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Influence of NADPH oxidase on inflammatory response in primary intestinal epithelial cells in patients with
Rima Ramonaite, Jurgita Skieceviciene, Gediminas Kiudelis
1Institute for Digestive Research, Academy of Medicine, Lithuanian University of Health Sciences, A, Mickeviciaus str, 9, LT-44307 Kaunas, Lithuania. l.kupcinskas@gmail.com.
Background:
The aim of this study is to evaluate the role of NADPH oxidase in primary intestinal epithelial cells during the active phase of UC.
Methods:
The primary human colonic epithelial cells were isolated from 19 patients with mild to moderate inflammatory activity of UC and 14 controls using chelation method. The cells were cultivated under the effect of mediators. Viability of cells was assessed by fluorescent microscopy. Production of reactive oxygen species (ROS) by the cells was measured fluorimetrically using Amplex Red. Production of TNF-α cytokine by the colonic epithelial cells was analysed by ELISA.
Results:
The results of our study showed that unstimulated cells of UC patients had a decreased viability, increased ROS production, but similar TNF-α level when compared to the controls. Stimulation with LPS increased hydrogen peroxide and TNF-α level in the UC group. Treatment of colonic epithelial cells with NADPH oxidase inhibitor increased cell viability decreased the levels of ROS and TNF-α in the LPS-treated cells isolated from UC patients.
Conclusions:
Our study showed that bacterial endotoxins induced NADPH oxidase activation in the colonic epithelial cells. Moreover, we revealed that treatment with NADPH oxidase inhibitors had a protective effect against pro-inflammatory action of LPS in human colonic epithelium cells during inflammation.
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