title: Physiological concentrations of insulin induce endothelin-mediated vasoconstriction during inhibition of NOS or PI3-kinase in skeletal muscle arterioles.
abstract: OBJECTIVE
  To determine the roles of nitric oxide, endothelin-1 and phosphatidylinositol 3-kinase (PI3-kinase) in acute responses of isolated rat skeletal muscle arterioles to insulin.
  
  
  METHODS
  Rat cremaster first order arterioles were separated from surrounding tissue, cannulated in a pressure myograph and responses to insulin (4 microU/ml-3.4 mU/ml) were studied without intraluminal blood or flow.
  
  
  RESULTS
  Insulin alone did not significantly affect arteriolar diameter. Non-selective antagonism of endothelin receptors, with PD-142893, uncovered insulin-induced vasodilatation (25+/-8% from baseline at 3.4 mU/ml), which was abolished by inhibition of NO synthesis with N(G)-nitro-L-arginine (L-NA). Inhibition of NO synthesis alone uncovered insulin-induced vasoconstriction at physiological concentrations (21+/-5% from baseline diameter at 34 microU/ml), which was abolished by PD-142893. The NO donor, S-nitroso-N-acetyl-penicillamine (SNAP) inhibited insulin-induced vasoconstriction during NOS inhibition, even at a concentration that did not elicit vasodilatation itself. Inhibition of PI3-kinase, an intracellular mediator of insulin-induced NO production, with wortmannin, also uncovered insulin-induced vasoconstriction (13+/-3% from baseline at 34 microU/ml) that was abolished by PD-142893.
  
  
  CONCLUSIONS
  Insulin induces both nitric oxide and endothelin-1 activity in rat cremaster first-order arterioles. This study demonstrates for the first time that vasoconstrictive effects of physiological concentrations of insulin during inhibition of NOS activity are mediated by endothelin and that insulin induces endothelin-1-mediated vasoconstriction in isolated skeletal muscle arterioles during inhibition of PI3-kinase. These findings support the hypothesis of altered microvascular reactivity to insulin in conditions of diminished PI3-kinase activity, a prominent feature of insulin resistance.
authors.affiliation: Laboratory for Physiology, VU Medical Centre, van der Boechorststraat 7, Amsterdam, The Netherlands.
authors.forename: Etto C
authors.initials: EC
authors.lastname: Eringa
authors.forename: Coen D A
authors.initials: CD
authors.lastname: Stehouwer
authors.forename: Thomas
authors.initials: T
authors.lastname: Merlijn
authors.forename: Nico
authors.initials: N
authors.lastname: Westerhof
authors.forename: Pieter
authors.initials: P
authors.lastname: Sipkema
mesh_terms: D000818:Animals; D001160:Arterioles / Q000187:drug effects / Q000502:physiology; D046508:Culture Techniques; D004305:Dose-Response Relationship, Drug; D019332:Endothelin-1 / Q000502:physiology*; D007328:Insulin / Q000494:pharmacology* / Q000502:physiology; D008297:Male; D008833:Microcirculation / Q000187:drug effects; D018482:Muscle, Skeletal / Q000098:blood supply*; D009569:Nitric Oxide / Q000502:physiology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000502:physiology*; D019869:Phosphatidylinositol 3-Kinases / Q000502:physiology; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction; D014661:Vasoconstriction / Q000187:drug effects* / Q000502:physiology
chemical_list: D019332:Endothelin-1; D007328:Insulin; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors