Expression of glutathione S-transferases Ya, Yb1, Yb2, Yc1 and Yc2 and microsomal epoxide hydrolase genes by thiazole, benzotbiazole and benzothiadiazole

Abstract

The effects of thiazole (TH), benzothiazole (BT) and benzothiadiazole (BZ) on the expression of hepatic glutathione S-transferases (GSTs) Ya, Yb1, Yb2, Yc1 and Yc2 and microsomal epoxide hydrolase (mEH) genes were compared in rats. TH treatment resulted in 4- to 24-fold increases in GST Ya mRNA levels at 24 hr posttreatment; the ED 50 value was 70 mg/kg. GST Ya mRNA levels were elevated 13-, 20-, 20- and 9-fold at 12, 24, 48 and 72 hr following 100 mg/kg of TH treatment, respectively, as compared with the control. BT was a moderate inducer of GST Ya with a maximal 18-fold increase observed, whereas BZ treatment caused a transient increase in GST Ya mRNA at 12 hr posttreatment, followed by a return to a 4-fold relative increase at 24 hr or afterward. Treatment of rats with TH at the dose of 100 mg/kg resulted in an ∼10-fold increase in either Yb1 or Yb2 mRNA levels at 24 hr posttreatment. BT-treated rats showed 7- and 3-fold increases in the GST subunit Yb1 and Yb2 mRNA levels at 24 hr posttreatment. BZ was the least effective in modulating either GST Ybl or Yb2 mRNA, resulting in < 2-fold changes. GST Yc1 and Yc2 mRNA levels were increased ∼8-fold at the dose of 200 mg/kg of TH. BT minimally affected GST subunit Yc1 and Yc2 mRNA levels, with a maximal 4-fold relative increase observed. BZ was the least effective in enhancing Yc1 and Yc2 mRNA levels. Protein levels for GST subunit Ya, Yb1, Yb2 and Yc were also elevated in response to TH by 3-, 2-, 2- and 2-fold, respectively. Thus, TH was effective in modulating both constitutive and inducible GST gene expression. BT or BZ was much less effective in increasing the expression of GST subunits. These RNA and Western blot analyses revealed that the levels of major GST were differentially increased after treatment with these thiazoles, exhibiting a rank order of GST expression of TH > BT > BZ. mEH expression by these compounds appeared to be consistent with that of GST Ya. The mRNA levels for GST Ya, Yb1, Yb2, Yc1 and Yc2 and mEH were also determined after treatment with triazole (TR), imidazole (IM), benzoxazole (BX), benzotriazole (BTR) or benzimidazole (BIM). TR, IM, BX or BTR caused increases in Ya, Yb1, Yc1 and Yc2 mRNA levels by 2- to 3-fold, whereas the agents failed to modulate the expression of GST Yb2. The fact that benzene, cyclohexane or n-hexane minimally affected the major GST or mEH mRNA levels provided evidence that certain heterocyclic compounds are more capable of modulating GST or mEH gene expression than hydrocarbons. These results corroborate evidence that the thiazoles differentially stimulate GST or mEH genes, with TH being the most efficacious; that thiazoles with carbocyclic ring are much less effective in increasing GST or mEH levels than is TH; and that the changes in these GST and mEH levels are primarily associated with increases in mRNA levels.