实验性肠梗阻中肠道稳态破坏的阶段性研究
Phasic study of intestinal homeostasis disruption in experimental intestinal obstruction.
作者信息
Yu Xiang-Yang, Zou Chang-Lin, Zhou Zhen-Li, Shan Tao, Li Dong-Hua, Cui Nai-Qiang
机构信息
Xiang-Yang Yu, Zhen-Li Zhou, Tao Shan, Department of Gastrointestinal Surgery, Nankai Hospital, Tianjin 300100, China.
出版信息
World J Gastroenterol. 2014 Jul 7;20(25):8130-8. doi: 10.3748/wjg.v20.i25.8130.
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.
目的
在肠梗阻实验模型中研究肠道内环境稳定的阶段性改变。
方法
通过将部分输液装置改造为体内牵拉式锁定夹，并在距回肠末端8 cm的肠系膜无血管区制造均匀可控的肠袢梗阻，建立兔肠梗阻模型。在肠梗阻后研究肠道内环境稳定的阶段性改变。从过碘酸-希夫染色切片中对杯状细胞、上皮内淋巴细胞、固有层淋巴细胞和肠上皮的变化进行定量分析。通过高效液相色谱法测量鸟氨酸脱羧酶（ODC）活性和血清瓜氨酸水平。通过实时聚合酶链反应分析检测闭合蛋白1（Claudin 1）mRNA表达。在肠梗阻后的多个时间点测定肠道微生物、湿重/干重比、pH值和内毒素水平。此外，通过流式细胞术测定CD3(+)、CD4(+)和CD8(+) T细胞的数量和比例，并用酶联免疫吸附测定法测量分泌型IgA水平。
结果
建立了合适的可控兔肠梗阻模型。肠梗阻导致杯状细胞损伤并减少细胞数量。观察到上皮细胞损伤的进一步指标为血清瓜氨酸水平降低和Claudin 1基因表达降低，以及ODC活性短暂升高。此外，肠腔的湿重/干重比和pH值也发生了显著变化。肠梗阻后双歧杆菌与肠杆菌的比例发生逆转。肠梗阻后派尔集合淋巴结的数量和面积先增加后急剧减少，同时CD4/CD8(+) T细胞比例发生改变，这是由CD3(+)和CD8(+) T细胞增加以及CD4(+) T细胞减少所致。随着梗阻时间延长，固有层淋巴细胞数量也逐渐减少。
结论
肠梗阻可导致肠道内环境稳定的破坏。
Zotero 插件