Morphological Condition of the Small Intestine after Prolonged Intake of Food Additive Carrageenan
About the author:
Gubinа-Vakulik G. I., Tkachenko A. S., Orlova M. A.
Type of article:
One of the potential etiological factors of chronic inflammatory bowel disease is known to be a food additive E407 (carrageenan), used in the food industry as a thickener. The aim of the investigation was to study the morphological status of the small intestine and features of inflam-mation and regeneration of small intestine enterocytes during prolonged systematic intake of the food additive carrageenan. Adult female Wistar rats were used for experiment. Rats of the basic group (10 animals) received a 1% solution of carrageenan in drinking water during 4 weeks. The control group consisted of 10 intact animals. Pieces of the small intestine (5 cm above the transition to the large intestine) were taken for morphological examination. Paraffin sections were stained with hematoxylin-eosin, Einarsson’s gallocyanin-chrome alum and van Gieson’s picrofuch-sin. PAS-reaction was done. Immunohistochemical reaction to Ki-67 was used for evaluation of proliferative activity of epithelial cells and immunohistochemical reaction to IL-1α was used to evaluate the intensivity of inflammatory process. Quantitative assessment of antigen Ki-67 was performed by determining the expression level in conventional units of lumines-cence, and quantitative assessment of IL-1α was performed by determining the optical density of the cytoplasmic IL-1α-producing cells in conventional units of optical density. Intake of carrageenan during 4 weeks caused damage to the mucosal epithelium and the development of chronic interstitial inflammation. Immunohistochemical study using antibodies to IL-1α revealed significant notice-able increase not only of IL-1α-synthesizing cells number in the small intestine, but also the level of IL-1α produc-tion. It was confirmed by significant increase in conventional optical density of the cytoplasm of IL-1α-synthesizing cells in the intestinal mucosa of the basic group compared to the control group (control group – 0. 17±0. 01, basic group – 0. 31±0. 02 conventional units of optical density, p < 0. 0001). Also increased expression of Ki-67 antigen in the proliferative zone of intestinal glands was found (control group – 0. 59±0. 02, basic group – 0. 66 ± 0. 01 con-ventional units of luminescence, p < 0. 01). It was experimentally established found that the intake of food additive carrageenan within 28 days resulted in accelerated injuring of epithelial cells and the development of interstitial inflammation in small intestine. Immuno-histochemically detected activation of IL-1α synthesis and release in the mucosa of the small intestine confirms the intensity of the current inflammatory process. Reinforced death of epithelial cells is accompanied by activation of proliferative processes, confirmed by the increase in the expression of Ki-67 antigen. Active proliferation of entero-cytes is a reparative reaction for the carrageenan intake that allows us to make a suggestion that further use of car-rageenan may lead to the exhaustion of reparative capacity with the formation of erosions and ulcers in the intestine.
carrageenan, rats, inflammation, Ki-67, IL-1α
- 1. Жуков, В. И. Система перекисного окисления липидов и активность апоптоза при экспериментальном хроническом гастроэнтероколите. / В. И. Жуков, А. С. Ткаченко // Науч. Вед. Бел. ун. Медицина. Фармация. – №18 (161), Вып. 23. – С. 138-141.
- 2. Загорский, С. Э. Хронические воспалительные заболевания у детей и подростков (современный подход) / С. Э. За-горский, Л. М. Беляева // Минск: БелМАПО. – 2007. – С. 4-28.
- 3. Клименко, Н. А. О значении лейкоцитов в повышенной сосудистой проницаемости при воспалении / Н. А. Клименко, Е. А. Павлова // Бюл. экс. биол. и мед. – 1999. – №8. – С. 165-167.
- 4. Патент на изобретение №97322 от 25. 01. 12 «Спосіб моделювання хронічного гастроентероколіту»
- 5. Bromley M. A comparison of proliferation markers (BrdUrd, Ki-67, PCNA) determined at each cell position in the crypts of normal human colonic mucosa / M. Bromley, D. Rew, A. Becciolini et al. // Eur. J. Histochem. – 1996. – Vol. 40. – P. 89–100.
- 6. Bruno S. Cell cycle dependent expression and stability of the nuclear protein detected by Ki-67 antibody in HL-60 cells / S. Bruno, Z. Darynkiewich // Cell. Prolif. – 1992. – №25. –P. 31–40.
- 7. Christopher J. Morris. Carrageenan-Induced Paw Edema in the Rat and Mouse / Christopher J. Morris // Inflammation Protocols. – 2003. – №225. – P. 115-121.
- 8. Holt P. R. Is Ki-67 a better proliferative marker in the colon then proliferating cell nuclear antigen? / P. R. Holt, S. F. Moss, A. M. Kapetanakis // Cancer. Epidimiol., Biomarkers. Prev. – 1997. – №6. – P. 131–135.
- 9. Pricolo V. E. Effects of lambda-carrageenan induced experimental enterocolitis on splenocyte function and nitric oxide pro-duction / V. E. Pricolo, S. M. Madhere, S. D. Finkelstein // J. Surg. Res. – 1996. – Vol. 66, №1. – P. 6-11.
- 10. Tobacman J. K. Review of harmful gastrointestinal effects of carrageenan in animal experiments / J. K. Tobacman // Environ-mental Health Perspectives. – 2001. Vol. 109, №10. – P. 983–994.
Publication of the article:
«Bulletin of problems biology and medicine» Issue 2 part 3 (109), 2014 year, 252-256 pages, index UDK 616. 341-091. 8-092. 9:577. 114. 4