title: Bradykinin enhances GLUT4 translocation through the increase of insulin receptor tyrosine kinase in primary adipocytes: evidence that bradykinin stimulates the insulin signalling pathway.
abstract: It has been suggested that bradykinin stimulates glucose uptake in experiments in vivo and in cultured cells. However, its mechanism has not yet been fully elucidated. In this study, the effects of bradykinin on the insulin signalling pathway were evaluated in isolated dog adipocytes. The bradykinin receptor binding study revealed that dog adipocytes possessed significant numbers of bradykinin receptors (Kd = 83 pmol/l, binding sites = 1.7 x 10(4) site/ cell). Reverse transcription-polymerase chain reaction amplification showed the mRNA specific for bradykinin B2 receptor in the adipocytes. Bradykinin alone did not increase 2-deoxyglucose uptake in adipocytes; however, in the presence of insulin (10(-7) mol/l) it significantly increased 2-deoxyglucose uptake in a dose-dependent manner. Bradykinin also enhanced insulin stimulated GLUT4 translocation from the intracellular fraction to the cell membrane, and insulin induced phosphorylation of the insulin receptor beta subunit and insulin receptor substrate-1 (IRS-1) without affecting the binding affinities or numbers of cell surface insulin receptors in dog adipocytes. The time-course of insulin stimulated phosphorylation of the insulin receptor beta subunit revealed that phosphorylation reached significantly higher levels at 10 min, and stayed at the higher levels until 120 min in the presence of bradykinin, suggesting that bradykinin delayed the dephosphorylation of the insulin receptor. It is concluded that bradykinin could potentiate insulin induced glucose uptake through GLUT4 translocation. This effect could be explained by the potency of bradykinin to upregulate the insulin receptor tyrosine kinase activity which stimulates phosphorylation of IRS-1, followed by GLUT4 translocation.
authors.affiliation: Department of Metabolic Medicine, Kumamoto University School of Medicine, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Isami
authors.forename: H
authors.initials: H
authors.lastname: Kishikawa
authors.forename: E
authors.initials: E
authors.lastname: Araki
authors.forename: M
authors.initials: M
authors.lastname: Uehara
authors.forename: K
authors.initials: K
authors.lastname: Kaneko
authors.forename: T
authors.initials: T
authors.lastname: Shirotani
authors.forename: M
authors.initials: M
authors.lastname: Todaka
authors.forename: S
authors.initials: S
authors.lastname: Ura
authors.forename: S
authors.initials: S
authors.lastname: Motoyoshi
authors.forename: K
authors.initials: K
authors.lastname: Matsumoto
authors.forename: N
authors.initials: N
authors.lastname: Miyamura
authors.forename: M
authors.initials: M
authors.lastname: Shichiri
mesh_terms: D017667:Adipocytes / Q000187:drug effects / Q000502:physiology*; D000273:Adipose Tissue / Q000502:physiology; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D001920:Bradykinin / Q000494:pharmacology*; D002478:Cells, Cultured; D017124:Conserved Sequence; D017931:DNA Primers; D003847:Deoxyglucose / Q000378:metabolism*; D004285:Dogs; D051275:Glucose Transporter Type 4; D006801:Humans; D055504:Insulin Receptor Substrate Proteins; D007700:Kinetics; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins / Q000378:metabolism*; D009124:Muscle Proteins; D010750:Phosphoproteins / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D043782:Receptor, Bradykinin B2; D011972:Receptor, Insulin / Q000187:drug effects / Q000378:metabolism*; D018002:Receptors, Bradykinin / Q000096:biosynthesis / Q000187:drug effects / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D017931:DNA Primers; D051275:Glucose Transporter Type 4; C526292:IRS1 protein, human; D055504:Insulin Receptor Substrate Proteins; C526295:Irs1 protein, mouse; D009004:Monosaccharide Transport Proteins; D009124:Muscle Proteins; D010750:Phosphoproteins; D043782:Receptor, Bradykinin B2; D018002:Receptors, Bradykinin; C495692:SLC2A4 protein, human; C495693:Slc2a4 protein, mouse; D003847:Deoxyglucose; D011972:Receptor, Insulin; D001920:Bradykinin