Aristolochic acid nephropathy induced by aristolochic acid type I in C57BL/6 mice

Aristolochic acid (AA) is a nephrotoxic carcinogen responsible for acute kidney injury, chronic renal failure, and associated urothelial cancers. This study aims to determine the genes in xenobiotic metabolism pathway regulated by AA and clarify the molecular mechanism underlying their action. Six-week-old female and male C57BL/6 mice were used. AAI (Sigma-Aldrich, St. Louis, MO, USA) was dissolved by corn oil and administered by i.p. injection at a dose of 2 mg/kg/day (male) or 5 mg/kg/day (female), 5 days per week for 3 weeks. Total liver RNA was isolated from four groups of animals, AAI- or corn oil-treated female and male WT mice, using RNeasy Mini Kits (Qiagen Inc., Valencia, CA, USA). In each group, pooled RNA was prepared by mixing the same amount total RNA from 5 mice. The microarray hybridization was performed by Welgen Biotech, Co., Ltd. (Taipei, Taiwan) using Agilent SurePrint Microarray (Agilent Technologies, USA). After microarrays were scanned with an Agilent microarray scanner (Agilent Technologies, USA) for fluorescent signal intensities, scanned images were analyzed by Feature extraction10.5.1.1 software (Agilent Technologies, USA). Raw signal data was normalized by quantile normalization for differentially expressed gene discovery.