Molecular Medicine

, Volume 21, Issue 1, pp 1–14 | Cite as

Regulation of Vascular Tone, Angiogenesis and Cellular Bioenergetics by the 3-Mercaptopyruvate Sulfurtransferase/H2S Pathway: Functional Impairment by Hyperglycemia and Restoration by dl-α-Lipoic Acid

  • Ciro Coletta
  • Katalin Módis
  • Bartosz Szczesny
  • Attila Brunyánszki
  • Gábor Oláh
  • Ester C. S. Rios
  • Kazunori Yanagi
  • Akbar Ahmad
  • Andreas Papapetropoulos
  • Csaba Szabo
Research Article


Hydrogen sulfide (H2S), as a reducing agent and an antioxidant molecule, exerts protective effects against hyperglycemic stress in the vascular endothelium. The mitochondrial enzyme 3-mercaptopyruvate sulfurtransferase (3-MST) is an important biological source of H2S. We have recently demonstrated that 3-MST activity is inhibited by oxidative stress in vitro and speculated that this may have an adverse effect on cellular homeostasis. In the current study, given the importance of H2S as a vasorelaxant, angiogenesis stimulator and cellular bioenergetic mediator, we first determined whether the 3-MST/H2S system plays a physiological regulatory role in endothelial cells. Next, we tested whether a dysfunction of this pathway develops during the development of hyperglycemia and diabetes-associated vascular complications. Intraperitoneal (IP) 3-MP (1 mg/kg) raised plasma H2S levels in rats. 3-MP (10 µmol/L to 1 mmol/L) promoted angiogenesis in vitro in bEnd3 microvascular endothelial cells and in vivo in a Matrigel assay in mice (0.3–1 mg/kg). In vitro studies with bEnd3 cell homogenates demonstrated that the 3-MP-induced increases in H2S production depended on enzymatic activity, although at higher concentrations (1–3 mmol/L) there was also evidence for an additional nonenzymatic H2S production by 3-MP. In vivo, 3-MP facilitated wound healing in rats, induced the relaxation of dermal microvessels and increased mitochondrial bioenergetic function. In vitro hyperglycemia or in vivo streptozotocin diabetes impaired angiogenesis, attenuated mitochondrial function and delayed wound healing; all of these responses were associated with an impairment of the proangiogenic and bioenergetic effects of 3-MP. The antioxidants DL-α-lipoic acid (LA) in vivo, or dihydrolipoic acid (DHLA) in vitro restored the ability of 3-MP to stimulate angiogenesis, cellular bioenergetics and wound healing in hyperglycemia and diabetes. We conclude that diabetes leads to an impairment of the 3-MST/H2S pathway, and speculate that this may contribute to the pathogenesis of hyperglycemic endothelial cell dysfunction. We also suggest that therapy with H2S donors, or treatment with the combination of 3-MP and lipoic acid may be beneficial in improving angiogenesis and bioenergetics in hyperglycemia.



This work was supported by the American Diabetes Association (7-12-BS-184) and the National Institutes of Health (R01GM107846) to C Szabo. C Coletta was supported by a fellowship by the American Heart Association. K Módis was supported by the James W. McLaughlin Fellowship Fund of the University of Texas. The editorial assistance of Ms. Li Li Szabo is appreciated.


