Skip to main content
SpringerLink
Log in

Testosterone-induced relaxation involves L-type and store-operated Ca2+ channels blockade, and PGE2 in guinea pig airway smooth muscle

  • Muscle physiology
  • Published:
Pflügers Archiv - European Journal of Physiology Aims and scope Submit manuscript

Abstract

In vascular smooth muscle, it has been described that testosterone (TES) produces relaxation by blocking L-type Ca2+ channels. Recently, we found that L-type Ca2+ and store-operated Ca2+ (SOC) channels are the main membranal structures that provide extracellular Ca2+ for carbachol (CCh)-induced contraction in airway smooth muscle (ASM). We studied the possible interactions between L-type and SOC channels in TES-induced relaxation in guinea pig ASM. TES (10, 32, 100, and 178 μM) induced a complete relaxation of CCh-precontracted tracheal smooth muscle, and indomethacin partially inhibited this response. In single myocytes, the KCl-induced intracellular Ca2+ increase ([Ca2+]i) was decreased by 32 and completely blocked by 100 nM TES. This androgen (32 and 100 μM) significantly diminished (~25 and 49 %, respectively) the capacitative Ca2+ entry. Myocytes stimulated with CCh produced a transient Ca2+ peak followed by a sustained plateau. D-600 was added during the plateau phase, and a partial diminution (~35 %) was observed. A greater decrease (~78 %) was seen when 2-aminoethyl diphenylborinate (2-APB, SOC antagonist) was used. The combination of both drugs completely abolished the Ca2+ plateau induced by CCh. TES (100 μM) also completely abolished the CCh-induced Ca2+ plateau. Indomethacin significantly diminished this effect of TES. PGE2 and butaprost proportionally decreased the Ca2+ plateau as indomethacin blocked it. Sarcoplasmic reticulum refilling was partially, dependently, and significantly diminished by TES. We concluded that TES-induced relaxation involves blockade of L-type Ca2+ channels at nanomolar and SOC channels at micromolar concentration and PGE2 seems to be also involved in this phenomenon.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

References

  1. Bazan-Perkins B, Flores-Soto E, Barajas-Lopez C, Montaño LM (2003) Role of sarcoplasmic reticulum Ca2+ content in Ca2+ entry of bovine airway smooth muscle cells. Naunyn Schmiedeberg’s Arch Pharmacol 368:277–283

  2. Bordallo J, de Boto MJ, Meana C, Velasco L, Bordallo C, Suarez L, Cantabrana B, Sanchez M (2008) Modulatory role of endogenous androgens on airway smooth muscle tone in isolated guinea-pig and bovine trachea; involvement of β2-adrenoceptors, the polyamine system and external calcium. Eur J Pharmacol 601:154–162

    Article  CAS  PubMed  Google Scholar 

  3. Bourreau JP, Abela AP, Kwan CY, Daniel EE (1991) Acetylcholine Ca2+ stores refilling directly involves a dihydropyridine-sensitive channel in dog trachea. Am J Physiol 261:C497–C505

    CAS  PubMed  Google Scholar 

  4. Cairrao E, Alvarez E, Santos-Silva AJ, Verde I (2008) Potassium channels are involved in testosterone-induced vasorelaxation of human umbilical artery. Naunyn Schmiedeberg’s Arch Pharmacol 376:375–383

    Article  CAS  Google Scholar 

  5. Carbajal V, Vargas MH, Flores-Soto E, Martinez-Cordero E, Bazan-Perkins B, Montaño LM (2005) LTD4 induces hyperresponsiveness to histamine in bovine airway smooth muscle: role of SR-ATPase Ca2+ pump and tyrosine kinase. Am J Physiol Lung Cell Mol Physiol 288:L84–L92

  6. Costarella CE, Stallone JN, Rutecki GW, Whittier FC (1996) Testosterone causes direct relaxation of rat thoracic aorta. J Pharmacol Exp Ther 277:34–39

    CAS  PubMed  Google Scholar 

  7. Deenadayalu VP, White RE, Stallone JN, Gao X, Garcia AJ (2001) Testosterone relaxes coronary arteries by opening the large-conductance, calcium-activated potassium channel. Am J Physiol Heart Circ Physiol 281:H1720–H1727

