Study of the mechanisms regulating human umbilical artery contractility
António José Santos-Silva, Elisa Cairrao, Ignacio Verde
DOI: 10.4236/health.2010.24049   PDF   HTML     5,365 Downloads   9,319 Views   Citations


We studied the involvement of different types of Ca2+ channels, cyclic nucleotides and different kinases in the regulation of human umbilical artery (HUA) contractility. The elucidation of the precise mechanisms regulating the contractility of this artery could be very important to reveal potential therapeutic targets to treat HUA disorders such as preeclampsia. The relevancy of different types of Ca2+ channels on the regulation of HUA tonus was analyzed. Among the different Ca2+ channel inhibitors used, only the L-type calcium channels (LTCC) inhibition induced relaxation of HUA in Ca2+ containing medium. The inhibition of T-type calcium channels (TTCC) or TRP channels did not significantly affect HUA contractility. In presence of Ca2+, the intracellular increase of a cyclic nucleotide (cAMP or cGMP) induces relaxation of HUA, which was almost complete in histamine-con- tracted HUA, and lower effect was observed in arteries contracted by KCl and serotonin (5-HT). Inhibition of PKA and PKG weakly reduced the relaxations induced by the increase of cAMP and cGMP respectively, suggesting that the relaxation induced by these nucleotides is not totally mediated by the activation of their respective kinases and that other mechanisms are involved. In calcium containing solution, PP2A inhibition produces relaxation of contracted HUA. In KCl contracted arteries, the OA and nifedipine relaxant effects are similar and not additive, suggesting that PP2A could activate LTCC. Besides, the increase of cyclic nucleotides significantly increased the OA effect, suggesting that the effect of PP2A inhibition is independent of the cyclic nucleotide pathways. The contractions induced by KCl, histamine and 5-HT in presence of Ca2+ were not significantly affected by ROCK, ERK1/2 or p38MAPK inhibitors. In absence of extracellular Ca2+, histamine and 5-HT elicited contractions of HUA characterized by two components, a rapid phasic contractile component followed by a decrease of the contraction until a tonic component. However, KCl elicited sustained contractions of HUA in absence of extracellular Ca2+. As in presence of calcium, the ERK1/2 and p38MAPK inhibitors did not influence the contractions induced by KCl, histamine or 5-HT in absence of extracellular Ca2+. However, in these conditions, ROCK inhibition significantly relaxed the contractions induced by KCl and reduced the phasic and tonic components of the contraction elicited either by histamine or 5-HT. Our results show that calcium-dependent contractions of HUA depend on Ca2+ entry by LTCC, and these chan- nels seems to be positive regulated by PP2A. Cyclic nucleotides mediate HUA vasodilatation but their dependent kinases are not the unique responsible of this effect. HUA is able to contract independently of Ca2+ influx by activating the ROCK pathway and/or due to intracellular Ca2+ release.

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Santos-Silva, A. , Cairrao, E. and Verde, I. (2010) Study of the mechanisms regulating human umbilical artery contractility. Health, 2, 321-331. doi: 10.4236/health.2010.24049.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Tufan, H., Ayan-Polat, B, Tecder-Unal, M, Polat, G, Kayhan, Z and Ogus, E. (2003) Contractile responses of the human umbilical artery to KCl and serotonin in Ca-free medium and the effects of levcromakalim. Life Sciences, 72, 1321-1329.
[2] Leung, S.W., Quan, A., Lao, T.T. and Man, R.Y. (2006) Efficacy of different vasodilators on human umbilical arterial smooth muscle under normal and reduced oxygen conditions. Early Human Development, 82, 457-462.
[3] Michael, S.K., Surks, H.K., Wang, Y., Zhu, Y., Blanton, R., Jamnongjit, M., Aronovitz, M., Baur, W., Ohtani, K., Wilkerson, M.K., Bonev, A.D., Nelson, M.T., Karas, R.H. and Mendelsohn, M.E. (2008) High blood pressure arising from a defect in vascular function. Proceedings of the National Academy of Sciences of the United States of America, 105, 6702-6707.
[4] McCarron, J.G., Craig, J.W., Bradley, K.N. and Muir, T.C. (2002) Agonist-induced phasic and tonic responses in smooth muscle are mediated by InsP(3). Journal of Cell Science, 115, 2207-2218.
