Macrolide antibiotics inhibit the degradation of the glucocorticoid-responsive cytochrome P-450p in rat hepatocytes in vivo and in primary monolayer culture.

Abstract Treatment of rats with macrolide antibiotics such as triacetyloleandomycin (TAO) dramatically increases the hepatic concentration of a cytochrome P-450 indistinguishable from P-450p, the major liver cytochrome induced by glucocorticoids such as dexamethasone (Wrighton, S. A., Maurel, P., Schuetz, E. G., Watkins, P. B., Young, B., and Guzelian, P. S. (1985) Biochemistry 24, 2171-2178). To investigate the mechanism of induction of P-450p, we treated rats for 4 days with these agents and found that dexamethasone and TAO induced the synthesis of P-450p at least 70- and 35-fold over control values, respectively, as estimated from measurements of P-450p mRNA translatable in a cell-free system. However, the accumulation of P-450p holocytochrome (measured as TAO-metabolite spectral complex) or P-450p protein (measured by quantitative immunoblotting) increased at least 150-fold by TAO but only 32-fold by dexamethasone. The possibility that TAO decreases the degradation of P-450p was supported by the observation that administration of TAO to dexamethasone-treated rats labeled with NaH[14C]O3 and [3H]-delta-aminolevulinic acid retarded the decay of radioactive immunoprecipitable P-450p protein (t1/2 = 60 versus 14 h) and heme (t1/2 = 73 versus 10 h). To confirm these results, P-450p protein synthesis was measured as radioactivity incorporated into immunoprecipitable P-450p in primary monolayer cultures of adult rat hepatocytes incubated with [3H]leucine. Dexamethasone treatment of the cultures stimulated P-450p synthesis by at least 30-fold whereas macrolides were without effect. However, macrolide antibiotics but not dexamethasone inhibited the disappearance of radiolabeled P-450p from cultured hepatocytes similar to the results obtained in vivo. We conclude that macrolide antibiotics induce P-450p, the most rapidly turning over cytochrome yet reported, by stimulating its synthesis indirectly and by blocking its degradation, significantly.