Influence of NADPH oxidase on inflammatory response in primary intestinal epithelial cells in patients with ulcerative colitis.
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.
Rima
R
Ramonaite
Jurgita
J
Skieceviciene
Gediminas
G
Kiudelis
Laimas
L
Jonaitis
Algimantas
A
Tamelis
Paulius
P
Cizas
Vilmante
V
Borutaite
Institute for Digestive Research, Academy of Medicine, Lithuanian University of Health Sciences, A, Mickeviciaus str, 9, LT-44307 Kaunas, Lithuania. l.kupcinskas@gmail.com.
Limas
L
Kupcinskas
D000328:Adult; D016022:Case-Control Studies; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003093:Colitis, Ulcerative / Q000276:immunology / Q000378:metabolism*; D003106:Colon / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D005260:Female; D006801:Humans; D006861:Hydrogen Peroxide / Q000276:immunology / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000502:physiology*; D009861:Onium Compounds / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000276:immunology / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D055815:Young Adult
D004791:Enzyme Inhibitors; D008070:Lipopolysaccharides; D009861:Onium Compounds; D017382:Reactive Oxygen Species; D014409:Tumor Necrosis Factor-alpha; C007517:diphenyleneiodonium; D006861:Hydrogen Peroxide; D019255:NADPH Oxidases