Browsing by Subject "colitis"

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Intake of semi-refined carrageenan causes low-grade colonic inflammation and alters expression of epithelial-mesenchymal transition markers
(2022) Onishchenko, Anatolii; Gubina-Vakulyck, Galina; Knigavko, Oleksandr; Sharashydze, Ketino; Pionova, Olena; Butov, Dmytro; Polikarpova, Hanna; Tkachenko, Anton
The aim of this research was to evaluate the effects of semi-refined carrageenan consumed orally on the amount of CD3 and CD68 positive cells residing in the colon and the expression of endothelial-mesenchymal transition (EMT) markers. Materials and methods. To assess colonic expression of CD3+, CD68+, fascin, vimentin and E-cadherin, sections obtained from 8 rats orally exposed to semi-refined carrageenan (E407a) at a dose of 140 mg / kg of weight during 14 consecutive days and 8 control animals were immunostained with the corresponding antibodies. Levels of expression were assessed quantitatively. Results. Oral exposure to semi-refined carrageenan resulted in an increase in CD3 and CD68 positive cells in the colonic lamina propria. Quantitative analysis of fascin, vimentin and E-cadherin immunostaining revealed changes in expression of these EMT markers both in the colonic stroma and epithelial cells. Vimentin and fascin were overexpressed in stromal and epithelial cells, while E-cadherin was upregulated in the stroma and downregulated in epithelia. Conclusions. Our observations suggest that oral intake of semi-refined carrageenan results in the development of low-grade colonic inflammation accompanied by infiltration with CD3 and CD68 positive cells and changes in the expression of EMT markers.
The significance of ischemia for the proliferative activity of the mucosa in inflammatory bowel diseases
(2022) Nakonechna, Oksana; Vyshnytska, I.; Vasylyeva, Irina; Babenko, Olha; Voitenko, Stanislav; Bondarenko, A.; Gargin, Vitaliy
The state of the microcirculatory bed remains one of the determining factors in course of inflammatory bowel diseases (IBD). The presence of small foci of ischemia could realize in dystrophic-necrobiotic consequences, which can also underlie the development or strengthening of the inflammatory process. Based on above, the goal of our study was to determine the impact of the development of mucosal ischemia in the colon on the activity of proliferative processes during inflammation. Materials and methods. The study was performed on 12 adult WAG rats with modeling IBD by oral administration of 2.5% solution Dextran Sulfate Sodium. Serial slides of the colon where made with stained with hematoxylin and eosin, according to Rego, immunohistochemical examination (IHC) to Ki67. Volume of ischemic area and Ki67 expression were detected. Statistical comparison was performed. Results. Inspection microscopy in the DSS experimental group determined alterative-desquamative changes in the surface epithelium and epithelium of intestinal glands (crypt); diffuse polymorphic cellular infiltration in the mucous membrane, which in some places spread to the submucosal base, that are morphological manifestations of IBD. Foci of ischemia had been detected in that group with 13.09±0.67% volume as just microfocal changes were observed in intact animals (p < 0.05). Detection of proliferative activity depending on ischemic signs was realized in different level of Ki67 expression. So, lowest level of Ki67 was estimated in mucosa above ischemia (18.06±3.33%). Most pronounced expression of Ki67 was observed in IBD group in area which no connected with ischemia and was even 57.71±4.68% (p < 0.05). Conclusions. Ki67 was strongly expressed in epithelial cells of the colon both in intact tissue and in modeling IBD with significant increasing expression more than twice in inflammatory group (p < 0.05) but spreading of activity process was uneven. Collation of slides with ICH and Rego staining realized in estimation of strong negative correlation between Ki67 expression and ischemia (r=-0.819).
The viability of leukocytes and reactive oxygen species generation by them in rats with chronic colitis
(2022) Babenko, Olha; Vasylyeva, Irina; Nakonechna, Oksana; Popova, Liudmyla; Voitenko, Stanislav; Pustova, Nataliia
The aim: To assess reactive oxygen species production by leukocytes and their viability in rats with chronic colitis. Materials and methods: Reactive oxygen species production was estimated in leukocytes, isolated from rats with Dextran Sulfate Sodium-induced chronic colitis and control rats, by flow cytometry using the fluorescent probe 2’,7’-dichlorodihydrofluorescein diacetate. Leukocyte viability and apoptosis stages were assessed by flow cytometry using annexin V and 7-aminoactinomycin D staining. White blood cell counting was carried out with using Hematology Analyzer. Results: The increased fluorescence intensity of 2’,7’-dichlorofluorescein in viable leukocytes by 36.7% was revealed in rats with chronic colitis compared control rats. A significant decrease in the percentage of viable cells and an increase in apoptotic cells were found compared to intact animals. Leukocytes, granulocytes, monocytes, lymphocytes counts in blood of experimental group animals were significantly higher compared to control those. Conclusions: Our findings indicate that Dextran Sulfate Sodium-induced chronic colitis increases an intracellular production of reactive oxygen species by leukocytes. Despite of increased leukopoiesis it reduces viability of white blood cells and promotes their apoptosis via stimulation of oxidative stress.