Increased activity of H 2O 2 in aorta isolated from chronically streptozotocin-diabetic rats: effects of antioxidant enzymes and enzyme inhibitors

Abstract

The effects of hydrogen peroxide (H 2O 2, 1 nM–5 mM) on the tone of the rings of aorta precontracted with phenylephrine (PE) were studied in 4–5 months streptozotocin (STZ)-diabetic rats and their age-matched controls. H 2O 2 induced brief contraction before relaxation in endothelium-containing rings that was more pronounced in diabetic rats. Removal of the endothelium or pretreatment of rings with N G-nitro- l-arginine methyl ester ( l-NAME, 100 μM) abolished H 2O 2-induced immediate and transient increase in tone, but preincubation with indomethacin (10 μM) had no effect on contractions induced by H 2O 2 in both group of animals. Pretreatment with l-NAME or indomethacin as well as absence of endothelium produced an inhibition of H 2O 2-induced relaxation that was more pronounced in diabetic rings. Chronically STZ-diabetes resulted in a significant increase in H 2O 2-induced maximum relaxation that was largely endothelium-dependent. Decreased sensitivity (pD 2) of diabetic vessels to vasorelaxant action of H 2O 2 was normalized by superoxide dismutase (SOD, 80 U/ml). Pretreatment with SOD had no effect on H 2O 2-induced maximum relaxations in both group of animals but led to an increase in H 2O 2-induced contractions in control rats. When the rings pretreated with diethyldithiocarbamate (DETCA, 5 mM), H 2O 2 produced only contraction in control rats, and H 2O 2-induced relaxations were markedly depressed in diabetic rats. H 2O 2 did not affect the tone of intact or endothelium-denuded rings in the presence of catalase (2000 U/ml). Aminotriazole (AT, 10 mM) failed to affect H 2O 2-induced contractions or relaxations in all rings. Our observations suggest that increased production of oxygen-derived free radicals (OFRs) in diabetic state leads to a decrease in SOD activity resulting an increase in endogenous superoxide anions (O 2 •−), that is limited cytotoxic actions, and an increase in catalase activity resulting a decrease in both H 2O 2 concentrations and the production of harmful hydroxyl radical ( OH) in diabetic aorta in long-term. Present results indicate that increased vascular activity of H 2O 2 may be an important factor in the development of vascular disorders associated with chronically diabetes mellitus. Enhanced formation of OH, that is a product of exogenous H 2O 2 and excess O 2 •−, seems to be contribute to increased relaxations to exogenously added H 2O 2 in chronically diabetic vessels.