Influence of alkaline buffers on cytoplasmic pH in myocardial cells exposed to hypoxia

Abstract

The fluorescent intracellular probe 2′,7′-bis-(carboxyethyl)-5,6-carboxyfluorescein acetoxymethyl ester was used in this experimental study to investigate the effects of different alkaline buffers on cytoplasmic pH in suspended myocardial cells under normal as well as hypoxic conditions. A dose-dependent intracellular acidification was achieved after addition of sodium bicarbonate or Tris buffer mixture (Tribonat®) to the myocardial cells under normal conditions. After this immediate decrease in cytoplasmic pH, a tendency for the pH to rise again was recorded during the observation period, but this elevation of pH occurred to variable degrees with the different agents and dosages. Addition of larger volumes of Tribonat® caused the cytoplasmic pH to return to the initial value during the observation time. Addition of Ringer's acetate produced a significant and persistent cytoplasmic acidification. Larger volumes of Carbicarb as well as pure trometamol (Tris) caused a lasting intracellular alkalinization. Hypoxia per se caused a marked intracellular acidosis in the cardiomyocytes. During hypoxia, addition of sodium bicarbonate caused a further decrease of cytoplasmic pH, turning into an increase during the observation period. Also, Tribonat® caused an immediate further acidification, but 15 min after the addition the intracellular pH-value had reached the normal level of normoxic cells. Addition of Ringer's acetate caused a further significant and lasting decrease of intracellular pH. The effect of Carbicarb was a persistent alkalinization of the cell interior. Trometamol produced the most pronounced rise of cytoplasmic pH. In conclusion, this in vitro study shows that Tris buffer mixture (Tribonat®) possesses important qualities for correction of metabolic acidosis due to hypoxia and may perhaps be preferred over other alkaline buffers in some situations.