title: Insulin suppresses LPS-induced iNOS and COX-2 expression and NF-kappaB activation in alveolar macrophages.
abstract: The development of septic shock is a common and frequently lethal consequence of gram-negative infection. Mediators released by lung macrophages activated by bacterial products such as lipopolysaccharide (LPS) contribute to shock symptoms. We have shown that insulin down-regulates LPS-induced TNF production by alveolar macrophages (AMs). In the present study, we investigated the effect of insulin on the LPS-induced production of nitric oxide (NO) and prostaglandin (PG)-E(2), on the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2, and on nuclear factor kappa B (NF-kappaB) activation in AMs. Resident AMs from male Wistar rats were stimulated with LPS (100 ng/mL) for 30 minutes. Insulin (1 mU/mL) was added 10 min before LPS. Enzymes expression, NF-kappaB p65 activation and inhibitor of kappa B (I-kappaB)alpha phosphorylation were assessed by immunobloting; NO by Griess reaction and PGE(2) by enzyme immunoassay (EIA). LPS induced in AMs the expression of iNOS and COX-2 proteins and production of NO and PGE(2), and, in parallel, NF-kappaB p65 activation and cytoplasmic I-kappaBalpha phosphorylation. Administration of insulin before LPS suppressed the expression of iNOS and COX-2, of NO and PGE(2) production and Nuclear NF-kappaB p65 activation. Insulin also prevented cytoplasmic I-kappaBalpha phosphorylation. These results show that in AMs stimulated by LPS, insulin prevents nuclear translocation of NF-kappaB, possibly by blocking I-kappaBalpha degradation, and supresses the production of NO and PGE(2), two molecules that contribute to septic shock.
authors.affiliation: Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, SP, Brazil. martinsj@usp.br
authors.forename: Joilson O
authors.initials: JO
authors.lastname: Martins
authors.forename: Matheus
authors.initials: M
authors.lastname: Ferracini
authors.forename: Natalia
authors.initials: N
authors.lastname: Ravanelli
authors.forename: Richardt G
authors.initials: RG
authors.lastname: Landgraf
authors.forename: Sonia
authors.initials: S
authors.lastname: Jancar