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Vasorelaxation elicited by endogenous and exogenous hydrogen sulfide in mouse mesenteric arteries

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Abstract

H2S causes vasorelaxation however there is considerable heterogeneity in the reported pharmacological mechanism of this effect. This study examines the contribution of endogenously released H2S in the regulation of vascular tone and the mechanism of H2S-induced vasorelaxation in small resistance-like arteries. Mesenteric arteries from C57 and eNOS−/− mice were mounted in myographs to record isometric force. Vasorelaxation responses to NaHS were examined in the presence of various inhibitors of vasorelaxation pathways. Expression and activity of the H2S-producing enzyme, cystathionine-γ-lyase (CSE), were also examined. CSE was expressed in vascular smooth muscle and perivascular adipose cells from mouse mesenteric artery. The substrate for CSE, l-cysteine, caused a modest vasorelaxation (35%) in arteries from C57 mice and poor vasorelaxation (10%) in arteries from eNOS−/− mice that was sensitive to the CSE inhibitor dl-propargylglycine. The fast H2S donor, NaHS, elicited a full and biphasic vasorelaxation response in mesenteric arteries (EC50 (1) 8.7 μM, EC50 (2) 0.6 mM), which was significantly inhibited in eNOS−/− vessels (P < 0.05), unaffected by endothelial removal, or blockers at any point in the NO via soluble guanylate cyclase and cGMP (NO-sGC-cGMP) vasorelaxation pathway. Vasorelaxation to NaHS was significantly inhibited by blocking K+ channels of the KCa and KV subtypes and the Cl−/HCO3− exchanger (P < 0.05). Further experiments showed that NaHS can significantly inhibit voltage-gated Ca2+ channel function (P < 0.05). The vasorelaxant effect of H2S in small resistance-like arteries is complex, involving eNOS, K+ channels, Cl−/HCO3− exchanger, and voltage-gated Ca2+ channels. CSE is present in the smooth muscle and periadventitial adipose tissue of these resistance-like vessels and can be activated to cause modest vasorelaxation under these in vitro conditions.

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References

  • Ali MY, Ping CY, Mok YY, Ling L, Whiteman M, Bhatia M, Moore PK (2006) Regulation of vascular nitric oxide in vitro and in vivo; a new role for endogenous hydrogen sulphide? Br J Pharmacol 149:625–634

    CAS  PubMed  PubMed Central  Google Scholar 

  • Al-Magableh MR, Hart JL (2011) Mechanism of vasorelaxation and role of endogenous hydrogen sulfide production in mouse aorta. Naunyn Schmiedeberg's Arch Pharmacol 383:403–413

    CAS  Google Scholar 

  • Al-Magableh MR, Kemp-Harper BK, Ng HH, Miller AA, Hart JL (2014) Hydrogen sulfide protects endothelial nitric oxide function under conditions of acute oxidative stress in vitro. Naunyn Schmiedeberg's Arch Pharmacol 387:67–74

    CAS  Google Scholar 

  • Al-Magableh MR, Kemp-Harper BK, Hart JL (2015) Hydrogen sulfide treatment reduces blood pressure and oxidative stress in angiotensin II-induced hypertensive mice. Hypertens Res 38:13–20

    CAS  PubMed  Google Scholar 

  • Bucci M, Papapetropoulos A, Vellecco V, Zhou Z, Pyriochou A, Roussos C, Roviezzo F, Brancaleone V, Cirino G (2010) Hydrogen sulfide is an endogenous inhibitor of phosphodiesterase activity. Arterioscler Thromb Vasc Biol 30:1998–2004

    CAS  PubMed  Google Scholar 

  • Bucci M, Papapetropoulos A, Vellecco V, Zhou Z, Zaid A, Giannogonas P, Cantalupo A, Dhayade S, Karalis KP, Wang R, Feil R, Cirino G (2012) cGMP-dependent protein kinase contributes to hydrogen sulfide-stimulated vasorelaxation. PLoS One 7:e53319

