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Molecular Medicine

, Volume 18, Issue 3, pp 497–506 | Cite as

Milk Fat Globule-Epidermal Growth Factor 8 Is Decreased in Intestinal Epithelium of Ulcerative Colitis Patients and Thereby Causes Increased Apoptosis and Impaired Wound Healing

  • Qiu-jie Zhao
  • Yan-bo Yu
  • Xiu-li Zuo
  • Yan-yan Dong
  • Yan-qing Li
Research Article

Abstract

Milk fat globule-epidermal growth factor 8 (MFG-E8) plays an important role in maintaining intestinal barrier homeostasis and accelerating intestinal restitution. However, studies of MFG-E8 expression in humans with ulcerative colitis are lacking. We examined MFG-E8 expression in colonic mucosal biopsies from ulcerative colitis patients and healthy controls (n = 26 each) by real-time quantitative polymerase chain reaction (PCR), Western blot analysis and immunohistochemistry. MFG-E8 mRNA and protein expression was lower in ulcerative colitis patients than in controls. MFG-E8 expression was inversely correlated with mucosal inflammatory activity and clinical disease activity in patients. MFG-E8 was present in human intestinal epithelial cells both in vivo and in vitro. Apoptosis induction was also detected in the intestinal epithelium of ulcerative colitis patients by terminal-deoxynucleoitidyl transferase mediated nick-end labeling assay. We used lentiviral vectors encoding human MFG-E8 targeting short hairpin RNA to obtain MFG-E8 knockdown intestinal epithelia cell clones. MFG-E8 knockdown could promote apoptosis in intestinal epithelial cell lines, accompanied by a decrease in level of the antiapoptotic protein B-cell lymphoma 2 (BCL-2) and induction of the proapoptotic protein BCL2-associated protein X (BAX). The addition of recombinant human MFG-E8 led to decreased BAX and cleaved caspase-3 levels and induction of BCL-2 level in intestinal epithelia cells. MFG-E8 knockdown also attenuated wound healing on scratch assay of intestinal epithelial cells. The mRNA level of intestinal trefoid factor 3, a pivotal factor in intestinal epithelial cell migration and restitution, was downregulated with MFG-E8 knockdown. In conclusion, we demonstrated that decreased colonic MFG-E8 expression in patients with ulcerative colitis may be associated with mucosal inflammatory activity and clinical disease activity through basal cell apoptosis and preventing tissue healing in the pathogenesis of ulcerative colitis.

Notes

Acknowledgments

This work was supported by a key program from Clinical Projects of the Ministry of Health of China (2010).

