Abstract
Background
In the vascular system, ATP-sensitive K+-channels are a target for H2 S. Recent evidence suggests that H2 S may also modulate Na+ — and Ca2+-permeable channels and intracellular Ca2+ stores, but the influence of H2S on endothelial Ca2+ dynamics and Ca2+-dependent activation of endothelial nitric oxide synthase (eNOS) is unclear. In this study, we investigated the effects of H2 S on Ca2+ signaling in endothelial and smooth muscle cells with special emphasis given to the role of H2 S in modulating endothelial NO formation.
Methods
Experiments were performed with endothelial cells from porcine aorta, the human endothelial cell line HMEC-1, and smooth muscle cells from rat aorta and trachea. Mobilization of intracellular Ca2+ and Ca2+ entry was monitored with Fura-2. Activity of eNOS was determined as conversion of incorporated l-[3H]arginine into l-[3H]citrulline.
Results
Incubation of endothelial cells with the H2 S donors sodium hydrogen sulfide (NaHS) and GYY4137 blocked activation of eNOS by the receptor agonist ATP but not by the Ca2+ ionophore A23187. Data revealed that H2 S inhibited ATP-induced release of Ca2+ from intracellular stores indicating that H2 S attenuates eNOS activity by blocking capacitative Ca2+ entry. A similar inhibitory effect of H2 S on ATP-induced Ca2+ release and Ca2+ entry was also observed in human microvascular endothelial cells and smooth muscle cells.
Conclusions
H2 S antagonized Ca2+ mobilization by receptor agonists and store-operated Ca2+ entry thereby limiting eNOS activation and NO formation. The effect of H2 S on Ca2+ stores was not restricted to endothelial cells but was also observed in vascular and tracheal smooth muscle cells.
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Abbreviations
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- eNOSk:
-
endothelial nitric oxide synthase
- IP3:
-
inositol (1,4,5)-trisphosphate
- H2S:
-
hydrogen sulfide
- HUVECs:
-
human umbilical vein endothelial cells
- NaHS:
-
sodium hydrogen sulfide
- PAECs:
-
porcine aortic endothelial cells
- SMCs:
-
smooth muscle cells
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Kloesch, B., Steiner, G., Mayer, B. et al. Hydrogen sulfide inhibits endothelial nitric oxide formation and receptor ligand-mediated Ca2+ release in endothelial and smooth muscle cells. Pharmacol. Rep 68, 37–43 (2016). https://doi.org/10.1016/j.pharep.2015.05.026
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DOI: https://doi.org/10.1016/j.pharep.2015.05.026