title: Overexpression of aldehyde dehydrogenase-2 (ALDH2) transgene prevents acetaldehyde-induced cell injury in human umbilical vein endothelial cells: role of ERK and p38 mitogen-activated protein kinase.
abstract: Acetaldehyde, the major ethanol metabolite that is far more toxic and reactive than ethanol, has been postulated to be responsible for alcohol-induced tissue and cell injury. This study was to examine whether facilitated acetaldehyde metabolism affects acetaldehyde-induced oxidative stress and apoptosis. Transgene-encoding human aldehyde dehydrogenase-2 (ALDH2), which converts acetaldehyde into acetate, was constructed under chicken beta-actin promoter and transfected into human umbilical vein endothelial cells (HUVECs). Efficacy of ALDH2 transfection was verified using green fluorescent protein and ALDH2 enzymatic assay. Generation of reactive oxygen species (ROS) was measured using chloromethyl-2',7'-dichlorodihydrofluorescein diacetate. Apoptosis was evaluated by 4',6'-diamidino-2'-phenylindoladihydrochloride fluorescence microscopy, quantitative DNA fragmentation, and caspase-3 assay. Acetaldehyde (0-200 microm) elicited ROS generation and apoptosis in HUVECs in a time- and concentration-dependent manner, associated with activation of the stress signal molecules ERK1/2 and p38 mitogen-activated protein (MAP) kinase. A close liner correlation was observed between the acetaldehyde-induced ROS generation and apoptosis. Interestingly, the acetaldehyde-induced ROS generation, apoptosis, activation of ERK1/2, and p38 MAP kinase were prevented by the ALDH2 transgene or antioxidant alpha-tocopherol. The involvement of ERK1/2 and p38 MAP kinase in acetaldehyde-induced apoptosis was confirmed by selective kinase inhibitors U0126, SB203580, and SB202190. Collectively, our data revealed that facilitation of acetaldehyde metabolism by ALDH2 transgene overexpression may prevent acetaldehyde-induced cell injury and activation of stress signals. These results indicated therapeutic potential of ALDH2 enzyme in the prevention and detoxification of acetaldehyde or alcohol-induced cell injury.
authors.affiliation: Division of Pharmaceutical Sciences, Department of Molecular Biology and Microscopic Facilities, University of Wyoming, Laramie, Wyoming 82071-3375.
authors.forename: Shi-Yan
authors.initials: SY
authors.lastname: Li
authors.forename: Mark
authors.initials: M
authors.lastname: Gomelsky
authors.forename: Jinhong
authors.initials: J
authors.lastname: Duan
authors.forename: Zhaojie
authors.initials: Z
authors.lastname: Zhang
authors.forename: Larissa
authors.initials: L
authors.lastname: Gomelsky
authors.forename: Xiaochun
authors.initials: X
authors.lastname: Zhang
authors.forename: Paul N
authors.initials: PN
authors.lastname: Epstein
authors.forename: Jun
authors.initials: J
authors.lastname: Ren
mesh_terms: D000079:Acetaldehyde / Q000633:toxicity*; D000444:Aldehyde Dehydrogenase / Q000235:genetics*; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis; D002460:Cell Line; D002478:Cells, Cultured; D004727:Endothelium / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D018384:Oxidative Stress; D017382:Reactive Oxygen Species; D013050:Spectrometry, Fluorescence; D019076:Transgenes; D014471:Umbilical Veins / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D024502:alpha-Tocopherol / Q000494:pharmacology
chemical_list: D000975:Antioxidants; D017382:Reactive Oxygen Species; D000444:Aldehyde Dehydrogenase; D020928:Mitogen-Activated Protein Kinases; D000079:Acetaldehyde; D024502:alpha-Tocopherol