Skip to main content
PMC home page
British Journal of Pharmacology logoLink to British Journal of Pharmacology
. 1996 Nov;119(5):1022–1028. doi: 10.1111/j.1476-5381.1996.tb15773.x

Effect of thiol modulators and Cu/Zn superoxide dismutase inhibition on nitrergic relaxations in the rat gastric fundus.

J G De Man 1, B Y De Winter 1, G E Boeckxstaens 1, A G Herman 1, P A Pelckmans 1
PMCID: PMC1915936  PMID: 8922754

Abstract

1. The effects of superoxide anion generators before and after treatment with inhibitors of Cu/Zn superoxide dismutase (Cu/Zn SOD) and the effects of thiol-modulating agents were investigated on nitrergic relaxations to electrical stimulation of non-adrenergic non-cholinergic (NANC) nerves of the rat gastric fundus and on relaxations to authentic nitric oxide (NO) and nitroglycerin. 2. The superoxide anion generators, pyrogallol (30 microM) and duroquinone (30-60 microM), significantly inhibited the relaxations to NO (0.03-3 microM) but not nitrergic relaxations to NANC nerve stimulation (0.5-8 Hz) or those to ATP (10 microM). Treatment of the rat gastric fundus with the inhibitors of Cu/Zn SOD, diethyldithiocarbamate (DETC, 1 mM for 2 h) or triethylenetetramine (TETA, 100 microM for 2 h) had no effect on the relaxations to NANC nerve stimulation (1-8 Hz), NO (0.03-3 microM) or on those to ATP (10 microM). 3. After treatment of the rat gastric fundus with DETC (1 mM) but not after treatment with TETA (100 microM), pyrogallol (30 microM) and duroquinone (30-60 microM) significantly inhibited the nitrergic relaxations to electrical stimulation (0.5-8 Hz) and those to NO (0.03-3 microM). This inhibitory effect of pyrogallol and duroquinone was prevented by addition of exogenous SOD (250 units ml-1). Pyrogallol but not duroquinone also inhibited the NO-independent relaxations to ATP (10 microM). 4. The thiol modulators, buthionine sulphoximine (1 mM for 2 h) and ethacrynic acid (30 microM for 2 h), significantly inhibited the relaxations to nitroglycerin (0.03-3 microM) but had no effect on the nitrergic relaxations to electrical stimulation (0.5-8 Hz) or on those to NO (0.03-10 microM) and ATP (10 microM). The thiol modulators, sulphobromophthalein (100 microM for 2 h) and diamide (30-100 microM for 2 h) did not affect the relaxations to nitroglycerin, or those to NANC nerve stimulation and NO. 5. In summary, thiol modulators significantly inhibited the thiol-dependent relaxations to nitroglycerin but not those to NANC nerve stimulation or NO. Relaxations to nitrergic stimulation were decreased by superoxide anion generators only after inhibition of Cu/Zn SOD. These results suggest that the nitrergic NANC neurotransmitter in the rat gastric fundus is not a nitrosothiol but more likely free NO, which is protected from breakdown by tissue SOD.

