Author(s): Link EM
The present paper is concerned with the influence of hydrogen ion concentration and composition of the medium on clonogenic survival of epithelial cells exposed to hydrogen peroxide in vitro. The survival of cells incubated with H2O2 in phosphate-buffered saline at pH 6.5 was 1 x 10(-2) and increased abruptly to 9 x 10(-2) at pH 7.0. The pH dependence of the cytocidal effect was particularly conspicuous when Eagle's minimum essential medium (SFMEM) was used for cell exposure to H2O2: the survival was characterized by exponential pH dependence and varied from 1 x 10(-1) to 9 x 10(-1) for pH 6.5 and 7.5, respectively, with a superimposed sharp peak value of 9 x 10(-1) at pH 7.0. The enhanced pH dependence of the H2O2 cytotoxicity in SFMEM was found to result from the additive action of glucose and histidine present in this medium. Glucose alone protected the cells with the efficiency decreasing with increasing hydrogen ion concentration. Histidine was responsible for the intermediate maximum in the pH-dependent survival spectrum. In addition, the changes in cell survival were accompanied by pH-dependent release of GSSG from the exposed cells. The GSSG efflux was inhibited by glucose in the medium. The influence of glucose on both the pattern of cell survival and the associated GSSG release indicate that the glutathione peroxidase activity supported by the pentose phosphate pathway is crucial in cell protection against extracellular H2O2 toxicity.