    CAS  PubMed  Google Scholar 

  8. Ding AQ, Stallone JN (2001) Testosterone-induced relaxation of rat aorta is androgen structure specific and involves K+ channel activation. J Appl Physiol 91:2742–2750

    CAS  PubMed  Google Scholar 

  9. Enfissi A, Prigent S, Colosetti P, Capiod T (2004) The blocking of capacitative calcium entry by 2-aminoethyl diphenylborate (2-APB) and carboxyamidotriazole (CAI) inhibits proliferation in Hep G2 and Huh-7 human hepatoma cells. Cell Calcium 36:459–467

    Article  CAS  PubMed  Google Scholar 

  10. Espinoza J, Montaño LM, Perusquia M (2013) Nongenomic bronchodilating action elicited by dehydroepiandrosterone (DHEA) in a guinea pig asthma model. J Steroid Biochem Mol Biol 138:174–182

  11. Fernandes VS, Barahona MV, Recio P, Martinez-Saenz A, Ribeiro AS, Contreras C, Martinez AC, Bustamante S, Carballido J, Garcia-Sacristan A, Prieto D, Hernandez M (2012) Mechanisms involved in testosterone-induced relaxation to the pig urinary bladder neck. Steroids 77:394–402

    Article  CAS  PubMed  Google Scholar 

  12. Flores-Soto E, Carbajal V, Reyes-Garcia J, Garcia-Hernandez LM, Figueroa A, Checa M, Barajas-Lopez C, Montaño LM (2011) In airways ATP refills sarcoplasmic reticulum via P2X smooth muscle receptors and induces contraction through P2Y epithelial receptors. Pflugers Arch 461:261–275

  13. Flores-Soto E, Reyes-Garcia J, Sommer B, Montaño LM (2013) Sarcoplasmic reticulum Ca2+ refilling is determined by L-type Ca2+ and store operated Ca2+ channels in guinea pig airway smooth muscle. Eur J Pharmacol 721:21–28

  14. Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450

    CAS  PubMed  Google Scholar 

  15. Hall J, Jones RD, Jones TH, Channer KS, Peers C (2006) Selective inhibition of L-type Ca2+ channels in A7r5 cells by physiological levels of testosterone. Endocrinology 147:2675–2680

    Article  CAS  PubMed  Google Scholar 

  16. Helli PB, Janssen LJ (2008) Properties of a store-operated nonselective cation channel in airway smooth muscle. Eur Respir J 32:1529–1539

    Article  CAS  PubMed  Google Scholar 

  17. Janssen LJ, Betti PA, Netherton SJ, Walters DK (1999) Superficial buffer barrier and preferentially directed release of Ca2+ in canine airway smooth muscle. Am J Physiol 276:L744–L753

    CAS  PubMed  Google Scholar 

  18. Jones RD, English KM, Jones TH, Channer KS (2004) Testosterone-induced coronary vasodilatation occurs via a non-genomic mechanism: evidence of a direct calcium antagonism action. Clin Sci (Lond) 107:149–158

    Article  CAS  Google Scholar 

  19. Kajita J, Yamaguchi H (1993) Calcium mobilization by muscarinic cholinergic stimulation in bovine single airway smooth muscle. Am J Physiol 264:L496–L503

    CAS  PubMed  Google Scholar 

  20. Kim MH, Seo JB, Burnett LA, Hille B, Koh DS (2013) Characterization of store-operated Ca2+ channels in pancreatic duct epithelia. Cell Calcium 54:266–275

    Article  CAS  PubMed  Google Scholar 

  21. Kouloumenta V, Hatziefthimiou A, Paraskeva E, Gourgoulianis K, Molyvdas PA (2006) Non-genomic effect of testosterone on airway smooth muscle. Br J Pharmacol 149:1083–1091

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Kubli-Garfias C, Medina-Jimenez M, Garcia-Yanez E, Vazquez-Alvarez A, Perusquia M, Gomez-Garcia N, Almanza J, Ibanez R, Rodriguez R (1987) Relaxant action of androgens, progestins and corticosteroids on the isolated ileum of the guinea pig. Acta Physiol Pharmacol Latinoam 37:357–364

    CAS  PubMed  Google Scholar 

  23. Malkin CJ, Jones RD, Jones TH, Channer KS (2006) Effect of testosterone on ex vivo vascular reactivity in man. Clin Sci (Lond) 111:265–274

