Digestive Diseases and Sciences

, Volume 54, Issue 5, pp 928–936 | Cite as

Taurocholate Potentiates Ethanol-Induced NF-κB Activation and Inhibits Caspase-3 Activity in Cultured Rat Gastric Mucosal Cells

  • Harri Mustonen
  • Pauli Puolakkainen
  • Esko Kemppainen
  • Tuula Kiviluoto
  • Eero Kivilaakso
Original Article


We have previously shown that ethanol (EtOH) induces protective NF-κB activation in gastric surface epithelial cells. This study investigates the defense systems in rat gastric mucosal cells (RGM-1) exposed simultaneously to EtOH and taurocholate (TC) or acetylsalicylic acid (ASA). Simultaneous exposure to ASA and EtOH increased EtOH-induced caspase-3 activity and decreased cell viability, indicating synergetic damaging action. Simultaneous exposure to TC (5 mM) with EtOH (5%) increased EtOH-induced NF-κB activation, opposing EtOH-induced decrease in cell membrane integrity and in cell viability as shown by decreasing RelA expression with siRNA technique. Low doses of TC decreased the EtOH (5%) induced caspase-3 activity independently from NF-κB pathway and inhibited EtOH-induced decrease in caspase-3 precursor protein levels, also indicating the inhibition of caspase-3 pathway. The TC (5 mM)-induced protection in EtOH exposed tissues seems to have two distinct pathways, inhibition of apoptosis and enhancement of NF-κB signaling.


NF-κB Acetylsalicylic acid Taurocholate gastric mucosa Apoptosis 



Nuclear factor-κB






Acetylsalicylic acid



Supported by Helsinki University Central Hospital Research Funds (EVO); Helsinki, Finland, Helsinki University Research Funds, Helsinki, Finland; Jenny and Antti Wihuri Foundation, Helsinki, Finland and Biomedicum Research Foundation, Helsinki, Finland. The authors thank Mrs. Sanna Vainionpää and Mrs. Paula Kokko for valuable technical assistance.


