Ece İSKENDER, Hülya CABADAK, AHMET AKICI, M. Zafer GÖREN, ATİLA KARAALP, Nefise B. ULUSOY, Beki KAN, Esam E. EL_FAKAHANY, Şule OKTAY

Carbachol induces nitric oxide generation in guinea-pig gallbladder

Anahtar Kelimeler:

Reseptörler, muskarinik, Karbakol, Safra kesesi, Kobaylar, Modeller, hayvan, Modeller, teorik, Nitrik oksit, NG-nitroarjinin metil ester, Omega-N-metilarjinin

Objective: Acetylcholine is one of the major contractile transmitters in gallbladder; whereas, it is also innervated by non-adrenergic non-cholinergic nerves which mediate relaxation. It was postulated that nitric oxide (NO) which activates soluble guanylate cyclase to increase cyclic GMP (cGMP) levels in the target cells may be involved in these processes. This study was designed to investigate whether muscarinic receptor stimulation via carbachol (CCh) induces NO-mediated cGMP synthesis in guinea-pig gallbladder. Methods: cGMP levels were measured via radioimmunoassay in gallbladder slices incubated with carbachol (CCh) (10-6-10-3 mol/l) in the presence and absence of NO synthase (NOS) inhibitor L-NAME or muscarinic antagonist scopolamine. The effect of L-NAME and another NOS inhibitor L-NMMA on carbachol-induced contractions were also investigated in an in vitro organ bath. Results: Carbachol stimulated cGMP formation in guinea-pig gallbladder slices significantly. CCh-induced cGMP formation was abolished by both L-NAME (1mmol/l) and scopolamine (10-8-10-6 mol/l). In contraction experiments, L-NAME (3x10-4 mol/l) did not produce any change in the resting tension of the strips and the oncentration-response curves to carbachol, whereas L-NMMA (3x10-4 mol/l) induced a slight, but significant contraction (6.3+1.9 % of carbachol (10-6 mol/l)-induced response). When carbachol was added to the bath after L-NMMA-induced increase in tension reached its maximum, the amplitude of the contractile response at the end was 108.0+4.0 % of the control. Conclusion: Therefore, it may be concluded that muscarinic receptor activation by carbachol stimulates NO production in guinea-pig gallbladder and the subsequent increase in cGMP counteracts with carbachol-induced contractions. However, it seems likely that an intact enteric nervous system is required for such an interaction to be prominent. The muscarinic receptor subtype(s) involved in this interaction remains to be elucidated.

Keywords:

Receptors, Muscarinic, Carbachol, Gallbladder, Guinea Pigs, Models, Animal, Models, Theoretical, Nitric Oxide, NG-Nitroarginine Methyl Ester, omega-N-Methylarginine,

___

1. Furchgott RF, Zawadski JV, Cherry PD. Role of endothelium in vasodilator response to acetylcholine. In: Vanhoutte P, Leusen I, eds. Vasodilatation. New York: Raven Press, 1981: 49-66. 2. Ignarro LJ, Lippton H, Edwards JC, et al. Mechanism of vascular smooth muscle relaxation by organic nitrates, nitrites, nitroprusside and nitric oxide: evidence for the involvement of S-nitrosothiols as active intermediates. J Pharmacol Exp Ther 1981; 218: 739-749. 3. Palmer RMJ, Ashton DS, Moncada S. Vascular enodthelial cells synthesize nitric oxide from L-arginine. Mature 1988; 333: 664-666. 4. Moncada S, Palmer RMJ, tiiggs A. Biosynthesis of nitric oxide from L-arginine. Biochem Pharmacol 1989; 38: 1709-1715. 5. Qruetter CA, Qruetter DY, Lyon JE, Kadowitz PJ, Ignarro LF. Relationship between cyclic guanosine 3'-5'-monophosphate formation and relaxation of coronary arterial smooth muscle by glyceryl trinitrate, nitroprusside, nitrite and nitric oxide: effects of methylene blue and methemoglobin. J Pharmacol Exp Ther 1981; 219; 181-186. 6. Kukovetz WR, Pöch G, fiolzmann S. Cyclic nucleotides and relaxation of vascular smooth muscle. In: Vanhoutte P, Leusen I, eds. Vasodilatation. Hew York: Raven Press, 1981: 339-353.

