Hydrogen sulfide-induced vasodilation mediated by endothelial TRPV4 channels

Hydrogen sulfide (H₂S) is a recently described gaseous vasodilator produced within the vasculature by the enzymes cystathionine γ-lyase and 3-mercaptopyruvate sulfurtransferase. Previous data demonstrate that endothelial cells (EC) are the source of endogenous H₂S production and are required for H₂S-induced dilation. However, the signal transduction pathway activated by H₂S within EC has not been elucidated. TRPV4 and large-conductance Ca²⁺-activated K channels (BK channels) are expressed in EC. H₂S-induced dilation is inhibited by luminal administration of iberiotoxin and disruption of the endothelium. Calcium influx through TRPV4 may activate these endothelial BK channels (eBK). We hypothesized that H₂S-mediated vasodilation involves activation of TRPV4 within the endothelium. In pressurized, phenylephrine-constricted mesenteric arteries, H₂S elicited a dose-dependent vasodilation blocked by inhibition of TRPV4 channels (GSK2193874A, 300 nM). H₂S (1 μM) increased TRPV4-dependent (1.8-fold) localized calcium events in EC of pressurized arteries loaded with fluo-4 and Oregon Green. In pressurized EC tubes, H₂S (1 μM) and the TRPV4 activator, GSK101679A (30 nM), increased calcium events 1.8- and 1.5-fold, respectively. H₂S-induced an iberiotoxin-sensitive outward current measured using whole cell patch-clamp techniques in freshly dispersed EC. H₂S increased K⁺ currents from 10 to 30 pA/pF at +150 mV. Treatment with Na₂S increased the level of sulfhydration of TRPV4 channels in aortic ECs. These results demonstrate that H₂S-mediated vasodilation involves activation of TRPV4-dependent Ca²⁺ influx and BK channel activation within EC. Activation of TRPV4 channels appears to cause calcium events that result in the opening of eBK channels, endothelial hyperpolarization, and subsequent vasodilation.