Marie-Laure Kleme1, Alain Théophile Sané2, Carole Garofalo2, Emile Levy3. 1. Research Centre, CHU Sainte-Justine, 3175 Sainte-Catherine Road, Montreal, Quebec H3T 1C4, Canada; Department of Nutrition, Université de Montréal, Montreal, Quebec H3T 1C5, Canada. 2. Research Centre, CHU Sainte-Justine, 3175 Sainte-Catherine Road, Montreal, Quebec H3T 1C4, Canada. 3. Research Centre, CHU Sainte-Justine, 3175 Sainte-Catherine Road, Montreal, Quebec H3T 1C4, Canada; Department of Nutrition, Université de Montréal, Montreal, Quebec H3T 1C5, Canada. Electronic address: emile.levy@recherche-ste-justine.qc.ca.
Abstract
UNLABELLED: Cystic fibrosis (CF) is a multisystemic pathology caused by mutations of the CF transmembrane conductance regulator (CFTR) gene. OBJECTIVES: As the intestine harbors the greatest number of CFTR transcripts after birth and since CFTR plays a role in glutathione transport, we hypothesized that CFTR deletion might produce oxidative stress (OxS) and inflammation in CF intestinal epithelial cell. METHODS: CFTR gene was abrogated in Caco-2/15 enterocytes through the zinc-finger nuclease system. Their oxidative and inflammatory characteristics were appreciated under basal conditions and after the treatment with the pro-oxidant iron-ascorbate (Fe/Asc) complex and pro-inflammatory lipopolysaccharide (LPS). RESULTS: Intestinal epithelial cells with CFTR knockout spontaneously exhibited an increased lipid peroxidation level, reflected by malondialdehyde overproduction and reduced antioxidant defense characterized by low enzymatic activities of glutathione peroxidase and catalase. CFTR silencing also resulted in elevated protein expression of pro-inflammatory tumor necrosis Factor-α, interleukin-6, cyclooxygenase-2, and the transcription factor nuclear factor-κB. Moreover, exaggerated OxS and inflammation processes occurred in CFTR(-/-) cells in response to the addition of Fe/Asc and LPS, respectively. CONCLUSIONS: Intestinal Caco-2/15 cells with CFTR deletion, display innate oxidative and inflammatory features while being more sensitive to pro-oxidant and pro-inflammatory stimuli. These two pathophysiological processes could be implicated in CF-related intestinal disorders.
UNLABELLED: Cystic fibrosis (CF) is a multisystemic pathology caused by mutations of the CF transmembrane conductance regulator (CFTR) gene. OBJECTIVES: As the intestine harbors the greatest number of CFTR transcripts after birth and since CFTR plays a role in glutathione transport, we hypothesized that CFTR deletion might produce oxidative stress (OxS) and inflammation in CF intestinal epithelial cell. METHODS:CFTR gene was abrogated in Caco-2/15 enterocytes through the zinc-finger nuclease system. Their oxidative and inflammatory characteristics were appreciated under basal conditions and after the treatment with the pro-oxidant iron-ascorbate (Fe/Asc) complex and pro-inflammatory lipopolysaccharide (LPS). RESULTS: Intestinal epithelial cells with CFTR knockout spontaneously exhibited an increased lipid peroxidation level, reflected by malondialdehyde overproduction and reduced antioxidant defense characterized by low enzymatic activities of glutathione peroxidase and catalase. CFTR silencing also resulted in elevated protein expression of pro-inflammatory tumor necrosis Factor-α, interleukin-6, cyclooxygenase-2, and the transcription factor nuclear factor-κB. Moreover, exaggerated OxS and inflammation processes occurred in CFTR(-/-) cells in response to the addition of Fe/Asc and LPS, respectively. CONCLUSIONS: Intestinal Caco-2/15 cells with CFTR deletion, display innate oxidative and inflammatory features while being more sensitive to pro-oxidant and pro-inflammatory stimuli. These two pathophysiological processes could be implicated in CF-related intestinal disorders.
Authors: Meike Hohwieler; Lukas Perkhofer; Stefan Liebau; Thomas Seufferlein; Martin Müller; Anett Illing; Alexander Kleger Journal: United European Gastroenterol J Date: 2016-09-21 Impact factor: 4.623
Authors: Victor Emanuel Miranda Soares; Thiago Inácio Teixeira do Carmo; Fernanda Dos Anjos; Jonatha Wruck; Sarah Franco Vieira de Oliveira Maciel; Margarete Dulce Bagatini; Débora Tavares de Resende E Silva Journal: Mol Cell Biochem Date: 2021-09-16 Impact factor: 3.396