Characterization of P2Y receptors mediating ATP induced relaxation in guinea pig airway smooth muscle: involvement of prostaglandins and K+ channels

Pflügers Archiv - Tập 462 - Trang 573-585 - 2011
Luis M. Montaño1, José E. Cruz-Valderrama1, Alejandra Figueroa1, Edgar Flores-Soto1, Luz M. García-Hernández1, Verónica Carbajal2, Patricia Segura2, Carmen Méndez3, Verónica Díaz3, Carlos Barajas-López4
1Departamento de Farmacología, Edificio de Investigación, sexto piso, laboratorio 3, Facultad de Medicina, Universidad Nacional Autónoma de México, Ciudad Universitaria, Mexico, Mexico
2Departamento de Investigación en Hiperreactividad Bronquial, Instituto Nacional de Enfermedades Respiratorias, Mexico, Mexico
3Departamento de Embriología, Facultad de Medicina, Universidad Nacional Autónoma de México, Mexico, Mexico
4División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológíca, San Luis Potosí, Mexico

Tóm tắt

In airway smooth muscle (ASM), adenosine 5′-triphosphate (ATP) induces a relaxation associated with prostaglandin production. We explored the role of K+ currents (I K) in this relaxation. ATP relaxed the ASM, and this effect was abolished by indomethacin. Removal of airway epithelium slightly diminished the ATP-induced relaxation at lower concentration without modifying the responses to ATP at higher concentrations. ATPγS and UTP induced a concentration-dependent relaxation similar to ATP; α,β-methylene-ATP was inactive from 1 to 100 μM. Suramin or reactive blue 2 (RB2), P2Y receptor antagonists, did not modify the relaxation, but their combination significantly reduced this effect of ATP. The relaxation was also inhibited by N-ethylmaleimide (NEM; which uncouples G proteins). In myocytes, the ATP-induced I K increment was not modified by suramin or RB2 but the combination of both drugs abolished it. This increment in the I K was also completely nullified by NEM and SQ 22,536. 4-Amynopyridine or iberiotoxin diminished the ATP-induced I K increment, and the combination of both substances diminished ATP-induced relaxation. The presence of P2Y2 and P2Y4 receptors in smooth muscle was corroborated by Western blot and confocal images. In conclusion, ATP: (1) produces relaxation by inducing the production of bronchodilator prostaglandins in airway smooth muscle, most likely by acting on P2Y4 and P2Y2 receptors; (2) induces I K increment through activation of the delayed rectifier K+ channels and the high-conductance Ca2+-dependent K+ channels, therefore both channels are implicated in the ATP-induced relaxation; and (3) this I K increment is mediated by prostaglandin production which in turns increase cAMP signaling pathway.

