Effects of glucagon-like peptide-2 on TNF-alpha/actinomycin D-induced intestinal epithelial injury: a scanning electron microscopic and immunohistochemical study

Year 2011, Volume: 70 Issue: 1, 27 - 38, 03.06.2011

Pelin Arda Pirincci , Pınar Turan Serap Arbak Sehnaz Bolkent

Abstract

Glucagon-like peptide-2 (GLP-2) is a peptide hormone with intestinotrophic activity, which is released from the intestinal endocrine cells. The aim of this study was to investigate the effects observed by scanning electron microscope of GLP-2 in intestinal epithelial injury induced by tumor necrosis factor-alpha (TNF-α)/actinomycin D (Act D). In addition, it aimed to elucidate whether the action mechanism of GLP-2 might be mediated by gastrointestinal hormones such as cholecystokinin and somatostatin. The intestinal epithelial injury was induced by intraperitoneal administration of 15 µg/kg TNF-α and 800 µg/kg Act D per mouse. Animals were injected subcutaneously 200 µg/kg GLP-2 analogue every 12 hr for 10 consecutive days prior to the administration of TNF-α and Act D. Scanning electron microscopic examination revealed severe epithelial damage in the small intestine of TNF-α/Act D-administered mice. The administration of  TNF-α/Act D was significantly increased the number of somatostatin-immunoreactive endocrine cells, but did not affect the number of cholecystokinin-immunoreactive endocrine cells in the intestinal mucosa. On the other hand, GLP-2 analogue pretreatment prevented TNF-α/Act D-induced intestinal epithelial injury by causing a marked decrease in the degenerative changes and the number of somatostatin-immunoreactive endocrine cells, and a significant increase in the number of  cholecystokinin-immunoreactive endocrine cells. As a result, the present study indicates that GLP-2 has a protective effect against TNF-α/Act D-induced intestinal epithelial injury. Morever protective effect of GLP-2 might be related to somatostatin and cholecystokinin.

Keywords

-

TNF-alfa/aktinomisin D ile oluşturulan ince bağırsak epitel hasarında glukagon-benzeri peptid-2’nin etkileri: taramalı elektron mikroskobik ve immunohistokimyasal bir çalışma

Abstract

Bağırsaktaki endokrin hücrelerden salınan glukagon-benzeri peptid-2 (GLP-2) intestinotrofik aktiviteye sahip peptid yapıda bir hormondur. Bu çalışmanın amacı, tümör nekroz faktör-alfa (TNF-α)/aktinomisin D (Act D) ile ince bağırsakta oluşturulan epitel hasarında GLP-2’nin taramalı elektron mikroskobu ile gözlenen etkilerini Arda Pirincci et.al. / IUFS Journal of Biology 2011, 70(1): 27-38 incelemekti. Bunun yanı sıra, GLP-2’nin etki mekanizmasının kolesistokinin ve somatostatin gibi gastrointestinal hormonlar aracılı olup olmadığını da aydınlatabilmek amaçlanmıştır. İnce bağırsak epitel hasarı, her fareye intraperitonal olarak 15 µg/kg TNF-α ve 800 µg/kg Act D uygulanması ile oluşturuldu. Hayvanlara TNF-α ve Act D uygulamasından önce 10 gün süreyle her 12 saatte bir subkutanöz olarak 200 µg/kg dozda GLP-2 analoğu enjekte edildi. Taramalı elektron mikroskobu ile yapılan incelemelerde TNF-α/Act D uygulanan farelerin ince bağırsak dokusunda şiddetli epitel hasarı tespit edildi. TNF-α/Act D uygulaması bağırsak mukozasındaki somatostatin immunreaktif endokrin hücre sayısını önemli ölçüde arttırdı, ancak kolesistokinin immunreaktif endokrin hücre sayısını değiştirmedi. Öte yandan, GLP-2 ön uygulaması TNF-α/Act D ile oluşturulan ince bağırsak epitel hasarını dejeneratif değişikliklerde ve somatostatin immunreaktif endokrin hücre sayısında belirgin bir azalmaya, kolesistokinin immunreaktif endokrin hücre sayısında ise belirgin bir artışa neden olarak önledi. Sonuç olarak bu çalışma, GLP-2’nin TNF-α/Act D ile ince bağırsakta oluşturulan epitel hasarına karşı koruyucu etkiye sahip olduğunu göstermiştir. Ayrıca, GLP-2’nin koruyucu etkisi kolesistokinin ve somatostatin ile ilişkilendirilebilir.

