Medizinische Universität Graz - Research portal
Selected Publication:
Diaz Perez, F.
Programming endothelial dysfunction in diabetic pregnancies
PhD-Studium (Doctor of Philosophy); Humanmedizin; [ Dissertation ] Graz Medical University; 2016. pp.
[OPEN ACCESS]
FullText
- Authors Med Uni Graz:
- Advisor:
- Desoye Gernot
- Hiden Ursula
- Altmetrics:
- Abstract:
- Gestational diabetes mellitus (GDM) is a maternal glucose disorder that leads to an increased risk of cardiovascular disease (CVD) for both, mother and the offspring. In GDM, the endothelium of the feto-placental vasculature is exposed to hyperglycaemia and a pro-inflammatory environment. We hypothesized that human placental arterial endothelial cells of healthy and GDM pregnancies differ in their phenotype as a result of epigenetic modifications caused by the low-grade pro-inflammatory environment of GDM. For this purpose, primary arterial endothelial cells were isolated from human term placentas of healthy (control AEC) and GDM pregnancies (GDM AEC) and cultured. Endothelial functions, such as cell adhesion molecule expression for macrophage attachment, nitric oxide synthesis for vascular tone regulation, and angiogenesis including proliferation were investigated. Transcriptome was analyzed by RT-qPCR. Target proteins were investigated by ELISA, immunoblot, immunofluorescence and/or immunohistochemistry. Angiogenic potential of the cells was evaluated by 3D tube-formation assay after seeding cells on top of fibrin matrices. The main findings of this study were: 1) eNOS mRNA expression and NO-bioavailability were unchanged in AEC from control and GDM placentas. 2) GDM causes decreased levels of ICAM-1 and unaltered levels of VCAM-1 and E-selectin in AEC. 3) GDM altered CDK4 and cyclin-D2 transcript and protein levels, together with decreased angiogenic potential of GDM AEC compared to control AEC. 4) GDM increased mRNA levels of versican (V0, V1, V2 and V3). ELISA showed decreased levels of all forms of versican in supernatant of GDM AEC and no change in cell lysate, while versican V0 was increased in cell lysate of GDM AEC and unaltered in supernatant. Immunoblot and immunofluorescence (IF) failed to detect versican V0 in cultured AEC. These data indicate changes in expression, translation and secretion of different isoforms of versican. Alterations found in extracellular matrix protein versican, in cell cycle regulators CDK4 and cyclin-D2, and in the angiogenic potential of AEC after GDM correlate with DNA methylation observed previously in our lab, suggesting in utero programming of AEC. Moreover, these changes seem to be compensated with unchanged NO bioavailability and eNOS transcript expression, together with decreased levels of ICAM-1 and unaltered levels of VCAM-1 and E-selectin, which appear to have a protective role of the placenta in GDM.