Vasculoprotective effect of insulin in the ischemic/reperfused canine heart: role of Akt-stimulated NO production

Cardiovasc Res. 2006 Jan;69(1):57-65. doi: 10.1016/j.cardiores.2005.08.019. Epub 2005 Oct 5.

Abstract

Objectives: The objectives of this study were to investigate the vasculoprotective effects of glucose-insulin-potassium (GIK) on ischemia/reperfusion-induced coronary endothelial functional injury and to elucidate the mechanism involved.

Methods: Dogs were subjected to 50 min of coronary occlusion and 4 h of reperfusion. Vehicle, GIK, or GK were intravenously infused 5 min before reperfusion, and the coronary vascular dysfunction and endothelial apoptosis were determined. In a separate study, cultured endothelial cells were subjected to simulated ischemia/reperfusion, and the signaling pathway involved in insulin's anti-apoptotic effect was investigated.

Results: In vivo ischemia/reperfusion caused significant coronary vascular endothelial dysfunction as evidenced by reduced endothelium-dependent vasorelaxation, decreased nitric oxide (NO) production, and endothelial cell apoptosis as determined by caspase 3 activation and TUNEL staining. Treatment with GIK, but not GK, markedly improved the endothelium-dependent coronary vasorelaxation (P<0.01 versus vehicle), increased total NO production (P<0.01), and attenuated endothelial apoptosis. In cultured endothelial cells, treatment with insulin also markedly increased NO production and reduced simulated ischemia/reperfusion-induced apoptosis. Moreover, pre-treatment with either Akt inhibitor or NO synthase inhibitor almost abolished the anti-apoptotic effect exerted by insulin but not by SNAP, an NO donor.

Conclusion: These results demonstrate that in vivo treatment with GIK at reperfusion attenuates ischemia/reperfusion-induced coronary endothelial dysfunction and endothelial apoptosis in an Akt-dependent and NO-mediated fashion. The coronary vasculoprotective effect elicited by insulin may contribute to the previously observed cardiac protective effect of GIK.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcholine / pharmacology
  • Animals
  • Apoptosis / drug effects
  • Caspase 3
  • Caspases / analysis
  • Cells, Cultured
  • Coronary Vessels / drug effects
  • Coronary Vessels / metabolism
  • Coronary Vessels / pathology*
  • Dogs
  • Endothelial Cells / drug effects
  • Endothelial Cells / metabolism
  • Endothelial Cells / pathology*
  • Endothelin-1 / pharmacology
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / metabolism
  • Endothelium, Vascular / pathology*
  • Female
  • Glucose / pharmacology
  • Humans
  • In Situ Nick-End Labeling
  • In Vitro Techniques
  • Insulin / pharmacology*
  • Male
  • Myocardial Reperfusion Injury / pathology
  • Myocardial Reperfusion Injury / prevention & control*
  • Nitrates / pharmacology
  • Nitric Oxide / analysis
  • Nitric Oxide / metabolism
  • Nitric Oxide Synthase / antagonists & inhibitors
  • Potassium / pharmacology
  • Proto-Oncogene Proteins c-akt / antagonists & inhibitors
  • Proto-Oncogene Proteins c-akt / metabolism
  • Vasodilator Agents / pharmacology

Substances

  • Endothelin-1
  • Insulin
  • Nitrates
  • Vasodilator Agents
  • Nitric Oxide
  • sodium nitrate
  • Nitric Oxide Synthase
  • Proto-Oncogene Proteins c-akt
  • CASP3 protein, human
  • Caspase 3
  • Caspases
  • Glucose
  • Acetylcholine
  • Potassium