Quercetin Inhibits the 5-Hydroxytryptamine Type 3 Receptor-mediated Ion Current by Interacting with Pre-Transmembrane Domain I

  • Lee, Byung-Hwan (Department of Physiology, College of Veterinary Medicine and Bio/Molecular Informatics Center, Konkuk University) ;
  • Jung, Sang-Min (Department of Physiology, College of Veterinary Medicine and Bio/Molecular Informatics Center, Konkuk University) ;
  • Lee, Jun-Ho (Department of Physiology, College of Veterinary Medicine and Bio/Molecular Informatics Center, Konkuk University) ;
  • Kim, Jong-Hoon (Department of Physiology, College of Veterinary Medicine and Bio/Molecular Informatics Center, Konkuk University) ;
  • Yoon, In-Soo (Department of Physiology, College of Veterinary Medicine and Bio/Molecular Informatics Center, Konkuk University) ;
  • Lee, Joon-Hee (Department of Physiology, College of Veterinary Medicine and Bio/Molecular Informatics Center, Konkuk University) ;
  • Choi, Sun-Hye (Department of Physiology, College of Veterinary Medicine and Bio/Molecular Informatics Center, Konkuk University) ;
  • Lee, Sang-Mok (Department of Physiology, College of Veterinary Medicine and Bio/Molecular Informatics Center, Konkuk University) ;
  • Chang, Choon-Gon (Department of Pharmacology, College of Pharmacy, Sungkyunkwan University) ;
  • Kim, Hyung-Chun (College of Pharmacy, Kangwon National University) ;
  • Han, YeSun (Department of Advanced Technology Fusion and Bio/Molecular Informatics Center, Konkuk University) ;
  • Paik, Hyun-Dong (Division of Animal Life Science and Bio/Molecular Informatics Center, Konkuk University) ;
  • Kim, Yangmee (Department of Chemistry and Bio/Molecular Informatics Center, Konkuk University) ;
  • Nah, Seung-Yeol (Department of Physiology, College of Veterinary Medicine and Bio/Molecular Informatics Center, Konkuk University)
  • Received : 2005.02.16
  • Accepted : 2005.05.03
  • Published : 2005.08.31

Abstract

The flavonoid, quercetin, is a low molecular weight substance found in apple, tomato and other fruit. Besides its antioxidative effect, quercetin, like other flavonoids, has a wide range of neuropharmacological actions including analgesia, and motility, sleep, anticonvulsant, sedative and anxiolytic effects. In the present study, we investigated its effect on mouse 5-hydroxytryptamine type 3 ($5-HT_{3A}$) receptor channel activity, which is involved in pain transmission, analgesia, vomiting, and mood disorders. The $5-HT_{3A}$ receptor was expressed in Xenopus oocytes, and the current was measured with the two-electrode voltage clamp technique. In oocytes injected with $5-HT_{3A}$ receptor cRNA, quercetin inhibited the 5-HT-induced inward peak current ($I_{5-HT}$) with an $IC_{50}$ of $64.7{\pm}2.2{\mu}M$. Inhibition was competitive and voltage-independent. Point mutations of pre-transmembrane domain 1 (pre-TM1) such as R222T and R222A, but not R222D, R222E and R222K, abolished inhibition, indicating that quercetin interacts with the pre-TM1 of the $5-HT_{3A}$ receptor.

Keywords

$5-HT_{3A}$ Receptor;Flavonoids;Ligand-gated Ion Channels;Quercetin;Serotonin;Site-directed Mutagenesis;Xenopus Oocytes

