Suppression of xenobiotic-metabolizing enzyme expression in rats by acriflavine, a protein kinase C inhibitor. Effects on epoxide hydrolase, glutathione S-transferases, and cytochromes p450.

摘要

The effects of acriflavine (ACF), a protein kinase C inhibitor, on the expression of hepatic microsomal epoxide hydrolase (mEH), glutathione S-transferases (GSTs), and cytochrome P450 (P450) were assessed in rat hepatic tissue. Northern blot analysis revealed that treatment of rats with thiazole, allyl disulfide (ADS), oltipraz, or clotrimazole at a single dose of 100 mg/kg resulted in 7-18-fold increases in mEH mRNA levels at 24 hr, whereas concomitant ACF treatment (20 mg/kg, im) caused 50-95% inhibition of the chemical-induced increases in hepatic mEH mRNA levels. rGSTA2, rGSTA3, and rGSTM1 mRNA levels were also significantly suppressed at 24 hr in response to a single dose of ACF (20 mg/kg, im). Animals treated with both ACF and ADS showed complete blockage of mEH and GST gene expression as early as 12 hr after treatment. ADS-inducible increases in mEH and rGSTA2 mRNA levels were suppressed at 24 hr after treatment with ACF, in a dose-related manner, with 50% inhibitory dose (ID50) values of 2.0-2.3 mg/kg, whereas glyceraldehyde-3-phosphate dehydrogenase mRNA levels were not altered. Immunoblot analysis revealed that ACF (15 mg/kg/day, im, for 3 days) inhibited induction of mEH or rGSTA2 protein by ADS (100 mg/kg/day, po, for 3 days). The levels of hepatic P450 2B1/2, P450 2C11, and P450 3A1/2 were decreased in rats treated with ACF (15 mg/kg/day, im, for 3 days), whereas P450 1A2 and P450 2E1 expression was not affected. Treatment of rats with ACF in combination with gadolinium chloride, which inhibits mEH and GST expression through calcium channel blocking, shifted the dose-inhibitory response curves for ACF to the left, with 7-15-fold decreases in the ID50 values, indicating that the active site for ACF for suppression of mEH and GST mRNA levels differs from that for gadolinium chloride. Proflavine and safranine O, which are structurally related to ACF, also caused suppression of ADS-induced increases in mRNA levels, in a dose-dependent manner, with ID50 values of 4-9 mg/kg. These results demonstrate that ACF and its related compounds effectively suppress the expression of a battery of hepatic xenobiotic-metabolizing enzymes, including mEH, GSTs, and certain P450 forms.