title: The effects of changes in glutathione levels through exogenous agents on intracellular cysteine content and protein adduct formation in chronic alcohol-treated VL17A cells.
abstract: Alcohol-mediated liver injury is associated with changes in the level of the major cellular antioxidant glutathione (GSH). It is interesting to investigate if the changes in intracellular GSH level through exogenous agents affect the intracellular cysteine content and the protein adduct formation indicative of oxidative insult in chronic alcohol treated liver cells. In VL-17A cells treated with 2 mM N-acetyl cysteine (NAC) or 0.1 mM ursodeoxycholic acid (UDCA) plus 100 mM ethanol, an increase in cysteine concentration which was accompanied by decreases in hydroxynonenal (HNE) and glutathionylated protein adducts were observed. Pretreatment of 100 mM ethanol treated VL-17A cells with 0.4 mM buthionine sulfoximine (BSO) or 1 mM diethyl maleate (DEM) had opposite effects. Thus, altered GSH level through exogenous agents may either potentiate or ameliorate chronic alcohol-mediated protein adduct formation and change the cysteine level in chronic alcohol treated VL-17A cells. The gene expression of non-treated and ethanol-treated hepatocytes in 2 microarray datasets was also compared to locate differentially expressed genes involved in cysteine metabolism. The study demonstrates that increased protein adducts formation and changes in cysteine concentration occur under chronic alcohol condition in liver cells which may increase alcohol-mediated oxidative injury.
authors.affiliation: a Life Science Division , AU-KBC Research Centre, MIT Campus of Anna University , Chennai , India.
authors.forename: S Mathan
authors.initials: SM
authors.lastname: Kumar
authors.affiliation: a Life Science Division , AU-KBC Research Centre, MIT Campus of Anna University , Chennai , India.
authors.forename: Madhumitha
authors.initials: M
authors.lastname: Haridoss
authors.affiliation: a Life Science Division , AU-KBC Research Centre, MIT Campus of Anna University , Chennai , India.
authors.forename: Kavitha
authors.initials: K
authors.lastname: Swaminathan
authors.affiliation: a Life Science Division , AU-KBC Research Centre, MIT Campus of Anna University , Chennai , India.
authors.forename: Ramesh Kumar
authors.initials: RK
authors.lastname: Gopal
authors.affiliation: b Department of Internal Medicine , University of Nebraska Medical Center , Omaha , NE , USA.|c Nebraska and Western Iowa Veterans Administration Medical Center , University of Nebraska Medical Center , Omaha , NE , USA.|d Fred and Pamela Buffet Cancer Center , University of Nebraska Medical Center , Omaha , NE , USA.
authors.forename: Dahn
authors.initials: D
authors.lastname: Clemens
authors.affiliation: a Life Science Division , AU-KBC Research Centre, MIT Campus of Anna University , Chennai , India.
authors.forename: Aparajita
authors.initials: A
authors.lastname: Dey
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics; D000447:Aldehydes / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism*; D000072467:Cytochrome P450 Family 2 / Q000235:genetics; D000431:Ethanol / Q000378:metabolism / Q000633:toxicity*; D005978:Glutathione / Q000378:metabolism*; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D018384:Oxidative Stress / Q000187:drug effects* / Q000235:genetics; D059467:Transcriptome / Q000187:drug effects
chemical_list: D000447:Aldehydes; C462359:tert-4-hydroxy-2-nonenal; D000431:Ethanol; D000426:Alcohol Dehydrogenase; C110332:CYP2F1 protein, human; D000072467:Cytochrome P450 Family 2; D005978:Glutathione; D003545:Cysteine
keywords: Alcohol; CYP2E1; GSH; intracellular cysteine; oxidative injury