  1. 1.
    Mustonen H, Kiviluoto T, Puolakkainen P, Kivilaakso E. Heat shock inhibits ethanol-induced NF-B activation in cultured rat gastric mucosal cells. Gastroenterology. 2005;128:A-599.Google Scholar
  2. 2.
    Mustonen H, Hietaranta A, Puolakkainen P, et al. Ethanol-induced NF-κB activation protects against cell injury in cultured rat gastric mucosal epithelium. Am J Physiol Gastrointest Liver Physiol. 2007;292:G1614–G1621. doi: 10.1152/ajpgi.00273.2006.PubMedCrossRefGoogle Scholar
  3. 3.
    Perkins ND. The Rel/NF-kappa B family: friend and foe. Trends Biochem Sci. 2000;25:434–440. doi: 10.1016/S0968-0004(00)01617-0.PubMedCrossRefGoogle Scholar
  4. 4.
    Jobin C, Sartor RB. The I kappa B/NF-kappa B system: a key determinant of mucosal inflammation and protection. Am J Physiol Cell Physiol. 2000;278:C451–C462.PubMedGoogle Scholar
  5. 5.
    Meng Y, Ma QY, Kou XP, Xu J. Effect of resveratrol on activation of nuclear factor kappa-B and inflammatory factors in rat model of acute pancreatitis. World J Gastroenterol. 2005;11:525–528.PubMedGoogle Scholar
  6. 6.
    Duane WC, Wiegand DM, Sievert CE. Bile acid and bile salt disrupt gastric mucosal barrier in the dog by different mechanisms. Am J Physiol. 1982;242:G95–G99.PubMedGoogle Scholar
  7. 7.
    Kiviluoto T, Mustonen H, Kivilaakso E. Effect of barrier-breaking agents on intracellular pH and epithelial membrane resistances: studies in isolated necturus antral mucosa exposed to luminal acid. Gastroenterology. 1989;96:1410–1418.PubMedGoogle Scholar
  8. 8.
    Thomas AJ, Nahrwold DL, Rose RC. Detergent action of sodium taurocholate on rat gastric mucosa. Biochim Biophys Acta. 1972;282:210–213. doi: 10.1016/0005-2736(72)90326-4.PubMedCrossRefGoogle Scholar
  9. 9.
    Bidel S, Mustonen H, Khalighi-Sikaroudi G, et al. Effect of the ulcerogenic agents ethanol, acetylsalicylic acid and taurocholate on actin cytoskeleton and cell motility in cultured rat gastric mucosal cells. World J Gastroenterol. 2005;11:4032–4039.PubMedGoogle Scholar
  10. 10.
    Konturek PC, Kania J, Gessner U, Konturek SJ, Hahn EG, Konturek JW. Effect of vitamin C-releasing acetylsalicylic acid on gastric mucosal damage before and after Helicobacter pylori eradication therapy. Eur J Pharmacol. 2004;506:169–177. doi: 10.1016/j.ejphar.2004.10.048.PubMedCrossRefGoogle Scholar
  11. 11.
    Sreedhar AS, Csermely P. Heat-shock proteins in the regulation of apoptosis: new strategies in tumor therapy: a comprehensive review. Pharmacol Ther. 2004;101:227–257. doi: 10.1016/j.pharmthera.2003.11.004.PubMedCrossRefGoogle Scholar
  12. 12.
    Schreiber E, Matthias P, Muller MM, Schaffner W. Rapid detection of octamer binding proteins with ‘mini-extracts’, prepared from a small number of cells. Nucleic Acids Res. 1989;17:6419. doi: 10.1093/nar/17.15.6419.PubMedCrossRefGoogle Scholar
  13. 13.
    Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248–254. doi: 10.1016/0003-2697(76)90527-3.PubMedCrossRefGoogle Scholar
  14. 14.
    Puurunen J, Huttunen P, Hirvonen J. Interactions between ethanol and acetylsalicylic acid in damaging the rat gastric mucosa. Acta Pharmacol Toxicol. 1983;52:321–327.Google Scholar
  15. 15.
    Saika M, Ueyama T, Senba E. Expression of immediate early genes, HSP70, and COX-2 mRNAs in rat stomach following ethanol ingestion. Dig Dis Sci. 2000;45:2455–2462. doi: 10.1023/A:1005615714451.PubMedCrossRefGoogle Scholar
  16. 16.
    Mustonen H, Kiviluoto T, Paimela H, Puolakkainen P, Kivilaakso E. Calcium signaling is involved in ethanol induced volume decrease and gap junction closure in cultured rat gastric mucosal cells. Dig Dis Sci. 2005;50:103–110. doi: 10.1007/s10620-005-1286-9.PubMedCrossRefGoogle Scholar
  17. 17.
    Mustonen H, Kiviluoto T, Puolakkainen P, Kivilaakso E. Ethanol induces volume changes and gap junction closure via intracellular Ca2+ signalling pathway in cultured rabbit gastric epithelial cells. Scand J Gastroenterol. 2004;39:104–110. doi: 10.1080/00365520310007855.PubMedCrossRefGoogle Scholar
  18. 18.
    Marzioni M, LeSage GD, Glaser S, et al. Taurocholate prevents the loss of intrahepatic bile ducts due to vagotomy in bile duct-ligated rats. Am J Physiol Gastrointest Liver Physiol. 2003;284:G837–G852.PubMedGoogle Scholar
  19. 19.
    Komichi D, Tazuma S, Nishioka T, Hyogo H, Une M, Chayama K. Unique inhibition of bile salt-induced apoptosis by lecithins and cytoprotective bile salts in immortalized mouse cholangiocytes. Dig Dis Sci. 2003;48:2315–2322. doi: 10.1023/B:DDAS.0000007869.67105.27.PubMedCrossRefGoogle Scholar
  20. 20.
    Que FG, Phan VA, Phan VH, LaRusso NF, Gores GJ. GUDC inhibits cytochrome c release from human cholangiocyte mitochondria. J Surg Res. 1999;83:100–105. doi: 10.1006/jsre.1999.5574.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Harri Mustonen
    • 1
  • Pauli Puolakkainen
    • 1
  • Esko Kemppainen
    • 1
  • Tuula Kiviluoto
    • 1
  • Eero Kivilaakso
    • 1
  1. 1.Department of SurgeryHelsinki University Central HospitalHelsinkiFinland

Personalised recommendations