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大鼠模型中小肠黏膜上皮结构和上皮紧密连接的年龄相关变化。

Age-related changes in small intestinal mucosa epithelium architecture and epithelial tight junction in rat models.

作者信息

Ren Wei-ying, Wu Ke-fen, Li Xi, Luo Man, Liu Hong-chun, Zhang Shun-cai, Hu Yu

机构信息

Department of Geriatrics, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China.

出版信息

Aging Clin Exp Res. 2014 Apr;26(2):183-91. doi: 10.1007/s40520-013-0148-0. Epub 2013 Nov 17.

Abstract

BACKGROUND AND AIMS

Functions of the intestinal mucosal barrier are often impaired in the elderly and are closely associated with many age-related diseases. However, mechanisms by which aging influences intestinal barrier function still remain unclear. The aim of this study was to investigate age-related changes in small intestinal morphology, bacteria contents and expression of epithelial tight junction (TJ) proteins.

METHODS

Thirty Sprague-Dawley rats were divided into groups: young (3 months), adult (12 months), and old (24 months). The small intestinal mucosal architecture and TJ of intestinal epithelial cells were examined by light microscopy and transmission electron microscopy. Jejunum and cecum contents were cultured to identify and measure bacterial species. mRNA expression of Zonula occludens-1 (ZO-1) and occludin were measured by semi-quantitative RT-PCR. Protein expression of ZO-1 and occludin were detected by immunohistochemistry and Western blot.

RESULTS

Normal ileum villi, which were thick and regularly arranged, though increasingly scattered and atrophic in character with shorter and narrower dimensions (P < 0.01), were observed in old rats, along with an elevated number of Gram-positive and Gram-negative bacteria in the jejunum. The TJs of intestinal epithelial cells, as detected by transmission electron microscopy, were wider and discontinuous in old rats. Age-induced down-regulation of mRNA expression and decreased protein expression of ZO-1 and occludin were observed in the ileum (P < 0.01).

CONCLUSIONS

Our study indicated that age-related intestinal barrier dysfunction may be associated with mucosal atrophy, damages to TJ structure, increased small intestine bacteria counts, and decreased epithelial TJ protein.

摘要

背景与目的

老年人肠道黏膜屏障功能常受损,且与许多年龄相关疾病密切相关。然而,衰老影响肠道屏障功能的机制仍不清楚。本研究旨在探讨小肠形态、细菌含量及上皮紧密连接(TJ)蛋白表达的年龄相关变化。

方法

将30只Sprague-Dawley大鼠分为三组:青年组(3个月)、成年组(12个月)和老年组(24个月)。通过光学显微镜和透射电子显微镜检查小肠黏膜结构和肠上皮细胞的紧密连接。培养空肠和盲肠内容物以鉴定和测量细菌种类。采用半定量RT-PCR检测闭合蛋白(occludin)和闭合小环蛋白1(ZO-1)的mRNA表达。通过免疫组织化学和蛋白质印迹法检测ZO-1和occludin的蛋白表达。

结果

老年大鼠回肠绒毛正常,原本厚且排列规则,但变得越来越分散且萎缩,尺寸变短变窄(P<0.01),同时空肠中革兰氏阳性菌和革兰氏阴性菌数量增加。透射电子显微镜检测显示,老年大鼠肠上皮细胞的紧密连接更宽且不连续。在回肠中观察到衰老导致的ZO-1和occludin的mRNA表达下调以及蛋白表达降低(P<0.01)。

结论

我们的研究表明,年龄相关的肠道屏障功能障碍可能与黏膜萎缩、紧密连接结构损伤、小肠细菌数量增加以及上皮紧密连接蛋白减少有关。

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