Skip to main content
SpringerLink
Log in

Bismuth Subsalicylate Increases Intracellular Ca2+, MAP-Kinase Activity, and Cell Proliferation in Normal Human Gastric Mucous Epithelial Cells

  • Published:
Digestive Diseases and Sciences Aims and scope Submit manuscript

Abstract

Clinical and laboratory studies have shown that bismuth subsalicylate (BSS) is helpful in the healing of gastric ulcers because of the bactericidal effects of bismuth (Bi3+) on H. pylori. Bismuth or BSS has also been reported to possess other nonbactericidal or “gastroprotective” effects in the stomach. It is known in other cell types that the effects of extracellular divalent or trivalent cations (e.g., Ca2+) can activate a plasma membrane-bound calcium-sensing receptor (CaSR). In a previous study, we found the existence of a CaSR which was activated by extracellular Ca2+ and found to increase intracellular Ca2+ [Ca2+]i, MAP-kinase activity, and gastric epithelial cell proliferation. In the present study, we were interested in determining whether the effects of the trivalent cation Bi3+ (in the form of BSS) on [Ca2+]i, MAP-kinase activity, and proliferation of gastric cells. We found that BSS dose dependently increased [Ca2+]i, p44/p42 and p38 MAP-kinase activites, and gastric mucous epithelial cell growth. The addition of BAPTA to chelate intracellular Ca2+ blocked BSS-induced p44/p42 MAP-kinase activities but not p38 MAP-kinase activity. The p44/p42 MAP-kinase inhibitor PD98059 and the p38 MAP-kinase inhibitor SB203580 dose dependently decreased gastric mucous cell growth over a 24 hr. All of the BSS-induced changes in [Ca2+]i, MAP-kinase activity, and gastric cell proliferation could be reproduced with the CaSR-agonist gadolinium (Gd3+). Our data suggest that BSS may possess additional novel effects by increasing gastric mucous epithelial cell growth through a Ca2+/MAP-kinase-dependent pathway.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Marshall BJ, Forbes GM, Warren JR, Glaser ME, Cullen DJ, Collins BJ, McGechie DB, Rogers PA, Glancy RJ and Armstrong JA: The future of Helicobacter pylori eradication: a personal perspective. Aliment Pharmacol Ther 11 (Suppl 1):109-115, 1997

    Google Scholar 

  2. Gorbach SL: Bismuth therapy in gastrointestinal diseases. Gastroenterology 99:863-875, 1990

    Google Scholar 

  3. Mosby: Bismuth subsalicylate. Mosby's Drug Consult, 13th ed., New York, Elsevier, 2003, p 2503

    Google Scholar 

  4. Holroyde M, Yeakle C, Pepple S: Gastric cytoprotection by bismuth subsalicylate. Gastroenterology 86:1116, 1984

    Google Scholar 

  5. Sox TE, Olson CA: Binding and killing of bacteria by bismuth subsalicylate. Antimicrob Agents Chemother 33:2075-2082, 1989

    Google Scholar 

  6. Munoz DJ, Tasman-Jones C, Pybus J: Effect of Helicobacter pylori infection on colloidal bismuth subcitrate concentration in gastric mucus. Gut 33:592-596, 1992

    Google Scholar 

  7. Lee SP: The mode of action of colloidal bismuth subcitrate. Scand J Gastroenterol Suppl 185:1-6, 1991

    Google Scholar 

  8. Ottlecz A, Romero JJ, Hazell SL, Graham DY, Lichtenberger LM: Phospholipase activity of Helicobacter pylori and its inhibition by bismuth salts. Biochemical and biophysical studies. Dig Dis Sci 38:2071-2080, 1993

    Google Scholar 

  9. Konturek SJ, Dembinski A, Warzecha Z, Bielanski W, Brzozowski T, Drozdowicz D: Epidermal growth factor (EGF) in the gastroprotective and ulcer healing actions of colloidal bismuth subcitrate (De-Nol) in rats. Gut 29:894-902, 1988

    Google Scholar 

  10. Bagchi D, McGinn TR, Ye X, Balmoori J, Bagchi M, Stohs SJ, Kuszynski CA, Carryl OR, Mitra S: Mechanism of gastroprotection by bismuth subsalicylate against chemically induced oxidative stress in cultured human gastric mucosal cells. Dig Dis Sci 44:2419-2428, 1999

    Google Scholar 

  11. Bagchi D, Carryl OR, Tran MX, Bagchi M, Garg A, Milnes MM, Williams CB, Balmoori J, Bagchi DJ, Mitra S, Stohs SJ: Acute and chronic stress-induced oxidative gastrointestinal mucosal injury in rats and protection by bismuth subsalicylate. Mol Cell Biochem 196:109-116, 1999

