Identification of epithelial gaps in human small and large intestine by confocal endomicroscopy
Identification of epithelial gaps in human small and large intestine by confocal endomicroscopy
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DOI：
10.1053/j.gastro.2007.09.011
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发表时间：
2007-12-01
期刊：
影响因子：
29.4
通讯作者：
Watson, Alastair J. M.
中科院分区：
文献类型：
--
作者：
Kiesslich, Ralf;Goetz, Martin;Watson, Alastair J. M.
背景和目标：共聚焦显微内镜是一种新兴的技术，它给内窥镜医生提出了识别人类肠道上皮结构的挑战。我们以前已经表明，小鼠肠上皮细胞是由细胞脱落引起的间隙间断。本研究的目的是确定共聚焦显微内镜是否可以解决人类上皮间隙的存在，以及促炎细胞因子是否可以增加细胞脱落。方法：采用吖啶黄染色法对肠粘膜进行成像。共聚焦显微内镜的17名患者产生了6277图像从人类末端回肠和直肠。通过使用刚性共聚焦探针显微镜、双光子/共聚焦显微镜和扫描电子显微镜对麻醉小鼠（野生型和Math 1（Delta Intestine））进行平行研究，验证了结果。结果如下：人类末端回肠和直肠上皮显示出具有单个上皮细胞直径的未染色区域，具有2种不同的形态。其中一个有一个“目标”的外观，老鼠的研究表明是杯状细胞。另一种形态没有细胞核，通过刚性共聚焦探针显微镜和扫描电子显微镜在Math 1（Delta Intestine）小鼠的绒毛中观察到，该小鼠缺乏杯状细胞。在小鼠中，肿瘤坏死因子a（0.33 μ g/g腹腔注射）使细胞脱落增加27倍，并导致20%的屏障功能丧失。结论：共聚焦显微内镜可以区分上皮间断（间隙）和杯状细胞在人类肠道。结果表明，上皮间隙的密封必须被认为是肠屏障的组成部分，并对人类疾病中的肠屏障功能障碍具有潜在的影响。
Background & Aims: Confocal endomicroscopy is an emerging technology that poses the endoscopist with challenges for identifying epithelial structures in the human intestine. We have shown previously that the murine intestinal epithelium is' punctuated by gaps caused by cell shedding. The goals of this study were to determine if confocal endomicroscopy could resolve the presence of human epithelial gaps and whether a proinflammatory cytokine could increase cell shedding. Methods: Intestinal mucosa was imaged after staining with acriflavine. Confocal endomicroscopy of 17 patients yielded 6277 images from the human terminal ileum and rectum. Results were validated by parallel studies of anesthetized mice (wild-type and Math1(Delta Intestine)) using rigid confocal probe microscopy, 2-photon/confocal microscopy, and scanning electron microscopy. Results: Human terminal ileal and rectal epithelium revealed unstained areas with the diameter of an individual epithelial cell, with 2 distinct morphologies. One had a "target" appearance, shown by mouse studies to be goblet cells. The other morphology had no nucleus and was observed by rigid confocal probe microscopy and scanning electron microscopy in the villi of Math1(Delta Intestine) mice, which lack goblet cells. In the mouse, tumor necrosis factor a (0.33 mu g/g intraperitoneally) increases cell shedding by 27-fold and caused loss of barrier function across 20% of resultant gaps. Conclusions: Confocal endomicroscopy can distinguish between epithelial discontinuities (gaps) and goblet cells in human intestine. Results suggest that the sealing of epithelial gaps must be considered as a component of the intestinal barrier and has potential implications for intestinal barrier dysfunction in human disease.