Involvement of protein kinase Cdelta and extracellular signal-regulated kinase-2 in the suppression of microglial inducible nitric oxide synthase expression by N-[3,4-dimethoxycinnamoyl]-anthranilic acid (tranilast).
Excess nitric oxide (NO) in the brain released by microglial cells contributes to neuronal damage in various pathologies of the central nervous system (CNS) including neurodegenerative diseases and multiple sclerosis. N-[3,4-Dimethoxycinnamoyl]-anthranilic acid (tranilast, TNL) is an anti-allergic compound which suppresses the activation of monocytes. We show that inducible nitric oxide synthase (iNOS) mRNA and protein expression and the release of NO from N9 microglial cells stimulated with the bacterial endotoxin lipopolysaccharide (LPS) are inhibited when the cells are exposed to TNL. TNL fails to modulate LPS-stimulated nuclear factor-kappaB (NF-kappaB) reporter gene activity and phosphorylation of inhibitory kappaB (IkappaB), indicating that NF-kappaB is not involved in the TNL-mediated suppression of LPS-induced iNOS expression. Moreover, TNL inhibits LPS-induced phosphorylation of extracellular signal-regulated kinase 2 (ERK-2). Finally, TNL abolishes translocation of protein kinase Cdelta (PKCdelta) to the nucleus and suppresses the phosphorylation of the PKCdelta substrate, myristoylated alanin-rich C kinase substrate (MARCKS). We conclude that the anti-allergic compound TNL suppresses microglial iNOS induction by LPS via inhibition of a signalling pathway involving PKCdelta and ERK-2.
D000818:Animals; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000187:drug effects; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D017628:Microglia / Q000187:drug effects* / Q000201:enzymology; D019950:Mitogen-Activated Protein Kinase 1 / Q000502:physiology*; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000096:biosynthesis* / Q000235:genetics; D052247:Nitric Oxide Synthase Type II; D009573:Nitrites / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011493:Protein Kinase C / Q000502:physiology*; D051745:Protein Kinase C-delta; D062367:ortho-Aminobenzoates / Q000494:pharmacology*; D048051:p38 Mitogen-Activated Protein Kinases
D008070:Lipopolysaccharides; D016328:NF-kappa B; D009573:Nitrites; D062367:ortho-Aminobenzoates; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; C497259:Prkcd protein, mouse; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; D019950:Mitogen-Activated Protein Kinase 1; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C012293:tranilast