Ischaemia-induced mucus barrier loss and bacterial penetration are rapidly counteracted by increased goblet cell secretory activity in human and rat colon
Ischaemia-induced mucus barrier loss and bacterial penetration are rapidly counteracted by increased goblet cell secretory activity in human and rat colon
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DOI：
10.1136/gutjnl-2011-301956
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发表时间：
2013-02-01
期刊：
影响因子：
24.5
通讯作者：
Buurman, Wim A.
中科院分区：
文献类型：
--
作者：
Grootjans, Joep;Hundscheid, Inca H. R.;Buurman, Wim A.
目的结肠缺血是临床上常见的疾病。这项研究提供了一个新的洞察结肠缺血/再灌注（IR）的病理生理学，使用一个新开发的人类和大鼠的实验model.Design在10例患者的一小部分结肠，不得不被删除的手术原因被隔离和暴露于60分钟的缺血（60 I）与/无不同时期的再灌注（30 R和60 R）。未暴露于IR的组织用作对照。在大鼠中，结肠暴露于60 I、60 I/30 R、60 I/120 R或60 I/240 R（n = 7/组）。将组织快速冷冻或固定在戊二醛、福尔马林或methacarn固定剂中。粘蛋白用过碘酸希夫/阿尔新蓝（PAS/AB）和MUC 2/双花扁豆凝集素（DBA）染色。细菌进行了研究，使用电子显微镜（EM）和荧光原位杂交（FISH）。使用髓过氧化物酶染色研究中性粒细胞。结果PAS/AB和MUC 2/DBA染色显示大鼠缺血时粘液层脱落，并伴有细菌穿透（EM和FISH）。人类和大鼠的研究表明，同时，杯状细胞分泌活性增加。这与再灌注240分钟时细菌从隐窝中排出和粘液层恢复有关。炎症仅限于轻微流入的中性粒细胞和增加的促炎细胞因子的表达在reperfusion.Conclusions结肠缺血导致破坏的粘液层促进细菌的渗透。这会被杯状细胞分泌活动的增加迅速抵消，导致细菌从隐窝中排出并恢复粘液屏障。
Objective Colonic ischaemia is frequently observed in clinical practice. This study provides a novel insight into the pathophysiology of colon ischaemia/reperfusion (IR) using a newly developed human and rat experimental model.Design In 10 patients a small part of colon that had to be removed for surgical reasons was isolated and exposed to 60 min of ischaemia (60I) with/without different periods of reperfusion (30R and 60R). Tissue not exposed to IR served as control. In rats, colon was exposed to 60I, 60I/30R, 60I/120R or 60I/240R (n = 7 per group). The tissue was snap-frozen or fixed in glutaraldehyde, formalin or methacarn fixative. Mucins were stained with Periodic Acid Schiff/Alcian Blue (PAS/AB) and MUC2/Dolichos biflorus agglutinin (DBA). Bacteria were studied using electron microscopy (EM) and fluorescent in situ hybridisation (FISH). Neutrophils were studied using myeloperoxidase staining. qPCR was performed for MUC2, interleukin (IL)-6, IL-1 beta and tumour necrosis factor a.Results In rats, PAS/AB and MUC2/DBA staining revealed mucus layer detachment at ischaemia which was accompanied by bacterial penetration (in EM and FISH). Human and rat studies showed that, simultaneously, goblet cell secretory activity increased. This was associated with expulsion of bacteria from the crypts and restoration of the mucus layer at 240 min of reperfusion. Inflammation was limited to minor influx of neutrophils and increased expression of proinflammatory cytokines during reperfusion.Conclusions Colonic ischaemia leads to disruption of the mucus layer facilitating bacterial penetration. This is rapidly counteracted by increased secretory activity of goblet cells, leading to expulsion of bacteria from the crypts as well as restoration of the mucus barrier.