    CAS  PubMed  PubMed Central  Google Scholar 

  • Cheang WS, Wong WT, Shen B, Lau CW, Tian XY, Tsang SY, Yao X, Chen ZY, Huang Y (2010) 4-aminopyridine-sensitive K+ channels contributes to NaHS-induced membrane hyperpolarization and relaxation in the rat coronary artery. Vasc Pharmacol 53:94–98

    CAS  Google Scholar 

  • Cheng Y, Ndisang J, Tang G, Cao K, Wang R (2004) Hydrogen sulfide-induced relaxation of resistance mesenteric artery beds of rats. Am J Physiol Heart Circ Physiol 287:2316–2323

    Google Scholar 

  • Coletta C, Papapetropoulos A, Erdelyi K, Olah G, Modis K, Panopoulos P, Asimakopoulou A, Gero D, Sharina I, Martin E, Szabo C (2012) Hydrogen sulfide and nitric oxide are mutually dependent in the regulation of angiogenesis and endothelium-dependent vasorelaxation. Proc Natl Acad Sci U S A 109:9161–9166

    CAS  PubMed  PubMed Central  Google Scholar 

  • Cox RH (2005) Molecular determinants of voltage-gated potassium currents in vascular smooth muscle. Cell Biochem Biophys 42:167–195

    CAS  PubMed  Google Scholar 

  • d’Emmanuele di Villa Bianca R, Sorrentino R, Coletta C, Mitidieri E, Rossi A, Vellecco V, Pinto A, Cirino G (2011) Hydrogen sulfide-induced dual vascular effect involves arachidonic acid cascade in rat mesenteric arterial bed. J Pharmacol Exp Ther 337:59–64

    PubMed  Google Scholar 

  • Dan P, Cheung JC, Scriven DR, Moore ED (2003) Epitope-dependent localization of estrogen receptor-alpha, but not -beta, in en face arterial endothelium. Am J Physiol Heart Circ Physiol 284:H1295–H1306

    CAS  PubMed  Google Scholar 

  • DeLeon ER, Stoy GF, Olson KR (2012) Passive loss of hydrogen sulfide in biological experiments. Anal Biochem 421:203–207

    CAS  PubMed  Google Scholar 

  • Delpire E, Lauf PK (1992) Kinetics of DIDS inhibition of swelling-activated K-Cl cotransport in low K sheep erythrocytes. J Membr Biol 126:89–96

    CAS  PubMed  Google Scholar 

  • Dombkowski RA, Russell MJ, Olson KR (2004) Hydrogen sulfide as an endogenous regulator of vascular smooth muscle tone in trout. Am J Phys Regul Integr Comp Phys 286:R678–R685

    CAS  Google Scholar 

  • Donovan J, Wong PS, Roberts RE, Garle MJ, Alexander SPH, Dunn WR, Ralevic V (2017) A critical role for cystathionine-beta-synthase in hydrogen sulfide-mediated hypoxic relaxation of the coronary artery. Vasc Pharmacol 93-95:20–32

    CAS  Google Scholar 

  • Fang L, Zhao J, Chen Y, Ma T, Xu G, Tang C, Liu X, Geng B (2009) Hydrogen sulfide derived from periadventitial adipose tissue is a vasodilator. J Hypertens 27:2174–2185

    CAS  PubMed  Google Scholar 

  • Favaloro J, McPherson G (2004) Vasorelaxation and hyperpolarisation of rat small mesenteric artery by the quaternary anion, tetraphenylboron. Naunyn-Schmeideberg’s Archives of Pharmacology 369:367–373

    CAS  Google Scholar 

  • Ford A, Al-Magableh M, Gaspari TA, Hart JL (2013) Chronic NaHS treatment is vasoprotective in high fat fed ApoE-/- mice. Int J Vasc Med 2013:915983

    PubMed  PubMed Central  Google Scholar 

  • Hart JL (2011) Role of sulfur-containing gaseous substances in the cardiovascular system. Front Biosci (Elite Ed) 3:736–749

