Mechanisms of the antispasmodic activity of 3,5-di-t-butyl catechol (DTCAT) on rat vascular smooth muscles

In skeletal muscle sarcoplasmic reticulum vesicles, 3,5-di-t-butyl catechol (DTCAT) promotes the release of Ca2+ through the activation of ryanodine receptor Ca2+ channels. DTCAT mechanical and electrophysiological effects have now been investigated in rat aorta rings and single tail artery myocytes. Rat aorta rings incubated with either 30 μM ryanodine or 100 μM DTCAT developed tension, which averaged 36% and 7%, respectively, of that induced by phenylephrine. DTCAT reduced concentration-dependently both aorta ring contractions to high K+ (IC50 = 13.5 μM) and L-type Ba2+ current (IC50 = 22.0 μM) in isolated myocytes. Tetraethylammonium did not reverse the antispasmodic effect of DTCAT in rings stimulated with either 25 or 60 mM K+. DTCAT relaxed concentration-dependently phenylephrine-pre-contracted rings with intact endothelium (IC50 = 10.9 μM). This effect was markedly reduced by pre-incubation of rings with 100 μM Nω-nitro-l-arginine methyl ester. DTCAT antagonised phenylephrine-induced contractions in endothelium-deprived rings, either in the presence or in the absence of ryanodine (IC50 = 18.7 μM and 39.8 μM, respectively). Furthermore, both DTCAT (IC50 = 53.3 μM) and ryanodine reduced significantly the response to phenylephrine in the absence of extracellular Ca2+. Phenylephrine-stimulated influx of extracellular Ca2+ was markedly inhibited when tissues were pre-treated with DTCAT (IC50 = 19.0 μM) as well as nifedipine. DTCAT (> 100 μM) was also able to antagonise the contractions induced by phorbol 12-myristate, 13-acetate. In conclusion, this is the first demonstration that DTCAT inhibits vascular smooth muscle voltage-operated Ca2+ channels and promotes the release of endothelial nitric oxide. Ryanodine receptor Ca2+ channels activation or the impairment of the contractile apparatus by DTCAT seem to play a secondary role in its vascular activity.