Resistance to the nitric oxide/cyclic guanosine 5′-monophosphate/ protein kinase G pathway in vascular smooth muscle cells from the obese zucker rat, a classical animal model of insulin resistance: Role of oxidative stress

I. Russo, P. Del Mese, G. Doronzo, L. Mattiello, M. Viretto, A. Bosia, G. Anfossi, Mariella Trovati

Research output: Contribution to journalArticle

Abstract

Some in vivo and ex vivo studies demonstrated a resistance to the vasodilating effects of nitric oxide (NO) in insulin-resistant states and, in particular, obese Zucker rats (OZR). To evaluate the biochemical basis of this phenomenon, we aimed to identify defects of the NO/cGMP/cGMP-dependent protein kinase (PKG) pathway in cultured vascular smooth muscle cells (VSMCs) from OZR and lean Zucker rats (LZR) by measuring: 1) NO donor ability to increase cGMP in the absence and presence of inhibitors of soluble guanylate cyclase (sGC) and phosphodiesterases (PDEs); 2) NO and cGMP ability to induce, via PKG, vasodilator-stimulated phosphoprotein (VASP) phosphorylation at serine 239 and PDE5 activity; 3) protein expression of sGC, PKG, total VASP, and PDE5; 4) superoxide anion concentrations and ability of antioxidants (superoxide dismutase-catalase and amifostine) to influence the NO/cGMP/PKG pathway activation; and 5) hydrogen peroxide influence on PDE5 activity and VASP phosphorylation. VSMCs from OZR vs. LZR showed: 1) baseline cGMP concentrations higher, at least in part owing to reduced catabolism by PDEs; 2) impairment of NO donor ability to increase cGMP, even in the presence of PDE inhibitors, suggesting a defect in the NO-induced sGC activation; 3) reduction of NO and cGMP ability to activate PKG, indicated by the impaired ability to phosphorylate VASP at serine 239 and to increase PDE5 activity via PKG; 4) similar baseline protein expression of sGC, PKG, total VASP, and PDE5; and 5) higher levels of superoxide anion. Antioxidants partially prevented the defects of the NO/cGMP/PKG pathway observed in VSMCs from OZR, which were reproduced by hydrogen peroxide in VSMCs from LZR, suggesting the pivotal role of oxidative stress.

Original languageEnglish
Pages (from-to)1480-1489
Number of pages10
JournalEndocrinology
Volume149
Issue number4
DOIs
Publication statusPublished - Apr 2008

Fingerprint

Guanosine Monophosphate
Zucker Rats
Cyclic GMP-Dependent Protein Kinases
Vascular Smooth Muscle
Smooth Muscle Myocytes
Insulin Resistance
Nitric Oxide
Oxidative Stress
Animal Models
Nitric Oxide Donors
Phosphoric Diester Hydrolases
Superoxides
Serine
Hydrogen Peroxide
Biochemical Phenomena
Antioxidants
Phosphorylation
Amifostine
Phosphodiesterase Inhibitors
Catalase

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Resistance to the nitric oxide/cyclic guanosine 5′-monophosphate/ protein kinase G pathway in vascular smooth muscle cells from the obese zucker rat, a classical animal model of insulin resistance : Role of oxidative stress. / Russo, I.; Del Mese, P.; Doronzo, G.; Mattiello, L.; Viretto, M.; Bosia, A.; Anfossi, G.; Trovati, Mariella.

In: Endocrinology, Vol. 149, No. 4, 04.2008, p. 1480-1489.

Research output: Contribution to journalArticle

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AU - Del Mese, P.

AU - Doronzo, G.

AU - Mattiello, L.

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