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Am J Physiol Heart Circ Physiol (January 14, 2005). doi:10.1152/ajpheart.01153.2004
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Submitted on November 17, 2004
Accepted on January 4, 2005

Defective Calcium Handling in Cardiomyocytes Isolated from Hearts Subjected to Ischemia-Reperfusion

Harjot K Saini1 and Naranjan S Dhalla1*

1 Department of Physiology, Faculty of Medicine, Institute of Cardiovascular Sciences, St. Boniface General Hospital research Centre, University of Manitoba, Winnipeg, Manitoba, Canada

* To whom correspondence should be addressed. E-mail: nsdhalla{at}sbrc.ca.

Although ischemia-reperfusion (I/R) has been shown to affect subcellular organelles which regulate the intracellular concentration of Ca2+ ([Ca2+]i), very little information regarding Ca2+- handling ability of cardiomyocytes obtained from I/R hearts is available. In order to investigate changes in [Ca2+]i due to I/R, rat hearts in vitro were subjected to 10 to 30 min of ischemia followed by 5 to 30 min of reperfusion. Cardiomyocytes from these hearts were isolated and purified; the [Ca2+]i was measured by employing Fura-2 microfluorometry. Reperfusion for 30 min of the 10 and 20 min ischemic hearts showed attenuated cardiac performance whereas basal [Ca2+]i as well as KCl-induced increase in [Ca2+]i and isoproterenol (ISO)-induced increase in [Ca2+]i in cardiomyocytes remained unaltered. On the other hand, reperfusion of the 30 min ischemic hearts for different periods revealed marked changes in cardiac function, basal [Ca2+]i and ISO-induced increase in [Ca2+]i without any alterations in KCl-induced increase in [Ca2+]i or S(-)-Bay K8644-induced increase in [Ca2+]i. The I/R-induced alterations in cardiac function, basal [Ca2+]i and ISO-induced increase in [Ca2+]i in cardiomyocytes were attenuated by an antioxidant mixture containing superoxide dismutase and catalase as well as by ischemic preconditioning. The observed changes due to I/R were simulated in hearts perfused with hydrogen peroxide for 30 min. These results suggest that abnormalities in basal [Ca2+]i as well as mobilization of [Ca2+]i upon {beta}-adrenoceptor stimulation in cardiomyocytes are dependent upon duration of ischemic injury to the myocardium. Furthermore, Ca2+-handling defects in cardiomyocytes appear to be mediated through oxidative stress in the I/R hearts.




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