Sex-Related Differences in Mouse Renal Metabolism and Toxicity of Acetaminophen

Abstract

The objective of this study is to elucidate the role of cytochrome P450 2E1 in the metabolic activation of acetaminophen (APAP) in mouse kidneys. With the kidney microsomes from C3H/HeJ mice, a significant sex-related difference was observed in the NADPH-dependent formation of a reactive APAP metabolite which was trapped as a glutathione conjugate. The enzyme activity in male mouse kidney was about 35- and 50-fold higher than that in the females at substrate concentrations 50 and 500 μM, respectively, and treatment of female mice with testosterone significantly induced the enzyme activity in the mouse kidney. No sex-related difference in this enzyme activity, however, was observed in the livers. The oxidative metabolism of APAP in control male mouse kidney microsomes displayed an apparent low Km of 43-45 μM and an apparent high Km of 603-702 μM. Studies using monoclonal antibodies against P450 2E1 showed that P450 2E1 accounted for about half of the activity in the metabolic activation of APAP in both mouse liver and kidney. Furthermore, there was a strong correlation between the renal P450 2E1 content (measured by Western blot analysis) and the rate of APAP oxidation by renal microsomes. To test the hypothesis that the susceptibility of renal tissue to APAP toxicity is determined by the distribution and level of P450 2E1, toxicity studies were conducted. A time-dependent depletion of hepatic and renal cortical glutathione was observed in both male and female mice following a dose of 1200 mg/kg APAP. At 4 hr after APAP dosing, the level of renal cortical glutathione depletion in male mice was significantly greater than that in the females. The level of blood creatinine elevation in male mice was higher than that in the females 8 hr after APAP dosing. Histopathology studies by light and electron microscopic assessments demonstrated that renal damage by APAP was restricted mostly to the epithelial cells of the proximal convoluted tubules where P450 2E1 was localized. The renal proximal tubular necrosis induced by APAP was more severe in males than that in the females. Results from this study suggest that P450 2E1 plays an important role in the metabolic activation of APAP and is a key factor in determining the sex-related difference of APAP-mediated toxicity in the mouse kidney.