7. Rapaport RM, Murad F. Agonist-induced endothelium dependent relaxation in rat thoracic aorta may be mediated through cQMP. Circ Res 1983; 52: 352-357. 8. Davison JS, Farasham A, Pearson GT, Peterson OH. Dibutyryl cQMP: a specific inhibitor of the action of CCK-like peptides on the pancreas, gallbladder and ileum. J Physiol (Lond) 1980; 305: 77-78. 9. Behar J, Biancani P. Pharmacology of biliary tract. In: Shultz SO, Wood JD, RaunerBB, eds. Handbook of Physiology, Section 6: The Gastorintestinal System. New York: Oxford University Press, 1989: 1103-1131.

10. Mourelle M, Guarner F, Molero X, Moncada S, Malagelada JR. Regulation of gallbladder motility by the arginine-nitric oxide pathway in guinea-pigs. Out 1993; 34: 911-915. 11. Göçer F, Yarış E, Tuncer M. The action ofamyl nitrite and isosorbide dinitrate on the contractility of sphincter of Oddi of guinea- pigs: Gen Pharmacol 1994; 25: 995-999.

12. Wotta DR, Birnbaum AK, Wilcox GL, Elde R, Law PY. u-Opioid receptor regulates CTFR coexpressed in Xenopus oocytes in a cAMP independent manner. Mol Brain Res 1997; 44: 55-65. 13. Afdhal HH, La Mont JT. Pathogenesis of gallstones. In: Gitnick G, ed. Principles and. Practices of Gastroenterology and Hepatology. New York: Elsevier, 1994: 561-571, ed2. 14. Kaufman HS, Shermak MA, May CA, Pitt HA, Lillemoe KD. nitric oxide inhibits resting sphincter of Oddi activity. Am J Surg 1993; 165: 74-80. 15. Grozdanovic Z, Mayer ZB, Baumgarten HG, Bruning G. nitric oxide synthase-containing nerve fibers and neurons in the gallbladder and biliary pathways of the guinea-pig, neuroreport 1994; 5: 837-840. 16. Mourelle M, Guarner F, Moncada S, Malagelada JR. The arginine/nitric oxide pathway modulates sphincter of Oddi motor activity in guinea-pigs and rabbits. Gastroenterology 1993; 105: 1299-1305. 17. Salomons H, Keaveny AP, Henihan R, et al. nitric oxide and gallbladder motility in prairie dogs. Am J Physiol 1997; 272: G770-G778. 18. Dufour JF, Turner TJ, Arias IM. Nitric oxide blocks bile canalicular contraction by inhibiting inositol triphosphate-dependent Ca2+ mobilization. Gastroenterology 1995; 108: 841-849.

