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首页> 外文期刊>World Journal of Gastroenterology >Protective action of NADPH oxidase inhibitors and role of NADPH oxidase in pathogenesis of colon inflammation in mice
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Protective action of NADPH oxidase inhibitors and role of NADPH oxidase in pathogenesis of colon inflammation in mice

机译：NADPH氧化酶抑制剂的保护作用和NADPH氧化酶在小鼠结肠炎症的发病机理中的作用

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AIM: To investigate the role of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in colon epithelial cells in the pathogenesis of acute and chronic colon inflammation in a mouse model of dextran sulphate sodium (DSS)-induced colitis. METHODS: Balb/c mice were divided into three groups: 8 mice with acute DSS-induced colitis (3.5% DSS solution; 7 d), 8 mice with chronic DSS-induced colitis (3.5% DSS solution for 5 d + water for 6 d; 4 cycles; total: 44 d) and 12 mice without DSS supplementation as a control group. Primary colonic epithelial cells were isolated using chelation method. The cells were cultivated in the presence of mediators (lipopolysaccharide (LPS), apocynin or diphenyleneiodonium). Viability of cells was assessed by fluorescent microscopy. Production of reactive oxygen species (ROS) by the cells was measured fluorometrically using Amplex Red. Production of tumour necrosis factor-alpha (TNF-α) by the colonic epithelial cells was analysed by ELISA. Nox1 gene expression was assessed by real-time PCR. RESULTS: Our study showed that TNF-α level was increased in unstimulated primary colonic cells both in the acute and chronic colitis groups, whereas decreased viability, increased ROS production, and expression of Nox1 was characteristic only for chronic DSS colitis mice when compared to the controls. The stimulation by LPS increased ROS generation via NADPH oxidase and decreased cell viability in mice with acute colitis. Treatment with NADPH oxidase inhibitors increased cell viability and decreased the levels of ROS and TNF-α in the LPS-treated cells isolated from mice of both acute and chronic colitis groups. CONCLUSION: Our study revealed the importance of NADPH oxidase in the pathogenesis of both acute and chronic inflammation of the colon.
机译：目的：在葡聚糖硫酸钠（DSS）诱导的结肠炎小鼠模型中，研究烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶在结肠上皮细胞在急性和慢性结肠炎症的发病机理中的作用。方法：Balb / c小鼠分为三组：8例急性DSS诱发结肠炎（3.5％DSS溶液; 7 d），8例慢性DSS诱发结肠炎（3.5％DSS溶液5 d +水6 d）。 d; 4个周期;总计：44d）和12只未补充DSS的小鼠作为对照组。使用螯合法分离原代结肠上皮细胞。在存在介体（脂多糖（LPS），载脂蛋白或二苯并碘鎓）的情况下培养细胞。通过荧光显微镜评估细胞的活力。使用Amplex Red荧光分析法测量细胞产生的活性氧（ROS）。通过ELISA分析结肠上皮细胞的肿瘤坏死因子-α（TNF-α）的产生。通过实时PCR评估Nox1基因表达。结果：我们的研究表明，急性和慢性结肠炎组中未刺激的原代结肠细胞中的TNF-α水平均升高，而与DSS结肠炎相比，存活率下降，ROS生成增加和Nox1表达仅是慢性DSS结肠炎小鼠的特征。控制。 LPS刺激在患有急性结肠炎的小鼠中增加了通过NADPH氧化酶产生的ROS，并降低了细胞活力。用NADPH氧化酶抑制剂处理可提高细胞活力，并降低从急性和慢性结肠炎组小鼠分离的LPS处理的细胞中ROS和TNF-α的水平。结论：我们的研究揭示了NADPH氧化酶在结肠急性和慢性炎症发病机制中的重要性。

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