HUVECs from newborns with a strong family history of diabetes show increased apoptosis by flow cytometry with annexin V.

Abstract

A family history of type 2 diabetesmellitus (DM) increases the probability to developDMand endothelial dysfunction. Human umbilical vein endothelial cells (HUVECs) isolated from healthy newborns with familiar background of diverse diseases show early alterations such as less resistance to shear stress. The aim of this study was to evaluate the apoptosis by flow cytometry in HUVECs obtained from healthy newborns with (experimental) and without (control) a strong family history of DM, exposed to different glucose concentrations. Methods: HUVECs were incubated in M-199 culturemedia (containing a 5 mmol/L physiological glucose concentration) or supraphysiological glucose concentrations (15 or 30 mmol/L), for 48 h. Apoptosis was quantified by flow cytometry with annexin V and a polycaspase assay kit (FLICA, Immunochemistry Technologies LLC), and cell death wasmeasurement by propidium iodide (PI, St Cruz Biotechnology Inc) positive stain. Results: Experimental HUVECs showed higher levels of apoptosis in the presence of increasing glucose concentration (p < 0.01), whereas control HUVECs showed low levels of apoptosis. Conclusions: Our results suggest that HUVECs, isolated from healthy newborns with a strong family history of DM, have an abnormal predisposition to apoptosis.