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J Thorac Cardiovasc Surg 1998;116:485-489
© 1998 Mosby, Inc.
Cardiopulmonary Support and Physiology |
Supported by the Heart and Stroke Foundation of Canada (grant T2683).V.R., F.M., and G.C. are Research Fellows of the HSFC. R.D.W. is a CareerInvestigator of the HSFO. R.K.L. is a Research Scholar of the HSFO.
Received for publication Dec. 4, 1997. Revisions requested Jan. 20, 1998; revisions received March 5, 1998. Accepted for publication April 15, 1998. Address for reprints: Richard D. Weisel, MD, EN 14-215, The TorontoHospital, 200 Elizabeth St., Toronto, Ontario M5G 2C4, Canada
Impaired myocardial metabolism after cardioplegic arrest results inpersistent anaerobic lactate production. Insulin may protect the heart fromischemia and reperfusion by enhancing myocardial metabolic recovery. However,the stimulation of glycolysis during ischemia may be detrimental because of anaccumulation of metabolic end-products. We examined the effect of insulin onquiescent human ventricular cardiomyocytes subjected to simulated cardioplegicischemia and reperfusion.
Methods: Primarycardiomyocyte cultures were established from patients undergoing correctiverepair of tetralogy of Fallot. Cells were exposed to varying concentrations ofglucose and insulin during 30 minutes of stabilization in 10 mL ofphosphate-buffered saline solution. Ischemia was simulated by exposing the cellsto a low volume (1.5 mL) of deoxygenated phosphate-buffered saline solution for90 minutes followed by 30 minutes of simulated reperfusion in 10 mL of normoxicphosphate-buffered saline solution. Cell viability was assessed by trypan blueexclusion. The activity of mitochondrial pyruvate dehydrogenase was measured in3 states: stabilization, ischemia, and reperfusion. In addition intracellularlactate, adenine nucleotides, extracellular lactate, pyruvate, and acid releasewere measured.
Results: Higher ambientglucose concentrations resulted in greater cellular injury althoughinsulin-treated cells displayed less injury after ischemia and reperfusion.Insulin increased the pyruvate dehydrogenase activity by 31% incardiomyocytes and reduced extracellular lactate production by 40%.Intracellular adenosine triphosphate was improved by 75% in cells exposedto high glucose concentrations in the presence of insulin.
Conclusions: Insulin protected human ventricularcardiomyocytes from ischemia and reperfusion. This protection may be due to astimulation of pyruvate dehydrogenase activity which resulted in improvedaerobic metabolism.
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