Full text

PDF
1023

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abdalla S., Will J. A. Potentiation of the hypoxic contraction of guinea-pig isolated pulmonary arteries by two inhibitors of superoxide dismutase. Gen Pharmacol. 1995 Jul;26(4):785–792. doi: 10.1016/0306-3623(94)00245-i. [DOI] [PubMed] [Google Scholar]
  2. Barbier A. J., Lefebvre R. A. Effect of LY 83583 on relaxation induced by non-adrenergic non-cholinergic nerve stimulation and exogenous nitric oxide in the rat gastric fundus. Eur J Pharmacol. 1992 Aug 25;219(2):331–334. doi: 10.1016/0014-2999(92)90315-u. [DOI] [PubMed] [Google Scholar]
  3. Barbier A. J., Lefebvre R. A. Influence of S-nitrosothiols and nitrate tolerance in the rat gastric fundus. Br J Pharmacol. 1994 Apr;111(4):1280–1286. doi: 10.1111/j.1476-5381.1994.tb14884.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boeckxstaens G. E., De Man J. G., De Winter B. Y., Herman A. G., Pelckmans P. A. Pharmacological similarity between nitric oxide and the nitrergic neurotransmitter in the canine ileocolonic junction. Eur J Pharmacol. 1994 Oct 13;264(1):85–89. doi: 10.1016/0014-2999(94)90640-8. [DOI] [PubMed] [Google Scholar]
  5. Boeckxstaens G. E., Pelckmans P. A., Bogers J. J., Bult H., De Man J. G., Oosterbosch L., Herman A. G., Van Maercke Y. M. Release of nitric oxide upon stimulation of nonadrenergic noncholinergic nerves in the rat gastric fundus. J Pharmacol Exp Ther. 1991 Feb;256(2):441–447. [PubMed] [Google Scholar]
  6. Boeckxstaens G. E., Pelckmans P. A., Bult H., De Man J. G., Herman A. G., Van Maercke Y. M. Non-adrenergic non-cholinergic relaxation mediated by nitric oxide in the canine ileocolonic junction. Eur J Pharmacol. 1990 Nov 6;190(1-2):239–246. doi: 10.1016/0014-2999(90)94132-h. [DOI] [PubMed] [Google Scholar]
  7. Bult H., Boeckxstaens G. E., Pelckmans P. A., Jordaens F. H., Van Maercke Y. M., Herman A. G. Nitric oxide as an inhibitory non-adrenergic non-cholinergic neurotransmitter. Nature. 1990 May 24;345(6273):346–347. doi: 10.1038/345346a0. [DOI] [PubMed] [Google Scholar]
  8. Cocco D., Calabrese L., Rigo A., Argese E., Rotilio G. Re-examination of the reaction of diethyldithiocarbamate with the copper of superoxide dismutase. J Biol Chem. 1981 Sep 10;256(17):8983–8986. [PubMed] [Google Scholar]
  9. De Man J. G., Boeckxstaens G. E., De Winter B. Y., Moreels T. G., Herman A. G., Pelckmans P. A. Inhibition of non-adrenergic non-cholinergic relaxations by nitric oxide donors. Eur J Pharmacol. 1995 Oct 24;285(3):269–274. doi: 10.1016/0014-2999(95)00420-p. [DOI] [PubMed] [Google Scholar]
  10. De Man J. G., Boeckxstaens G. E., De Winter B. Y., Moreels T. G., Misset M. E., Herman A. G., Pelckmans P. A. Comparison of the pharmacological profile of S-nitrosothiols, nitric oxide and the nitrergic neurotransmitter in the canine ileocolonic junction. Br J Pharmacol. 1995 Mar;114(6):1179–1184. doi: 10.1111/j.1476-5381.1995.tb13331.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. De Man J. G., Pelckmans P. A., Boeckxstaens G. E., Bult H., Oosterbosch L., Herman A. G., Van Maercke Y. M. The role of nitric oxide in inhibitory non-adrenergic non-cholinergic neurotransmission in the canine lower oesophageal sphincter. Br J Pharmacol. 1991 May;103(1):1092–1096. doi: 10.1111/j.1476-5381.1991.tb12305.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fang S., Christensen J. Manganese superoxide dismutase and reduced nicotinamide adenine dinucleotide diaphorase colocalize in the rat gut. Gastroenterology. 1995 Nov;109(5):1429–1436. doi: 10.1016/0016-5085(95)90627-4. [DOI] [PubMed] [Google Scholar]
  13. Gibson A., Babbedge R., Brave S. R., Hart S. L., Hobbs A. J., Tucker J. F., Wallace P., Moore P. K. An investigation of some S-nitrosothiols, and of hydroxy-arginine, on the mouse anococcygeus. Br J Pharmacol. 1992 Nov;107(3):715–721. doi: 10.1111/j.1476-5381.1992.tb14512.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gillespie J. S., Sheng H. The effects of pyrogallol and hydroquinone on the response to NANC nerve stimulation in the rat anococcygeus and the bovine retractor penis muscles. Br J Pharmacol. 1990 Jan;99(1):194–196. doi: 10.1111/j.1476-5381.1990.tb14677.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hecker M., Siegle I., Macarthur H., Sessa W. C., Vane J. R. Role of intracellular thiols in release of EDRF from cultured endothelial cells. Am J Physiol. 1992 Mar;262(3 Pt 2):H888–H896. doi: 10.1152/ajpheart.1992.262.3.H888. [DOI] [PubMed] [Google Scholar]
  16. Hobbs A. J., Tucker J. F., Gibson A. Differentiation by hydroquinone of relaxations induced by exogenous and endogenous nitrates in non-vascular smooth muscle: role of superoxide anions. Br J Pharmacol. 1991 Nov;104(3):645–650. doi: 10.1111/j.1476-5381.1991.tb12483.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ignarro L. J., Lippton H., Edwards J. C., Baricos W. H., Hyman A. L., Kadowitz P. J., Gruetter C. A. 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 Sep;218(3):739–749. [PubMed] [Google Scholar]
