Bile salts disrupt human esophageal squamous epithelial barrier function by modulating tight junction proteins
Bile salts disrupt human esophageal squamous epithelial barrier function by modulating tight junction proteins
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DOI：
10.1152/ajpgi.00454.2011
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发表时间：
2012-07-01
影响因子：
4.5
通讯作者：
Miwa, Hiroto
中科院分区：
文献类型：
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作者：
Chen, Xin;Oshima, Tadayuki;Miwa, Hiroto
Chen X，Oshima T，Shan J，Fukui H，Watari J，Miwa H。胆汁盐通过调节紧密连接蛋白破坏人食管鳞状上皮屏障功能。 Am J Physiol Gastrointest Liver Physiol 303：G199-G208，2012。首次发表于 2012 年 5 月 10 日； doi:10.1152/ajpgi.00454.2011.-酸和胆汁酸的反流导致胃食管反流病的上皮组织损伤。然而，回流物质对人食管分层上皮屏障功能和紧密连接（TJ）蛋白的影响尚未完全阐明。在这里，我们使用新开发的基于原代人食管上皮细胞（HEEC）的复层鳞状上皮的气液界面（ALI）体外培养模型，研究了酸和胆汁酸对屏障功能和TJ蛋白分布的影响。在 ALI 条件下，培养 7 天后，HEEC 在 Transwell 插入物上形成明显的上皮层。上皮层形成TJ，通过免疫荧光染色检测claudin-1、claudin-4和occludin的存在。这些 TJ 蛋白的 NP-40 不溶部分在 ALI 培养第 7 天时显着升高。将 HEEC 暴露于 pH 2、牛磺胆酸 (TCA) 和甘胆酸 (GCA) pH 3（而非 pH 4）中 1 小时，会降低跨上皮电阻 (TEER) 并增加细胞旁通透性。将细胞层暴露于 GCA (pH 3) 和 TCA (pH 3) 1 小时也显着减少了claudin-1 和-4 的不溶部分。我们发现脱氧胆酸（pH 7.4 或 6，1 小时）和胃蛋白酶（pH 3，24 小时）显着降低 TEER 并增加通透性。基于这些发现，ALI培养的HEEC代表了一种新的人食管复层上皮体外模型，适合研究食管上皮屏障功能。使用该模型，我们证明酸、胆汁酸和胃蛋白酶部分通过调节 TJ 蛋白来破坏鳞状上皮屏障功能。这些结果为理解 TJ 蛋白在食管炎中的作用提供了新的见解。
Chen X, Oshima T, Shan J, Fukui H, Watari J, Miwa H. Bile salts disrupt human esophageal squamous epithelial barrier function by modulating tight junction proteins. Am J Physiol Gastrointest Liver Physiol 303: G199-G208, 2012. First published May 10, 2012; doi:10.1152/ajpgi.00454.2011.-Reflux of acid and bile acids contributes to epithelial tissue injury in gastro-esophageal reflux disease. However, the influence of refluxed material on human esophageal stratified epithelial barrier function and tight junction (TJ) proteins has not been fully elucidated. Here, we investigated the influence of acid and bile acids on barrier function and TJ protein distribution using a newly developed air-liquid interface (ALI) in vitro culture model of stratified squamous epithelium based on primary human esophageal epithelial cells (HEECs). Under ALI conditions, HEECs formed distinct epithelial layers on Transwell inserts after 7 days of culture. The epithelial layers formed TJ, and the presence of claudin-1, claudin-4, and occludin were detected by immunofluorescent staining. The NP-40-insoluble fraction of these TJ proteins was significantly higher by day 7 of ALI culture. Exposure of HEECs to pH 2, and taurocholic acid (TCA) and glycocholic acid (GCA) at pH 3, but not pH 4, for 1 h decreased transepithelial electrical resistance (TEER) and increased paracellular permeability. Exposure of cell layers to GCA (pH 3) and TCA (pH 3) for 1 h also markedly reduced the insoluble fractions of claudin-1 and -4. We found that deoxycholic acid (pH 7.4 or 6, 1 h) and pepsin (pH 3, 24 h) significantly decreased TEER and increased permeability. Based on these findings, ALI-cultured HEECs represent a new in vitro model of human esophageal stratified epithelium and are suitable for studying esophageal epithelial barrier functions. Using this model, we demonstrated that acid, bile acids, and pepsin disrupt squamous epithelial barrier function partly by modulating TJ proteins. These results provide new insights into understanding the role of TJ proteins in esophagitis.