Mechanism of nitric oxide-induced vasodilatation - Refilling of intracellular stores by sarcoplasmic reticulum Ca2+ ATPase and inhibition of store-operated Ca2+ influx

The precise mechanisms by which nitric oxide (NO) decreases free [Ca2+](i), inhibits Ca2+ influx, and relaxes vascular smooth muscle are poorly unders tood. In rabbit and mouse aorta, agonist-induced contractions and increases in [Ca2+](i) were resistant to nifedipine, suggesting Ca2+ entry through n on-L type Ca2+ channels. Relaxations to NO were inhibited by thapsigargin ( TG) or cyclopiazonic acid (CPA) indicating the involvement of sarcoplasmic reticulum ATPase (SERCA). Studies of the effect of NO on [Ca2+](i) and the rate of Mn2+ influx with fura-2 fluorometry in rabbit aortic smooth muscle cells in primary culture were designed to test how SERCA is involved in med iating the response to NO. When cells were stimulated with angiotensin II ( AII), NO accelerated the removal of Ca2+ from the cytoplasm, decreased [Ca2 +](i), and inhibited Ca2+ and Mn2+ influx. Inhibition of SERCA abolished al l the effects of NO. In contrast, inhibition of the Na+/Ca2+ exchanger or t he plasma membrane Ca2+ ATPase had no influence on the ability of NO to dec rease [Ca2+](i). NO maximally decreased [Ca2+](i) within 5 s, whereas signi ficant inhibition of All-induced Ca2+ and Mn2+ influx required more than 15 s. The inhibition of cation influx strictly depended on [Ca2+](0) and func tional SERCA, suggesting that during the delay before NO inhibits Ca2+ infl ux, the influx of Ca2+ and the uptake into intracellular stores are require d. In the absence of [Ca2+](0), NO diminished the All-induced [Ca2+](i) tra nsient by a SERCA-dependent mechanism and increased the amount of Ca2+ in t he stores subsequently released by ionomycin. The present study indicates t hat the initial rapid decrease in [Ca2+](i) caused by NO in vascular smooth muscle is accounted for by the uptake of Ca2+ by SERCA into intracellular stores. It is proposed that the refilling of the stores inhibits store-oper ated Ca2+ influx through non-L-type Ca2+ conducting ion channels and that t his maintains the decrease in [Ca2+](i) and NO-induced relaxation.