Kleme Marie-Laure, Sané Alain Théophile, Garofalo Carole, Levy Emile
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.
Research Centre, CHU Sainte-Justine, 3175 Sainte-Catherine Road, Montreal, Quebec H3T 1C4, Canada.
Int J Biochem Cell Biol. 2016 May;74:84-94. doi: 10.1016/j.biocel.2016.02.022. Epub 2016 Feb 26.
Cystic fibrosis (CF) is a multisystemic pathology caused by mutations of the CF transmembrane conductance regulator (CFTR) gene.
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.
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).
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.
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.
囊性纤维化（CF）是一种由囊性纤维化跨膜传导调节因子（CFTR）基因突变引起的多系统病理疾病。
由于出生后肠道中CFTR转录本数量最多，且CFTR在谷胱甘肽转运中发挥作用，我们推测CFTR缺失可能会在CF肠道上皮细胞中产生氧化应激（OxS）和炎症。
通过锌指核酸酶系统在Caco-2/15肠上皮细胞中敲除CFTR基因。在基础条件下以及用促氧化剂铁 - 抗坏血酸（Fe/Asc）复合物和促炎脂多糖（LPS）处理后，评估它们的氧化和炎症特征。
CFTR基因敲除的肠道上皮细胞自发地表现出脂质过氧化水平升高，表现为丙二醛产生过多，以及抗氧化防御能力降低，表现为谷胱甘肽过氧化物酶和过氧化氢酶的酶活性较低。CFTR沉默还导致促炎肿瘤坏死因子-α、白细胞介素-6、环氧化酶-2和转录因子核因子-κB的蛋白表达升高。此外，分别添加Fe/Asc和LPS后，CFTR（-/-）细胞中出现了过度的氧化应激和炎症过程。
CFTR缺失的肠道Caco-2/15细胞表现出先天性氧化和炎症特征，同时对促氧化剂和促炎刺激更敏感。这两个病理生理过程可能与CF相关的肠道疾病有关。
