title: Insulin-induced Akt activation is inhibited by angiotensin II in the vasculature through protein kinase C-alpha.
abstract: Insulin resistance is an important risk factor in the development of cardiovascular diseases such as hypertension and atherosclerosis. However, the specific role of insulin resistance in the etiology of these diseases is poorly understood. Angiotensin (Ang) II is a potent vasculotrophic and vasoconstricting factor. We hypothesize that in vascular smooth muscle cells (VSMCs), Ang II interferes with insulin action by inhibiting Akt, a major signaling molecule implicated in the biological actions of insulin. By immunoblotting with a phospho-specific antibody for Akt, we found that Ang II inhibits insulin-induced Akt phosphorylation in a time- and concentration-dependent manner. The inhibitory effect of Ang II was blocked by a Ang II type 1 receptor antagonist, RNH6270. A protein kinase C (PKC) activator, phorbol 12-myristate 13-acetate, also inhibited insulin-induced Akt phosphorylation. PKC inhibitors, including Go6976 (specific for alpha- and beta-isoforms), blocked the Ang II- and PMA-induced inhibition of Akt phosphorylation by insulin. Moreover, overexpression of PKC-alpha but not PKC-beta isoform by adenovirus inhibited insulin-induced Akt phosphorylation. By contrast, an epidermal growth factor receptor inhibitor (AG1478), a p42/44 mitogen-activated protein kinase (MAPK) kinase inhibitor (PD 598,059), and a p38 MAPK inhibitor (SB 203,580) did not block the Ang II-induced inhibition of Akt phosphorylation. From these data, we conclude that Ang II negatively regulates the insulin signal, Akt, in the vasculature specifically through PKC-alpha activation, providing an alternative molecular mechanism that may explain the association of hyperinsulinemia with cardiovascular diseases.
authors.affiliation: Department of Anatomy and Physiology, Meharry Medical College, Nashville, TN 37208, USA. emotley@mmc.edu
authors.forename: Evangeline D
authors.initials: ED
authors.lastname: Motley
authors.forename: Kunie
authors.initials: K
authors.lastname: Eguchi
authors.forename: Carla
authors.initials: C
authors.lastname: Gardner
authors.forename: Adrienne L
authors.initials: AL
authors.lastname: Hicks
authors.forename: Cherilynn M
authors.initials: CM
authors.lastname: Reynolds
authors.forename: Gerald D
authors.initials: GD
authors.lastname: Frank
authors.forename: Mizuo
authors.initials: M
authors.lastname: Mifune
authors.forename: Motoi
authors.initials: M
authors.lastname: Ohba
authors.forename: Satoru
authors.initials: S
authors.lastname: Eguchi
mesh_terms: D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D002478:Cells, Cultured; D004789:Enzyme Activation; D007329:Insulin Antagonists / Q000494:pharmacology*; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000201:enzymology*; D011493:Protein Kinase C / Q000502:physiology*; D051571:Protein Kinase C-alpha; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D007329:Insulin Antagonists; D011518:Proto-Oncogene Proteins; D000804:Angiotensin II; C494920:Akt1 protein, rat; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha