Email updates

Keep up to date with the latest news and content from BMC Gastroenterology and BioMed Central.

Open Access Research article

Influence of NADPH oxidase on inflammatory response in primary intestinal epithelial cells in patients with ulcerative colitis

Rima Ramonaite1, Jurgita Skieceviciene1, Gediminas Kiudelis2, Laimas Jonaitis2, Algimantas Tamelis3, Paulius Cizas4, Vilmante Borutaite4 and Limas Kupcinskas12*

Author Affiliations

1 Institute for Digestive Research, Academy of Medicine, Lithuanian University of Health Sciences, A. Mickeviciaus str. 9, LT-44307 Kaunas, Lithuania

2 Department of Gastroenterology, Academy of Medicine, Lithuanian University of Health Sciences, A. Mickeviciaus str. 9, LT-44307 Kaunas, Lithuania

3 Department of Surgery, Academy of Medicine, Lithuanian University of Health Science, A. Mickeviciaus str. 9, LT-44307 Kaunas, Lithuania

4 Institute for Neurosciences, Academy of Medicine, Lithuanian University of Health Sciences, A. Mickeviciaus str. 9, LT-44307 Kaunas, Lithuania

For all author emails, please log on.

BMC Gastroenterology 2013, 13:159  doi:10.1186/1471-230X-13-159

Published: 14 November 2013

Abstract

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.

Keywords:
Cell-biology; Cytokines; Ulcerative colitis; NADPH oxidase