Protection of rat hepatocytes from tert-butyl hydroperoxide-induced injury by catechol.
Metabolism of tert-butyl hydroperoxide (TBHP, 2.0 mM) by glutathione peroxidase within isolated rat hepatocytes caused a rapid oxidation of intracellular reduced glutathione and ultimately NADPH through glutathione reductase. TBHP also caused the formation of surface blebs in the hepatocyte plasma membrane followed by the leakage of cytosolic enzymes, such as lactate dehydrogenase, into the incubation medium. Catechol (0.1 mM) protected hepatocytes from the cytotoxic effects of TBHP but did not prevent the rapid oxidation of glutathione indicating normal metabolism of TBHP through glutathione reductase. In contrast, addition of catechol to the hepatocyte incubations prevented TBHP-induced depletion of intracellular NADPH and increased the total NADP+ + NADPH concentration without altering significantly the intracellular NADP+ content or the NADPH/NADP + NADPH ratio. Catechol did not alter TBHP stimulation of the pentose phosphate pathway. Hepatocytes incubated with sublethal concentrations of TBHP (1.0 mM) did not leak lactate dehydrogenase into the medium but did lose intracellular potassium. In these experiments, TBHP caused a sustained increase in phosphorylase alpha activity suggesting that TBHP metabolism may be associated with a sustained increase in cytosolic free Ca2+. In the presence of catechol, phosphorylase alpha activity was increased by 5 min but returned toward control by 20 min. These data suggest that catechol may be protecting hepatocytes from TBHP-induced injury by preventing a sustained rise in cytosolic free Ca2+ concentration.
D000818:Animals; D002396:Catechols / Q000494:pharmacology*; D002470:Cell Survival; D005947:Glucose / Q000378:metabolism; D066298:In Vitro Techniques; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction; D010545:Peroxides / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000633:toxicity; D010762:Phosphorylase a / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D020122:tert-Butylhydroperoxide
D002396:Catechols; D010545:Peroxides; D009249:NADP; D020122:tert-Butylhydroperoxide; D010762:Phosphorylase a; D005947:Glucose; C034221:catechol