    Google Scholar 

  • Hart J (2019) Vascular myography to examine functional responses of isolated blood vessels. In: Beltowski J (ed.) Vascular effects of hydrogen sulfide: methods and protocols. Springer Nature, pp. 205-217

  • Hedegaard ER, Nielsen BD, Kun A, Hughes AD, Kroigaard C, Mogensen S, Matchkov VV, Frobert O, Simonsen U (2014) KV7 channels are involved in hypoxia-induced vasodilatation of porcine coronary arteries. Br J Pharmacol 171:69–82

    CAS  PubMed  Google Scholar 

  • Hedegaard ER, Gouliaev A, Winther AK, Arcanjo DDR, Aalling M, Renaltan NS, Wood ME, Whiteman M, Skovgaard N, Simonsen U (2016) Involvement of potassium channels and calcium-independent mechanisms in hydrogen sulfide-induced relaxation of rat mesenteric small arteries. J Pharmacol Exp Ther 356:53–63

    CAS  PubMed  Google Scholar 

  • Hosoki R, Matsuki N, Kimura H (1997) The possible role of hydrogen sulfide as an endogenous smooth muscle relaxant in synergy with nitric oxide. Biochem Biophys Res Commun 237:527–531

    CAS  PubMed  Google Scholar 

  • Huang A, Sun D, Smith CJ, Connetta JA, Shesely EG, Koller A, Kaley G (2000) In eNOS knockout mice skeletal muscle arteriolar dilation to acetylcholine is mediated by EDHF. Am J Physiol Heart Circ Physiol 278:H762–H768

    CAS  PubMed  Google Scholar 

  • Hughes MN, Centelles MN, Moore KP (2009) Making and working with hydrogen sulfide: the chemistry and generation of hydrogen sulfide in vitro and its measurement in vivo: a review. Free Radic Biol Med 47:1346–1353

    CAS  PubMed  Google Scholar 

  • Jackson-Weaver O, Paredes DA, Gonzalez Bosc LV, Walker BR, Kanagy NL (2011) Intermittent hypoxia in rats increases myogenic tone through loss of hydrogen sulfide activation of large-conductance Ca2+-activated potassium channels. Circ Res 108:1439–1447

    CAS  PubMed  PubMed Central  Google Scholar 

  • Jackson-Weaver O, Osmond J, Riddle MA, Naik JS, Gonzalez Bosc LV, Walker BR, Kanagy NL (2013) Hydrogen sulfide dilates rat mesenteric arteries by activating endothelial large-conductance Ca2+-activated K+ channels and smooth muscle Ca2+ sparks. Am J Physiol Heart Circ Physiol

  • Jackson-Weaver O, Osmond JM, Naik JS, Gonzalez Bosc LV, Walker BR, Kanagy NL (2015) Intermittent hypoxia in rats reduces activation of Ca2+ sparks in mesenteric arteries. Am J Physiol Heart Circ Physiol 309:H1915–H1922

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kanagy NL, Szabo C, Papapetropoulos A (2017) Vascular biology of hydrogen sulfide. Am J Phys Cell Phys 312:C537–C549

    Google Scholar 

  • Kiss L, Deitch EA, Szabo C (2008) Hydrogen sulfide decreases adenosine triphosphate levels in aortic rings and leads to vasorelaxation via metabolic inhibition. Life Sci 83:589–594

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kitamura K, Yamazaki J (2001) Chloride channels and their functional roles in smooth muscle tone in the vasculature. Jpn J Pharmacol 85:351–357

    CAS  PubMed  Google Scholar 

  • Kloesch B, Steiner G, Mayer B, Schmidt K (2016) Hydrogen sulfide inhibits endothelial nitric oxide formation and receptor ligand-mediated Ca2+ release in endothelial and smooth muscle cells. Pharmacol Rep 68:37–43

    CAS  PubMed  Google Scholar 

  • Ko WH, Law VW, Yip WC, Yue GG, Lau CW, Chen ZY, Huang Y (2002) Stimulation of chloride secretion by baicalein in isolated rat distal colon. Am J Physiol Gastrointest Liver Physiol 282:G508–G518

