, Volume 91, Issue 5, pp 374-381

Loss of glycogen during preconditioning is not a prerequisite for protection of the rabbit heart

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Abstract

Depletion of gycogen has been proposed as the mechanism of protection from ischemic preconditioning. The hypothesis was tested by seeing whether pharmacological manipublation of preconditioning causes parallel changes in cardiac glycogen content. Five groups of isolated rabbit hearts were studied. Group 1 experienced 30 min of ischemia only. Group 2 (PC) was preconditioned with 5 min of global ischemia followed by 10 min of reperfusion. Group 3 was preconditioned with 5 min exposure to 400 nM bradykinin followed by a 10 min washout period. Group 4 experienced exposure to 10 μM adenosine followed by a 10 min washout period, and the fifth group was also preconditioned with 5 min ischemia and 10 min reperfusion but 100 μM8-(p-sulfophenyl) theophylline (SPT), which blocks adenosine receptors, was included in the buffer to block preconditioning's protection. Transmural biopsies were taken before treatment, just prior to the 30 min period of global ischemia, and after 30 min of global ischemia. Glycogen in the samples was digested with amyloglucosidase and the resulting glucose was assayed. Baseline glycogen averaged 17.3±0.6 μmol glucose/g wet weight. After preconditioning glycogen decreased to 13.3±1.3 μmol glucose/g wet weight (p<0.005 vs. baseline). Glycogen was similarly depleted after pharmacological preconditioning with adenosine (14.0±1.0 μmol glucose/g wet weight, p<0.05 vs. baseline) suggesting a correlation. However, when proconditioning was performed in the pressence of SPT, which blocks protection, glycogen was also depleted by the same amount (13.3±0.7 μmol glucose/g wet weight, p=ns vs. PC). Bradykinin, which also mimics preconditioning, caused no depletion of glycogen (16.3±0.8 μmol glucoseig wet weight, p=ns vs. baseline). Because preconditioning with bradykinin did not deplete glycogen and because glycogen continued to be low when protection from preconditioning was blocked with SPT, we conclude that loss of glycogen per se does not cause the protection of preconditioning.