BACKGROUND Mast cells (MCs) have been found to play a critical role during development of inflammatory bowel disease (IBD) that characterized by dysregulation of inflammation and impaired intestinal barrier function. However, the function of MCs in IBD remains to be fully elucidated. RESULTS In our study, we used exosomes isolated from human mast cells-1 (HMCs-1) to culture with NCM460, HT-29 or CaCO2 of intestinal epithelial cells (IECs) to investigate the communication between MCs and IECs. We found that MCs-derived exosomes significantly increased intestinal epithelial permeability and destroyed intestinal barrier function, which is attributed to exosome-mediated functional miRNAs were transferred from HMCs-1 into IECs, leading to inhibit tight junction-related proteins expression, including tight junction proteins 1 (TJP1, ZO-1), Occludin (OCLN), Claudin 8 (CLDN8). Microarray and bioinformatic analysis have further revealed that a panel of miRNAs target different tight junction-related proteins. Interestingly, miR-223 is enriched in mast cell-derived exosome, which inhibit CLDN8 expression in IECs, while treatment with miR-223 inhibitor in HT-29 cells significantly reversed the inhibitory effect of HMCs-1-derived exosomes on CLDN 8 expression. Most importantly, enrichment of MCs accumulation in intestinal mucosa of patients with IBD compared with those healthy control. CONCLUSIONS These results indicated that enrichment of exosomal miR-223 from HMCs-1 inhibited CLDN8 expression, leading to destroy intestinal barrier function. These finding provided a novel insight of MCs as a new target for therapeutic treatment of IBD.

中文翻译：

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肥大细胞来源的MiR-223通过抑制肠上皮细胞中CLDN8的表达破坏肠屏障功能。

背景技术已经发现肥大细胞（MC）在炎症性肠病（IBD）的发展过程中起关键作用，所述炎症性肠病的特征在于炎症失调和肠屏障功能受损。然而，MC在IBD中的功能仍有待充分阐明。结果在我们的研究中，我们使用从人肥大细胞-1（HMCs-1）分离的外泌体与肠上皮细胞（IECs）的NCM460，HT-29或CaCO2培养，以研究MC与IECs之间的通讯。我们发现，MCs衍生的外泌体显着增加了肠上皮通透性并破坏了肠屏障功能，这归因于外泌体介导的功能性miRNA从HMCs-1转移到IECs中，从而抑制了紧密连接相关蛋白的表达，包括紧密连接蛋白1（TJP1，ZO-1），闭合蛋白（OCLN），克劳丁8（CLDN8）。微阵列和生物信息学分析进一步揭示了一组miRNA靶向不同的紧密连接相关蛋白。有趣的是，miR-223富含肥大细胞来源的外来体，可抑制IECs中CLDN8的表达，而在HT-29细胞中用miR-223抑制剂处理可显着逆转HMCs-1来源的外来体对CLDN 8表达的抑制作用。最重要的是，与健康对照组相比，IBD患者肠粘膜中MC的积累富集。结论这些结果表明，从HMCs-1中富集外泌体miR-223抑制了CLDN8的表达，从而破坏了肠屏障功能。这些发现为MCs作为IBD治疗的新靶标提供了新颖的见解。微阵列和生物信息学分析进一步揭示了一组miRNA靶向不同的紧密连接相关蛋白。有趣的是，miR-223富含肥大细胞来源的外来体，可抑制IECs中CLDN8的表达，而在HT-29细胞中用miR-223抑制剂处理可显着逆转HMCs-1来源的外来体对CLDN 8表达的抑制作用。最重要的是，与健康对照组相比，IBD患者肠粘膜中MC的积累富集。结论这些结果表明从HMCs-1富集外泌体miR-223抑制了CLDN8的表达，从而破坏了肠屏障功能。这些发现为MCs作为IBD治疗的新靶标提供了新颖的见解。微阵列和生物信息学分析进一步揭示了一组miRNA靶向不同的紧密连接相关蛋白。有趣的是，miR-223富含肥大细胞来源的外来体，可抑制IECs中CLDN8的表达，而在HT-29细胞中用miR-223抑制剂处理可显着逆转HMCs-1来源的外来体对CLDN 8表达的抑制作用。最重要的是，与健康对照组相比，IBD患者肠粘膜中MC的积累富集。结论这些结果表明从HMCs-1富集外泌体miR-223抑制了CLDN8的表达，从而破坏了肠屏障功能。这些发现为MCs作为IBD治疗的新靶标提供了新颖的见解。