NF-κB信号通路的激活在胃食管反流破坏食管黏膜上皮屏障功能机制中的作用

发布时间：2018-04-27 04:17

本文选题：小鼠模型 + 胃食管反流　；参考：《重庆医科大学学报》2017年09期

【摘要】：目的:本研究旨在探讨肿瘤坏死因子κB(nuclear factorκB,NF-κB)在反流破坏食管黏膜屏障发病机制中的作用,为寻求有效的干预措施提供理论依据。方法:建立单纯十二指肠和混合反流小鼠模型,检测食管黏膜跨上皮电阻抗以及细胞间间隙,使用基因芯片和生物信息学的方法筛选出反流食管黏膜差异表达的基因。运用免疫组化检测食管黏膜NF-κB亚基p50、p65和紧密连接蛋白Cldn1、Cldn4的表达,运用qRT-PCR和Western blot检测食管黏膜上皮Cldn1、Cldn4的表达,运用ELISA检测食管黏膜NF-κB靶基因白细胞介素(interleukin,IL)-1β、IL-6和IL-8的表达,通过特殊细胞染色检测并计数食管黏膜中性粒细胞、肥大细胞以及嗜酸性粒细胞。反流小鼠模型予以腹腔注射NFκB抑制剂Bay11-7085[20 mg/(kg·d)10 d,观察用药组以及未用药组小鼠食管黏膜跨上皮电阻抗以及IL-1β、IL-6和IL-8的表达差异。结果:反流食管黏膜跨上皮电阻抗下降,细胞间间隙增宽;基因富集分析和显著性分析指出反流食管黏膜NF-κB及其下游靶基因表达被激活;基因表达实验指出反流食管黏膜内NF-κB亚基p50和p65以及NF-κB下游靶基因IL-1β、IL-6和IL-8表达上调,紧密连接Cldn1和Cldn4的表达和分布发生失调;反流食管黏膜中性粒细胞、肥大细胞以及嗜酸性粒细胞计数均明显增加;NF-κB抑制实验指出给药组反流黏膜的跨上皮电阻抗值较未给药组显著上调,而炎症因子的表达水平则显著下调。结论:胃食管反流激活小鼠食管黏膜NF-κB信号通路,导致上皮屏障功能的损伤。抑制NF-κB通路具有潜在增强食管黏膜抗反流的能力。
[Abstract]:Objective: to investigate the role of tumor necrosis factor-魏 B(nuclear factor 魏-魏 B in the pathogenesis of esophageal mucosal barrier disruption by reflux, and to provide theoretical basis for effective intervention. Methods: a simple duodenum and mixed reflux mouse model was established to detect the transdermal impedance and intercellular space of esophageal mucosa. The differentially expressed genes were screened by gene chip and bioinformatics. The expression of NF- 魏 B subunit p50 p65 and tight junction protein (Cldn1) Cldn4 in esophageal mucosa was detected by immunohistochemistry, the expression of Cldn4 in esophageal mucosal epithelium was detected by qRT-PCR and Western blot, and the expression of IL-6 and IL-8 was detected by ELISA. Neutrophils, mast cells and eosinophils were detected and counted by special cell staining. The mouse model of reflux was injected intraperitoneally with NF 魏 B inhibitor Bay11-7085 [20 mg/(kg d] for 10 days. The transdermal electrical impedance and the expression of IL-1 尾 -IL-6 and IL-8 in the esophageal mucosa of the treated and untreated mice were observed. Results: the transdermal electrical impedance of reflux esophagus decreased and the intercellular gap widened, the gene enrichment analysis and significant analysis indicated that NF- 魏 B and its downstream target gene expression were activated in reflux esophageal mucosa. Gene expression experiment indicated that the expression of NF- 魏 B subunit p50 and p65 and the downstream target gene IL-1 尾, IL-6 and IL-8 of NF- 魏 B were up-regulated, and the expression and distribution of Cldn1 and Cldn4 were closely related to the expression and distribution of reflux esophageal mucosa neutrophils. Mast cells and eosinophilic granulocytes count increased significantly in NF- 魏 B inhibition experiment. The results showed that the transepithelial electrical impedance of reflux mucosa in the administration group was significantly higher than that in the control group, while the expression of inflammatory factors was significantly down-regulated. Conclusion: gastroesophageal reflux activates NF- 魏 B signaling pathway in mouse esophageal mucosa, resulting in injury of epithelial barrier function. Inhibition of NF- 魏 B pathway has the potential to enhance esophageal mucosal anti-reflux ability.
【作者单位】：重庆医科大学附属第一医院胸外科心脏大血管外科;
【基金】：国家自然科学基金青年科学基金资助项目(编号:81400590) 重庆医科大学附属第一医院博士生启动基金
【分类号】：R571

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