  1. 1.
    Fiorucci S, Distrutti E, Cirino G, Wallace JL. (2006) The emerging roles of hydrogen sulfide in the gastrointestinal tract and liver. Gastroenterology. 131:259–71.CrossRefGoogle Scholar
  2. 2.
    Szabo C. (2007) Hydrogen sulphide and its therapeutic potential. Nat. Rev. Drug Discov. 6:917–35.CrossRefGoogle Scholar
  3. 3.
    Whiteman M, Winyard PG. (2011) Hydrogen sulfide and inflammation: the good, the bad, the ugly and the promising. Expert Rev. Clin. Pharmacol. 4:13–32.CrossRefGoogle Scholar
  4. 4.
    Bucci M, Cirino G. (2011) Hydrogen sulphide in heart and systemic circulation. Inflamm. Allergy Drug Targets. 10:103–8.CrossRefGoogle Scholar
  5. 5.
    Wang R. (2012) Physiological implications of hydrogen sulfide: a whiff exploration that blossomed. Physiol. Rev. 92:791–896.CrossRefGoogle Scholar
  6. 6.
    Módis K, Wolanska K, Vozdek R. (2013) H2S in cell signaling, signal transduction, cellular bioenergetics and physiology in C. Elegans. Gen. Physiol. Biophys. 32:1–22.CrossRefGoogle Scholar
  7. 7.
    Polhemus DJ, Lefer DJ. (2014) Emergence of hydrogen sulfide as an endogenous gaseous signaling molecule in cardiovascular disease. Circ. Res. 114:730–7.CrossRefGoogle Scholar
  8. 8.
    Jain SK, et al. (2010) Low levels of hydrogen sulfide in the blood of diabetes patients and streptozotocin-treated rats causes vascular inflammation? Antioxid. Redox. Signal. 12:1333–7.CrossRefGoogle Scholar
  9. 9.
    Whiteman M, et al. (2010) Adiposity is a major determinant of plasma levels of the novel vasodilator hydrogen sulphide. Diabetologia. 53:1722–6.CrossRefGoogle Scholar
  10. 10.
    Suzuki K, et al. (2011) Hydrogen sulfide replacement therapy protects the vascular endothelium in hyperglycemia by preserving mitochondrial function. Proc. Natl. Acad. Sci. U. S. A. 108:13829–34.CrossRefGoogle Scholar
  11. 11.
    Ahmad FU, et al. (2012) Exogenous hydrogen sulfide (H2S) reduces blood pressure and prevents the progression of diabetic nephropathy in spontaneously hypertensive rats. Ren. Fail. 34:203–10.CrossRefGoogle Scholar
  12. 12.
    Xue H, et al. (2013) H2S inhibits hyperglycemia-induced intrarenal renin-angiotensin system activation via attenuation of reactive oxygen species generation. PLoS One. 8:e74366.CrossRefGoogle Scholar
  13. 13.
    Manna P, Jain SK. (2013) L-cysteine and hydrogen sulfide increase PIP3 and AMPK/PPARγ expression and decrease ROS and vascular inflammation markers in high glucose treated human U937 monocytes. J. Cell. Biochem. 114:2334–45.CrossRefGoogle Scholar
  14. 14.
    Si YF, et al. (2013) Treatment with hydrogen sulfide alleviates streptozotocin-induced diabetic retinopathy in rats. Br. J. Pharmacol. 169:619–31.CrossRefGoogle Scholar
  15. 15.
    Yamamoto J, et al. (2013) Distribution of hydrogen sulfide (H2S)-producing enzymes and the roles of the H2S donor sodium hydrosulfide in diabetic nephropathy. Clin. Exp. Nephrol. 17:32–40.CrossRefGoogle Scholar
  16. 16.
    Wei WB, Hu X, Zhuang XD, Liao LZ, Li WD. (2014) GYY4137, a novel hydrogen sulfide-releasing molecule, likely protects against high glucose-induced cytotoxicity by activation of the AMPK/mTOR signal pathway in H9c2 cells. Mol. Cell. Biochem. 389:249–56.CrossRefGoogle Scholar
  17. 17.
    Shibuya N, Mikami Y, Kimura Y, Nagahara N, Kimura H. (2009) Vascular endothelium expresses 3-mercaptopyruvate sulfurtransferase and produces hydrogen sulfide. J. Biochem. 146:623–6.CrossRefGoogle Scholar
  18. 18.
    Sen U, et al. (2012) Increased endogenous H2S generation by CBS, CSE, and 3MST gene therapy improves ex vivo renovascular relaxation in hyperhomocysteinemia. Am. J. Physiol. Cell Physiol. 303:C41–51.CrossRefGoogle Scholar
  19. 19.
