Abstract
Intracellular catalase activity was measured in isolated rat hepatocytes by adding H 2O 2 under anaerobic conditions and measuring O 2 evolution. Hydrogen peroxide was introduced either by continuous infusion or by pulse injection. Continuous infusion at a rate similar to the endogenous H 2O 2 production rate provided results that 60–70% of the H 2O 2 was metabolized by the catalatic reaction. Comparison of rates of O 2 evolution to estimated rates of H 2O 2 metabolism obtained by the methanol-titration method ( H. Sies and B. Chance, 1970, FEBS Lett. 11, 172–176) indicated that the contribution of the peroxidatic reaction of catalase was small. The intracellular activity of glutathione peroxidase was estimated as the catalase-independent metabolism and used to determine the rate of intracellular H 2O 2 metabolism by the peroxidase. The results provide a quantitative basis for analysis of the physiological and toxicological aspects of H 2O 2 metabolism by liver.
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