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短期禁食诱导鸡肠道通透性增加相关的肠道黏膜基因表达及形态学变化

Gene expression and morphological changes in the intestinal mucosa associated with increased permeability induced by short-term fasting in chickens.

作者信息

Gilani S, Howarth G S, Nattrass G, Kitessa S M, Barekatain R, Forder R E A, Tran C D, Hughes R J

机构信息

School of Animal and Veterinary Sciences, University of Adelaide, Adelaide, SA, Australia.

Poultry CRC, University of New England, Armidale, NSW, Australia.

出版信息

J Anim Physiol Anim Nutr (Berl). 2018 Apr;102(2):e653-e661. doi: 10.1111/jpn.12808. Epub 2017 Oct 16.

DOI:10.1111/jpn.12808
PMID:29034530
Abstract

Short-term fasting for 4.5 and 9 hr has been demonstrated to increase intestinal permeability (IP) in chickens. This study aimed to investigate the effects of 0, 4.5, 9 and 19.5 hr fasting on intestinal gene expression and villus-crypt architecture of enterocytes in jejunal and ileal samples. On day 38, Ross-308 male birds were fasted according to their group and then euthanised. Two separate intestinal sections (each 2 cm long, jejunum and ileum) were collected. One section was utilised for villus height and crypt depth measurements. The second section was snap-frozen in liquid nitrogen for quantitative polymerase chain reaction (qPCR) analysis of tight junction proteins (TJP) including claudin-1, claudin-3, occludin, zonula occludens (ZO-1, ZO-2), junctional adhesion molecules (JAM) and E-cadherin. Additionally genes involved in enterocyte protection including glucagon-like peptide (GLP-2), heat-shock protein (HSP-70), intestinal alkaline phosphatase (IAP), mammalian target of rapamycin (mTOR), toll-like receptors (TLR-4), mucin (MUC-2), cluster differentiation (CD-36) and fatty acid-binding protein (FABP-6) were also analysed. Normally distributed data were analysed using one-way analysis of variance ANOVA. Other data were analysed by non-parametric one-way ANOVA. Villus height and crypt depth were increased (p < .05) only in the ileum after fasting for 4.5 and 9 hr compared with non-fasting group. mRNA expression of claudin-3 was significantly reduced in the ileum of birds fasted for 9 and 19.5 hr, suggesting a role in IP modulation. However, all other TJP genes examined were not statistically different from control. Nevertheless, ileal FABP-6 of all fasted groups was significantly reduced, which could possibly be due to reduced bile acid production during fasting.

摘要

已证明,4.5小时和9小时的短期禁食会增加鸡的肠道通透性(IP)。本研究旨在调查0、4.5、9和19.5小时禁食对空肠和回肠样本中肠细胞的肠道基因表达及绒毛-隐窝结构的影响。在第38天,将罗斯308雄性鸡按组禁食,然后实施安乐死。采集两段独立的肠段(每段长2厘米,分别为空肠和回肠)。一段用于测量绒毛高度和隐窝深度。另一段在液氮中速冻,用于对紧密连接蛋白(TJP)进行定量聚合酶链反应(qPCR)分析,这些蛋白包括闭合蛋白-1、闭合蛋白-3、闭锁蛋白、闭合小环蛋白(ZO-1、ZO-2)、连接黏附分子(JAM)和E-钙黏蛋白。此外,还分析了参与肠细胞保护的基因,包括胰高血糖素样肽(GLP-2)、热休克蛋白(HSP-70)、肠碱性磷酸酶(IAP)、雷帕霉素靶蛋白(mTOR)、Toll样受体(TLR-4)、黏蛋白(MUC-2)、分化簇(CD-36)和脂肪酸结合蛋白(FABP-6)。对呈正态分布的数据使用单因素方差分析(ANOVA)进行分析。其他数据通过非参数单因素方差分析进行分析。与非禁食组相比,仅在禁食4.5小时和9小时后的回肠中,绒毛高度和隐窝深度增加(p<0.05)。在禁食9小时和19.5小时的鸡的回肠中,闭合蛋白-3的mRNA表达显著降低,表明其在调节肠道通透性中发挥作用。然而,所检测的所有其他TJP基因与对照组相比无统计学差异。尽管如此,所有禁食组的回肠FABP-6均显著降低,这可能是由于禁食期间胆汁酸生成减少所致。

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