    Madden JA, Ahlf SB, Dantuma MW, Olson KR, Roerig DL. (2012) Precursors and inhibitors of hydrogen sulfide synthesis affect acute hypoxic pulmonary vasoconstriction in the intact lung. J. Appl. Physiol (1985). 112:411–8.CrossRefGoogle Scholar
  20. 20.
    Modis K, Asimakopoulou A, Coletta C, Papapetropoulos A, Szabo C. (2013) Oxidative stress suppresses the cellular bioenergetic effect of the 3-mercaptopyruvate sulfurtransferase/hydrogen sulfide pathway. Biochem. Biophys. Res. Commun. 433:401–7.CrossRefGoogle Scholar
  21. 21.
    Modis K, et al. (2014) Regulation of mitochondrial bioenergetic function by hydrogen sulfide. Part II. Pathophysiological and therapeutic aspects. Br. J. Pharmacol. 171:2123–46.CrossRefGoogle Scholar
  22. 22.
    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. 237:527–31.CrossRefGoogle Scholar
  23. 23.
    Zhao W, Wang R. (2002) H2S-induced vasorelaxation and underlying cellular and molecular mechanisms. Am. J. Physiol. Heart Circ. Physiol. 283:H474–80.CrossRefGoogle Scholar
  24. 24.
    Bucci M, et al. (2010) Hydrogen sulfide is an endogenous inhibitor of phosphodiesterase activity. Arterioscler. Thromb. Vasc. Biol 30:1998–2004.CrossRefGoogle Scholar
  25. 25.
    d’Emmanuele di Villa Bianca R, et al. (2011) Hydrogen sulfide-induced dual vascular effect involves arachidonic acid cascade in rat mesenteric arterial bed. J. Pharmacol. Exp. Ther. 337:59–64.CrossRefGoogle Scholar
  26. 26.
    Mustafa AK, et al. (2011) Hydrogen sulfide as endothelium-derived hyperpolarizing factor sulfhydrates potassium channels. Circ. Res. 109:1259–68.CrossRefGoogle Scholar
  27. 27.
    Coletta C, et al. (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. 109:9161–6.CrossRefGoogle Scholar
  28. 28.
    Bucci M, et al. (2012) cGMP-dependent protein kinase contributes to hydrogen sulfide-stimulated vasorelaxation. PLoS One.7:e53319.CrossRefGoogle Scholar
  29. 29.
    Bettowski J, Jamroz-Wiśniewska A. (2014) Hydrogen sulfide and endothelium-dependent vasorelaxation. Molecules 19:21183–99.CrossRefGoogle Scholar
  30. 30.
    Cai WJ, et al. (2007) The novel proangiogenic effect of hydrogen sulfide is dependent on Akt phosphorylation. Cardiovasc. Res. 76:29–40.CrossRefGoogle Scholar
  31. 31.
    Papapetropoulos A, et al. (2009) Hydrogen sulfide is an endogenous stimulator of angiogenesis. Proc. Natl. Acad. Sci. U. S. A. 106:21972–7.CrossRefGoogle Scholar
  32. 32.
    Wang MJ, et al. (2010) The hydrogen sulfide donor NaHS promotes angiogenesis in a rat model of hind limb ischemia. Antioxid. Redox. Signal. 12:1065–77.CrossRefGoogle Scholar
  33. 33.
    Szabo C, Papapetropoulos A. (2011) Hydrogen sulphide and angiogenesis: mechanisms and applications. Br. J. Pharmacol. 164:853–65.CrossRefGoogle Scholar
  34. 34.
    Polhemus DJ, et al. (2013) Hydrogen sulfide attenuates cardiac dysfunction after heart failure via induction of angiogenesis. Circ. Heart Fail. 6:1077–86.CrossRefGoogle Scholar
  35. 35.
    Szabo C, et al. (2013) Tumor-derived hydrogen sulfide, produced by cystathionine-β-synthase, stimulates bioenergetics, cell proliferation, and angiogenesis in colon cancer. Proc. Natl. Acad. Sci. U. S. A. 110:12474–9.CrossRefGoogle Scholar
  36. 36.
    Liu F, et al. (2014) Hydrogen sulfide improves wound healing via restoration of endothelial progenitor cell functions and activation of angiopoietin-1 in type 2 diabetes. Diabetes. 63:1763–78.CrossRefGoogle Scholar
  37. 37.
    Goubern M, Andriamihaja M, Nübel T, Blachier F, Bouillaud F. (2007) Sulfide, the first inorganic substrate for human cells. FASEB J. 21:1699–706.CrossRefGoogle Scholar
  38. 38.
    Modis K, Coletta C, Erdélyi K, Papapetropoulos A, Szabo C. (2013) Intramitochondrial hydrogen sulfide production by 3-mercaptopyruvate sulfurtransferase maintains mitochondrial electron flow and supports cellular bioenergetics. FASEB J. 27:601–11.CrossRefGoogle Scholar
  39. 39.