    Article  CAS  Google Scholar 

  24. Maruyama T, Kanaji T, Nakade S, Kanno T, Mikoshiba K (1997) 2APB, 2-aminoethoxydiphenyl borate, a membrane-penetrable modulator of Ins(1,4,5)P3-induced Ca2+ release. J Biochem 122:498–505

    Article  CAS  PubMed  Google Scholar 

  25. Montaño LM, Calixto E, Figueroa A, Flores-Soto E, Carbajal V, Perusquia M (2008) Relaxation of androgens on rat thoracic aorta: testosterone concentration dependent agonist/antagonist L-type Ca2+ channel activity, and 5β-dihydrotestosterone restricted to L-type Ca2+ channel blockade. Endocrinology 149:2517–2526

  26. Montaño LM, Cruz-Valderrama JE, Figueroa A, Flores-Soto E, Garcia-Hernandez LM, Carbajal V, Segura P, Mendez C, Diaz V, Barajas-Lopez C (2011) Characterization of P2Y receptors mediating ATP induced relaxation in guinea pig airway smooth muscle: involvement of prostaglandins and K+ channels. Pflugers Arch 462:573–585

  27. Montaño LM, Espinoza J, Flores-Soto E, Chavez J, Perusquia M (2014) Androgens are bronchoactive drugs by relaxing airway smooth muscle and preventing bronchospasm. J Endocrinol. doi:10.1530/JOE-14-0074

  28. Navarro-Dorado J, Orensanz LM, Recio P, Bustamante S, Benedito S, Martinez AC, Garcia-Sacristan A, Prieto D, Hernandez M (2008) Mechanisms involved in testosterone-induced vasodilatation in pig prostatic small arteries. Life Sci 83:569–573

    Article  CAS  PubMed  Google Scholar 

  29. Ong HL, Brereton HM, Harland ML, Barritt GJ (2003) Evidence for the expression of transient receptor potential proteins in guinea pig airway smooth muscle cells. Respirology 8:23–32

    Article  PubMed  Google Scholar 

  30. Peel SE, Liu B, Hall IP (2008) ORAI and store-operated calcium influx in human airway smooth muscle cells. Am J Respir Cell Mol Biol 38:744–749

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  31. Perusquia M (2003) Androgen-induced vasorelaxation: a potential vascular protective effect. Exp Clin Endocrinol Diabetes 111:55–59

    Article  CAS  PubMed  Google Scholar 

  32. Perusquia M, Espinoza J, Montaño LM, Stallone JN (2012) Regional differences in the vasorelaxing effects of testosterone and its 5-reduced metabolites in the canine vasculature. Vasc Pharmacol 56:176–182

  33. Perusquia M, Garcia-Yanez E, Ibanez R, Kubli-Garfias C (1990) Non-genomic mechanism of action of delta-4 and 5-reduced androgens and progestins on the contractility of the isolated rat myometrium. Life Sci 47:1547–1553

    Article  CAS  PubMed  Google Scholar 

  34. Perusquia M, Hernandez R, Morales MA, Campos MG, Villalon CM (1996) Role of endothelium in the vasodilating effect of progestins and androgens on the rat thoracic aorta. Gen Pharmacol 27:181–185

    Article  CAS  PubMed  Google Scholar 

  35. Perusquia M, Navarrete E, Gonzalez L, Villalon CM (2007) The modulatory role of androgens and progestins in the induction of vasorelaxation in human umbilical artery. Life Sci 81:993–1002

    Article  CAS  PubMed  Google Scholar 

  36. Perusquia M, Navarrete E, Jasso-Kamel J, Montaño LM (2005) Androgens induce relaxation of contractile activity in pregnant human myometrium at term: a nongenomic action on L-type calcium channels. Biol Reprod 73:214–221

  37. Perusquia M, Stallone JN (2010) Do androgens play a beneficial role in the regulation of vascular tone? Nongenomic vascular effects of testosterone metabolites. Am J Physiol Heart Circ Physiol 298:H1301–H1307

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  38. Rowell KO, Hall J, Pugh PJ, Jones TH, Channer KS, Jones RD (2009) Testosterone acts as an efficacious vasodilator in isolated human pulmonary arteries and veins: evidence for a biphasic effect at physiological and supra-physiological concentrations. J Endocrinol Investig 32:718–723