    Google Scholar 

  12. Copeman M, Matuz J, Leonard AJ, Pearson JP, Dettmar PW, Allen A: The gastroduodenal mucus barrier and its role in protection against luminal pepsins: The effect of 16,16 dimethyl prostaglandin E2, carbopol-polyacrylate, sucralfate and bismuth subsalicylate. J Gastroenterol Hepatol 9:S55-S59, 1994

    Google Scholar 

  13. Tanaka S, Guth PH, Carryl OR, Kaunitz JD: Cytoprotective effect of bismuth subsalicylate in indomethacin-treated rats is associated with enhanced mucus bismuth concentration. Aliment Pharmacol Ther 11:605-612, 1997

    Google Scholar 

  14. Sidebotham RL, Batten JJ, Li K, Spencer J, Baron JH: Some effects of tripotassium dicitrato bismuthate on gastric secretion and the mucus barrier. Aliment Pharmacol Ther 4:535-546, 1990

    Google Scholar 

  15. Tanaka S, Guth PH, Paulsen G, Kaunitz JD: Gastroprotective effect of ranitidine bismuth citrate is associated with increased mucus bismuth concentration in rats. Gut 39:164-171, 1996

    Google Scholar 

  16. Suarez FL, Furne J, Stiehm J, Garten C, Levitt MD: Site of bismuth absorption from bismuth subsalicylate: implications for treatment of colonic conditions. Dig Dis Sci 45:1444-1446, 2000

    Google Scholar 

  17. Sandha GS, LeBlanc R, Van Zanten SJ, Sitland TD, Agocs L, Burford N, Best L, Mahoney D, Hoffman P, Leddin DJ: Chemical structure of bismuth compounds determines their gastric ulcer healing efficacy and anti-Helicobacter pylori activity. Dig Dis Sci 43:2727-2732, 1998

    Google Scholar 

  18. Dahlgren A, Glogard C, Gammelsaether M, Aasen AJ, Klaveness J, Berdal BP, Bergan T: Organobismuth compounds: activity against Helicobacter pylori. Scand J Gastroenterol 34:135-137, 1999

    Google Scholar 

  19. Hinsull SM: Effect of colloidal bismuth subcitrate on age related gastric lesions in the rat. Gut 32:355-360, 1991

    Google Scholar 

  20. Chiba N, Hunt RH, Thomson AB: Ranitidine bismuth citrate. Can J Gastroenterol 15:389-398, 2001

    Google Scholar 

  21. Wagstaff AJ, Benfield P, Monk JP: Colloidal bismuth subcitrate. A review of its pharmacodynamic and pharmacokinetic properties, and its therapeutic use in peptic ulcer disease. Drugs 36:132-157, 1988

    Google Scholar 

  22. Quarles LD, Hartle JE, 2nd, Middleton JP, Zhang J, Arthur JM and Raymond JR: Aluminum-induced DNA synthesis in osteoblasts: Mediation by a G-protein coupled cation sensing mechanism. J Cell Biochem 56:106-117, 1994

    Google Scholar 

  23. Brown EM, MacLeod RJ: Extracellular calcium sensing and extracellular calcium signaling. Physiol Rev 81:239-297, 2001

    Google Scholar 

  24. Brown EM, Gamba G, Riccardi D, Lombardi M, Butters R, Kifor O, Sun A, Hediger MA, Lytton J, Hebert SC: Cloning and characterization of an extracellular Ca2+-sensing receptor from bovine parathyroid. Nature 366:575-580, 1993

    Google Scholar 

  25. Buchan AM, Squires PE, Ring M, Meloche RM: Mechanism of action of the calcium-sensing receptor in human antral gastrin cells. Gastroenterology 120:1128-1139, 2001

    Google Scholar 

  26. Gama L, Baxendale-Cox LM, Breitwieser GE: Ca2+-sensing receptors in intestinal epithelium. Am J Physiol 273:C1168-C1175, 1997

    Google Scholar 

  27. Chattopadhyay N, Cheng I, Rogers K, Riccardi D, Hall A, Diaz R, Hebert SC, Soybel DI, Brown EM: Identification and localization of extracellular Ca2+-sensing receptor in rat intestine. Am J Physiol 274:G122-G130, 1998

    Google Scholar 

  28. Chakrabarty S, Radjendirane V, Appelman H, Varani J: Extracellular calcium and calcium sensing receptor function in human colon carcinomas: promotion of E-cadherin expression and suppression of beta-catenin/TCF activation. Cancer Res 63:67-71, 2003

    Google Scholar 

  29. Kallay E, Bajna E, Wrba F, Kriwanek S, Peterlik M, Cross HS: Dietary calcium and growth modulation of human colon cancer cells: role of the extracellular calcium-sensing receptor. Cancer Detect Prev 24:127-136, 2000

