Abstract
This study aimed to elucidate the molecular mechanism of H2S-induced vasorelaxation. Vasorelaxation responses to the H2S donor NaHS and the H2S precursor l-cysteine were examined by measuring isometric tone of mouse aortic rings in a small vessel myograph. H2S concentrations in Krebs' solution were determined with a polarographic sensor. H2S expression was examined by Western blot, and H2S production from CSE was assayed using a spectroscopic method. In pre-constricted mouse aorta, NaHS (1 μM–3 mM) elicited vasorelaxation of 95 ± 7%, EC50 189 ± 69 μM. This response was unaffected by removal of the endothelium. Maximum vasorelaxation was significantly attenuated by global blockade of K+ channels (50 mM K+) and the KATP channel blocker glibenclamide (10 μM) alone (P < 0.01, ANOVA). Specific inhibition of KCa, KIR, or KV channels elicited a significant shift to the right in the concentration–response curve to NaHS (P < 0.01, ANOVA) without affecting maximum relaxation. NaHS-mediated vasorelaxation was inhibited by the Cl− channel inhibitor DIDS (1 mM, P < 0.05, t test), and NaHS caused a significant concentration-dependent inhibition of voltage-gated Ca2+ channels (P < 0.001, two-way ANOVA). The H2S-producing enzyme cystathionine-γ-lyase (CSE) was expressed in mouse aorta and had activity of 7 ± 3 μmol H2S/g/min. l-cysteine (1 μM–3 mM) elicited a CSE-dependent vasorelaxation of mouse aorta with intact endothelium (20 ± 7%), but not when the endothelium was removed. CSE inhibitors dl-propargylglycine (20 mM) and β-cyanoalanine (1 mM) caused concentration-dependent contraction of mouse aorta. In mouse aorta, H2S elicits endothelium-independent vasorelaxation involving several different ion channels and seems to converge at the vascular smooth muscle cell voltage-gated Ca2+ channel. The l-cysteine-CSE-H2S pathway contributes to vasorelaxation and appears to modulate basal vessel tone.
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Acknowledgements
We gratefully acknowledge the use of the facilities of the Molecular Biology Laboratories (Monash University, Department of Pharmacology) and the generous assistance of their staff.
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Dr. Hart was supported by an Australian NHMRC Peter Doherty Research Fellowship, and Dr. Al-Magableh was supported by Hashemite University of Jordan scholarship. This work was also supported by grants from the William Buckland Foundation, the Ramaciotti Foundation, and the School of Medical Sciences, RMIT University.
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Al-Magableh, M.R., Hart, J.L. Mechanism of vasorelaxation and role of endogenous hydrogen sulfide production in mouse aorta. Naunyn-Schmied Arch Pharmacol 383, 403–413 (2011). https://doi.org/10.1007/s00210-011-0608-z
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DOI: https://doi.org/10.1007/s00210-011-0608-z