title: The Role of Maternal Gestational Diabetes in Inducing Fetal Endothelial Dysfunction.
abstract: Gestational diabetes mellitus (GDM) is known to be associated with fetal endothelial dysfunction, however, the mechanisms are not fully understood. This study examines the effect of maternal diabetes on fetal endothelial function and gene expression under physiological glucose conditions (5 mM). 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 versus 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 results. Additional confirmatory immunofluorescence staining revealed increased protein expression of ICAM-1 compared with c.HUVEC which was reduced by vitamin C treatment (100 μM). Thus, maternal diabetes induces persistent alterations in fetal endothelial function and gene expression following glucose normalization and antioxidant treatment could help reverse endothelium dysfunction.
authors: Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia.
authors: Samar A
authors: SA
authors: Sultan
authors: School of Computing, Informatics and Media, University of Bradford, Bradford, UK.
authors: Wanting
authors: W
authors: Liu
authors: School of Computing, Informatics and Media, University of Bradford, Bradford, UK.
authors: Yonghong
authors: Y
authors: Peng
authors: School of Medical Sciences, University of Bradford, Bradford, UK.
authors: W
authors: W
authors: Roberts
authors: Department of Diabetes and Endocrinology, Bradford Royal Infirmary, Bradford, UK.
authors: Donald
authors: D
authors: Whitelaw
authors: School of Medical Sciences, University of Bradford, Bradford, UK.
authors: Anne M
authors: AM
authors: Graham
mesh_terms: D049109:Cell Proliferation / Q000235:genetics*; D016640:Diabetes, Gestational / Q000235:genetics* / Q000473:pathology; D042783:Endothelial Cells / Q000378:metabolism* / Q000473:pathology; D005260:Female; D005786:Gene Expression Regulation; D055786:Gene Knockout Techniques; D005947:Glucose / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis; D011247:Pregnancy; D020033:Protein Isoforms; D014471:Umbilical Veins / Q000254:growth & development / Q000378:metabolism* / Q000473:pathology; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D020033:Protein Isoforms; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D018799:Intercellular Adhesion Molecule-1; D005947:Glucose