    Google Scholar 

  30. Kallay E, Kifor O, Chattopadhyay N, Brown EM, Bischof MG, Peterlik M, Cross HS: Calcium-dependent c-myc proto-oncogene expression and proliferation of Caco-2 cells: a role for a luminal extracellular calcium-sensing receptor. Biochem Biophys Res Commun 232:80-83, 1997

    Google Scholar 

  31. Kato M, Doi R, Imamura M, Furutani M, Hosotani R, Shimada Y: Calcium-evoked insulin release from insulinoma cells is mediated via calcium-sensing receptor. Surgery 122:1203-1211, 1997

    Google Scholar 

  32. Bapty BW, Dai LJ, Ritchie G, Jirik F, Canaff L, Hendy GN, Quamme GA: Extracellular Mg2+-and Ca2+-sensing in mouse distal convoluted tubule cells. Kidney Int 53:583-592, 1998

    Google Scholar 

  33. Tu CL, Oda Y, Bikle DD: Effects of a calcium receptor activator on the cellular response to calcium in human kerationocytes. J Invest Dermatol 113:340-345, 1999

    Google Scholar 

  34. McNeil SE, Hobson SA, Nipper V, Rodland KD: Functional calcium-sensing receptors in rat fibroblasts are required for activation of SRC kinase and mitogen-activated protein kinase in response to extracellular calcium. J Biol Chem 273:1114-1120, 1998

    Google Scholar 

  35. Hobson SA, Wright J, Lee F, McNeil SE, Bilderback T, Rodland KD: Activation of the MAP kinase cascade by exogenous calcium-sensing receptor. Mol Cell Endocrinol 200:189-198, 2003

    Google Scholar 

  36. Handlogten ME, Shiraishi N, Awata H, Huang C, Miller RT: Extracellular Ca2+-sensing receptor is a promiscuous divalent cation sensor that responds to lead. Am J Physiol Renal Physiol 279:F1083-F1091, 2000

    Google Scholar 

  37. Kifor O, MacLeod RJ, Diaz R, Bai M, Yamaguchi T, Yao T, Kifor I, Brown EM: Regulation of MAP kinase by calcium-sensing receptor in bovine parathyroid and CaR-transfected HEK293 cells. Am J Physiol Renal Physiol 280:F291-F302, 2001

    Google Scholar 

  38. Yamaguchi T, Chattopadhyay N, Kifor O, Sanders JL, Brown EM: Activation of p42/44 and p38 mitogen-activated protein kinases by extracellular calcium-sensing receptor agonists induces mitogenic responses in the mouse osteoblastic MC3T3-E1 cell line. Biochem Biophys Res Commun 279:363-368, 2000

    Google Scholar 

  39. Johnson GL, Lapadat R: Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science 298:1911-1912, 2002

    Google Scholar 

  40. Elzi DJ, Bjornsen AJ, MacKenzie T, Wyman TH, Silliman CC: Ionomycin causes activation of p38 and p42/44 mitogen-activated protein kinases in human neutrophils. Am J Physiol Cell Physiol 281:C350-C360, 2001

    Google Scholar 

  41. Rutten MJ, Bacon KD, Marlink KL, Stoney M, Meichsner CL, Lee FP, Hobson SA, Rodland KD, Sheppard BC, Trunkey DD, Deveney KE, Deveney CW: Identification of a functional Ca2+-sensing receptor in normal human gastric mucous epithelial cells. Am J Physiol 277:G662-G670, 1999

    Google Scholar 

  42. Rutten MJ, Campbell DR, Luttropp CA, Fowler WM, Hawkey MA, Boland CR, Krauss ER, Sheppard BC, Crass RA, Deveney KE, Deveney CW: A method for the isolation of human gastric mucous epithelial cells for primary cell culture; A comparison of biopsy vs. surgical tissue. Methods Cell Sci 18:269-281, 1996

    Google Scholar 

  43. Lin K, Sadee W, Quillan JM: Rapid measurements of intracellular calcium using a fluorescence plate reader. Biotechniques 26:318-326, 1999

    Google Scholar 

  44. Thomas D, Tovey SC, Collins TJ, Bootman MD, Berridge MJ, Lipp P: A comparison of fluorescent Ca2+ indicator properties and their use in measuring elementary and global Ca2+ signals. Cell Calcium 28:213-223, 2000

    Google Scholar 

  45. Hellman P, Hellman B, Juhlin C, Juppner H, Rastad J, Ridefelt P, Akerstrom G: Regulation of proliferation in JEG-3 cells by a 500-kDa Ca2+ sensor and parathyroid hormone-related protein. Arch Biochem Biophys 307:379-385, 1993

    Google Scholar 

  46. Chattopadhyay N, Ye CP, Yamaguchi T, Kerner R, Vassilev PM, Brown EM: Extracellular calcium-sensing receptor induces cellular proliferation and activation of a nonselective cation channel in U373 human astrocytoma cells. Brain Res 851:116-124, 1999

