International Journal of Colorectal Disease

, Volume 25, Issue 3, pp 335–341 | Cite as

Carbachol induces TGF-alpha expression and colonic epithelial cell proliferation in sensory-desensitised rats

  • Kerem Bulut
  • Peter Felderbauer
  • Karoline Hoeck
  • Wolfgang E. Schmidt
  • Peter Hoffmann
Original Article



Signals for the expression of the peptide growth factors epidermal growth factor and transforming growth factor-α (TGFα) in the gastrointestinal mucosa are largely unknown. We have shown earlier that extrinsic afferents in the gastrointestinal tract induce TGFα expression in colonic mucosa via the deliberation of neurotransmitters substance P and calcitonin gene-related peptide. The aim of our present study was to determine the effects of carbachol on mucosal TGFα expression and epithelial cell proliferation in vivo.


Rats were divided in three groups. Group 1 was treated with vehicle only, group 2 received one single subcutaneous injection of 250 μg/kg of carbachol and animals in group 3 were sensory-desensitised prior to the injection of 250 μg/kg carbachol. TGFα expression and epithelial cell proliferation was evaluated by polymerase chain reaction, Western blot analysis and bromodeoxyuridine staining.


Carbachol induced a significant increase in mucosal epithelial cell proliferation and TGFα expression. Sensory desensitisation did neither abolish the increased TGFα expression nor the increase in epithelial cell proliferation.


Parasympathetic pathways are involved in the control of TGFα expression in gastrointestinal mucosa as well as in epithelial cell proliferation.


Gastrointestinal nervous system TGFα EGF Experimental model of colitis Carbachol Parasympathetic nerves Extrinsic primary afferents 


