sFlt-1 protein was detected by ELISA. Fluorescently-labeled bovine serum albumin infiltrating through lower chamber of Transwell was measured by multifunction microplate reader. Morphologic change of ciGENC was observed under inverted phase contrast microscope. The concentration of sflt-1 in preeclampsia groups was significantly increased as compared with control group (P<0.01). Permeability in preeclampsia groups was significantly increased as compared with control group (P<0.01). By contrast with severe preeclampsia group, the permeability of ciGENC monolayer in mild preeclampsia group was decreased significantly (P<0.05). Intervention of exogenous VEGF significantly decreased permeability of ciGENC in preeclampsia groups. It was concluded that sFlt-1 increased ciGENC permeability by damaging integrity of endothelial barrier function.

Low molecular weight heparin decreases the permeability of glomerular endothelial cells when exposed to pre‐eclampsia serum in vitro

“…The conditionally immortalized human GEnC line (ciGEnC line) was provided by Dr Simon Satchell of the University of Bristol (Bristol, UK) and cultured in endothelial growth medium2‐microvascular (EGM2‐MV, Lonza, Switzerland) containing 5% foetal calf serum and growth factors such as human epidermal growth factor (hEGF), human fibroblast growth factor‐basic (hFGF‐B) and insulin‐like growth factor‐1 with the substitution of Arg for Glu at position 3 (R3‐IGF‐1). Endothelium was identified in tissues by staining for endothelium specific markers vascular endothelial growth factor (VEGF), Factor VIII and CD31, as previously described . The ciGEnC line was incubated at 33°C in a 5% CO 2 incubator until grown to confluency.…”

“…Endothelium 13 was identified in tissues by staining for endothelium specific markers vascular endothelial growth factor (VEGF), Factor VIII and CD31, as previously described. 14 The ciGEnC line was incubated at 33°C in a 5% CO2 incubator until grown to confluency. Then we transferred the cell culture plates into a 37°C, 5% CO2 incubator to induce differentiation.…”

Upregulation of sFlt-1 by trophoblasts induces the barrier dysfunction of glomerular endothelial cells

“…The ciGEnCs barrier function was evaluated according to the method described previously [14][15][16] . CiGEnCs were incubated at a high density (5×10 7 per well) on the fibronectin-coated polycarbonate filters of the Transwell system (0.4-μm aperture, 6-well plate; Corning Costar, USA) until highly confluent monolayers were obtained.…”