The protective effects of berberine and phentolamine against anoxia and reoxygenation damage in isolated rat hearts have been investigated. Incorporation of berberine (24.5 mumol/L) in both anoxic and aerobic perfusion media resulted in a significant reduction of CPK release during the reoxygenation period, and the ultrastructural damage was reduced as compared with the control group; the myocytes in the berberine-treated group displayed mild intracellular edema, well-registered myofibrils without contracted bands, and swollen mitochondria with partially broken cristae but without dense bodies. Berberine did not inhibit calcium and sodium accumulation or magnesium and potassium loss. Treatment with phentolamine (6.6 mumol/L), an alpha-adrenoceptor antagonist, had similar effects, though the CPK release profile was shifted to the right and downwards. These results suggest that although berberine and phentolamine have some beneficial effects on myocardial reoxygenation injury, they may not abolish the injury. Therefore alpha-adrenoceptor stimulation may not be the major mechanism behind the injury.