title: Acute effects of phorbol ester and insulin on insulin-induced glucose uptake and protein kinase C activation in rat adipocytes.
abstract: We examined the acute effect of pretreatment with phorbol ester and insulin on insulin-induced glucose uptake and protein kinase C (PKC) translocation from cytosol to the membrane in rat adipocytes. Adipocytes were preincubated with 1 microM tetradecanoylphorbol 13-acetate (TPA) and 10 nM insulin for 60 min and then stimulated with 10 nM insulin for 10 and 30 min to measure PKC activity in cytosol and membrane fractions using a Mono Q column connected onto an HPLC system and [3H]2-deoxyglucose (DOG) uptake, respectively. Pretreatment with 1 microM TPA and 10 nM insulin for 60 min resulted in the marked decreases of insulin-induced [3H]2-DOG uptake. Translocation of Mono Q column-purified cytosolic PKC enzyme activity and PKC beta immunoreactivity from cytosol to the membrane was suppressed by pretreatment with TPA and insulin for 60 min. These results indicate that acute treatment with TPA and insulin which are PKC activators suppress translocation/activation of PKC, and accordingly inhibit insulin-induced glucose uptake. We suggest that a decrease of cytosolic PKC activity may mainly-contribute to the impaired responsiveness of the glucose transport system after acute TPA and insulin treatment.
authors.affiliation: Third Department of Internal Medicine, Gifu University School of Medicine, Japan. tishizuk-gif@umin.u-tokyo.ac.jp
authors.forename: T
authors.initials: T
authors.lastname: Ishizuka
authors.forename: T
authors.initials: T
authors.lastname: Nagashima
authors.forename: K
authors.initials: K
authors.lastname: Kajita
authors.forename: M
authors.initials: M
authors.lastname: Yamamoto
authors.forename: H
authors.initials: H
authors.lastname: Wada
authors.forename: S
authors.initials: S
authors.lastname: Itaya
authors.forename: K
authors.initials: K
authors.lastname: Yamada
authors.forename: A
authors.initials: A
authors.lastname: Miura
authors.forename: Y
authors.initials: Y
authors.lastname: Kanoh
authors.forename: M
authors.initials: M
authors.lastname: Ishizawa
authors.forename: K
authors.initials: K
authors.lastname: Yasuda
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D003600:Cytosol / Q000201:enzymology; D003847:Deoxyglucose / Q000378:metabolism*; D004789:Enzyme Activation; D004822:Epididymis; D007328:Insulin / Q000494:pharmacology*; D008297:Male; D011493:Protein Kinase C / Q000378:metabolism*; D064546:Protein Kinase C beta; D051381:Rats; D017208:Rats, Wistar; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
chemical_list: D007328:Insulin; D003847:Deoxyglucose; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D013755:Tetradecanoylphorbol Acetate