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Defective calcium handling in cardiomyocytes isolated from hearts subjected to ischemia-reperfusion

Institute of Cardiovascular Sciences, St. Boniface General Hospital Research Centre, and Department of Physiology, University of Manitoba, Winnipeg, Canada

Submitted 16 November 2004 ; accepted in final form 4 January 2005

Although ischemia-reperfusion (I/R) has been shown to affect subcellular organelles that regulate the intracellular Ca²⁺ concentration ([Ca²⁺]_i), very little information regarding the Ca²⁺ handling ability of cardiomyocytes obtained from I/R hearts is available. To investigate changes in [Ca²⁺]_i due to I/R, rat hearts in vitro were subjected to 10–30 min of ischemia followed by 5–30 min of reperfusion. Cardiomyocytes from these hearts were isolated and purified; [Ca²⁺]_i was measured by employing fura-2 microfluorometry. Reperfusion for 30 min of the 20-min ischemic hearts showed attenuated cardiac performance, whereas basal [Ca²⁺]_i as well as the 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 the KCl-induced increase in [Ca²⁺]_i or S(–)-BAY K 8644-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 H₂O₂ for 30 min. These results suggest that abnormalities in basal [Ca²⁺]_i as well as mobilization of [Ca²⁺]_i upon -adrenoceptor stimulation in cardiomyocytes are dependent on the duration of ischemic injury to the myocardium. Furthermore, Ca²⁺ handling defects in cardiomyocytes appear to be mediated through oxidative stress in I/R hearts.

Ca²⁺ mobilization; oxidative stress; ischemic preconditioning

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