    Google Scholar 

  47. Hobson SA, McNeil SE, Lee F, Rodland KD: Signal transduction mechanisms linking increased extracellular calcium to proliferation in ovarian surface epithelial cells. Exp Cell Res 258:1-11, 2000

    Google Scholar 

  48. Tu CL, Chang W, Bikle DD: The extracellular calcium-sensing receptor is required for calciuminduced differentiation in human keratinocytes. J Biol Chem 276:41079-41085, 2001

    Google Scholar 

  49. Racz GZ, Kittel A, Riccardi D, Case RM, Elliott AC, Varga G: Extracellular calcium sensing receptor in human pancreatic cells. Gut 51:705-711, 2002

    Google Scholar 

  50. Yamaguchi T, Yamauchi M, Sugimoto T, Chauhan D, Anderson KC, Brown EM, Chihara K: The extracellular calcium Ca2+ o-sensing receptor is expressed in myeloma cells and modulates cell proliferation. Biochem Biophys Res Commun 299:532-538, 2002

    Google Scholar 

  51. Mailland M, Waelchli R, Ruat M, Boddeke HG, Seuwen K: Stimulation of cell proliferation by calcium and a calcimimetic compound. Endocrinology 138:3601-3605, 1997

    Google Scholar 

  52. Chattopadhyay N, Ye CP, Yamaguchi T, Kifor O, Vassilev PM, Nishimura R, Brown EM: Extracellular calcium-sensing receptor in rat oligodendrocytes: expression and potential role in regulation of cellular proliferation and an outward K+ channel. Glia 24:449-458, 1998

    Google Scholar 

  53. Cheng I, Klingensmith ME, Chattopadhyay N, Kifor O, Butters RR, Soybel DI, Brown EM: Identification and localization of the extracellular calcium-sensing receptor in human breast. J Clin Endocrinol Metab 83:703-707, 1998

    Google Scholar 

  54. McNeil L, Hobson S, Nipper V, Rodland KD: Functional calcium-sensing receptor expression in ovarian surface epithelial cells. Am J Obstet Gynecol 178:305-313, 1998

    Google Scholar 

  55. Yamaguchi T, Chattopadhyay N, Kifor O, Butters RR Jr, Sugimoto T, Brown EM: Mouse osteoblastic cell line (MC3T3-E1) expresses extracellular calcium (Ca2+ o)-sensing receptor and its agonists stimulate chemotaxis and proliferation of MC3T3-E1 cells. J Bone Min Res 13:1530-1538, 1998

    Google Scholar 

  56. Quarles LD, Hartle JE 2nd, Siddhanti SR, Guo R, Hinson TK: A distinct cation-sensing mechanism in MC3T3-E1 osteoblasts functionally related to the calcium receptor. J Bone Min Res 12:393-402, 1997

    Google Scholar 

  57. Bai M, Quinn S, Trivedi S, Kifor O, Pearce SHS, Pollak MR, Krapcho K, Hebert SC, Brown EM: Expression and characterization of inactivating and activating mutations in the human Ca2+ o-sensing receptor. J Biol Chem 271:19537-19545, 1996

    Google Scholar 

  58. Motoyama HI, Friedman PA: Calcium-sensing receptor regulation of PTH-dependent calcium absorption by mouse cortical ascending limbs. Am J Physiol Renal Physiol 283:F399-F406, 2002

    Google Scholar 

  59. Galaris D, Evangelou A: The role of oxidative stress in mechanisms of metal-induced carcinogenesis. Crit Rev Oncol Hematol 42:93-103, 2002

    Google Scholar 

  60. Goldman R, Moshonov S, Chen X, Berchansky A, Furstenberger G, Zor U: Crosstalk between elevation of [Ca2+]i, reactive oxygen species generation and phospholipase A2 stimulation in a human keratinocyte cell line. Adv Exp Med Biol 433:41-45, 1997

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. The Department of Surgery, Oregon Health and Science University, Portland, Oregon, 97201, USA

    Jason Gilster, Kathy Bacon, Katie Marlink, Brett Sheppard, Clifford Deveney & Michael Rutten

Authors
  1. Jason Gilster
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kathy Bacon
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Brett Sheppard
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Clifford Deveney
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Michael Rutten
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gilster, J., Bacon, K., Marlink, K. et al. Bismuth Subsalicylate Increases Intracellular Ca2+, MAP-Kinase Activity, and Cell Proliferation in Normal Human Gastric Mucous Epithelial Cells. Dig Dis Sci 49, 370–378 (2004). https://doi.org/10.1023/B:DDAS.0000020488.55854.99

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/B:DDAS.0000020488.55854.99

Search

Navigation