17β-OESTRADIOL ENHANCES NITRIC OXIDE SYNTHASE ACTIVITY IN ENDOTHELIUM-DENUDED RAT AORTA

1. It has been suggested that oestrogen-produced vasodilatation is due to induction of endothelial nitric oxide synthase (NOS), but there are many reports of direct effects on vascular smooth muscle. In the present study, these processes were investigated in rat aorta isolated from ovariectomized rats. 2. Short-term treatment (10min) with 17β-oestradiol (10 μmol/L) produced a small attenuation of the phenylephrine (PE)-induced constriction, which was unaffected by the nitric oxide synthase inhibitor l-N 5(-1-iminoethyl)ornithine (NIO; 100μmol/L). Long-term treatment (6h) with 17β-oestradiol (10 μmol/L) did not affect acetylcholine-mediated vasorelaxation in endothelium-intact aortic rings, but did attenuate PE-induced constriction. This attenuation was also observed in endotheliumdenuded preparations after 17β-oestradiol (10 μmol/L for 6h) and was far greater than the acute effect of 17β-oestradiol (10 μmol/L). 3. The attenuation produced by 17β-oestradiol (10 μmol/L for 6 h) was significantly inhibited by concomitant treatment with cycloheximide (1 μmol/L), suggesting that protein synthesis was involved. NIO (100 μmol/L) also attenuated the effect, which suggests that the anti-constrictor effect of 17β-oestradiol occurs through the increased production of nitric oxide (NO). 17β-Oestradiol increased NO production, as assessed by the conversion of [³H]-arginine to [³H]-citrulline in rat aorta. These effects were prevented by cycloheximide and NIO. The anti-constrictor effect of oestrogen was blocked by the oestrogen receptor antagonist ICI 182780 (100nmol/L). 4. Western blotting using an antibody specific for inducible nitric oxide synthase (NOS) revealed that 17β-oestradiol (10 μmol/L for 24 h) treatment induced the formation of inducible NOS protein in the aorta, an effect blocked by cycloheximide. The results indicate that 17β-oestradiol can attenuate the vasoconstrictor effect of PE by a specific receptor-mediated process that involves induction of inducible NOS.