Vasoconstriction triggered by hydrogen sulfide: Evidence for Na⁺,K⁺,2Cl^-cotransport and L-type Ca²⁺ channel-mediated pathway

Objectives: This study examined the dose-dependent actions of hydrogen sulfide donor sodium hydrosulphide (NaHS) on isometric contractions and ion transport in rat aorta smooth muscle cells (SMC).

Methods: Isometric contraction was measured in ring aortas segments from male Wistar rats. Activity of Na⁺/K⁺-pump and Na⁺,K⁺,2Cl^-cotransport was measured in cultured endothelial and smooth muscle cells from the rat aorta as ouabain-sensitive and ouabain-resistant, bumetanide-sensitive components of the ⁸⁶Rb influx, respectively.

Results: NaHS exhibited the bimodal action on contractions triggered by modest depolarization ([K⁺]_o=30 mM). At 10^-4 M, NaHS augmented contractions of intact and endothelium-denuded strips by ~ 15% and 25%, respectively, whereas at concentration of 10^-3 M it decreased contractile responses by more than two-fold. Contractions evoked by 10^-4 M NaHS were completely abolished by bumetanide, a potent inhibitor of Na⁺,K⁺,2Cl^-cotransport, whereas the inhibition seen at 10^-3 M NaHS was suppressed in the presence of K⁺ channel blocker TEA. In cultured SMC, 5×10^-5 M NaHS increased Na⁺,K⁺,2Cl^- - cotransport without any effect on the activity of this carrier in endothelial cells. In depolarized SMC, ⁴⁵Ca influx was enhanced in the presence of 10^-4 M NaHS and suppressed under elevation of [NaHS] up to 10^-3 M. ⁴⁵Ca influx triggered by 10^-4 M NaHS was abolished by bumetanide and L-type Ca²⁺ channel blocker nicardipine.

Conclusions: Our results strongly suggest that contractions of rat aortic rings triggered by low doses of NaHS are mediated by activation of Na⁺,K⁺,2Cl^-cotransport and Ca²⁺ influx via L-type channels.