Importance of bicarbonate transport for protection of cardiomyocytes against reoxygenation injury

Abstract

Isolated cardiomyocytes from adult rats were incubated in anoxic bicarbonate-buffered media at extracellular pH (pHo) 6.4 until a cytosolic Ca2+ overload and intracellular pH (pHi) of 6.4 were reached. On reoxygenation, the pH of the medium was changed to 7.4 to activate the Na+/H+exchanger (NHE) and the Na+-[Formula: see text] symporter (NBS). The reoxygenation was performed in the absence or presence of the NHE inhibitor HOE-642 (3 μmol/l) and/or the NBS inhibitor DIDS (0.5 mmol/l ), as in bicarbonate-free media. In reoxygenated control cells pHi rapidly recovered to the preanoxic level, and a burst of spontaneous oscillations of cytosolic Ca2+ occurred, accompanied by the development of hypercontracture. When NBS and NHE were simultaneously inhibited during reoxygenation, pHi recovery was prevented, Ca2+oscillations were attenuated, and hypercontracture was abolished. Sole inhibition of NBS or NHE showed no protection against hypercontracture. In the absence of cytosolic acidosis, HOE-642 or DIDS did not prevent hypercontracture induced by Ca2+ overload. The results demonstrate that simultaneous inhibition of NHE and NBS is needed to protect myocardial cells against reoxygenation-induced hypercontracture.