[5] Hilgers, R.H. and Webb, R.C. (2005) Molecular aspects of arterial smooth muscle contraction: Focus on Rho. Experimental Biology and Medicine (Maywood), 230, 829-835.
[6] Somara, S. and Bitar, K.N. (2006) Phosphorylated HSP27 modulates the association of phosphorylated cal- desmon with tropomyosin in colonic smooth muscle. American Journal of Physiology - Gastrointestinal and Liver Physiology, 291, G630-639.
[7] Hall, J., Jones, T.H., Channer, K.S. and Jones, R.D. (2006) Mechanisms of agonist-induced constriction in isolated human mesenteric arteries. Vascular Pharmacology, 44, 427-433.
[8] Mikkelsen, E.O., Sakr, A.M. and Jespersen, L.T. (1983) Effects of nifedipine on contractile responses to potassium, histamine, and 5-hydroxytryptamine in isolated human pulmonary vessels. Journal of Cardiovascular Pharmacology, 5, 317-320.
[9] Potocnik, S.J., Murphy, T.V., Kotecha, N. and Hill, M.A. (2000) Effects of mibefradil and nifedipine on arteriolar myogenic responsiveness and intracellular Ca(2+). British Journal of Pharmacology, 131, 1065-1072.
[10] Liu, M., Large, W.A. and Albert, A.P. (2005) Stimulation of beta-adrenoceptors inhibits store-operated channel currents via a cAMP-dependent protein kinase mechanism in rabbit portal vein myocytes. Journal of Physiology, 562, 395-406.
[11] Muraki, K., Iwata, Y., Katanosaka, Y., Ito, T., Ohya, S., Shigekawa, M. and Imaizumi, Y. (2003) TRPV2 is a component of osmotically sensitive cation channels in murine aortic myocytes. Circulation Research, 93, 829- 838.
[12] Park, K.S., Lee, H.A., Earm, K.H., Ko, J.H., Earm, Y.E. and Kim, S.J. (2006) Differential distribution of mechanosensitive nonselective cation channels in systemic and pulmonary arterial myocytes of rabbits. Journal of Vascular Research, 43, 347-354.
[13] Dietrich, A., Chubanov, V., Kalwa, H., Rost, B.R. and Gudermann, T. (2006) Cation channels of the transient receptor potential superfamily: Their role in physiological and pathophysiological processes of smooth muscle cells. Pharmacology & Therapeutics.
[14] Beech, D.J., Muraki, K. and Flemming, R. (2004) Non- selective cationic channels of smooth muscle and the mammalian homologues of Drosophila TRP. J Physiol, 559, 685-706. Li, S., Gosling, M. and Poll, C. (2005) Determining the functional role of TRPC channels in primary cells. Pflugers Archiv (European Journal of Physiology), 451, 43-52.
[15] Cribbs, L.L. (2006) T-type Ca2+ channels in vascular smooth muscle: Multiple functions. Cell Calcium, 40, 221-230.
[16] Akaike, N., Kanaide, H., Kuga, T., Nakamura, M., Sadoshima, J. and Tomoike, H. (1989) Low-voltage-activated calcium current in rat aorta smooth muscle cells in primary culture. Journal of Physiology, 416, 141-160.
[17] Salemme, S., Rebolledo, A., Speroni, F., Petruccelli, S. and Milesi, V. (2007) L, P-/Q- and T-type Ca2+ channels in smooth muscle cells from human umbilical artery. Cellular Physiology and Biochemistry, 20, 55-64.
[18] Belfort, M.A., Saade, G.R., Suresh, M., Johnson, D. and Vedernikov, Y.P. (1995) Human umbilical vessels: responses to agents frequently used in obstetric patients. American Journal of Obstetrics and Gynecology, 172, 1395-1403.
[19] Obara, K., Takai, A., Ruegg, J.C. and de Lanerolle, P. (1989) Okadaic acid, a phosphatase inhibitor, produces a Ca2+ and calmodulin-independent contraction of smooth muscle. Pflugers Archiv (European Journal of Physiology), 414, 134-138.
[20] Obara, K. and Yabu, H. (1993) Dual effect of phosphatase inhibitors on calcium channels in intestinal smooth muscle cells. American Journal of Physiology, 264, C296-301.
