title: Contribution of inducible and neuronal nitric oxide synthases to mitochondrial damage and melatonin rescue in LPS-treated mice.
abstract: NOS isoform activation is related to liver failure during sepsis, but the mechanisms driving mitochondrial impairment remain unclear. We induced sepsis by LPS administration to inducible nitric oxide synthase (iNOS-/-) and neuronal nitric oxide synthase (nNOS-/-) mice and their respective wild-type controls to examine the contribution of iNOS to mitochondrial failure in the absence of nNOS. To achieve this goal, the determination of messenger RNA (mRNA) expression and protein content of iNOS in cytosol and mitochondria, the mitochondrial respiratory complex content, and the levels of nitrosative and oxidative stress (by measuring 3-nitrotyrosine residues and carbonyl groups, respectively) were examined in the liver of control and septic mice. We detected strongly elevated iNOS mRNA expression and protein levels in liver cytosol and mitochondria of septic mice, which were related to enhanced oxidative and nitrosative stress, and with fewer changes in respiratory complexes. The absence of the iNOS, but not nNOS, gene absolutely prevented mitochondrial impairment during sepsis. Moreover, the nNOS gene did not modify the expression and the effects of iNOS here shown. Melatonin administration counteracted iNOS activation and mitochondrial damage and enhanced the expression of the respiratory complexes above the control values. These effects were unrelated to the presence or absence of nNOS. iNOS is a main target to prevent liver mitochondrial impairment during sepsis, and melatonin represents an efficient antagonist of these iNOS-dependent effects whereas it may boost mitochondrial respiration to enhance liver survival.
authors.affiliation: Centro de Investigación Biomédica, Parque Tecnológico de Ciencias de la Salud, Instituto de Biotecnología, Universidad de Granada, Granada, Spain.|Departamento de Fisiología, Facultad de Medicina, Universidad de Granada, Granada, Spain.
authors.forename: José Antonio
authors.initials: JA
authors.lastname: García
authors.affiliation: Centro de Investigación Biomédica, Parque Tecnológico de Ciencias de la Salud, Instituto de Biotecnología, Universidad de Granada, Granada, Spain.|Departamento de Fisiología, Facultad de Medicina, Universidad de Granada, Granada, Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: Ortiz
authors.affiliation: Departamento de Farmacología y Fisiología, Facultad de Medicina, Universidad de Zaragoza, Zaragoza, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Miana
authors.affiliation: Centro de Investigación Biomédica, Parque Tecnológico de Ciencias de la Salud, Instituto de Biotecnología, Universidad de Granada, Granada, Spain.|Departamento de Fisiología, Facultad de Medicina, Universidad de Granada, Granada, Spain.
authors.forename: Carolina
authors.initials: C
authors.lastname: Doerrier
authors.affiliation: Centro de Investigación Biomédica, Parque Tecnológico de Ciencias de la Salud, Instituto de Biotecnología, Universidad de Granada, Granada, Spain.|Departamento de Fisiología, Facultad de Medicina, Universidad de Granada, Granada, Spain.
authors.forename: Marisol
authors.initials: M
authors.lastname: Fernández-Ortiz
authors.affiliation: Centro de Investigación Biomédica, Parque Tecnológico de Ciencias de la Salud, Instituto de Biotecnología, Universidad de Granada, Granada, Spain.|Departamento de Fisiología, Facultad de Medicina, Universidad de Granada, Granada, Spain.
authors.forename: Iryna
authors.initials: I
authors.lastname: Rusanova
authors.affiliation: Centro de Investigación Biomédica, Parque Tecnológico de Ciencias de la Salud, Instituto de Biotecnología, Universidad de Granada, Granada, Spain.|Departamento de Fisiología, Facultad de Medicina, Universidad de Granada, Granada, Spain.|CIBERfes, Ibs.Granada, 18016, Granada, Spain.
authors.forename: Germaine
authors.initials: G
authors.lastname: Escames
authors.affiliation: Departamento de Farmacología y Fisiología, Facultad de Medicina, Universidad de Zaragoza, Zaragoza, Spain.
authors.forename: José Joaquín
authors.initials: JJ
authors.lastname: García
authors.affiliation: Centro de Investigación Biomédica, Parque Tecnológico de Ciencias de la Salud, Instituto de Biotecnología, Universidad de Granada, Granada, Spain. dacuna@ugr.es.|Departamento de Fisiología, Facultad de Medicina, Universidad de Granada, Granada, Spain. dacuna@ugr.es.|CIBERfes, Ibs.Granada, 18016, Granada, Spain. dacuna@ugr.es.|Unidad de Gestión Clínica de Laboratorios, Complejo Hospitalario de Granada, Granada, Spain. dacuna@ugr.es.
authors.forename: Darío
authors.identifier: http://orcid.org/0000-0002-9680-1560
authors.initials: D
authors.lastname: Acuña-Castroviejo
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000008:administration & dosage / Q000627:therapeutic use*; D015415:Biomarkers / Q000097:blood; D004195:Disease Models, Animal; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D048550:Hepatic Insufficiency / Q000209:etiology / Q000517:prevention & control*; D007274:Injections, Intraperitoneal; D008070:Lipopolysaccharides / Q000633:toxicity; D008099:Liver / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D008550:Melatonin / Q000008:administration & dosage / Q000627:therapeutic use*; D057507:Mice, 129 Strain; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008930:Mitochondria, Liver / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D052248:Nitric Oxide Synthase Type I / Q000235:genetics / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects; D050050:Protein Carbonylation / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D018805:Sepsis / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism / Q000503:physiopathology
chemical_list: D000975:Antioxidants; D015415:Biomarkers; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D052248:Nitric Oxide Synthase Type I; D052247:Nitric Oxide Synthase Type II; C496321:Nos1 protein, mouse; C496317:Nos2 protein, mouse; D008550:Melatonin
keywords: Free radicals; LPS; Liver; Melatonin; Mitochondria; Nitric oxide synthase