© 1996 Oxford University Press
research-article |
Protein Arylation Precedes Acetaminophen Toxicity in a Dynamic Organ Slice Culture of Mouse Kidney
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*Department of Pharmaceutical Sciences, University of Connecticut Storrs, Connecticut 06269
Department of Molecular and Cell Biology, University of Connecticut Storrs, Connecticut 06269
Department of Pharmacology, University of Arizona Tucson, Arizona 85724
Department of Anesthesiology, University of Arizona Tucson, Arizona 85724
Received April 12, 1996; accepted August 7, 1996
Acetaminophen (APAP) is an analgesic and antipyretic agent which may cause hepatotoxicity and nephrotoxicity with overdose in man and laboratory animals. In vivo studies suggest that in situ activation of APAP contributes to the development of nephrotoxicity. Associated with target organ toxicity is selective arylation of proteins, with a 58-kDa acetaminophen binding protein (58-ABP) being the most prominent cytosolic target. In this study a mouse kidney slice model was developed to further evaluate the contribution of in situ activation of APAP to the development of nephrotoxicity and to determine the selectivity of protein arylation. Precision cut kidney slices from male CD-1 mice were incubated with selected concentrations of APAP (0–25 mM) for 2 to 24 hr. APAP caused a dose- and time-dependent decrease in nonprotein sulfhy-dryls (NPSH), ATP content, and K+ retention. Preceding toxicity was arylation of cytosolic proteins, the most prominent one being the 58-ABP. The association of 58-ABP arylation with APAP toxicity in this mouse kidney slice model is consistent with earlier, in vivo results and demonstrates the importance of in situ activation of APAP for the development of nephrotoxicity. Precision cut renal slices and dynamic organ culture are a good model for further mechanistic studies of APAP-induced renal toxicity.