Applicability of the Ussing chamber technique to permeability determinations in functionally distinct regions of the gastrointestinal tract in the rat
Applicability of the Ussing chamber technique to permeability determinations in functionally distinct regions of the gastrointestinal tract in the rat
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DOI：
10.1080/00365520310003101
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发表时间：
2003-07-01
影响因子：
1.9
通讯作者：
Howarth, GS
中科院分区：
文献类型：
--
作者：
Bajka, BH;Gillespie, CM;Howarth, GS
背景：Ussing室通常用于胃肠道通透性的体外研究。然而，它们对小肠的敏感性和适用性尚未得到很好的表征。研究方法：为了研究实验诱导的损伤的影响和粘膜和肌层的相对贡献的透壁通透性在小肠，胃和结肠，正常大鼠肠组织安装在Ussing室与或不去除肌层或粘膜层。体内胃组织因暴露于消炎痛（100 mg kg（-1））而受损，而体外回肠组织因0.4 M NaCl而受损。组织损伤进行了组织学评估，而渗透性参数包括电导（G），电位差（PD）和辣根过氧化物酶（HRP）的粘膜到浆膜通量。结果如下：位于组织边缘的损伤（边缘损伤）占回肠中暴露上皮长度的25%-50%，而小于20%的胃和结肠上皮受到边缘损伤的影响。在受损的胃中，上皮形成减少20%伴随着G（P < 0.001）和HRP（P < 0.01）通量的增加。去除肠外肌层后，虽然结肠内HRP流量增加，但对回肠无影响（P < 0.01），但对未损伤的回肠或结肠粘膜活力无影响。去除回肠粘膜产生的G和HRP流量增加，而PD维持。结论：Ussing小室技术适用于大鼠胃、结肠通透性的研究。然而，由于小肠边缘损伤的普遍性和程度，我们将谨慎使用该技术进行大鼠胃肠道该区域的渗透性研究。
Background: Ussing chambers are commonly utilized for in vitro investigations into gastrointestinal permeability. However, their sensitivity and applicability to the small intestine have not been well characterized. Methods: In order to investigate the effects of experimentally induced damage and the relative contribution of the mucosa and muscularis externa layers to transmural permeability in the small intestine, stomach and colon, normal rat intestinal tissues were mounted in Ussing chambers with or without removal of the muscularis externa or mucosal layers. Gastric tissues were damaged in vivo by exposure to indomethacin (100 mg kg(-1)), while ileal tissues were damaged in vitro by 0.4 M NaCl. Tissue damage was assessed histologically, while permeability parameters included conductance (G), potential difference (PD) and mucosal to serosal flux of horseradish peroxidase (HRP). Results: Damage localized to the tissue edges (edge damage) accounted for 25%-50% of the exposed epithelial length in the ileum, while less than 20% of stomach and colon epithelium was affected by edge damage. In the damaged stomach, a 20% reduction in epithelialization was accompanied by increases in G (P < 0.001) and HRP (P < 0.01) flux. Removal of the muscularis externa did not affect mucosal viability in the undamaged ileum or colon although HRP flux in the colon, but not ileum, was increased (P < 0.01). Removal of the ileal mucosa produced increases in G and HRP flux, while PD was maintained. Conclusion: We conclude that the Ussing chamber technique is suitable for application to studies of gastric and colonic permeability in rats. However, owing to the prevalence and extent of edge damage in the small intestine, we would caution against the use of this technique for permeability studies in this region of the gastrointestinal tract in the rat.