19. Murthy KS, Makhlouf GM. Interaction of cA- kinase and cG-kinase in mediating relaxation of dispersed smooth muscle cells. Am J Physiol 1995;268: C171-C180. 20. Stark ME, Szurszewski JH. Role of nitric oxide in gastrointestinal and hepatic function and disease. Gastroen terology 1992; 103:1928- 1949. 21. Özkutlu U, Alican İ, Karahan F, et al. Are m- cholinoceptors of guinea-pig gallbladder smooth muscle ofm^ subtype? Pharmacology 1993,-46:308-314. 22. Von Schrenck T, SieversJ, Mirau S, Raedier A, Qreten H. Characterization of muscarinic receptors on guinea-pig gallbladder smooth muscle. Gastroenterology 1993; 105:1341-1349. 23.Von Schrenck T, Mackensen B, Mende U, et al. Signal transduction pathway of the muscarinic receptors mediating gallbladder contraction. naunyn-Schmied Arch Pharmacol 1994:349:346-354. 24.Takahashi T, Kurosawa S, Owyang C.Regulation of PI hydrolysis and cAMP formation by muscarinic M3 receptor in guinea-pig gallbladder. Am J Physiol 1994;267:O523-Q528. 25. Barocelli E, Ballabeni V, Chiavarini M, Molina E, Impicciatore M. Functional comparison between nuvenzepine and pirenzepine on different guinea pig isolated smooth muscle preparations. Pharmacol Res 1994:30:161- 170. 26. Eltze M, König fi, Ullrich B, Grebe T. Contraction of guinea-pig gallbladder: muscarinic M$ or M4, receptors? Eur J Pharmacol 199 7;332:7 7-8 7. 27. Oktay Ş, Cabadak H, İskender E, et al. Evidence for the presence of muscarinic M2 and M4, receptors in guinea-pig gallbladder smooth muscle. J Auton Pharmacol 1998; 18:195-204. 28. Karaalp A, Akıcı A, Akbulut H, Ulusoy MB, Oktay Ş. Distinct functional muscarinic receptors in guinea-pig gallbladder, ileum and atria. Pharmacol Res 1999:39:389-395. 29. Parkman UP, Pagano AP, Ryan JP. Subtypes of muscarinic receptors regulating gallbladder cholinergic contractions. Am J Physiol 1999;276:G1243-G 1250. 30. Felder CC. Muscarinic acetylcholine receptors: signal transduction through multiple effectors. FASEB J 1995:9:619-625, 31. Eglen RM, fiegde SS, Watson Pi. Muscarinic receptor subtypes and smooth muscle function. Pharmacol Rev 1996:48:531-565. 32. Ignarro LJ, Byrns RE, Buga QM, Wood KS, Chaudhuri G. Pharmacological evidence that endothelium-derived relaxing factor is nitric oxide: use of pyrogallol and superoxide dismutase to study endothelium-dependent and nitric oxide-elicited vascular smooth muscle relaxation. J Pharmacol Exp Ther 1988,-244:181-189.

33. Furchgott RF, Vanhoutte P. Endothelium- derived relaxing and contracting factors. FASEB J 1989:3:2007-2018. 34. George WJ, Wilkerson RD, Kadowitz PJ. Influence of acetylcholine on contractile force and cyclic nucleotide levels in the isolated perfused rat heart. J Pharmacol Exp Ther 1973:184:228-235. 35. Balligand JL, Kelly RA, Marsden PA, Smith TW, Michel T. Control of cardiac muscle cell function by an endogenous nitric oxide signalling system. Proc riatl Acad Sci USA 1993:90:347-351. 36. Castoldi AF, Manzo L, Costa LQ. Cyclic QMP formation induced by muscarinic receptors is mediated by nitric oxide synthesis in rat cortical primary cultures. Brain Res 1993:610:57-61. 37. liuJ, El-Fakahany EE. Role of intercellular and intracellular communication by nitric oxide in coupling of muscarinic receptors to activation of guanylate cyclase in neuronal cells. J rieurochem 1993:61:578-585. 38. Dauphin F, Linville DQ, Hamel E. Cholinergic dilatation and constriction of feline cerebral blood vessels are mediated by stimulation of phosphoinositide metabolism via two different muscarinic receptor subtypes. J rieurochem 1994:63:544-551. 39. Eglen RM, Whiting RL. Heterogeneity of vascular muscarinic receptors. J Auton Pharmacol 1990:10:233-245. 40. Dodds WJ, Hogan WJ, Geenen JE. Motility of the biliary system. In: Schultz SQ, Wood JD, Rauner BB, eds. Handbook of Physiology, Section 6: The Gastrointestinal System. Mew York: Oxford University Press, 1989:1055- 1101. 41. Mourelle M, Guarner F, Moncada S, Malagelada JR. Regulation of sphincter of Oddi motor activity by nitric oxide. Eur J Gastroenterol Hepatol 1991;3:S64. 42. Hellmich B, Gyermek L. PiG -riitro-L^arginine methyl ester:a muscarinic receptor antagonist? Fundam Clin Pharmacol 1997:11:305-314. 43. Buxton ILO, Cheek DJ, Eckman D, Westfall DP, Sanders KM, Keef KD. riG-riitro L-arginine methyl ester and other alkyl esters of arginine are muscarinic receptor antagonists. Circ Res 1993:72:387-395.