title: PPARα induced NOS1 phosphorylation via PI3K/Akt in guinea pig antral mucous cells: NO-enhancement in Ca(2+)-regulated exocytosis.
abstract: A PPARα (peroxisome proliferation activation receptor α) agonist (GW7647) activates nitric oxide synthase 1 (NOS1) to produce NO leading to cGMP accumulation in antral mucous cells. In this study, we examined how PPARα activates NOS1. The NO production stimulated by GW7647 was suppressed by inhibitors of PI3K (wortmannin) and Akt (AKT 1/2 Kinase Inhibitor, AKT-inh), although it was also suppressed by the inhibitors of PPARα (GW6471) and NOS1 (N-PLA). GW7647 enhanced the ACh (acetylcholine)-stimulated exocytosis (Ca(2+)-regulated exocytosis) mediated via NO, which was abolished by GW6471, N-PLA, wortmannin, and AKT-inh. The Western blotting revealed that GW7647 phosphorylates NOS1 via phosphorylation of PI3K/Akt in antral mucous cells. The immunofluorescence examinations demonstrated that PPARα existing with NOS1 co-localizes with PI3K and Akt in the cytoplasm of antral mucous cells. ACh alone and AACOCF3, an analogue of arachidonic acid (AA), induced the NOS1 phosphorylation via PI3K/Akt to produce NO, which was inhibited by GW6471. Since AA is a natural ligand for PPARα, ACh stimulates PPARα probably via AA. In conclusion, PPARα activates NOS1 via PI3K/Akt phosphorylation to produce NO in antral mucous cells during ACh stimulation.
authors.affiliation: Laboratory of Pharmacotherapy, Osaka University of Pharmaceutical Sciences.
authors.forename: Saori
authors.initials: S
authors.lastname: Tanaka
authors.forename: Shigekuni
authors.initials: S
authors.lastname: Hosogi
authors.forename: Yukinori
authors.initials: Y
authors.lastname: Sawabe
authors.forename: Chikao
authors.initials: C
authors.lastname: Shimamoto
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Matsumura
authors.forename: Toshio
authors.initials: T
authors.lastname: Inui
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Marunaka
authors.forename: Takashi
authors.initials: T
authors.lastname: Nakahari
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D002087:Butyrates / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D005089:Exocytosis / Q000187:drug effects; D005753:Gastric Mucosa / Q000378:metabolism; D020397:Goblet Cells / Q000187:drug effects / Q000378:metabolism*; D006168:Guinea Pigs; D008297:Male; D009569:Nitric Oxide; D052248:Nitric Oxide Synthase Type I / Q000378:metabolism*; D010080:Oxazoles / Q000494:pharmacology; D047493:PPAR alpha / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010671:Phenylurea Compounds / Q000494:pharmacology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D010766:Phosphorylation; D021381:Protein Transport; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D002087:Butyrates; C449302:GW 6471; C453899:GW 7647; D010080:Oxazoles; D047493:PPAR alpha; D010671:Phenylurea Compounds; D009569:Nitric Oxide; D014443:Tyrosine; D052248:Nitric Oxide Synthase Type I; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D000109:Acetylcholine; D002118:Calcium