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 Ca(2+) through the activation of ryanodine receptor Ca(2+) 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 microM ryanodine or 100 microM 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(+) (IC(50)=13.5 microM) and L-type Ba(2+) current (IC(50)=22.0 microM) 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 (IC(50)=10.9 microM). This effect was markedly reduced by pre-incubation of rings with 100 microM Nomega-nitro-l-arginine methyl ester. DTCAT antagonised phenylephrine-induced contractions in endothelium-deprived rings, either in the presence or in the absence of ryanodine (IC(50)=18.7 microM and 39.8 microM, respectively). Furthermore, both DTCAT (IC(50)=53.3 microM) and ryanodine reduced significantly the response to phenylephrine in the absence of extracellular Ca(2+). Phenylephrine-stimulated influx of extracellular Ca(2+) was markedly inhibited when tissues were pre-treated with DTCAT (IC(50)=19.0 microM) as well as nifedipine. DTCAT (>100 microM) 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 Ca(2+) channels and promotes the release of endothelial nitric oxide. Ryanodine receptor Ca(2+) channels activation or the impairment of the contractile apparatus by DTCAT seem to play a secondary role in its vascular activity.
Fusi
Fabio
F
Dipartimento di Scienze Biomediche, Università degli Studi di Siena, via A. Moro 2, 53100 Siena, Italy. usif@unisi.it
Ferrara
Antonella
A
Sgaragli
Giampietro
G
Saponara
Simona
S
D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000187:drug effects* / Q000378:metabolism; D002396:Catechols / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects*; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012433:Ryanodine / Q000494:pharmacology; D019837:Ryanodine Receptor Calcium Release Channel / Q000187:drug effects / Q000378:metabolism; D013035:Spasm / Q000188:drug therapy
D015220:Calcium Channels; D002396:Catechols; D019837:Ryanodine Receptor Calcium Release Channel; C054068:3,5-di-tert-butylcatechol; D012433:Ryanodine; D009569:Nitric Oxide; D002118:Calcium