03. Науково-дослідний інститут експериментальної та клінічної медицини ХНМУ

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Browsing 03. Науково-дослідний інститут експериментальної та клінічної медицини ХНМУ by Subject "food additives"

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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.
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    Semi-refined carrageenan induces eryptosis in a Ca2+-dependent manner
    (2022) Tkachenko, Anton; Prokopiuk, Volodymyr; Onishchenko, Anatolii
    Background: Semi-refined carrageenan (food additive E407a) is a widely used thickener, which has been reported to exert toxic and pro-inflammatory effects. In particular, there is accumulating evidence that it induces eryptosis, i.e. a programmed cell death of eryptocytes, via ROS-mediated pathways. However, the role of Ca2+-dependent mechanisms in E407a-induced eryptosis is not elucidated. Material and methods: Semi-refined carrageenan at concentrations of 0 mg/ml, 1 mg/ml, 5 mg/ml and 10 mg/ml was incubated with blood of intact female WAG rats (n=9) for 24 h in RPMI and fetal bovine serum. After 24 h, the samples were used to obtain erythrocyte suspensions. The obtained suspensions were stained with a Ca2+-sensitive FLUO4 AM probe (30 min, 2.5 µM). The fluorescence of FLUO4 in erythrocytes was detected by a BD FACSCanto II flow cytometer. Results: The intracellular Ca2+ levels are proportional to the fluorescence of FLUO4. The mean fluorescence intensities (MFI) were compared. Low levels (1 mg/ml) of E407a had no impact on Ca2+ concentrations in erythrocytes (p>0.05). On the contrary, high concentrations (5 mg/ml and 10 mg/ml) of this food additive promoted an increase in the intracellular Ca2+ levels. The MFI values were 2.3- and 2.5-fold higher, respectively (p<0.0001). In addition, the exposure to E407a at concentrations of 5 mg/ml and 10 mg/ml (p<0.0001) increased the percentage of cells with high FLUO4 fluorescence. Conclusion: Food additive E407a induces eryptosis in a Ca2+-dependent manner.