    Módis K, Panopoulos P, Coletta C, Papapetropoulos A, Szabo C. (2013) Hydrogen sulfide-mediated stimulation of mitochondrial electron transport involves inhibition of the mitochondrial phosphodiesterase 2A, elevation of cAMP and activation of protein kinase A. Biochem. Pharmacol 86:1311–9.CrossRefGoogle Scholar
  40. 40.
    Szczesny B, et al. (2014) AP39, a novel mitochondria-targeted hydrogen sulfide donor, stimulates cellular bioenergetics, exerts cytoprotective effects and protects against the loss of mitochondrial DNA integrity in oxidatively stressed endothelial cells in vitro. Nitric Oxide.41:120–30.CrossRefGoogle Scholar
  41. 41.
    Helmy N, et al. (2014) Oxidation of hydrogen sulfide by human liver mitochondria. Nitric Oxide. 41:105–12.CrossRefGoogle Scholar
  42. 42.
    Szabo C, et al. (2014) Regulation of mitochondrial bioenergetic function by hydrogen sulfide. Part I. Biochemical and physiological mechanisms. Br. J. Pharmacol. 171:2099–122.CrossRefGoogle Scholar
  43. 43.
    Mikami Y, et al. (2011) Thioredoxin and dihydrolipoic acid are required for 3-mercaptopyruvate sulfurtransferase to produce hydrogen sulfide. Biochem. J. 439:479–85.CrossRefGoogle Scholar
  44. 44.
    Cameron NE, Jack AM, Cotter MA. (2001) Effect of alpha-lipoic acid on vascular responses and nociception in diabetic rats. Free Radic. Biol. Med. 31:125–35.CrossRefGoogle Scholar
  45. 45.
    Nebbioso M, Pranno F, Pescosolido N. (2013) Lipoic acid in animal models and clinical use in diabetic retinopathy. Expert Opin. Pharmacother. 14:1829–38.CrossRefGoogle Scholar
  46. 46.
    Committee for the Update of the Guide for the Care and Use of Laboratory Animals, Institute for Laboratory Animal Research, Division on Earth and Life Studies, National Research Council of the National Academies. (2011) Guide for the Care and Use of Laboratory Animals. 8th edition. Washington (DC): National Academies Press.Google Scholar
  47. 47.
    Warner TD. (1990) Simultaneous perfusion of rat isolated superior mesenteric arterial and venous beds: comparison of their vasoconstrictor and vasodilator responses to agonists. Br. J. Pharmacol. 99:427–33.CrossRefGoogle Scholar
  48. 48.
    Ferreira FM, Palmeira CM, Seiça R, Moreno AJ, Santos MS. (2003) Diabetes and mitochondrial bioenergetics: alterations with age. J. Biochem. Mol. Toxicol. 17:214–22.CrossRefGoogle Scholar
  49. 49.
    Szabo C. (2009) Role of nitrosative stress in the pathogenesis of diabetic vascular dysfunction. Br. J. Pharmacol. 156:713–27.CrossRefGoogle Scholar
  50. 50.
    Sivitz WI, Yorek MA. (2010) Mitochondrial dysfunction in diabetes: from molecular mechanisms to functional significance and therapeutic opportunities. Antioxid. Redox. Signal. 12:537–77.CrossRefGoogle Scholar
  51. 51.
    Giacco F, Brownlee M. (2010) Oxidative stress and diabetic complications. Circ. Res. 107:1058–70.CrossRefGoogle Scholar
  52. 52.
    Papanas N, Ziegler D. (2014) Efficacy of α-lipoic acid in diabetic neuropathy. Expert Opin. Pharmacother. 15:2721–31.CrossRefGoogle Scholar
  53. 53.
    Nebbioso M, Pranno F, Pescosolido N. (2013) Lipoic acid in animal models and clinical use in diabetic retinopathy. Expert Opin. Pharmacother. 14:1829–38.CrossRefGoogle Scholar
  54. 54.
    Kiguchi S. (1983) Metabolism of 3-mercaptopyruvate in rat tissues. Acta Med. Okayama. 37:85–91.PubMedGoogle Scholar
  55. 55.
    Shibuya N, et al. (2009) 3-Mercaptopyruvate sulfurtransferase produces hydrogen sulfide and bound sulfane sulfur in the brain. Antioxid. Redox. Signal. 11:703–14.CrossRefGoogle Scholar
  56. 56.
    Nagahara N, Katayama A. (2005) Post-translational regulation of mercaptopyruvate sulfurtransferase via a low redox potential cysteine-sulfenate in the maintenance of redox homeostasis. J. Biol. Chem. 280:34569–76.CrossRefGoogle Scholar
  57. 57.