    CAS  PubMed  Google Scholar 

  • Kohn C, Dubrovska G, Huang Y, Gollasch M (2012) Hydrogen sulfide: potent regulator of vascular tone and stimulator of angiogenesis. Int J Biomed Sci 8:81–86

    PubMed  PubMed Central  Google Scholar 

  • Kubo S, Kajiwara M, Kawabata A (2007) Dual modulation of the tension of isolated gastric artery and gastric mucosal circulation by hydrogen sulfide in rats. Inflammopharmacology 15:288–292

    CAS  PubMed  Google Scholar 

  • Lee SW, Cheng Y, Moore PK, Bian JS (2007) Hydrogen sulphide regulates intracellular pH in vascular smooth muscle cells. Biochem Biophys Res Commun 358:1142–1147

    CAS  PubMed  Google Scholar 

  • Li L, Whiteman M, Guan YY, Neo KL, Cheng Y, Lee SW, Zhao Y, Baskar R, Tan CH, Moore PK (2008) Characterization of a novel, water-soluble hydrogen sulfide-releasing molecule (GYY4137): new insights into the biology of hydrogen sulfide. Circulation 117:2351–2360

    CAS  PubMed  Google Scholar 

  • Li Y, Mihara K, Saifeddine M, Krawetz A, Lau DC, Li H, Ding H, Triggle CR, Hollenberg MD (2011) Perivascular adipose tissue-derived relaxing factors: release by peptide agonists via proteinase-activated receptor-2 (PAR2) and non-PAR2 mechanisms. Br J Pharmacol 164:1990–2002

    CAS  PubMed  PubMed Central  Google Scholar 

  • Liang GH, Adebiyi A, Leo MD, McNally EM, Leffler CW, Jaggar JH (2011) Hydrogen sulfide dilates cerebral arterioles by activating smooth muscle cell plasma membrane KATP channels. Am J Physiol Heart Circ Physiol 300:H2088–H2095

    CAS  PubMed  PubMed Central  Google Scholar 

  • Liang GH, Xi Q, Leffler CW, Jaggar JH (2012) Hydrogen sulfide activates Ca2+ sparks to induce cerebral arteriole dilatation. J Physiol 590:2709–2720

    CAS  PubMed  PubMed Central  Google Scholar 

  • Liu YH, Bian JS (2010) Bicarbonate-dependent effect of hydrogen sulfide on vascular contractility in rat aortic rings. Am J Phys Cell Phys 299:C866–C872

    CAS  Google Scholar 

  • Manicam C, Ginter N, Li H, Xia N, Goloborodko E, Zadeh JK, Musayeva A, Pfeiffer N, Gericke A (2017) Compensatory vasodilator mechanisms in the ophthalmic artery of endothelial nitric oxide synthase gene knockout nice. Sci Rep 7:7111

    PubMed  PubMed Central  Google Scholar 

  • Martelli A, Testai L, Breschi MC, Lawson K, McKay NG, Miceli F, Taglialatela M, Calderone V (2013) Vasorelaxation by hydrogen sulphide involves activation of Kv7 potassium channels. Pharmacol Res 70:27–34

    CAS  PubMed  Google Scholar 

  • Mustafa AK, Gadalla MM, Sen N, Kim S, Mu W, Gazi SK, Barrow RK, Yang G, Wang R, Snyder SH (2009) H2S signals through protein S-sulfhydration. Sci Signal 2:ra72

    PubMed  PubMed Central  Google Scholar 

  • Ng H, Yildiz G, Ku J, Miller A, Woodman O, Hart J (2017) Chronic NaHS treatment decreases oxidative stress and improves endothelial function in diabetic mice. Diab Vasc Dis Res 14:246–253

    CAS  PubMed  Google Scholar 

  • Patel S, Fedinec AL, Liu J, Weiss MA, Pourcyrous M, Harsono M, Parfenova H, Leffler CW (2018) H2S mediates the vasodilator effect of endothelin-1 in the cerebral circulation. Am J Physiol Heart Circ Physiol 315:H1759–H1764