References

  1. 1.
    Xavier RJ, Podolsky DK. (2007) Unravelling the pathogenesis of inflammatory bowel disease. Nature. 448:427–34.CrossRefGoogle Scholar
  2. 2.
    Strober W, Fuss I, Mannon P. (2007) The fundamental basis of inflammatory bowel disease. J. Clin. Invest. 117:514–21.CrossRefPubMedCentralGoogle Scholar
  3. 3.
    Podolsky DK. (2002) Inflammatory bowel disease. N. Engl. J. Med. 347:417–29.CrossRefGoogle Scholar
  4. 4.
    Yamamoto-Furusho JK, Podolsky DK. (2007) Innate immunity in inflammatory bowel disease. World J. Gastroenterol. 13:5577–80.CrossRefPubMedCentralGoogle Scholar
  5. 5.
    Matsuda R, et al. (2009) Quantitive cytokine mRNA expression profiles in the colonic mucosa of patients with steroid naive ulcerative colitis during active and quiescent disease. Inflamm. Bowel Dis. 15:328–34.CrossRefGoogle Scholar
  6. 6.
    Hagiwara C, Tanaka M, Kudo H. (2002) Increase in colorectal epithelial apoptotic cells in patients with ulcerative colitis ultimately requiring surgery. J. Gastroenterol. Hepatol. 17:758–64.CrossRefGoogle Scholar
  7. 7.
    Strater J, et al. (1997) CD95 (APO-1/Fas)-mediated apoptosis in colon epithelial cells: a possible role in ulcerative colitis. Gastroenterology. 113:160–7.CrossRefGoogle Scholar
  8. 8.
    Roda G, et al. (2010) Intestinal epithelial cells in inflammatory bowel diseases. World J. Gastroenterol. 16:4264–71.CrossRefPubMedCentralGoogle Scholar
  9. 9.
    Hanayama R, et al. (2002) Identification of a factor that links apoptotic cells to phagocytes. Nature. 417:182–7.CrossRefGoogle Scholar
  10. 10.
    Ensslin MA, Shur BD. (2007) The EGF repeat and discoidin domain protein, SED1/MFG-E8, is required for mammary gland branching morphogenesis. Proc. Natl. Acad. Sci. U S A. 104:2715–20.CrossRefPubMedCentralGoogle Scholar
  11. 11.
    Watanabe T, et al. (2005) Production of the long and short forms of MFG-E8 by epidermal keratinocytes. Cell Tissue Res. 321:185–93.CrossRefGoogle Scholar
  12. 12.
    Hanayama R, et al. (2004) Autoimmune disease and impaired uptake of apoptotic cells in MFG-E8-deficient mice. Science. 304:1147–50.CrossRefGoogle Scholar
  13. 13.
    Miksa M, Amin D, Wu R, Ravikumar TS, Wang P. (2007) Fractalkine-induced MFG-E8 leads to enhanced apoptotic cell clearance by macrophages. Mol. Med. 13:553–60.CrossRefPubMedCentralGoogle Scholar
  14. 14.
    Yamaguchi H, et al. (2008) Milk fat globule EGF factor 8 in the serum of human patients of systemic lupus erythematosus. J. Leukoc. Biol. 83:1300–7.CrossRefGoogle Scholar
  15. 15.
    Lyng R, Shur BD. (2007) Sperm-egg binding requires a multiplicity of receptor-ligand interactions: new insights into the nature of gamete receptors derived from reproductive tract secretions. Soc. Reprod. Fertil. Suppl. 65:335–51.PubMedGoogle Scholar
  16. 16.
    Bu HF, et al. (2007) Milk fat globule-EGF factor 8/lactadherin plays a crucial role in maintenance and repair of murine intestinal epithelium. J. Clin. Invest. 117:3673–83.PubMedPubMedCentralGoogle Scholar
  17. 17.
    Aziz MM, et al. (2009) MFG-E8 attenuates intestinal inflammation in murine experimental colitis by modulating osteopontin-dependent alphav-beta3 integrin signaling. J. Immunol. 182:7222–32.CrossRefGoogle Scholar
  18. 18.
    Chogle A, et al. (2011) Milk fat globule-EGF factor 8 is a critical protein for healing of dextran sodium sulfate-induced acute colitis in mice. Mol. Med. 17:502–7.CrossRefPubMedCentralGoogle Scholar
  19. 19.
    Matts SG. (1961) The value of rectal biopsy in the diagnosis of ulcerative colitis. Q. J. Med. 30:393–407.PubMedGoogle Scholar
  20. 20.
    Rutgeerts P, et al. (2005) Infliximab for induction and maintenance therapy for ulcerative colitis. N. Engl. J. Med. 353:2462–76.CrossRefGoogle Scholar
  21. 21.
    Livak KJ, Schmittgen TD. (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–8.CrossRefGoogle Scholar
  22. 22.
    Larocca D, et al. (1991) A Mr 46,000 human milk fat globule protein that is highly expressed in human breast tumors contains factor VIII-like domains. Cancer Res. 51:4994–8.Google Scholar
  23. 23.
    Jinushi M, et al. (2009) Milk fat globule epidermal growth factor-8 blockade triggers tumor destruction through coordinated cell-autonomous and immunemediated mechanisms. J. Exp. Med. 206:1317–26.CrossRefPubMedCentralGoogle Scholar