  1. 1.
    Luck MS, Bass P (1993) Effect of epidermal growth factor on experimental colitis in the rat. J Pharmacol Exp Ther 264:984–990PubMedGoogle Scholar
  2. 2.
    Procaccino F, Reinshagen M, Hoffmann P, Zeeh JM, Lakshmanan J, McRoberts JA, Patel A, French S, Eysselein VE (1994) Protective effect of epidermal growth factor in an experimental model of colitis in rats. Gastroenterology 107:12–17PubMedGoogle Scholar
  3. 3.
    Wu SQ, Zhu WY, Yu JR (1993) Submaxillary gland epidermal growth factor accelerated healing of chronic gastric mucosal lesions in rats. Sheng Li Xue Bao 45:575–580PubMedGoogle Scholar
  4. 4.
    Konturek SJ, Dembinski A, Warzecha Z, Brzozowski T, Gregory H (1988) Role of epidermal growth factor in healing of chronic gastroduodenal ulcers in rats. Gastroenterology 94:1300–1307PubMedGoogle Scholar
  5. 5.
    Tepperman BL, Kiernan JA, Soper BD (1989) The effect of sialoadenectomy on gastric mucosal integrity in the rat: roles of epidermal growth factor and prostaglandin E2. Can J Physiol Pharmacol 67:1512–1519PubMedGoogle Scholar
  6. 6.
    Egger B, Procaccino F, Lakshmanan J, Reinshagen M, Hoffmann P, Patel A, Reuben W, Gnanakkan S, Liu L, Barajas L, Eysselein VE (1997) Mice lacking transforming growth factor alpha have an increased susceptibility to dextran sulfate-induced colitis. Gastroenterology 113:825–832CrossRefPubMedGoogle Scholar
  7. 7.
    Egger B, Buchler MW, Lakshmanan J, Moore P, Eysselein VE (2000) Mice harboring a defective epidermal growth factor receptor (waved-2) have an increased susceptibility to acute dextran sulfate-induced colitis. Scand J Gastroenterol 35:1181–1187CrossRefPubMedGoogle Scholar
  8. 8.
    Egger B, Carey HV, Procaccino F, Chai NN, Sandgren EP, Lakshmanan J, Buslon VS, French SW, Buchler MW, Eysselein VE (1998) Reduced susceptibility of mice overexpressing transforming growth factor alpha to dextran sodium sulphate induced colitis. Gut 43:64–70PubMedGoogle Scholar
  9. 9.
    Sinha A, Nightingale J, West KP, Berlanga-Acosta J, Playford RJ (2003) Epidermal growth factor enemas with oral mesalamine for mild-to-moderate left-sided ulcerative colitis or proctitis. N Engl J Med 349:350–357CrossRefPubMedGoogle Scholar
  10. 10.
    Polk WH Jr, Dempsey PJ, Russell WE, Brown PI, Beauchamp RD, Barnard JA, Coffey RJ Jr (1992) Increased production of transforming growth factor alpha following acute gastric injury. Gastroenterology 102:1467–1474PubMedGoogle Scholar
  11. 11.
    Hoffmann P, Zeeh JM, Lakshmanan J, Wu VS, Procaccino F, Reinshagen M, McRoberts JA, Eysselein VE (1997) Increased expression of transforming growth factor alpha precursors in acute experimental colitis in rats. Gut 41:195–202PubMedCrossRefGoogle Scholar
  12. 12.
    Sottili M, Sternini C, Brecha NC, Lezoche E, Walsh JH (1992) Transforming growth factor alpha receptor binding sites in the canine gastrointestinal tract. Gastroenterology 103:1427–1436PubMedGoogle Scholar
  13. 13.
    Sottili M, Sternini C, Reinshagen M, Brecha NC, Nast CC, Walsh JH, Eysselein VE (1995) Up-regulation of transforming growth factor alpha binding sites in experimental rabbit colitis. Gastroenterology 109:24–31CrossRefPubMedGoogle Scholar
  14. 14.
    Hoffmann P, Reinshagen M, Zeeh JM, Lakshmanan J, Wu VS, Goebell H, Gerken G, Eysselein VE (2000) Increased expression of epidermal growth factor-receptor in an experimental model of colitis in rats. Scand J Gastroenterol 35:1174–1180CrossRefPubMedGoogle Scholar
  15. 15.
    Mendelsohn J (1990) Anti-epidermal growth factor receptor monoclonal antibodies as potential anti-cancer agents. J Steroid Biochem Mol Biol 37:889–892CrossRefPubMedGoogle Scholar
  16. 16.
    Carpenter G, Cohen S (1979) Epidermal growth factor. Annu Rev Biochem 48:193–216CrossRefPubMedGoogle Scholar
  17. 17.
    Anderson BK, Karnes WE, Pearson RK (1994) Bile acids regulate the growth rate of transformed colonic epithelium and increase TGF-alpha mRNA levels. Gastroenterology 54:A384Google Scholar
  18. 18.
    Hoffmann P, Mazurkiewicz J, Holtmann G, Gerken G, Eysselein VE, Goebell H (2002) Capsaicin-sensitive nerve fibres induce epithelial cell proliferation, inflammatory cell immigration and transforming growth factor-alpha expression in the rat colonic mucosa in vivo. Scand J Gastroenterol 37:414–422CrossRefPubMedGoogle Scholar
  19. 19.
    Bulut K, Felderbauer P, Hoeck K, Schmidt WE, Hoffmann P (2008) Increased duodenal expression of transforming growth factor-alpha and epidermal growth factor during experimental colitis in rats. Eur J Gastroenterol Hepatol 20:989–994CrossRefPubMedGoogle Scholar
  20. 20.
    Horikawa Y, Shatos MA, Hodges RR, Zoukhri D, Rios JD, Chang EL, Bernardino CR, Rubin PA, Dartt DA (2003) Activation of mitogen-activated protein kinase by cholinergic agonists and EGF in human compared with rat cultured conjunctival goblet cells. Invest Ophthalmol Vis Sci 44:2535–2544CrossRefPubMedGoogle Scholar
  21. 21.
    McCole DF, Keely SJ, Coffey RJ, Barrett KE (2002) Transactivation of the epidermal growth factor receptor in colonic epithelial cells by carbachol requires extracellular release of transforming growth factor-alpha. J Biol Chem 277:42603–42612CrossRefPubMedGoogle Scholar
  22. 22.
