title: Increased oxidative stress and toxicity in ADH and CYP2E1 overexpressing human hepatoma VL-17A cells exposed to high glucose.
abstract: High glucose mediated oxidative stress and cell death is a well documented phenomenon. Using VL-17A cells which are HepG2 cells over-expressing alcohol dehydrogenase (ADH) and cytochrome P450 2E1 (CYP2E1) and control HepG2 cells, the association of ADH and CYP2E1 with high glucose mediated oxidative stress and toxicity in liver cells was investigated. Cell viability was measured and apoptosis or necrosis was determined through caspase-3 activity, Annexin V-propidium iodide staining and detecting decreases in mitochondrial membrane potential. Reactive oxygen species, lipid peroxidation and the formation of advanced glycated-end products were assessed. The levels of several antioxidants which included glutathione, glutathione peroxidase, catalase and superoxide dismutase were altered in high glucose treated VL-17A cells. Greater toxicity was observed in VL-17A cells exposed to high glucose when compared to HepG2 cells. Oxidative stress parameters were greatly increased in high glucose exposed VL-17A cells and apoptotic cell death was observed. Inhibition of CYP2E1 or caspase 3 or addition of the antioxidant trolox led to significant decreases in high glucose mediated oxidative stress and toxicity. Thus, the over-expression of ADH and CYP2E1 in liver cells is associated with increased high glucose mediated oxidative stress and toxicity.
authors.affiliation: Life Science Division, AU-KBC Research Centre, MIT Campus of Anna University, Chromepet, Chennai-600044, India.
authors.forename: Karthikeyan
authors.initials: K
authors.lastname: Chandrasekaran
authors.forename: Kavitha
authors.initials: K
authors.lastname: Swaminathan
authors.forename: S Mathan
authors.initials: SM
authors.lastname: Kumar
authors.forename: Dahn L
authors.initials: DL
authors.lastname: Clemens
authors.forename: Aparajita
authors.initials: A
authors.lastname: Dey
mesh_terms: D000426:Alcohol Dehydrogenase / Q000378:metabolism*; D017209:Apoptosis / Q000187:drug effects; D006528:Carcinoma, Hepatocellular / Q000473:pathology* / Q000503:physiopathology*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D000072467:Cytochrome P450 Family 2; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D018384:Oxidative Stress / Q000187:drug effects*; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D003577:Cytochrome P-450 Enzyme System; D000426:Alcohol Dehydrogenase; C110332:CYP2F1 protein, human; D000072467:Cytochrome P450 Family 2; D005947:Glucose