Urine acetaminophen (APAP) and acetaminophen metabolite concentrations and APAP metabolizing enzymes expression in male C57BKS and db/db mice. A) Urinary levels of APAP and its conjugation metabolites glucuronide, sulfate, and N-acetyl cysteine levels in male C57BKS and db/db mice. Acetaminophen (150 mg/kg, po) was administered to C57BKS and db/db male mice (n = 5), mice were housed in metabolic cages and urine was collected for 24 hrs. Urine proteins were precipitated by methanol precipitation and the extracted samples analyzed by HPLC. Asterisks (*) represent a statistically significant concentration difference between C57BKS and db/db mice (p≤0.05). APAP-glucuronide (APAP-G), sulfate (APAP-S), and N-acetyl L-cysteine were detected in higher amounts in urine of db/dB mice as compared to C57BKS. B) Messenger RNA expression of Ugt1a6 and Sult1a1 in livers of male C57BKS and db/db mice. Total RNA was isolated from livers of adult db/db and C57BKS male mice, and mRNA expression was quantified using the branched DNA signal amplification assay. The data plotted as average RLU per 10 μg total RNA ± SEM. Asterisks (*) represent a statistically significant expression difference between db/db mice and C57BKS mice of the same gender (p≤0.05). Both Ugt1a6 and Sult1a1 mRNA expression was increased significantly in livers of male db/db mice as compared to C57BKS mice.