  24. 24.
    Rescigno M. (2011) The intestinal epithelial barrier in the control of homeostasis and immunity. Trends. Immunol. 32:256–64.CrossRefGoogle Scholar
  25. 25.
    Catalioto RM, Maggi CA, Giuliani S. (2010) Intestinal epithelial barrier dysfunction in disease and possible therapeutical interventions. Curr. Med. Chem. 18:398–426.CrossRefGoogle Scholar
  26. 26.
    Edelblum KL, Yan F, Yamaoka T, Polk DB. (2006) Regulation of apoptosis during homeostasis and disease in the intestinal epithelium. Inflamm. Bowel. Dis. 12:413–24.CrossRefGoogle Scholar
  27. 27.
    Qiu W, et al. (2011) PUMA-mediated intestinal epithelial apoptosis contributes to ulcerative colitis in humans and mice. J. Clin. Invest. 121:1722–32.CrossRefPubMedCentralGoogle Scholar
  28. 28.
    Jinushi M, et al. (2008) Milk fat globule EGF-8 promotes melanoma progression through coordinated Akt and twist signaling in the tumor microenvironment. Cancer Res. 68:8889–98.CrossRefGoogle Scholar
  29. 29.
    Iizuka M, Konno S. (2011) Wound healing of intestinal epithelial cells. World J. Gastroenterol. 17:2161–71.CrossRefPubMedCentralGoogle Scholar
  30. 30.
    Sturm A, Dignass AU. (2008) Epithelial restitution and wound healing in inflammatory bowel disease. World J. Gastroenterol. 14:348–53.CrossRefPubMedCentralGoogle Scholar
  31. 31.
    Ciacci C, Lind SE, Podolsky DK. (1993) Transforming growth factor beta regulation of migration in wounded rat intestinal epithelial monolayers. Gastroenterology. 105:93–101.CrossRefGoogle Scholar
  32. 32.
    Beck PL, et al. (2003) Transforming growth factor-beta mediates intestinal healing and susceptibility to injury in vitro and in vivo through epithelial cells. Am. J. Pathol. 162:597–608.CrossRefPubMedCentralGoogle Scholar
  33. 33.
    Dignass AU, Podolsky DK. (1993) Cytokine modulation of intestinal epithelial cell restitution: central role of transforming growth factor beta. Gastroenterology. 105:1323–32.CrossRefGoogle Scholar
  34. 34.
    Bulut K, et al. (2006) Vascular endothelial growth factor (VEGF164) ameliorates intestinal epithelial injury in vitro in IEC-18 and Caco-2 monolayers via induction of TGF-beta release from epithelial cells. Scand. J. Gastroenterol. 41:687–92.CrossRefGoogle Scholar
  35. 35.
    Dignass A, Lynch-Devaney K, Kindon H, Thim L, Podolsky DK. (1994) Trefoil peptides promote epithelial migration through a transforming growth factor beta-independent pathway. J. Clin. Invest. 94:376–83.CrossRefPubMedCentralGoogle Scholar
  36. 36.
    Mashimo H, Wu DC, Podolsky DK, Fishman MC. (1996) Impaired defense of intestinal mucosa in mice lacking intestinal trefoil factor. Science. 274:262–5.CrossRefGoogle Scholar
  37. 37.
    Durer U, Hartig R, Bang S, Thim L, Hoffmann W. (2007) TFF3 and EGF induce different migration patterns of intestinal epithelial cells in vitro and trigger increased internalization of E-cadherin. Cell. Physiol. Biochem. 20:329–46.CrossRefGoogle Scholar
  38. 38.
    Song M, Xia B, Li J. (2006) Effects of topical treatment of sodium butyrate and 5-aminosalicylic acid on expression of trefoil factor 3, interleukin 1beta, and nuclear factor kappaB in trinitrobenzene sulphonic acid induced colitis in rats. Postgrad. Med. J. 82:130–5.CrossRefPubMedCentralGoogle Scholar
  39. 39.
    Longman RJ, et al. (2006) Alterations in the composition of the supramucosal defense barrier in relation to disease severity of ulcerative colitis. J. Histochem. Cytochem. 54:1335–48.CrossRefPubMedCentralGoogle Scholar
  40. 40.
    Satsangi J, Silverberg MS, Vermeire S, Colombel JF. (2006) The Montreal classification of inflammatory bowel disease: controversies, consensus, and implications. Gut. 55:749–53.CrossRefPubMedCentralGoogle Scholar

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Authors and Affiliations

  • Qiu-jie Zhao
    • 1
    • 2
  • Yan-bo Yu
    • 1
    • 2
  • Xiu-li Zuo
    • 1
  • Yan-yan Dong
    • 1
  • Yan-qing Li
    • 1
  1. 1.Department of Gastroenterology, Qilu HospitalShandong UniversityJinan, ShandongChina
  2. 2.The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Qilu HospitalShandong UniversityJinan, ShandongChina

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