    Article  CAS  Google Scholar 

  39. Sanchez Aparicio JA, Gutierrez M, Hidalgo A, Cantabrana B (1993) Effects of androgens on isolated rat uterus. Life Sci 53:269–274

    Article  CAS  PubMed  Google Scholar 

  40. Scragg JL, Dallas ML, Peers C (2007) Molecular requirements for L-type Ca2+ channel blockade by testosterone. Cell Calcium 42:11–15

    Article  CAS  PubMed  Google Scholar 

  41. Scragg JL, Jones RD, Channer KS, Jones TH, Peers C (2004) Testosterone is a potent inhibitor of L-type Ca2+ channels. Biochem Biophys Res Commun 318:503–506

    Article  CAS  PubMed  Google Scholar 

  42. Seyrek M, Yildiz O, Ulusoy HB, Yildirim V (2007) Testosterone relaxes isolated human radial artery by potassium channel opening action. J Pharmacol Sci 103:309–316

    Article  CAS  PubMed  Google Scholar 

  43. Tep-areenan P, Kendall DA, Randall MD (2002) Testosterone-induced vasorelaxation in the rat mesenteric arterial bed is mediated predominantly via potassium channels. Br J Pharmacol 135:735–740

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  44. Tep-areenan P, Kendall DA, Randall MD (2003) Mechanisms of vasorelaxation to testosterone in the rat aorta. Eur J Pharmacol 465:125–132

    Article  CAS  PubMed  Google Scholar 

  45. Tilley SL, Hartney JM, Erikson CJ, Jania C, Nguyen M, Stock J, McNeisch J, Valancius C, Panettieri RA Jr, Penn RB, Koller BH (2003) Receptors and pathways mediating the effects of prostaglandin E2 on airway tone. Am J Physiol Lung Cell Mol Physiol 284:L599–L606

    Article  CAS  PubMed  Google Scholar 

  46. van Breemen C, Chen Q, Laher I (1995) Superficial buffer barrier function of smooth muscle sarcoplasmic reticulum. Trends Pharmacol Sci 16:98–105

    Article  PubMed  Google Scholar 

  47. Xiao JH, Zheng YM, Liao B, Wang YX (2010) Functional role of canonical transient receptor potential 1 and canonical transient receptor potential 3 in normal and asthmatic airway smooth muscle cells. Am J Respir Cell Mol Biol 43:17–25

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  48. Yildiz O, Seyrek M, Gul H, Un I, Yildirim V, Ozal E, Uzun M, Bolu E (2005) Testosterone relaxes human internal mammary artery in vitro. J Cardiovasc Pharmacol 45:580–585

    Article  CAS  PubMed  Google Scholar 

  49. Zhou H, Iwasaki H, Nakamura T, Nakamura K, Maruyama T, Hamano S, Ozaki S, Mizutani A, Mikoshiba K (2007) 2-Aminoethyl diphenylborinate analogues: selective inhibition for store-operated Ca2+ entry. Biochem Biophys Res Commun 352:277–282

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This study was partly supported by grants from the Dirección General de Asuntos del Personal Académico-Universidad Nacional Autónoma de México (IN200613-3 and IN205511-3).

Author information

Authors and Affiliations

  1. Departamento de Biología Celular y Fisiología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, México, DF, México

    Mercedes Perusquía

  2. Departamento de Farmacología, Facultad de Medicina, Universidad Nacional Autónoma de México, México, DF, México

    Edgar Flores-Soto, Elias Campuzano-González, Inocencio Martínez-Villa, Aldo I. Martínez-Banderas & Luis M. Montaño

  3. Departamento de Investigación en Hiperreactividad Bronquial, Instituto Nacional de Enfermedades Respiratorias, México, DF, México

    Bettina Sommer

Authors
  1. Mercedes Perusquía
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Edgar Flores-Soto
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bettina Sommer
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Elias Campuzano-González
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Inocencio Martínez-Villa
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Aldo I. Martínez-Banderas
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Luis M. Montaño
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Luis M. Montaño.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Perusquía, M., Flores-Soto, E., Sommer, B. et al. Testosterone-induced relaxation involves L-type and store-operated Ca2+ channels blockade, and PGE2 in guinea pig airway smooth muscle. Pflugers Arch - Eur J Physiol 467, 767–777 (2015). https://doi.org/10.1007/s00424-014-1534-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00424-014-1534-y

Keywords

Search

Navigation