Keywords

Glukagon-benzeri peptid-2, TNF-α/aktinomisin D, İnce bağırsak epitel hasarı, Kolesistokinin, Somatostatin

References

• Arda-Pirincci P. and Bolkent S. (2011) The role of glucagon-like peptide-2 on apoptosis, cell proliferation, and oxidant-antioxidant system at a mouse model of intestinal injury induced by tumor necrosis factor-alpha /actinomycin D. Molecular and Cellular Biochemistry, 350:13-27.
• Benali N., Ferjoux G., Puente E., Buscail L., Susuni C. (2000) Somatostatin receptors. Digestion, 62: 27-32.
• Boushey R.P., Yusta B., Drucker D.J. (1999) Glucagon-like mortality and reduces the severity of indomethacin-induced American Endocrinology and Metabolism,277(40): 937-47. decreases murine of Journal Physiology
• Buchman A.L., Katz S., Fang J.C., Bernstein C.N., Abou-Assi S.G., for the Teduglutide Study Group. (2010) Teduglutide, a novel mucosally active analog of glucagon-like peptide-2 (GLP-2) for the treatment of moderate to severe Crohn's disease. Inflammatory Bowel Diseases, 16: 962-73. Chandra R. and Liddle Cholecystokinin. Curr Opin Endocrinol Diabetes Obes, 14: 63-7. R.A. (2007)
• Chang Q. and Teppermann B.L. (2001) The role of protein kinase C isozymes in TNF- α-induced cytotoxicity to a rat intestinal epithelial cell line. American Journal of Physiology-Gastrointestinal Physiology, 280: 572-83. and Liver
• Chang Q. and Teppermann B.L. (2003) Effect of selective PKC İsoform activation and inhibition on TNF-α-induced injury and apoptosis in human intestinal epithelial cells. British Journal of Pharmacology, 140: 41-52.
• De Haan J.J., Lubbers T., Hadfoune M., Luyer M.D., Dejong C.H., Greve JW. (2008) Postshock intervention with high-lipid enteral nutrition reduces inflammation and tissue damage. Annals of Surgery, 248: 842-8.
• De Jonge F., Van Nassauw L., De Man J.G., De Winter B.Y., Van Meir F., Depoortere I., Peeters T.L., Pelckmans P.A., Van Marcks E., Timmermans J.P. (2003) Effects of schistosoma somatostatin and somatostatin receptor 2A expression Neurogastroenterology and Motility, 15: 149-59. ınfection on in mouse ileum.
• Drucker D.J., Ehrlich P., Asa S.L., Brubaker P.L. (1996) Induction of intestinal epithelial proliferation by glucagon-like peptide-2. Proceedings of The National Academy of Sciences of The United States of America, 3: 7911-6.
• Drucker D.J. (2005) Biologic actions and therapeutic potential of the proglucagon- derived peptides. Nature Clinical Practice Endocrinology & Metabolism, 1: 22-31.
• Estall J.L. and Drucker D.J. (2003) Dual regulation of cell proliferation and survival via activation of glucagon-like peptide-2 receptor signaling. Journal of Nutrition, 133: 3708-11.
• Estall J.L and Drucker D.J. (2005) Tales beyond the crypt: glucagon-like peptide-2 and cytoprotection in the intestinal mucosa. Endocrinology, 146: 19-21.
• Ferjoux G., Bousquet C., Cordelier P., Benali N., Lopez F., Rochaix P., Buscail L., Susini C. somatostatin controlling cell proliferation. Journal of Physiology-Paris, 94: 205-10. of receptors negatively D Actinomycin deoxyribonucleic acid-dependent synthesis of ribonucleic acid. Science, 136: 315-6.
• Green D.W., Gomez G., Greeley G.H. (1989) Gastrointestinal peptides. Gastroenterology Clinics of North America, 18: 695-733.
• Ivory C.P., Wallace , Mccafferty D.M., Sigalet D.L. (2008) Interleukin-10-independent anti-inflammatory actions of glucagon-like peptide Physiology-Gastrointestinal Physiology, 295: G1202-10. Journal and Liver
• Jeppesen P.B., Sanguinetti E.L., Buchman A., Howard L., Scolapio J.S., Ziegler T.R., Gregory J., Tappenden K.A., Holst J., Mortensen P.B. (2005) Teduglutide (ALX- 0600), a dipeptidyl peptidase IV resistant glucagon-like peptide 2 analogue, improves intestinal function in short bowel syndrome patients. Gut, 54: 1224-31.