Acknowledgement

Supported by : Konkuk University

References

  1. Dascal, N. (1987) The use of Xenopus oocytes for the study of ion channels. CRC Crit. Rev. Biochem. 22, 317-387 https://doi.org/10.3109/10409238709086960
  2. Fortney, J. T., Gan, T. J., Graczyk, S., Wetchler, B., Melson, T., et al. (1998) A comparison of the efficacy, safety, and patient satisfaction of ondansetron versus droperidol as antiemetics for elective outpatient surgical procedures. S3A-409 and S3A-410 Study Groups. Anesth. Analg. 86, 731-738 https://doi.org/10.1097/00000539-199804000-00011
  3. Griebel, G., Perrault, G., Tan, S., Schoemaker, H., and Sanger, D. J. (1999) Pharmacological studies on synthetic flavonoids: comparison with diazepam. Neuropharmacology 38, 965–977 https://doi.org/10.1016/S0028-3908(99)00026-X
  4. Jackson, M. B. and Yakel, J. L. (1995) The 5-HT3 receptor channel. Annu. Rev. Physiol. 57, 447-468 https://doi.org/10.1146/annurev.ph.57.030195.002311
  5. Mariq, A. V., Peterson, A. S., Brake, A. J., Myers, R. M., and Julius, D. (1991) Primary structure and functional expression of the 5-$HT_3$ receptor, a serotonin-gated ion channel. Science 254, 432-437 https://doi.org/10.1126/science.1718042
  6. Medina, J. H., Viola, H., Wolfman, C., Marder, M., Wasowski, C., et al. (1998) Neuroactive flavonoids: new ligands for the benzodiazepine receptors. Phytomedicine 5, 235–243
  7. Middleton, E. and Kandaswami, C. (1994) The impact of plant flavonoids on mammalian biology; in Implications for Immunity, Inflammation and Cancer, Harbone, J. B. (ed.), pp. 619– 645, The Flavonoids, Chapman & Hall, London
  8. Ortells, M. O. and Lunt, G. G. (1995) Evolutionary history of the ligand-gated ion channel superfamily of receptors. Trends Neurosci. 18, 121–127 https://doi.org/10.1016/0166-2236(95)93887-4
  9. Picq, M., Cheav, S. V., and Prigent, A. F. (1991). Effect of two flavonoid compounds on the central nervous system. Analgesic activity. Life Sci. 49, 1979–1988 https://doi.org/10.1016/0024-3205(91)90640-W
  10. Hu, X. Q., Zhang, L., Stewart, R. R., and Weight, F. F. (2003) Arginine 222 in the pre-transmembrane domain 1 of 5-HT3A receptors links agonist binding to channel gating. J. Biol. Chem. 278, 46583-46589 https://doi.org/10.1074/jbc.M308974200
  11. Marder, M., Viola, H., Wasowski, C., Wolfman, C., Waterman, P. G., et al. (1995) 6,3′-Dinitroflavone, a novel high affinity ligand for the benzodiazepine receptor with potent anxiolytic properties. Bioorg. Med. Chem. Lett. 5, 2717–2720 https://doi.org/10.1016/0960-894X(95)00464-5
  12. Dang, H., England, P. M., Farivar, S. S., Dougherty, D. A., and Lester, H. A. (2000) Probing the role of a conserved M1 proline residue in 5-hyroxytryptamine3 receptor gating. Mol. Pharmacol. 57, 1114-1122
  13. Harborne, J. B. and Williams, C. A. (2000) Advances in flavonoid research since 1992. Phytochemistry 55, 481–504 https://doi.org/10.1016/S0031-9422(00)00235-1
  14. Arias, H. R. (1996) Luminal and non-luminal non-competitive inhibitor binding sites on the nicotinic acetylcholine receptor. Mol. Membr. Biol. 13, 1-17
  15. Miksicek, R. J. (1993) Commonly occurring plant flavonoids have estrogenic activity. Mol. Pharmacol. 44, 37–43
  16. Goutman, J. D., Waxemberg, M. D., Donate-Oliver, F., Pomata, P. E., and Calvo, D. J. (2003) Flavonoid modulation of ionic currents mediated $GABA_A$ and $GABA_C$ receptors. Eur. J. Pharmacol. 461, 79-87 https://doi.org/10.1016/S0014-2999(03)01309-8
  17. Havsteen, B. H. (2002) The biochemistry and medical significance of the flavonoids. Pharmacol. Ther. 96, 67-202 https://doi.org/10.1016/S0163-7258(02)00298-X
  18. Speroni, E. and Minghetti, A. (1988) Neuropharmacological activity of extracts from Passiflora incarnata. Planta Med. 54, 488–491 https://doi.org/10.1055/s-2006-962525
  19. Lee, B. H., Jeong, S. M., Lee, J. H., Kim, D. H., Kim, J. H., et al. (2004) Differential effect of ginsenoside metabolites on the 5-$HT_{3A}$ receptor-mediated ion current in Xenopus oocytes. Mol. Cells 17, 51-56
  20. Oyama, Y., Fuchs, P. A., Katayama, N., and Noa, K. (1994) Myricetin and quercetin, the flavonoid constituents of Ginkgo biloba extract, greatly reduce oxidative metabolism in both resting and $Ca^{2+}$-loaded brain neurons. Brain Res. 635, 125– 129 https://doi.org/10.1016/0006-8993(94)91431-1
  21. Choi, S., Lee, J. H., Oh, S., Rhim, H., Lee, S. M., et al. (2003) Effects of ginsenoside $Rg_2$ on the 5-$HT_{3A}$ receptor-mediated ion current in Xenopus oocytes. Mol. Cells 15, 108-113
  22. Sine, S. M. and Taylor, P. (1982) Local anesthetic and histrionicotoxin are allosteric inhibitors of the acetylcholine receptor. J. Biol. Chem. 257, 8106–8114
  23. Polati, E., Verlato G., Finco, G., Monsaner, W., Grosso, S., et al. (1997) Ondansetron versus metoclopramide in the treatment of postoperative nausea and vomiting. Anesth. Analg. 85, 395– 399 https://doi.org/10.1097/00000539-199708000-00027
  24. Heidman, T., Oswald, R. E., and Changeux, J. P. (1983) Multiple sites of action for noncompetitive blockers on acetylcholine receptor rich membrane fragments from Torpedo marmorata. Biochemistry 22, 3112-3127 https://doi.org/10.1021/bi00282a014