zaprinast and Insulin-Resistance

Topics: Adipocytes; Animals; Blotting, Western; Bradykinin; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Dual-Specificity Phosphatases; Glucose; Guanylate Cyclase; Immunoprecipitation; Insulin Resistance; JNK Mitogen-Activated Protein Kinases; Male; Mitogen-Activated Protein Kinase Phosphatases; Nitric Oxide; Nitric Oxide Synthase Type III; Phosphodiesterase Inhibitors; Proto-Oncogene Proteins c-akt; Purinones; Rats; Rats, Sprague-Dawley; Real-Time Polymerase Chain Reaction; RNA, Messenger; Signal Transduction

Nitric oxide activates guanylate cyclase to form cGMP, comprising a signalling system that is believed to be a distinct mechanism for increasing glucose transport and metabolism in skeletal muscle. The effects of a selective cGMP phosphodiesterase inhibitor, zaprinast, on basal glucose utilization was investigated in incubated rat soleus muscle preparations isolated from both insulin-sensitive (lean Zucker; Fa/?) and insulin-resistant (obese Zucker; fa/fa) rats. Zaprinast at 27 microM significantly increased cGMP levels in incubated soleus muscle isolated from lean, but not obese, Zucker rats. Muscles were incubated with 14C-labelled glucose and various concentrations of zaprinast (3, 27 and 243 microM). Zaprinast (at 27 and 243 microM) significantly increased rates of net and 14C-labelled lactate release and of glycogen synthesis in lean Zucker rat soleus muscle; glucose oxidation was also increased by 27 microM zaprinast. In addition, regardless of concentration, the phosphodiesterase inhibitor failed to increase any aspect of 14C-labelled glucose utilization in soleus muscles isolated from obese Zucker rats. The maximal activity of nitric oxide synthase (NOS) was significantly decreased in insulin-resistant obese Zucker muscles. Thus the lack of effect of zaprinast in insulin-resistant skeletal muscle is consistent with decreased NOS activity. To test whether there is a defect in insulin-resistant skeletal muscle for endogenous activation of guanylate cyclase, soleus muscles were isolated from both insulin-sensitive and insulin-resistant Zucker rats and incubated with various concentrations of the NO donor sodium nitroprusside (SNP; 0.1, 1, 5 and 15 mM). SNP significantly increased rates of net and 14C-labelled lactate release, as well as glucose oxidation in muscles isolated from both insulin-sensitive and insulin-resistant rats. A decreased response to SNP was observed in the dose-dependent generation of cGMP within isolated soleus muscles from insulin-resistant rats. A possible link between impaired NO/cGMP signalling and abnormal glucose utilization by skeletal muscle is discussed.

Topics: Animals; Cell Fractionation; Cyclic GMP; Female; Glucose; Glycogen; Insulin; Insulin Resistance; Lactic Acid; Muscle, Skeletal; Nitric Oxide; Nitric Oxide Synthase; Nitroprusside; Obesity; Phosphodiesterase Inhibitors; Purinones; Rats; Rats, Zucker; Signal Transduction

Topics: Animals; Biological Transport; Glucose; Glycogen; Glycolysis; Insulin Resistance; Lactates; Muscle, Skeletal; Phosphodiesterase Inhibitors; Purinones; Rats; Rats, Inbred SHR