    Miyamoto R, Otsuguro K, Yamaguchi S, Ito S. (2014) Contribution of cysteine aminotransferase and mercaptopyruvate sulfurtransferase to hydrogen sulfide production in peripheral neurons. J. Neurochem. 130:29–40.CrossRefGoogle Scholar
  58. 58.
    Nagahara N, Nirasawa T, Yoshii T, Niimura Y. (2012) Is novel signal transducer sulfur oxide involved in the redox cycle of persulfide at the catalytic site cysteine in a stable reaction intermediate of mercaptopyruvate sulfurtransferase? Antioxid. Redox. Signal. 16:747–53.CrossRefGoogle Scholar
  59. 59.
    Nagahara N. (2013) Regulation of mercaptopyruvate sulfurtransferase activity via intrasubunit and intersubunit redox-sensing switches. Antioxid. Redox. Signal. 19:1792–802.CrossRefGoogle Scholar
  60. 60.
    Yadav PK, Yamada K, Chiku T, Koutmos M, Banerjee R. (2013) Structure and kinetic analysis of H2S production by human mercaptopyruvate sulfurtransferase. J. Biol. Chem. 288:20002–13.CrossRefGoogle Scholar
  61. 61.
    Flannigan KL, Ferraz JG, Wang R, Wallace JL. (2013) Enhanced synthesis and diminished degradation of hydrogen sulfide in experimental colitis: a site-specific, pro-resolution mechanism. PLoS One. 8:e71962.CrossRefGoogle Scholar
  62. 62.
    Nie LH, et al. (2013) Effects of intrauterine cigarette smoking exposure on expression of 3-mercaptopyruvate sulfurtransferase in medulla oblongata of neonatal rats [in Chinese]. Sichuan Da Xue Xue Bao Yi Xue Ban. 44:526–30.PubMedGoogle Scholar
  63. 63.
    Li M, et al. (2013) Chronic intermittent hypoxia promotes expression of 3-mercaptopyruvate sulfurtransferase in adult rat medulla oblongata. Auton. Neurosci. 179:84–9.CrossRefGoogle Scholar
  64. 64.
    Zhao H, Chan SJ, Ng YK, Wong PT. (2013) Brain 3-mercaptopyruvate sulfurtransferase (3MST): cellular localization and downregulation after acute stroke. PLoS One. 8:e67322.CrossRefGoogle Scholar
  65. 65.
    Nagahara N, et al. (2013) Antioxidant enzyme, 3-mercaptopyruvate sulfurtransferase-knockout mice exhibit increased anxiety-like behaviors: a model for human mercaptolactate-cysteine disulfiduria. Sci. Rep. 3:1986.CrossRefGoogle Scholar
  66. 66.
    Yusuf M, et al. (2005) Streptozotocin-induced diabetes in the rat is associated with enhanced tissue hydrogen sulfide biosynthesis. Biochem. Biophys. Res. Commun. 333:1146–52.CrossRefGoogle Scholar
  67. 67.
    Szabo C. (2012) Roles of hydrogen sulfide in the pathogenesis of diabetes mellitus and its complications. Antioxid. Redox. Signal. 17:68–80.CrossRefGoogle Scholar
  68. 68.
    Langouche L, Mesotten D, Vanhorebeek I. (2010) Endocrine and metabolic disturbances in critical illness: relation to mechanisms of organ dysfunction and adverse outcome. Verh. K. Acad Geneeskd. Belg. 72:149–63.PubMedGoogle Scholar
  69. 69.
    Rizk M, Witte M, Barbul A. (2004) Nitric oxide and wound healing. World J. Surg. 28:301–6.CrossRefGoogle Scholar
  70. 70.
    Buganza Tepole A, Kuhl E. (2013) Systems-based approaches toward wound healing. Pediatr Res. 73:553–63.CrossRefGoogle Scholar

Copyright information

© The Author(s) 2015

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, and provide a link to the Creative Commons license. You do not have permission under this license to share adapted material derived from this article or parts of it.

The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this license, visit (

Authors and Affiliations

  • Ciro Coletta
    • 1
  • Katalin Módis
    • 1
  • Bartosz Szczesny
    • 1
  • Attila Brunyánszki
    • 1
  • Gábor Oláh
    • 1
  • Ester C. S. Rios
    • 1
  • Kazunori Yanagi
    • 1
  • Akbar Ahmad
    • 1
  • Andreas Papapetropoulos
    • 2
  • Csaba Szabo
    • 1
  1. 1.Department of AnesthesiologyUniversity of Texas Medical BranchGalvestonUSA
  2. 2.Faculty of PharmacyUniversity of AthensAthensGreece

Personalised recommendations