Abstract

Glutathione (GSH) conjugation of trichloroethylene (Tri) to formS-(1,2-dichlorovinyl)glutathione (DCVG) has been implicated in the nephrotoxicity and nephrocarcinogenicity of Tri. Marked sex- and species-dependent differences exist, however, in the susceptibility to Tri-induced renal toxicity, with the male rat being the most susceptible. The present study, therefore, focuses on potential differences in the initial step of the GSH pathway. Rates of DCVG formation were measured in suspensions of isolated renal cortical cells and isolated hepatocytes from male and female Fischer 344 rats and in kidney and liver microsomes and cytosol from male and female Fischer 344 rats and B6C3F1 mice to determine if sex- and species-dependent differences in GSH conjugation correlate with susceptibility to renal toxicity from Tri. Rates of γ-glutamyltransferase (GGT) with γ-glutamyl-p-nitroanilide and glycylglycine as substrates and GSH S-transferase (GST) with 1-chloro-2,4-dinitrobenzene as substrate were also measured in liver and kidney subcellular fractions to provide further information on the biochemical basis of susceptibility to Tri. Rates of DCVG formation in rat kidney cells and kidney subcellular fractions were 5- to 20-fold lower than those in rat hepatocytes and liver subcellular fractions. Rates of DCVG formation in kidney cells and subcellular fractions were comparable in male and female rats with the exception of male rat kidney microsomes, where DCVG formation was below the limit of detection, and those in liver cells and subcellular fractions were >3-fold higher in male rats than in female rats. Rates of DCVG formation in mouse kidney subcellular fractions were approximately 10-fold higher than in corresponding fractions from the rat, whereas those in mouse liver subcellular fractions were 4- to 8-fold higher than in corresponding rat tissues, with rates in male mouse liver cytosol and microsomes being modestly higher than in corresponding fractions from female mice. GGT activity was barely detectable in livers, was about 20-fold higher in rat kidneys than in mouse kidneys, and was slightly higher in female rat kidneys than in male rat kidneys. GST activity with 1-chloro-2,4-dinitrobenzene as substrate exhibited tissue-, sex-, and species-dependent patterns that were generally similar to those with Tri as the substrate. These results suggest that the higher susceptibility to Tri-induced renal toxicity of male rats as compared with female rats correlates with rates of DCVG formation. The high rates of DCVG formation in mice, however, indicate that other factors, possibly including differences in activities of cysteine conjugate β-lyase or N-acetyltransferase, may also be important determinants of the susceptibility to Tri.