title: Cell bioenergetics in Leghorn male hepatoma cells and immortalized chicken liver cells in response to 4-hydroxy 2-nonenal-induced oxidative stress.
abstract: The major objectives of this study were to compare cell bioenergetics in 2 avian liver cell lines under control conditions and in response to oxidative stress imposed by 4-hydroxy 2-nonenal (4-HNE). Cells in this study were from a chemically immortalized Leghorn male hepatoma (LMH) cell line and a spontaneously immortalized chicken liver (CELi) cell line. Oxygen consumption rate (OCR) was monitored in specialized microtiter plates using an XF24 Flux Analyzer (Seahorse Bioscience, Billerica, MA). Cell bioenergetics was assessed by sequential additions of oligomycin, carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP), and antimycin-A that enables the determination of a) OCR linked to adenosine triphosphate (ATP) synthase activity, b) mitochondrial oxygen reserve capacity, c) proton leak, and d) nonmitochondrial cytochrome c oxidase activity. Under control (unchallenged) conditions, LMH cells exhibited higher basal OCR and higher OCR attributed to each of the bioenergetic components listed above compared with CELi cells. When expressed as a percentage of maximal OCR (following uncoupling with FCCP), LMH cells exhibited higher OCR due to ATP synthase and proton leak activity, but lower mitochondrial oxygen reserve capacity compared with CELi cells; there were no differences in OCR associated with nonmitochondrial cytochrome c oxidase activity. Whereas the LMH cells exhibited robust ATP synthase activity up to 50 μM 4-HNE, CELi cells exhibited a progressive decline in ATP synthase activity with 10, 20, and 30 μM 4-HNE. The CELi cells exhibited higher mitochondrial oxygen reserve capacity compared with LMH cells with 0 and 20 μM 4-HNE but not with 30 μM 4-HNE. Both cell lines exhibited inducible proton leak in response to increasing levels of 4-HNE that was evident with 30 μM 4-HNE for CELi cells and with 40 and 50 μM 4-HNE in LMH cells. The results of these studies demonstrate fundamental differences in cell bioenergetics in 2 avian liver-derived cell lines under control conditions and in response to oxidative challenge due to 4-HNE.
authors.affiliation: Department of Poultry Science, Center of Excellence for Poultry Science, University of Arkansas, Fayetteville 72701.
authors.forename: A L
authors.initials: AL
authors.lastname: Piekarski
authors.affiliation: Department of Poultry Science, Center of Excellence for Poultry Science, University of Arkansas, Fayetteville 72701.
authors.forename: B-W
authors.initials: BW
authors.lastname: Kong
authors.affiliation: Department of Poultry Science, Center of Excellence for Poultry Science, University of Arkansas, Fayetteville 72701.
authors.forename: K
authors.initials: K
authors.lastname: Lassiter
authors.affiliation: Department of Poultry Science, Center of Excellence for Poultry Science, University of Arkansas, Fayetteville 72701.
authors.forename: B M
authors.initials: BM
authors.lastname: Hargis
authors.affiliation: Department of Poultry Science, Center of Excellence for Poultry Science, University of Arkansas, Fayetteville 72701 wbottje@uark.edu.
authors.forename: W G
authors.initials: WG
authors.lastname: Bottje
mesh_terms: D000447:Aldehydes / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D002645:Chickens; D015853:Cysteine Proteinase Inhibitors / Q000378:metabolism / Q000494:pharmacology; D004734:Energy Metabolism; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D064688:Metabolic Flux Analysis / Q000662:veterinary; D018384:Oxidative Stress / Q000187:drug effects*
chemical_list: D000447:Aldehydes; D015853:Cysteine Proteinase Inhibitors; C027576:4-hydroxy-2-nonenal
keywords: 4-hydroxy 2-nonenal; avian liver cell; bioenergetics; oxidative stress