Tài liệu tham khảo

Abbracchio MP, Burnstock G, Boeynaems JM, Barnard EA, Boyer JL, Kennedy C, Knight GE, Fumagalli M, Gachet C, Jacobson KA, Weisman GA (2006) International Union of Pharmacology LVIII: update on the P2Y G protein-coupled nucleotide receptors: from molecular mechanisms and pathophysiology to therapy. Pharmacol Rev 58:281–341 Aiello EA, Malcolm AT, Walsh MP, Cole WC (1998) β-Adrenoceptor activation and pKa regulate delayed rectifier K+ channels of vascular smooth muscle cells. Am J Physiol 275:H448–459 Aksoy MO, Kelsen SG (1994) Relaxation of rabbit tracheal smooth muscle by adenine nucleotides: mediation by P2-purinoceptors. Am J Respir Cell Mol Biol 10:230–236 Bergner A, Sanderson MJ (2002) ATP stimulates Ca2+ oscillations and contraction in airway smooth muscle cells of mouse lung slices. Am J Physiol Lung Cell Mol Physiol 283:L1271–1279 Bramley AM, Samhoun MN, Piper PJ (1990) The role of the epithelium in modulating the responses of guinea-pig trachea induced by bradykinin in vitro. Br J Pharmacol 99:762–766 Breslin JW, Pappas PJ, Cerveira JJ, Hobson RW 2nd, Duran WN (2003) VEGF increases endothelial permeability by separate signaling pathways involving ERK-1/2 and nitric oxide. Am J Physiol Heart Circ Physiol 284:H92–H100 Burnstock G (2007) Physiology and pathophysiology of purinergic neurotransmission. Physiol Rev 87:659–797 Cole WC, Clement-Chomienne O, Aiello EA (1996) Regulation of 4-aminopyridine-sensitive, delayed rectifier K+ channels in vascular smooth muscle by phosphorylation. Biochem Cell Biol 74:439–447 Flores-Soto E, Carbajal V, Reyes-Garcia J, Garcia-Hernandez LM, Figueroa A, Checa M, Barajas-Lopez C, Montano 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 Fortner CN, Breyer RM, Paul RJ (2001) EP2 receptors mediate airway relaxation to substance P, ATP, and PGE2. Am J Physiol Lung Cell Mol Physiol 281:L469–474 Govindaraju V, Martin JG, Maghni K, Ferraro P, Michoud MC (2005) The effects of extracellular purines and pyrimidines on human airway smooth muscle cells. J Pharmacol Exp Ther 315:941–948 Kamikawa Y, Shimo Y (1976) Mediation of prostaglandin E2 in the biphasic response to ATP of the isolated tracheal muscle of guinea-pigs. J Pharm Pharmacol 28:294–297 King BF, Townsend-Nicholson A (2008) Involvement of P2Y1 and P2Y11 purinoceptors in parasympathetic inhibition of colonic smooth muscle. J Pharmacol Exp Ther 324:1055–1063 Koh SD, Dick GM, Sanders KM (1997) Small-conductance Ca2+-dependent K+ channels activated by ATP in murine colonic smooth muscle. Am J Physiol 273:C2010–2021 Koh SD, Sanders KM, Carl A (1996) Regulation of smooth muscle delayed rectifier K+ channels by protein kinase A. Pflugers Arch 432:401–412 Liu B, Freyer AM, Hall IP (2007) Bradykinin activates calcium-dependent potassium channels in cultured human airway smooth muscle cells. Am J Physiol Lung Cell Mol Physiol 292:L898–907 Lyubchenko T, Woodward H, Veo KD, Burns N, Nijmeh H, Liubchenko GA, Stenmark KR, Gerasimovskaya EV (2011) P2Y1 and P2Y13 purinergic receptors mediate Ca2+ signaling and proliferative responses in pulmonary artery vasa vasorum endothelial cells. Am J Physiol Cell Physiol 300:C266–275 Miyake R, Gross RW (1992) Multiple phospholipase A2 activities in canine vascular smooth muscle. Biochim Biophys Acta 1165:167–176 Mounkaila B, Marthan R, Roux E (2005) Biphasic effect of extracellular ATP on human and rat airways is due to multiple P2 purinoceptor activation. Respir Res 6:143 North RA (2002) Molecular physiology of P2X receptors. Physiol Rev 82:1013–1067 Piper AS, Hollingsworth M (1996) ATP and β, γ-methylene ATP produce relaxation of guinea-pig isolated trachealis muscle via actions at P1 purinoceptors. Eur J Pharmacol 307:183–189 Ralevic V (2001) Mechanism of prolonged vasorelaxation to ATP in the rat isolated mesenteric arterial bed. Br J Pharmacol 132:685–692 Ruan YC, Wang Z, Du JY, Zuo WL, Guo JH, Zhang J, Wu ZL, Wong HY, Chung YW, Chan HC, Zhou WL (2008) Regulation of smooth muscle contractility by the epithelium in rat vas deferens: role of ATP-induced release of PGE2. J Physiol 586:4843–4857 Serir K, Hayoz S, Fanchaouy M, Beny JL, Bychkov R (2006) A delayed ATP-elicited K+ current in freshly isolated smooth muscle cells from mouse aorta. Br J Pharmacol 147:45–54 Shapiro MS, Wollmuth LP, Hille B (1994) Modulation of Ca2+ channels by PTX-sensitive G-proteins is blocked by N-ethylmaleimide in rat sympathetic neurons. J Neurosci 14:7109–7116 Smith MM, Harden TK (1984) Modification of receptor-mediated inhibition of adenylate cyclase in NG108-15 neuroblastoma X glioma cells by n-ethylmaleimide. J Pharmacol Exp Ther 228:425–433 Snetkov VA, Ward JP (1999) Ion currents in smooth muscle cells from human small bronchioles: presence of an inward rectifier K+ current and three types of large conductance K+ channel. Exp Physiol 84:835–846 Tanaka Y, Yamashita Y, Yamaki F, Horinouchi T, Shigenobu K, Koike K (2003) MaxiK channel mediates β2-adrenoceptor-activated relaxation to isoprenaline through cAMP-dependent and -independent mechanisms in guinea-pig tracheal smooth muscle. J Smooth Muscle Res 39:205–219 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:L599–606 Van Crombruggen K, Van Nassauw L, Timmermans JP, Lefebvre RA (2007) Inhibitory purinergic P2 receptor characterisation in rat distal colon. Neuropharmacology 53:257–271 von Kugelgen I (2006) Pharmacological profiles of cloned mammalian P2Y-receptor subtypes. Pharmacol Ther 110:415–432 Wihlborg AK, Wang L, Braun OO, Eyjolfsson A, Gustafsson R, Gudbjartsson T, Erlinge D (2004) ADP receptor P2Y12 is expressed in vascular smooth muscle cells and stimulates contraction in human blood vessels. Arterioscler Thromb Vasc Biol 24:1810–1815 Wildman SS, Unwin RJ, King BF (2003) Extended pharmacological profiles of rat P2Y2 and rat P2Y4 receptors and their sensitivity to extracellular H+ and Zn2+ ions. Br J Pharmacol 140:1177–1186 Zhao LM, Xu YJ, Zhang ZX, Ni W, Chen SX (2004) Effect of passive sensitization by serum from allergic asthmatic patients on the activity and expression of voltage-dependent delayed rectifier potassium channel in human bronchial smooth muscle cells. Chin Med J (Engl) 117:1630–1636