  18. Jenkinson K. M., Reid J. J., Rand M. J. Hydroxocobalamin and haemoglobin differentiate between exogenous and neuronal nitric oxide in the rat gastric fundus. Eur J Pharmacol. 1995 Mar 6;275(2):145–152. doi: 10.1016/0014-2999(94)00762-v. [DOI] [PubMed] [Google Scholar]
  19. Kelm M., Feelisch M., Spahr R., Piper H. M., Noack E., Schrader J. Quantitative and kinetic characterization of nitric oxide and EDRF released from cultured endothelial cells. Biochem Biophys Res Commun. 1988 Jul 15;154(1):236–244. doi: 10.1016/0006-291x(88)90675-4. [DOI] [PubMed] [Google Scholar]
  20. Kerr S. W., Buchanan L. V., Bunting S., Mathews W. R. Evidence that S-nitrosothiols are responsible for the smooth muscle relaxing activity of the bovine retractor penis inhibitory factor. J Pharmacol Exp Ther. 1992 Oct;263(1):285–292. [PubMed] [Google Scholar]
  21. Kim S. J., Han D., Moon K. D., Rhee J. S. Measurement of superoxide dismutase-like activity of natural antioxidants. Biosci Biotechnol Biochem. 1995 May;59(5):822–826. doi: 10.1271/bbb.59.822. [DOI] [PubMed] [Google Scholar]
  22. Kitamura K., Lian Q., Carl A., Kuriyama H. S-nitrosocysteine, but not sodium nitroprusside, produces apamin-sensitive hyperpolarization in rat gastric fundus. Br J Pharmacol. 1993 Jun;109(2):415–423. doi: 10.1111/j.1476-5381.1993.tb13585.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Li C. G., Brosch S. F., Rand M. J. Inhibition by ethacrynic acid of NO-mediated relaxations of the rat anococcygeus muscle. Clin Exp Pharmacol Physiol. 1994 Apr;21(4):293–299. doi: 10.1111/j.1440-1681.1994.tb02515.x. [DOI] [PubMed] [Google Scholar]
  24. Lilley E., Gibson A. Inhibition of relaxations to nitrergic stimulation of the mouse anococcygeus by duroquinone. Br J Pharmacol. 1995 Dec;116(8):3231–3236. doi: 10.1111/j.1476-5381.1995.tb15129.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Liu X., Gillespie J. S., Martin W. Non-adrenergic, non-cholinergic relaxation of the bovine retractor penis muscle: role of S-nitrosothiols. Br J Pharmacol. 1994 Apr;111(4):1287–1295. doi: 10.1111/j.1476-5381.1994.tb14885.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Martin W., McAllister K. H., Paisley K. NANC neurotransmission in the bovine retractor penis muscle is blocked by superoxide anion following inhibition of superoxide dismutase with diethyldithiocarbamate. Neuropharmacology. 1994 Nov;33(11):1293–1301. doi: 10.1016/0028-3908(94)90029-9. [DOI] [PubMed] [Google Scholar]
  27. Misra H. P. Reaction of copper-zinc superoxide dismutase with diethyldithiocarbamate. J Biol Chem. 1979 Nov 25;254(22):11623–11628. [PubMed] [Google Scholar]
  28. Murphy M. E., Piper H. M., Watanabe H., Sies H. Nitric oxide production by cultured aortic endothelial cells in response to thiol depletion and replenishment. J Biol Chem. 1991 Oct 15;266(29):19378–19383. [PubMed] [Google Scholar]
  29. Needleman P., Jakschik B., Johnson E. M., Jr Sulfhydryl requirement for relaxation of vascular smooth muscle. J Pharmacol Exp Ther. 1973 Nov;187(2):324–331. [PubMed] [Google Scholar]
  30. Paisley K., Martin W. Blockade of nitrergic transmission by hydroquinone, hydroxocobalamin and carboxy-PTIO in bovine retractor penis: role of superoxide anion. Br J Pharmacol. 1996 Apr;117(8):1633–1638. doi: 10.1111/j.1476-5381.1996.tb15333.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pelckmans P. A., Boeckxstaens G. E., Van Maercke Y. M., Herman A. G., Verbeuren T. J. Acetylcholine is an indirect inhibitory transmitter in the canine ileocolonic junction. Eur J Pharmacol. 1989 Nov 7;170(3):235–242. doi: 10.1016/0014-2999(89)90544-x. [DOI] [PubMed] [Google Scholar]
  32. Rand M. J., Li C. G. Discrimination by the NO-trapping agent, carboxy-PTIO, between NO and the nitrergic transmitter but not between NO and EDRF. Br J Pharmacol. 1995 Sep;116(2):1906–1910. doi: 10.1111/j.1476-5381.1995.tb16681.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Thomas R. M., Fang S., Leichus L. S., Oberley L. W., Christensen J., Murray J. A., Ledlow A., Conklin J. L. Antioxidant enzymes in intramural nerves of the opossum esophagus. Am J Physiol. 1996 Jan;270(1 Pt 1):G136–G142. doi: 10.1152/ajpgi.1996.270.1.G136. [DOI] [PubMed] [Google Scholar]
  34. Thornbury K. D., Ward S. M., Dalziel H. H., Carl A., Westfall D. P., Sanders K. M. Nitric oxide and nitrosocysteine mimic nonadrenergic, noncholinergic hyperpolarization in canine proximal colon. Am J Physiol. 1991 Sep;261(3 Pt 1):G553–G557. doi: 10.1152/ajpgi.1991.261.3.G553. [DOI] [PubMed] [Google Scholar]
  35. Wood J., Garthwaite J. Models of the diffusional spread of nitric oxide: implications for neural nitric oxide signalling and its pharmacological properties. Neuropharmacology. 1994 Nov;33(11):1235–1244. doi: 10.1016/0028-3908(94)90022-1. [DOI] [PubMed] [Google Scholar]
  36. Yeates R. A., Schmid M., Leitold M. Antagonism of glycerol trinitrate activity by an inhibitor of glutathione S-transferase. Biochem Pharmacol. 1989 Jun 1;38(11):1749–1753. doi: 10.1016/0006-2952(89)90408-5. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Pharmacology are provided here courtesy of The British Pharmacological Society

RESOURCES