Mutagenicity and DNA adduct formation by aristolochic acid in the spleen of Big Blue® rats.

Abstract

Aristolochic acid (AA) is a potent human nephrotoxin and carcinogen. We previously reported that AA treatment resulted in DNA damage and mutation in the kidney and liver of rats. In this study, we have determined the DNA adducts and mutations induced by AA in rat spleen. Big Blue® transgenic rats were gavaged with 0, 0.1, 1.0, and 10.0 mg AA/kg body weight five-times/week for 3 months. Three DNA adducts, [7-(deoxyadenosin-N(6)-yl)-aristolactam I, 7-(deoxyadenosin-N(6)-yl)-aristolactam II and 7-(deoxyguanosin-N(2)-yl)-aristolactam I], were identified by (32)P-postlabeling. Over the dose range studied, there were strong linear dose-responses for AA-DNA adduct formation in the treated rat spleens, ranging from 4.6 to 217.6 adducts/10(8) nucleotides. Spleen cII mutant frequencies also increased in a dose-dependent manner, ranging from 32.7 to 286.2 × 10(-6) in the treated animals. Mutants isolated from the different treatment groups were sequenced; analysis of the resulting spectra indicated that there was a significant difference between the pattern of mutation in the 10 mg/kg AA-treated and the vehicle control rats. A:T → T:A transversion was the major type of mutation in AA-treated rats, whereas G:C → A:T transition was the main type of mutation in the vehicle controls. These results indicate that AA is genotoxic in the spleen of rats exposed under conditions that result in DNA adduct formation and mutation induction in kidney and liver.