[21] Groschner, K., Schuhmann, K., Mieskes, G., Baumgart- ner, W. and Romanin, C. (1996) A type 2A phosphatase- sensitive phosphorylation site controls modal gating of L-type Ca2+ channels in human vascular smooth-muscle cells. Biochemical Journal, 318, 513-517.
[22] Ark, M., Ozveren, E., Yazici, G., Korkmaz, B., Buyukafsar, K., Arikan, O., Kubat, H. and Songu-Mize, E. (2004) Effects of HA-1077 and Y-27632, two rho-kinase inhibitors, in the human umbilical artery. Cell Biochemistry and Biophysics, 41, 331-342
[23] Sakurada, S., Takuwa, N., Sugimoto, N., Wang, Y., Seto, M., Sasaki, Y. and Takuwa, Y. (2003) Ca2+-dependent activation of Rho and Rho kinase in membrane depolarization-induced and receptor stimulation-induced vascular smooth muscle contraction. Circulation Research, 93, 548-556.
[24] Ratz, P.H. and Miner, A.S. (2009) Role of protein kinase Czeta and calcium entry in KCl-induced vascular smooth muscle calcium sensitization and feedback control of cellular calcium levels. Journal of Pharmacology and Experimental Therapeutics, 328, 399-408.
[25] Zhao, Y., Long, W., Zhang, L. and Longo, L.D. (2003) Extracellular signal-regulated kinases and contractile responses in ovine adult and fetal cerebral arteries. Journal of Physiology, 551, 691-703.
[26] Muslin, A.J. (2008) MAPK signalling in cardiovascular health and disease: molecular mechanisms and therapeutic targets. Clinical Science (London), 115, 203-218.
[27] Somlyo, A.P. and Somlyo, A.V. (1994) Signal transduction and regulation in smooth muscle. Nature, 372, 231-236.
[28] Watts, S.W. (1996) Serotonin activates the mitogen-activated protein kinase pathway in vascular smooth muscle: Use of the mitogen-activated protein kinase inhibitor PD098059. Journal of Pharmacology and Experimental Therapeutics, 279, 1541-1550.
[29] Kim, B., Kim, J., Bae, Y.M., Cho, S.I., Kwon, S.C., Jung, J.Y., Park, J.C. and Ahn, H.Y. (2004) p38 mitogen-activated protein kinase contributes to the diminished aortic contraction by endothelin-1 in DOCA-salt hypertensive rats. Hypertension, 43, 1086-1091.
[30] Tasaki, K., Hori, M., Ozaki, H., Karaki, H. and Wakabayashi, I. (2003) Difference in signal transduction mechanisms involved in 5-hydroxytryptamine- and U46619-induced vasoconstrictions. Journal of Smooth Muscle Research, 39, 107-117.
[31] Banes, A., Florian, J.A. and Watts, S.W. (1999) Mechanisms of 5-hydroxytryptamine(2A) receptor activation of the mitogen-activated protein kinase pathway in vascular smooth muscle. Journal of Pharmacology and Experimental Therapeutics, 291, 1179-1187.
[32] Matsumoto, T., Kobayashi, T. and Kamata, K. (2003) Phosphodiesterases in the vascular system. Journal of Smooth Muscle Research, 39, 67-86.
[33] Bentley, J.K. and Beavo, J.A. (1992) Regulation and function of cyclic nucleotides. Current Opinion in Cell Biology, 4, 233-240.
[34] Segarra, G., Medina, P., Vila, J.M., Martinez-Leon, J.B., Domenech, C., Prieto, F. and Lluch, S. (2002) Relaxation induced by milrinone and rolipram in human penile arteries and veins. European Journal of Pharmacology, 444, 103-106.
[35] Rabe, K.F., Tenor, H., Dent, G., Schudt, C., Nakashima, M. and Magnussen, H. (1994) Identification of PDE isozymes in human pulmonary artery and effect of selective PDE inhibitors. American Journal of Physiology, 266, L536-543.
[36] Figueroa, R., Martinez, E., Fayngersh, R.P., Tejani, N., Mohazzab, H.K. and Wolin, M.S. (2000) Alterations in relaxation to lactate and H2O2 in human placental vessels from gestational diabetic pregnancies. American Journal of Physiology—Heart and Circulatory Physiology, 278, H706-713.
