Ethanol induces oxidative stress and apoptosis in human umbilical vein endothelial cells

We investigated the cytotoxicity of ethanol on human umbilical vein endothelial cell (HUVEC). Human umbilical vein endothelial cells were treated with ethanol (50, 100, or 200 mM) for 24 hours. Cell proliferation was determined by a methyl thiazolyl tetrazolium (MTT) assay. In addition, cell supernatants were collected to determine levels of malondialdehyde (by a thiobarbituric acid kit), superoxide dismutase activity (by a xanthine oxidase kit), nitric oxide release (by a nitrate reductase kit), and expression and release of soluble ICAM-1 (by an ELISA assay). In addition, apoptosis was determined using an Annexin V/PI kit and flow cytometry. High concentrations of ethanol inhibited HUVEC proliferation, increase levels of malondialdehyde, decrease superoxide dismutase activity, and inhibited secretion of nitric oxide. Human umbilical vein endothelial cells treated with ethanol also exhibited up-regulated expression or release of ICAM-1. Furthermore, ethanol-treated HUVECs exhibited a dose-dependent increase in apoptosis. Ethanol can damage endothelial cells by promoting oxidative stress and inflammatory responses, leading to apoptosis.