    CAS  PubMed  PubMed Central  Google Scholar 

  • Powell CR, Dillon KM, Matson JB (2018) A review of hydrogen sulfide (H2S) donors: chemistry and potential therapeutic applications. Biochem Pharmacol 149:110–123

    CAS  PubMed  Google Scholar 

  • Schleifenbaum J, Kohn C, Voblova N, Dubrovska G, Zavarirskaya O, Gloe T, Crean CS, Luft FC, Huang Y, Schubert R, Gollasch M (2010) Systemic peripheral artery relaxation by KCNQ channel openers and hydrogen sulfide. J Hypertens 28:1875–1882

    CAS  PubMed  Google Scholar 

  • Selley E, Kun S, Szijarto IA, Laczy B, Kovacs T, Fulop F, Wittmann I, Molnar GA (2014) Exenatide induces aortic vasodilation increasing hydrogen sulphide, carbon monoxide and nitric oxide production. Cardiovasc Diabetol 13:69

    PubMed  PubMed Central  Google Scholar 

  • Shibuya N, Mikami Y, Kimura Y, Nagahara N, Kimura H (2009) Vascular endothelium expresses 3-mercaptopyruvate sulfurtransferase and produces hydrogen sulfide. J Biochem 146:623–626

    CAS  PubMed  Google Scholar 

  • Streeter E, Hart J, Badoer E (2012) An investigation of the mechanisms of hydrogen sulfide-induced vasorelaxation in rat middle cerebral arteries. Naunyn Schmiedeberg's Arch Pharmacol 385:991–1002

    CAS  Google Scholar 

  • Sun YG, Cao YX, Wang WW, Ma SF, Yao T, Zhu YC (2008) Hydrogen sulphide is an inhibitor of l-type calcium channels and mechanical contraction in rat cardiomyocytes. Cardiovasc Res 79:632–641

    CAS  PubMed  Google Scholar 

  • Sun Y, Huang Y, Yu W, Chen S, Yao Q, Zhang C, Bu D, Tang C, Du J, Jin H (2017) Sulfhydration-associated phosphodiesterase 5A dimerization mediates vasorelaxant effect of hydrogen sulfide. Oncotarget 8:31888–31900

    PubMed  PubMed Central  Google Scholar 

  • Szabo C (2017) Hydrogen sulfide, an enhancer of vascular nitric oxide signaling: mechanisms and implications. Am J Phys Cell Phys 312:C3–C15

    Google Scholar 

  • Takir S, Ortakoylu GZ, Toprak A, Uydes-Dogan BS (2015) NaHS induces relaxation response in prostaglandin F(2alpha) precontracted bovine retinal arteries partially via Kv and Kir channels. Exp Eye Res 132:190–197

    CAS  PubMed  Google Scholar 

  • Tang G, Wu L, Liang W, Wang R (2005) Direct stimulation of KATP channels by exogenous and endogenous hydrogen sulfide in vascular smooth muscle cells. Mol Pharmacol 68:1757–1764

    CAS  PubMed  Google Scholar 

  • Tang G, Yang G, Jiang B, Ju Y, Wu L, Wang R (2013) H2S is an endothelium-derived hyperpolarizing factor. Antioxid Redox Signal 19:1634–1646

    CAS  PubMed  Google Scholar 

  • van der Sterren S, Kleikers P, Zimmermann LJ, Villamor E (2011) Vasoactivity of the gasotransmitters hydrogen sulfide and carbon monoxide in the chicken ductus arteriosus. Am J Phys Regul Integr Comp Phys 301:R1186–R1198

    Google Scholar 

  • Webb GD, Lim LH, Oh VM, Yeo SB, Cheong YP, Ali MY, El Oakley R, Lee CN, Wong PS, Caleb MG, Salto-Tellez M, Bhatia M, Chan ES, Taylor EA, Moore PK (2008) Contractile and vasorelaxant effects of hydrogen sulfide and its biosynthesis in the human internal mammary artery. J Pharmacol Exp Ther 324:876–882