• Jones B.E., Lo C.R., Liu H., Srinivasan A., Streetz K., Valentino K.L., Czaja M.J. (2000) Hepatocytes sensitized to tumor necrosis factor-alpha cytotoxicity undergo apoptosis through caspase-dependent and caspase-independent pathways. Journal of Biological Chemistry, 275: 705-12.
• Kunstle G., Leist M., Uhlig S., Revesz L., Feifel R., Mackenzie A., Wendel A. (1997) Ice-protease inhibitors block murine liver injury and apoptosis caused by CD95 or by TNF-α. Immunology Letters, 55: 5-10.
• Leist M., Gantner F., Bohlinger I., Germann P.G., Tiegs G., Wendel A.(1994) Murine hepatocyte apoptosis induced in vitro and in vivo by TNF-α requires transcriptional arrest. Journal of Immunology, 153: 1778- 88.
• Leist M., Gantner F., Naumann H., Bluethmann H., Vogt K., Brigelius-Flohe R., Nicotera P., Volk H. (1997) Tumor necrosis factor- induced apoptosis during the poisoning of mice with hepatotoxins. Gastroenterology, 112: 923-34.
• L’Heureux M.C. and Brubaker P.L. (2003) Glucagon-like peptide-2 and common therapeutics in a murine model of ulcerative colitis. Journal of Pharmacology and Experimental Therapeutics, 306:347-354.
• Martin G.R., Wallace L.E., Sigalet D.L. (2004) Glucagon-like peptide-2 induces intestinal adaptation in parenterally fed rats with short bowel syndrome. American Journal of Physiology-Gastrointestinal Physiology, 286: G964-72. and Liver
• Mueller C. (2002) Tumour necrosis factor in mouse inflammation. Immunology, 105: 1-8.
• Panteris V., Kouroumalis E. Somatostatin and inflammation. (2007) In: Rarley EP, ed. Progress in neuropeptide research. New York: Nova science publishers, 642-48.
• Paran D. and Paran H. (2003) Somatostatin analogs in rheumatoid arthritis and other inflammatory conditions. Investigational Drugs, 4: 578-82. Opinion in
• Pastorino J.G., Simbula G., Yamamoto K., Glascott P.A., Rothman R.J., Farber J.L. (1996) The cytotoxicity of tumor necrosis factor depends on induction of the mitochondrial Journal 271:29792-8. transition. Chemistry, of
• Remick D.G. and Kunkel S.L. (1993) Pathophysiologic alterations induced by tumor necrosis factor. International Review of Experimental Pathology, 3: 7-25.
• Sawicki S.G. and Godman G.C. (1971) On the differential cytotoxicity of actinomycin D. Journal of Cell Biology, 50: 746-61.
• Schwartz H.S. and Sodergren J.E. (1968) Actinomycin D effects on nucleic acids during tumor regression. Cancer Research, 28: 445-51
• Schwartz H.S., Sternberg S.S., Philips F.S. (1963) Pharmacology of mitomycin C. IV. Effects in vivo on nucleic acid synthesis; comparison with actinomycin D. Cancer Research, 23: 1125-37.
• Sinclair E.M. and Drucker D.J. (2005) Proglucagon-derived peptides: mechanisms of Physiology, 20: 357-65. potential.
• Tavakkolizadeh A., Shen R., Abraham P., Kormi N., Seifert P., Edelman E.R., Jacops D.O., Zinner M.J., Ashley S.W., Whang E.E. (2000) Glucagon-like peptide-2: a new treatment enteritis. Journal of Surgical Research, 91: 77-82.
• chemotherapy-induced
• Thomas R.P., Hellmich M.R., Townsend C.M., Evers B.M. (2003) Role of gastrointestinal hormones in the proliferation of normal and neoplastic tissues. Endocrine Reviews, 24: 571-99.
• Van Horssen R., Ten Hagen T.L.M., Eggermont A.M.M. (2006) TNF-α in cancer treatment: molecular insights, antitumor effects, and clinical utility. Oncologist, 11: 397-408.
• Yusta B., Huang L., Munroe D., Wolff G., Fantaske R., Sharma S., Demchyshyn L., Asa Enteroendocrine localization of GLP-2 receptor expression in humans and rodents. Gastroenterology, 119: 744-55. (2000)