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Phasic study of intestinal homeostasis disruption in experimental intestinal obstruction

机译:实验性肠梗阻肠内稳态破坏的阶段性研究

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摘要

AIM: To investigate the phasic alteration of intestinal homeostasis in an experimental model of intestinal obstruction.METHODS: A rabbit model of intestinal obstruction was established by transforming parts of an infusion set into an in vivo pulled-type locking clamp and creating a uniform controllable loop obstruction in the mesenteric non-avascular zone 8 cm from the distal end of the ileum. The phasic alteration of intestinal homeostasis was studied after intestinal obstruction. The changes in goblet cells, intraepithelial lymphocytes, lamina propria lymphocytes, and intestinal epithelium were quantified from periodic acid-Schiff-stained sections. Ornithine decarboxylase (ODC) activity and serum citrulline levels were measured by high-performance liquid chromatography. Claudin 1 mRNA expression was examined by real-time polymerase chain reaction analysis. Intestinal microorganisms, wet/dry weight ratios, pH values, and endotoxin levels were determined at multiple points after intestinal obstruction. Furthermore, the number and ratio of CD3+, CD4+ and CD8+ T cells were determined by flow cytometry, and secretory IgA levels were measured with an enzyme-linked immunosorbent assay.RESULTS: A suitable controllable rabbit model of intestinal obstruction was established. Intestinal obstruction induced goblet cell damage and reduced cell number. Further indicators of epithelial cell damage were observed as reduced serum citrulline levels and claudin 1 gene expression, and a transient increase in ODC activity. In addition, the wet/dry weight ratio and pH of the intestinal lumen were also dramatically altered. The ratio of Bacillus bifidus and enterobacteria was reversed following intestinal obstruction. The number and area of Peyer’s patches first increased then sharply decreased after the intestinal obstruction, along with an alteration in the ratio of CD4/CD8+ T cells, driven by an increase in CD3+ and CD8+ T cells and a decrease in CD4+ T cells. The number of lamina propria lymphocytes also gradually decreased with prolonged obstruction.CONCLUSION: Intestinal obstruction can induce disruption of intestinal homeostasis.
机译:目的:探讨在肠梗阻实验模型中肠稳态的阶段性变化。方法:通过将输液器的一部分转变成活体拉式锁定夹并建立均匀可控环,建立兔肠梗阻模型。在距回肠远端8 cm的肠系膜非血管区域阻塞。研究了肠梗阻后肠道稳态的阶段性变化。从高碘酸-希夫染色切片中定量测定杯状细胞,上皮内淋巴细胞,固有层淋巴细胞和肠上皮的变化。鸟氨酸脱羧酶(ODC)活性和血清瓜氨酸水平通过高效液相色谱法测定。通过实时聚合酶链反应分析检查了克劳丁1 mRNA的表达。在肠梗阻后的多个时间点确定肠道微生物,干/湿比,pH值和内毒素水平。此外,通过流式细胞术确定CD3 + ,CD4 + 和CD8 + T细胞的数量和比例,并测量分泌型IgA水平。结果:建立了合适的可控兔肠梗阻模型。肠梗阻引起杯状细胞损伤并减少细胞数量。观察到的上皮细胞损伤的其他指标是血清瓜氨酸水平和claudin 1基因表达降低,以及ODC活性短暂升高。另外,肠腔的干/湿重比和pH也发生了显着变化。肠梗阻后双歧杆菌和肠杆菌的比例发生逆转。肠道阻塞后,派伊尔氏淋巴集结的数量和面积先增加然后急剧减少,而CD4 / CD8 + T细胞比例的改变是由CD3 + < / sup>和CD8 + T细胞,而CD4 + T细胞减少。结论:肠梗阻可引起肠内稳态的破坏,并随梗阻时间的延长逐渐减少固有层淋巴细胞的数量。

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