Influence of alkaline buffers on cytoplasmic pH in myocardial cells exposed to metabolic acidosis.
The influence of different clinically used alkaline buffers on cytoplasmic pH in normal as well as acidotic rat myocardial cells was investigated in this study by means of the fluorescent intracellular probe 2',7'-bis-(carboxyethyl)-5,6-carboxyfluorescein acetoxymethyl ester (BCECF-AM). It was shown that both sodium bicarbonate and Tris buffer mixture (Tribonat) caused a significant and dose-dependent acidification of the cytoplasm of suspended myocardial cells with normal initial intracellular pH. This decrease was followed by a slow increase during the observation period. The initial cytoplasmic pH value was more easily reached when Tris buffer mixture was used. Ringer's acetate also caused a decrease of intracellular pH, but this change persisted and was further amplified during the experiment. Carbicarb in larger dosages as well as pure trometamol (Tris) caused a pronounced dose-dependent and lasting intracellular alkalinization. Intracellular acidosis was achieved by preincubating the cells in sodium acetate. Addition of sodium bicarbonate caused an initial and dose-dependent acidification of the cytoplasm followed by a slow increase to values slightly above the induced acidosis. In contrast, Tris buffer mixture showed a tendency towards an initial acidification only when larger dosages were used, and correction of the induced acidosis was possible by use of moderate to large volumes. Ringer's acetate produced a lasting and dose-dependent decrease of cytoplasmic pH, while Carbicarb and pure trometamol caused an immediate, pronounced and persistent alkalinization. Myocardial cells with low initial cytoplasmic pH due to preincubation in an acid buffer also showed an early decrease of intracellular pH after addition of sodium bicarbonate and Tris buffer mixture. In the case of sodium bicarbonate correction of the acid-base disturbance was not achieved during the observation period, while this was accomplished by use of larger volumes of Tris buffer mixture. Carbicarb in larger volumes caused an increase in intracellular pH. The most significant and persistent increases of cytoplasmic pH was achieved by use of pure trometamol. In conclusion, the present in vitro study implies that Tris buffer mixture (Tribonat) is well-suited for correction of intracellular acidosis since it acts without causing a pronounced initial intracellular acidosis or a later potentially hazardous huge cytoplasmic alkalinization.