    CAS  PubMed  Google Scholar 

  • Wen JY, Wang M, Li YN, Jiang HH, Sun XJ, Chen ZW (2018) Vascular protection of hydrogen sulfide on cerebral ischemia/reperfusion injury in rats. Front Neurol 9:779

    PubMed  PubMed Central  Google Scholar 

  • White BJ, Smith PA, Dunn WR (2013) Hydrogen sulphide-mediated vasodilatation involves the release of neurotransmitters from sensory nerves in pressurized mesenteric small arteries isolated from rats. Br J Pharmacol 168:785–793

    CAS  PubMed  PubMed Central  Google Scholar 

  • Wulff H (2008) New light on the “old” chloride channel blocker DIDS. ACS Chem Biol 3:399–401

    CAS  PubMed  Google Scholar 

  • Zhao W, Wang R (2002) H2S-induced vasorelaxation and underlying cellular and molecular mechanisms. Am J Physiol Heart Circ Physiol 283:H474–H480

    CAS  PubMed  Google Scholar 

  • Zhao W, Zhang J, Lu Y, Wang R (2001) The vasorelaxant effect of H2S as a novel endogenous gaseous KATP channel opener. EMBO J 20:6008–6016

    CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

JLH was the recipient of an Australian National Health and Medical Research Council Peter Doherty Fellowship. We wish to thank Prof C Sobey and Assoc Prof B Kemp-Harper, Monash University for the eNOS−/− mouse tissue and use of laboratory facilities at Monash University. We also acknowledge the expert advice and assistance of Dr Simon Potocnik, RMIT University, for the confocal imaging and the provision of the linopirdine. We thank Prof Owen Woodman and his research group along with the technical staff at the RMIT School of Health and Biomedical Sciences for their support and assistance.

Funding

This study was supported by grants from the Ramaciotti Foundation and the William Buckland Foundation.

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  1. School of Medicine, Faculty of Medicine and Health, University of Sydney, 111B Edward Ford Building, Camperdown, Sydney, NSW, 2006, Australia

    Joanne L Hart

  2. School of Health and Biomedical Sciences, RMIT University, Melbourne, Australia

    Joanne L Hart

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JLH conceived and designed the research, conducted experiments, analysed data and wrote the manuscript.

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Correspondence to Joanne L Hart.

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This study was approved by the Monash University Department of Pharmacology Animal Ethics Committee and the RMIT University Animal Ethics Committee, Australia. This investigation conforms with the National Health and Medical Research Council (NHMRC) Australian code of practice for the care and use of animals for scientific purposes.

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Supplementary Figure 1

Concentration-response curves to NaHS in C57 mouse mesenteric arteries A: in the absence (●) and presence (♦) of TEA (1mM) B: in the absence (●) and presence (▲) of Iberiotoxin (10nM) and C: in the absence (●) and presence (■) of Glibenclamide (10μM). Values are expressed as a % relaxation and given as mean ± SEM, n=6-9. (PDF 48 kb)

Supplementary Figure 2

Concentration-response curves to NaHS in C57 mouse mesenteric arteries A: in the absence (●) and presence (▲) of L-NAME (100μM) B: in the absence (●) and presence (○) of ODQ (10μM) C: in the absence (●) and presence (■) of L-NAME (100μM) and ODQ (10μM) D: in the absence (●) and presence (♦) of Cb-PTIO (200μM) E: in the absence (●) and presence () of zaprinast (1μM) and F: in the absence (●) and presence (▼) of KT5823 (1μM). Values are expressed as a % relaxation and given as mean ± SEM, n=6-9. (PDF 36 kb)

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Hart, J.L. Vasorelaxation elicited by endogenous and exogenous hydrogen sulfide in mouse mesenteric arteries. Naunyn-Schmiedeberg's Arch Pharmacol 393, 551–564 (2020). https://doi.org/10.1007/s00210-019-01752-w

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