NADPH氧化酶NOX1、NOX2和DUOX2在溃疡性结肠炎中的表达分析

发布时间：2018-11-27 09:47

【摘要】：目的探讨NADPH氧化酶家族(nicotinamide adenine dinucleotide phosphate oxidases,NOXs)的主要成员NOX1、NOX2和双氧化物酶(dual oxidase,DUOX)2(人:NOX1、NOX2和DUOX2;小鼠:Nox1、Nox2和Duox2)在人炎症性肠病和小鼠肠炎模型结肠中的表达。方法人结肠标本选自于通过结肠镜活检或手术切除的炎症性肠病组织及距离病变组织边缘5 cm以上正常组织;同时选用8~10周龄的C57BL/6雌性小鼠建立结肠炎模型,采用掷硬币法将其随机分为对照组和慢性肠炎组,慢性肠炎组先自由饮用1.0%(质量分数)葡聚糖硫酸钠7 d,然后自由饮水14 d,循环3次;通过体质量变化和HE染色方法评估肠道炎性反应程度。采用实时定量PCR技术和免疫组织化学方法分别检测结肠组织中NOX1、NOX2及DUOX2 mRNA和蛋白表达情况,并分析在人炎症性肠病中三者表达情况与临床特征之间的关系。结果 NOX1、NOX2和DUOX2 mRNA在人炎症性肠病结肠组织中的表达量均显著高于自身正常对照组织,分别增高2.8、2.0和3.3倍(P均0.01);与人不同,Nox1和Duox2 mRNA在小鼠慢性肠炎结肠组织中的表达量差异无统计学意义,Nox2 mRNA增加2.4倍(P0.01)。Nox1蛋白在正常结肠上皮细胞刷状缘和胞质中表达;Nox2蛋白主要表达于浸润的吞噬细胞和中性粒细胞;Duox2蛋白在正常结肠黏膜组织中低表达;三者在人炎症性肠病结肠组织中表达均上调(均P0.01);在小鼠慢性肠炎模型中,Nox2和Duox2蛋白上调(均P0.01),而Nox1无变化。除了NOX2 mRNA在男性病人表达高于女性病人外,未发现这些氧化酶与病人临床特征之间有关联。结论NOX1、NOX2和DUOX2不但在维持机体正常生理功能过程中发挥重要作用,而且在炎症性肠病初期阶段的发病中也起一定的作用。
[Abstract]:Objective to investigate the main members of NADPH oxidase family (nicotinamide adenine dinucleotide phosphate oxidases,NOXs (NOX1,NOX2 and dual oxidase,DUOX) 2 (human: NOX1,NOX2 and DUOX2;) Mouse: Nox1,Nox2 and Duox2) expression in colon of human inflammatory bowel disease and mouse enteritis model. Methods Human colon specimens were collected from inflammatory bowel disease tissues and normal tissues which were more than 5 cm from the margin of the lesion by colonoscopic biopsy or surgical resection. At the same time, female C57BL/6 mice aged 8 ~ 10 weeks were used to establish colitis model. The mice were randomly divided into control group and chronic enteritis group by coin toss method. The chronic enteritis group drank 1.0% sodium dextran sulfate for 7 days. Then drinking water freely for 14 days and then circulating for 3 times; The degree of intestinal inflammatory response was evaluated by body mass change and HE staining. The expression of NOX1,NOX2, DUOX2 mRNA and protein in colon tissue was detected by real-time quantitative PCR and immunohistochemistry, and the relationship between the expression of NOX1,NOX2, DUOX2 mRNA and protein in human inflammatory bowel disease was analyzed. Results the expression of NOX1,NOX2 and DUOX2 mRNA in human inflammatory bowel disease colon was significantly higher than that in normal control group (2.8U 2.0 and 3.3fold, respectively) (P 0.01). There was no significant difference in the expression of Nox1 and Duox2 mRNA in colonic tissues of mice with chronic enteritis, and Nox2 mRNA increased 2.4-fold (P0.01). Nox1 protein was expressed in brushes and cytoplasm of normal colonic epithelial cells. Nox2 protein was mainly expressed in infiltrating phagocytes and neutrophils, Duox2 protein was low expressed in normal colonic mucosa, all three proteins were up-regulated in human inflammatory bowel disease colon tissue (P0.01). In mouse chronic enteritis model, Nox2 and Duox2 proteins were up-regulated (P0.01), but Nox1 did not change. There was no correlation between the expression of NOX2 mRNA and clinical features except in males and females. Conclusion NOX1,NOX2 and DUOX2 not only play an important role in maintaining the normal physiological function of the body, but also play a certain role in the early stage of inflammatory bowel disease.
【作者单位】：河南科技大学临床医学院河南科技大学第一附属医院消化内科;首都医科大学附属北京康复医院消化内科;
【基金】：国家自然科学基金(81370487)~~
【分类号】：R574.62

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