    Phillips TE (1992) Both crypt and villus intestinal goblet cells secrete mucin in response to cholinergic stimulation. Am J Physiol 262:G327–G331PubMedGoogle Scholar
  23. 23.
    Reinshagen M, Patel A, Sottili M, French S, Sternini C, Eysselein VE (1996) Action of sensory neurons in an experimental at colitis model of injury and repair. Am J Physiol 270:G79–G86PubMedGoogle Scholar
  24. 24.
    Holzer P (1991) Capsaicin: cellular targets, mechanisms of action, and selectivity for thin sensory neurons. Pharmacol Rev 43:143–201PubMedGoogle Scholar
  25. 25.
    Liu L, Barajas L (1993) The rat renal nerves during development. Anat Embryol (Berl) 188:345–361Google Scholar
  26. 26.
    Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159CrossRefPubMedGoogle Scholar
  27. 27.
    Ishidoya S, Morrissey J, McCracken R, Klahr S (1996) Delayed treatment with enalapril halts tubulointerstitial fibrosis in rats with obstructive nephropathy. Kidney Int 49:1110–1119CrossRefPubMedGoogle Scholar
  28. 28.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefPubMedGoogle Scholar
  29. 29.
    Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354CrossRefPubMedGoogle Scholar
  30. 30.
    Reinshagen M, Geerling I, Lakshmanan J, Rohm H, Lutz MP, Soinila S, Eysselein VE, Adler G (1997) Commercial mouse and human nerve growth factors contain nerve growth factor prohormone isoforms. J Neurosci Methods 76:75–81CrossRefPubMedGoogle Scholar
  31. 31.
    Kiba T, Tanaka K, Hoshino M, Numata K, Okano K, Inoue S (1995) Ventromedial hypothalamic lesions induce the proliferation of gastrointestinal mucosal cells in the rat. Life Sci 57:827–832CrossRefPubMedGoogle Scholar
  32. 32.
    Yoshimura R, Omori H, Somekawa S, Osaka T, Ito R, Inoue S, Endo Y (2006) Continuous carbachol infusion promotes peripheral cell proliferation and mimics vagus hyperactivity in a rat model of hypothalamic obesity. Biomed Res 27:81–88CrossRefPubMedGoogle Scholar
  33. 33.
    Frucht H, Jensen RT, Dexter D, Yang WL, Xiao Y (1999) Human colon cancer cell proliferation mediated by the M3 muscarinic cholinergic receptor. Clin Cancer Res 5:2532–2539PubMedGoogle Scholar
  34. 34.
    Ukegawa JI, Takeuchi Y, Kusayanagi S, Mitamura K (2003) Growth-promoting effect of muscarinic acetylcholine receptors in colon cancer cells. J Cancer Res Clin Oncol 129:272–278PubMedGoogle Scholar
  35. 35.
    Derynck R (1986) Transforming growth factor-alpha: structure and biological activities. J Cell Biochem 32:293–304CrossRefPubMedGoogle Scholar
  36. 36.
    Taylor P (1990) Cholinergic agonists. In: Goodman Gilman A, Rall TW, Bues AS, Taylor P (eds) The pharmacological basis of therapeutics. Pergamon, New York, pp 122–130Google Scholar
  37. 37.
    Kelly SM, Hunter JO (1990) Epidermal growth factor stimulates synthesis and secretion of mucus glycoproteins in human gastric mucosa. Clin Sci (Lond) 79:425–427Google Scholar
  38. 38.
    McLeay LM, Comeskey MA, Waters MJ (1990) Effects of epidermal growth factor on gastrointestinal electromyographic activity of conscious sheep. J Endocrinol 124:109–115CrossRefPubMedGoogle Scholar
  39. 39.
    Rhodes JA, Tam JP, Finke U, Saunders M, Bernanke J, Silen W, Murphy RA (1986) Transforming growth factor alpha inhibits secretion of gastric acid. Proc Natl Acad Sci USA 83:3844–3846CrossRefPubMedGoogle Scholar
  40. 40.
    Dembinski A, Drozdowicz D, Gregory H, Konturek SJ, Warzecha Z (1986) Inhibition of acid formation by epidermal growth factor in the isolated rabbit gastric glands. J Physiol 378:347–357PubMedGoogle Scholar
  41. 41.
    Hui WM, Chen BW, Kung AW, Cho CH, Luk CT, Lam SK (1993) Effect of epidermal growth factor on gastric blood flow in rats: possible role in mucosal protection. Gastroenterology 104:1605–1610PubMedGoogle Scholar
  42. 42.
    Code RA, Seroogy KB, Fallon JH (1987) Some transforming growth factor-alpha connections and their colocalization with enkephalin in the rat central nervous system. Brain Res 421:401–405CrossRefPubMedGoogle Scholar
  43. 43.
    Hoffmann P, Zeeh JM, Lakshmanan J, Liu L, Cooray D, Barajas L, Eysselein VE (1997) Transforming growth factor alpha-immunoreactivity in neural tissues of the rat stomach. Regul Pept 69:121–125CrossRefPubMedGoogle Scholar
  44. 44.
    Lazar LM, Blum M (1992) Regional distribution and developmental expression of epidermal growth factor and transforming growth factor-alpha mRNA in mouse brain by a quantitative nuclease protection assay. J Neurosci 12:1688–1697PubMedGoogle Scholar
  45. 45.
    Yasui W, Ji ZQ, Kuniyasu H, Ayhan A, Yokozaki H, Ito H, Tahara E (1992) Expression of transforming growth factor alpha in human tissues: immunohistochemical study and Northern blot analysis. Virchows Arch A Pathol Anat Histopathol 421:513–519CrossRefPubMedGoogle Scholar
  46. 46.
    Fisher DA, Lakshmanan J (1990) Metabolism and effects of epidermal growth factor and related growth factors in mammals. Endocr Rev 11:418–442CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Kerem Bulut
    • 1
  • Peter Felderbauer
    • 1
  • Karoline Hoeck
    • 2
  • Wolfgang E. Schmidt
    • 1
  • Peter Hoffmann
    • 2
  1. 1.Department of Internal Medicine I, St. Josef HospitalRuhr-University of BochumBochumGermany
  2. 2.Department of Internal Medicine IKliniken Essen-Mitte, Huyssens-StiftungEssenGermany

Personalised recommendations