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大黄素对盲肠结扎穿刺致脓毒症大鼠肠上皮紧密连接屏障完整性的保护作用

Protective effect of emodin on intestinal epithelial tight junction barrier integrity in rats with sepsis induced by cecal ligation and puncture.

作者信息

Li Yanjun, Guo Ruimin, Zhang Mengying, Chen Peng, Li Jingxin, Sun Yanni

机构信息

Department of Emergency, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200062, P.R. China.

Department of Emergency, Shanghai Putuo Central School of Clinical Medicine, Anhui Medical University, Shanghai 200062, P.R. China.

出版信息

Exp Ther Med. 2020 Jun;19(6):3521-3530. doi: 10.3892/etm.2020.8625. Epub 2020 Mar 24.

Abstract

The present study investigated the protective effects of emodin on intestinal epithelial tight junction (TJ) barrier integrity in cecal ligation and puncture (CLP)-induced septic rats and its possible mechanisms of action. Healthy male Sprague-Dawley rats were randomly divided into three groups (n=20 per group): Sham group, CLP group and CLP + emodin group. Animals were sacrificed at 12 and 24 h after the model was established. Abdominal aortic blood and specimens of the ileum were harvested for analysis. The histopathological changes in intestinal mucosa and the ultrastructures of intestinal epithelial cells were investigated using light microscopy and transmission electron microscopy. The integrity of the intestinal barrier was assessed by examining plasma diamine oxidase (DAO) levels and the ratio of urine lactulose to mannitol (L/M). The levels of the intestinal TJ proteins claudin-3, zonula occludens (ZO)-1 and occludin were detected using immunohistochemistry, western blotting and reverse transcription-quantitative PCR. The results showed that the pathological damage to intestinal mucosa and the intestinal tissue injury score in the CLP + emodin group were significantly reduced compared to those of the CLP group, and the differences were more obvious at 24 h compared with 12 h. DAO activity and the L/M ratio in the emodin pre-treatment group decreased significantly at 24 h compared with the CLP groups. The protein and mRNA levels of the TJ proteins claudin-3, ZO-1 and occludin in the emodin pre-treatment groups at 12 and 24 h were increased, while occludin mRNA level was found to be decreased compared with the CLP groups. The present study suggested that emodin may significantly reduce the damage to the intestinal epithelial barrier in sepsis, inhibit intestinal barrier permeability and protect intestinal barrier integrity. Emodin may protect intestinal barrier integrity by elevating expression levels of the TJ proteins claudin-3, ZO-1 and occludin in CLP rats.

摘要

本研究探讨了大黄素对盲肠结扎穿刺(CLP)诱导的脓毒症大鼠肠上皮紧密连接(TJ)屏障完整性的保护作用及其可能的作用机制。将健康雄性Sprague-Dawley大鼠随机分为三组(每组n = 20):假手术组、CLP组和CLP +大黄素组。在模型建立后12小时和24小时处死动物。采集腹主动脉血和回肠标本进行分析。采用光学显微镜和透射电子显微镜观察肠黏膜的组织病理学变化和肠上皮细胞的超微结构。通过检测血浆二胺氧化酶(DAO)水平和尿乳果糖与甘露醇比值(L/M)评估肠屏障的完整性。采用免疫组织化学、蛋白质印迹法和逆转录-定量PCR检测肠TJ蛋白claudin-3、闭合蛋白(ZO)-1和闭锁蛋白的水平。结果显示,与CLP组相比,CLP +大黄素组肠黏膜的病理损伤和肠组织损伤评分显著降低,且24小时时的差异比12小时时更明显。与CLP组相比,大黄素预处理组在24小时时DAO活性和L/M比值显著降低。大黄素预处理组在12小时和24小时时TJ蛋白claudin-3、ZO-1和闭锁蛋白的蛋白质和mRNA水平均升高,而与CLP组相比,闭锁蛋白mRNA水平降低。本研究表明,大黄素可能显著减轻脓毒症时肠上皮屏障的损伤,抑制肠屏障通透性,保护肠屏障完整性。大黄素可能通过提高CLP大鼠TJ蛋白claudin-3、ZO-1和闭锁蛋白的表达水平来保护肠屏障完整性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/00c4/7185184/2366bec4b8a7/etm-19-06-3521-g00.jpg

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