title: Role of protein kinase C in BSA-AGE-mediated inducible nitric oxide synthase expression in RAW 264.7 macrophages.
abstract: In the present study, the roles of protein kinase C (PKC) in BSA-derived advanced glycosylation end products (BSA-AGEs)-induced nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression were investigated. Treatment of RAW 264.7 cells with BSA-AGEs caused dose- and time-dependent increases in NO release and iNOS expression in RAW 264.7 cells, whereas BSA alone had no effect on iNOS induction. The tyrosine kinase inhibitor (genistein), the phosphatidylinositol-specific phospholipase C inhibitor (U-73122), the phosphatidylcholine-specific phospholipase C inhibitor (D-609), and the PKC inhibitors (staurosporine, Ro 31-8220, and Go 6976) all inhibited BSA-AGE-induced NO release and iNOS expression in RAW 264.7 cells. Stimulation of RAW 264.7 cells with BSA-AGEs resulted in the formation of inositol monophosphate; the response was attenuated by U-73122 and genistein. BSA-AGEs stimulated PKC-alpha, -betaI, -delta, and -eta but not -zeta translocation from the cytosol to the membrane. However, incubation of RAW 264.7 cells with BSA-AGEs increased phosphorylation of PKC-zeta at threonine-410, which reflects activation of PKC-zeta, indicating the possible involvement of these PKC isoforms in AGE-mediated effects. Pretreatment of RAW 264.7 cells with U-73122, D-609, and genistein reduced the AGE-stimulated translocation of PKC-alpha, -betaI, -delta, and -eta and activation of PKC-zeta. Taken together, these data suggest that BSA-AGEs might activate PKC and subsequently induce iNOS expression and NO release.
authors.affiliation: School of Medicine, Taipei Medical University, Taipei, Taiwan, ROC.
authors.forename: Chih-Hsiung
authors.initials: CH
authors.lastname: Wu
authors.forename: Chien-Hsi
authors.initials: CH
authors.lastname: Chang
authors.forename: Hsiu-Chen
authors.initials: HC
authors.lastname: Lin
authors.forename: Chien-Ming
authors.initials: CM
authors.lastname: Chen
authors.forename: Chien-Huang
authors.initials: CH
authors.lastname: Lin
authors.forename: Horng-Mo
authors.initials: HM
authors.lastname: Lee
mesh_terms: D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D001952:Bridged-Ring Compounds / Q000494:pharmacology; D002460:Cell Line; D004347:Drug Interactions; D004963:Estrenes / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects*; D019833:Genistein / Q000494:pharmacology; D017127:Glycation End Products, Advanced / Q000494:pharmacology*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D008264:Macrophages / Q000187:drug effects* / Q000201:enzymology; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000235:genetics / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II; D009636:Norbornanes; D011493:Protein Kinase C / Q000502:physiology*; D011505:Protein-Tyrosine Kinases / Q000502:physiology; D011760:Pyrrolidinones / Q000494:pharmacology; D013859:Thiocarbamates; D013871:Thiones / Q000494:pharmacology; D010738:Type C Phospholipases / Q000502:physiology
chemical_list: D001952:Bridged-Ring Compounds; D004963:Estrenes; D017127:Glycation End Products, Advanced; D007527:Isoenzymes; D009636:Norbornanes; D011760:Pyrrolidinones; D013859:Thiocarbamates; D013871:Thiones; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; D009569:Nitric Oxide; C046498:tricyclodecane-9-yl-xanthogenate; D019833:Genistein; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; D011505:Protein-Tyrosine Kinases; D011493:Protein Kinase C; D010738:Type C Phospholipases; C105598:phosphatidylcholine-specific phospholipase C