The Role of Maternal Gestational Diabetes in Inducing Fetal Endothelial Dysfunction

This study examines the effect of maternal diabetes on fetal endothelial function and gene expression under physiological glucose conditions (5mM). Human umbilical vein endothelial cell (HUVEC) isolated from diabetic mothers (d.HUVEC) grew more slowly than HUVEC isolated from healthy mothers (c.HUVEC) and had delayed doubling time despite increased levels of total vascular endothelial growth factor (VEGF) expression and protein production as determined by real time PCR and ELISA respectively. Using western blot, the levels of antiproliferative VEGF165b isoform were increased in d.HUVEC relative to c.HUVEC. Successful VEGF165b knockdown by small interfering RNA (siRNA) resulted in increased proliferation of d.HUVEC measured by MTT, compared with negative siRNA control, to similar levels measured in c.HUVEC. In addition, d.HUVEC generated excess levels of ROS as revealed by 2′,7′ Dichlorodihydrofluorescein Diacetate (DCFH‐DA) and Nitrotetrazolium blue (NBT). Using microarray, 102 genes were differentially overexpressed between d.HUVEC vs. c.HUVEC (>1.5 fold change; p < 0.05). Functional clustering analysis of these differentially expressed genes revealed participation in inflammatory responses (including adhesion) which may be related to pathological outcomes. Of these genes, ICAM‐1 was validated as upregulated, confirming microarray result. Additional confirmatory immunofluorescence staining revealed increased protein level of ICAM‐1 compared with c.HUVEC a...
Source: Journal of Cellular Physiology - Category: Cytology Authors: Tags: Original Research Article Source Type: research