[37] Santos-Silva, A.J., Cairrão, E., Morgado, M., Álvarez, E. and Verde, I. (2008) PDE4 and PDE5 regulate cyclic nucleotides relaxing effects in human umbilical arteries. European Journal of Pharmacology, 582, 102-109.
[38] Kristek, F., Koprdova, R. and Cebova, M. (2007) Long- term effects of early administered sildenafil and NO donor on the cardiovascular system of SHR. Journal of Physiology and Pharmacology, 58, 33-43.
[39] Lovren, F. and Triggle, C. (2000) Nitric oxide and sodium nitroprusside-induced relaxation of the human umbilical artery. British Journal of Pharmacology, 131, 521-529.
[40] Wu, C.C., Chen, S.J. and Yen, M.H. (1999) Cyclic GMP regulates cromakalim-induced relaxation in the rat aortic smooth muscle: role of cyclic GMP in KATP-channels. Life Sciences, 64, 2471-2478.
[41] Li, P.L., Jin, M.W. and Campbell, W.B. (1998) Effect of selective inhibition of soluble guanylyl cyclase on the KCa channel activity in coronary artery smooth muscle. Hypertension, 31, 303-308.
[42] Lincoln, T.M., Cornwell, T.L. and Taylor, A.E. (1990) cGMP-dependent protein kinase mediates the reduction of Ca2+ by cAMP in vascular smooth muscle cells. American Journal of Physiology, 258, C399-407; Murray, K.J. (1990) Cyclic AMP and mechanisms of vasodilation. Pharmacology & Therapeutics, 47, 329-345.
[43] Keef, K.D., Hume, J.R. and Zhong, J. (2001) Regulation of cardiac and smooth muscle Ca2+ channels (Ca(V) 1.2a,b) by protein kinases. American Journal of Physiology, 281, C1743-1756.
[44] Sausbier, M., Schubert, R., Voigt, V., Hirneiss, C., Pfeifer, A., Korth, M., Kleppisch, T., Ruth, P. and Hofmann, F. (2000) Mechanisms of NO/cGMP-dependent vasorelaxation. Circulation Research, 87, 825-830.
[45] Jensen, J. (2007) More PKA independent beta-adrenergic signalling via cAMP: Is Rap1-mediated glucose uptake in vascular smooth cells physiologically important? British Journal of Pharmacology, 151, 423-425.
[46] Cairrao, E., Alvarez, E., Santos-Silva, A.J. and Verde, I. (2008) Potassium channels are involved in testosterone-induced vasorelaxation of human umbilical artery. Naunyn-Schmiedeber’s Archives of Pharmacology, 376, 375-383.
[47] Tosun, M., Paul, R.J. and Rapoport, R.M. (1998) Coupling of store-operated Ca2+ entry to contraction in rat aorta. Journal of Pharmacology and Experimental Therapeutics, 285, 759-766.
[48] Santos-Silva, A.J., Cairrao, E., Marques, B. and Verde, I. (2009) Regulation of human umbilical artery contractility by different serotonin and histamine receptors. Reproductive Sciences, 16, 1175-1185.
[49] Jarajapu, Y.P., Oomen, C., Uteshev, V.V. and Knot, H.J. (2006) Histamine decreases myogenic tone in rat cerebral arteries by H2-receptor-mediated KV channel activation, independent of endothelium and cyclic AMP. European Journal of Pharmacology, 547, 116-124.
[50] Cogolludo, A., Moreno, L., Lodi, F., Frazziano, G., Cobeno, L., Tamargo, J. and Perez-Vizcaino, F. (2006) Serotonin inhibits voltage-gated K+ currents in pulmonary artery smooth muscle cells: Role of 5-HT2A receptors, caveolin-1, and KV1.5 channel internalization. Circulation Research, 98, 931-938.
[51] Bae, Y.M., Kim, A., Kim, J., Park, S.W., Kim, T.K., Lee, Y.R., Kim, B. and Cho, S.I. (2006) Serotonin depolarizes the membrane potential in rat mesenteric artery myocytes by decreasing voltage-gated K+ currents. Biochemical and Biophysical Research Communications, 347, 468- 476.
[52] Hirano, K., Derkach, D.N., Hirano, M., Nishimura, J. and Kanaide, H. (2003) Protein kinase network in the regulation of phosphorylation and dephosphorylation of smooth muscle myosin light chain. Molecular and Cellular Biochemistry, 248, 105-114.

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