Vascular Effects of H2S-Donors: Fluorimetric Detection of H2S Generation and Ion Channel Activation in Human Aortic Smooth Muscle Cells

  • Alma MartelliEmail author
  • Valentina Citi
  • Vincenzo Calderone
Part of the Methods in Molecular Biology book series (MIMB, volume 2007)


Hydrogen sulfide (H2S) evokes vascular effects through several mechanisms including in wide part the activation of some ion channels such as ATP-sensitive potassium (KATP) channels and voltage-gated Kv7 potassium channels. Electrophysiological methods are very accurate, but they require high expertise and high specialized equipment. A more manageable fluorimetric technique which allows to record the membrane potential variations by the employment of an anionic bis-oxonol dye named DiBac4(3) with the administration of different blockers of several potassium channels could be useful to discover the targets of H2S-induced vascular hyperpolarization. Coupled with this technique, a fluorimetric detection (by the use of WSP-1 dye) of H2S generation in human vascular smooth muscle cells after H2S-donor administration could confirm the ability of these molecules to evoke the hyperpolarizing effect through the H2S release.

Key words

Human aortic smooth muscle cells (HASMCs) DiBac4(3) WSP-1 Hydrogen sulfide detection Hyperpolarizing effect Potassium channels 


  1. 1.
    Martelli A, Testai L, Breschi MC, Blandizzi C, Virdis A, Taddei S, Calderone V (2012) Hydrogen sulphide: novel opportunity for drug discovery. Med Res Rev 32:1093–1130CrossRefGoogle Scholar
  2. 2.
    Martelli A, Testai L, Marino A, Breschi MC, Da Settimo F, Calderone V (2012) Hydrogen sulphide: biopharmacological roles in the cardiovascular system and pharmaceutical perspectives. Curr Med Chem 19:3325–3336CrossRefGoogle Scholar
  3. 3.
    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 70:27–34CrossRefGoogle Scholar
  4. 4.
    Martelli A, Testai L, Citi V, Marino A, Pugliesi I, Barresi E, Nesi G, Rapposelli S, Taliani S, Da Settimo F, Breschi MC, Calderone V (2013) Arylthioamides as H2S donors: L-cysteine-activated releasing properties and vascular effects in vitro and in vivo. ACS Med Chem Lett 4:904–908CrossRefGoogle Scholar
  5. 5.
    Martelli A, Testai L, Citi V, Marino A, Bellagambi FG, Ghimenti S, Breschi MC, Calderone V (2014) Pharmacological characterization of the vascular effects of aryl isothiocyanates: is hydrogen sulfide the real player? Vasc Pharmacol 60:32–41CrossRefGoogle Scholar
  6. 6.
    Gopalakrishnan M, Whiteaker KL, Molinari EJ, Davis-Taber R, Scott VE, Shieh CC, Buckner SA, Milicic I, Cain JC, Postl S, Sullivan JP, Brioni JD (1999) Characterization of the ATP-sensitive potassium channels (KATP) expressed in guinea pig bladder smooth muscle cells. J Pharmacol Exp Ther 289:551–558PubMedGoogle Scholar
  7. 7.
    Miller TR, Taber RD, Molinari EJ, Whiteaker KL, Monteggia LM, Scott VE, Brioni JD, Sullivan JP, Gopalakrishnan M (1999) Pharmacological and molecular characterization of ATP-sensitive K+ channels in the TE671 human medulloblastoma cell line. Eur J Pharmacol 370:179–185CrossRefGoogle Scholar
  8. 8.
    Martelli A, Manfroni G, Sabbatini P, Barreca ML, Testai L, Novelli M, Sabatini S, Massari S, Tabarrini O, Masiello P, Calderone V, Cecchetti V (2013) 1,4-Benzothiazine ATP-sensitive potassium channel openers: modifications at the C-2 and C-6 positions. J Med Chem 56:4718–4728CrossRefGoogle Scholar
  9. 9.
    Liu C, Pan j LS, Zhao Y, Wu LY, Berkman CE, Whorton AR, Xian M (2011) Capture and visualization of hydrogen sulfide via a fluorescent probe. Angew Chem Int Ed Engl 50:10327–10329CrossRefGoogle Scholar
  10. 10.
    Martelli A (2016) Vascular effects of p-carboxyphenyl-isothiocyanate, a novel H2S-donor. In: Abstracts book of the 4th international conference on the biology of hydrogen sulfide, Naples, 3–5 June 2016Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Alma Martelli
    • 1
    Email author
  • Valentina Citi
    • 1
  • Vincenzo Calderone
    • 1
  1. 1.Department of PharmacyUniversity of PisaPisaItaly

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