Heme oxygenase induction by CoCl 2, Co-protoporphyrin IX, phenylhydrazine, and diamide: Evidence for oxidative stress involvement

Abstract

The induction of heme oxygenase in rat liver by cobaltous chloride (CoCl 2) and Co-protoporphyrin IX is entirely prevented by the administration of α-tocopherol and allopurinol. CoCl 2 was converted in the liver into Co-protoporphyrin IX before it induced heme oxygenase activity. Actinomycin and cycloheximide affected to a similar degree the induction of heme oxygenase by both CoCl 2 and Co-protoporphyrin IX. Administration of either CoCl 2 or Co-protoporphyrin strongly decreased the intrahepatic GSH pool, a decrease which was completely prevented by the administration of either α-tocopherol or allopurinol. The latter compounds prevented heme oxygenase induction as well as the decrease in hepatic GSH when administered 2 h before, together with, or 2 h after CoCl 2. However, when given 5 h after administration of CoCl 2, α-tocopherol and allopurinol showed no preventive effect. Similar results were obtained when Co-protoporphyrin IX was used, with the difference that when α-tocopherol and allopurinol were given 2 h after administration of the inducer, they showed no protective effect. Phenylhydrazine and diamide also induced heme oxygenase activity in rat liver. This inductive effect was preceded by a decrease in the intrahepatic GSH pool, which took place several hours before induction of the oxygenase. Administration of α-tocopherol and allopurinol prevented induction of the oxygenase but had no effect on the decrease in GSH levels. These results suggest that the induction of heme oxygenase by phenylhydrazine and the diamide is preceded by an oxidative stress which very likely originates in the depletion of GSH. The induction of heme oxygenase by hemin was not prevented by administration of α-tocopherol or allopurinol. Co-protoporphyrin IX did not affect the pattern of the molecular forms of hepatic biliverdin reductase, at variance with CoCl 2, which is known to convert molecular form 1 of the enzyme into molecular form 3.