肠三叶因子对非酒精性脂肪性肝炎大鼠回肠黏液屏障的影响

梁凯; 孔维宗; 陈娟; 王迎春

doi:10.3969/j.issn.1001-5256.2017.08.029

肠三叶因子对非酒精性脂肪性肝炎大鼠回肠黏液屏障的影响

DOI: 10.3969/j.issn.1001-5256.2017.08.029

1. 大连大学
2. 大连大学附属中山医院消化二科

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中图分类号: R575
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出版历程
- 收稿日期: 2017-02-22
- 出版日期: 2017-08-20

Effect of trefoil factor 3 on intestinal mucous barrier in rats with nonalcoholic steatohepatitis

Dalian University

摘要

摘要:
目的研究非酒精性脂肪性肝炎（NASH）大鼠肠道黏液屏障改变,探讨肠三叶因子（TFF3）对NASH大鼠肠道黏液屏障的影响,及其对NASH的治疗作用。方法将清洁级雄性SD大鼠60只随机分为正常组、模型组和治疗组,每组20只。正常组给予普通饲料,模型组和治疗组给予高脂饲料12周,诱导NASH模型后,治疗组给予rh TFF3 1 ml·kg-1·d-1（浓度0.1 mg/ml）腹腔内注射,正常组与模型组给予1 ml·kg-1·d-1生理盐水,连续3周。第15周末,FITC标记的右旋糖酐灌胃检测大鼠回肠通透性后,处死大鼠,检测血清AST、ALT、TC、TG、内毒素（ET）水平及二胺氧化酶（DAO）活性。HE染色观察肝脏和回肠末端组织病理变化,PAS染色观察回肠末端杯状细胞并计数,免疫组化方法检测回肠末端紧密连接蛋白Occludin和TFF3蛋白表达水平,实时荧光定量PCR检测TFF3 mRNA转录水平。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果大鼠血清AST、ALT、TG、T...
- 脂肪肝 /
- 肠三叶因子 /
- 杯状细胞 /
- 大鼠,Sprague-Dawley
Abstract:
Objective To investigate the change in intestinal mucous barrier in rats with nonalcoholic steatohepatitis ( NASH) , the effect of trefoil factor 3 ( TFF3) on intestinal mucous barrier in NASH rats, and the therapeutic effect of TFF3 on NASH. Methods A total of 60 clean male Sprague-Dawley rats were randomly divided into normal group, model group, and treatment group, with 20 rats in each group.The rats in the normal group were given normal diet, and those in the model group and the treatment group were given high-fat diet to induce NASH. The rats in the treatment group were given intraperitoneal injection of rh TFF3 at a dose of 1 ml·kg-1·d-1 ( a concentration of 0. 1 mg/ml) , and those in the normal group and the model group were given normal saline at a dose of 1 ml·kg-1·d-1; the course of treatment was 3 weeks for all groups. At the end of week 15, fluorescein isothiocyanate-labeled dextran was given by gavage to evaluate intestinal permeability, and after the rats were sacrificed, serum levels of aspartate aminotransferase ( AST) , alanine aminotransferase ( ALT) , total cholesterol ( TC) , triglyceride ( TG) , and endotoxin ( ET) and diamine oxidase ( DAO) activity were measured. HE staining was performed to observe the histopathological changes of the liver and the terminal ileum, PAS staining was performed to observe and count the goblet cells of the terminal ileum, immunohistochemistry was used to measure the expression of the tight junction protein Occludin and TFF3 in the terminal ileum, and quantitative real-time PCR was used to measure the mRNA transcription level of TFF3. A one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between any two groups. Results The model group had significant increases in serum levels of AST, ALT, TC, TG, and ET and DAO activity, and the treatment group had significant reductions compared with the model group ( all P < 0. 01) . The model group had a significant increase in NAFLD activity score compared with the normal group ( P < 0. 01) , and the treatment group had significant improvement in liver inflammation and a significant reduction in NAFLD activity score compared with the model group ( P < 0. 01) . The model group had cell necrosis, damage of the intestinal villi, and a significant reduction in goblet cells in the terminal ileum under a light microscope; in the treatment group, damage of the intestinal villi was repaired and there was an increase in goblet cells. The model group had a significant increase in intestinal permeability compared with the normal group, and the treatment group had a significant reduction compared with the model group ( both P < 0. 01) . The model group had a significant reduction in the expression of Occludin and TFF3, and the treatment group had a significant increase compared with the model group ( all P < 0. 01) . The model group had a downregulated TFF3 mRNA transcription level in the terminal ileum compared with the normal group, and the treatment group had an upregulated level compared with the model group ( both P < 0. 01) . Conclusion NASH rats have damaged goblet cells and mucous barrier dysfunction. TFF3 can repair the damaged terminal ileum, promote the regeneration of goblet cells and mucus secretion, restore intestinal barrier function, reduce intestinal permeability, and thus exert its therapeutic effect on NASH.
- fatty liver /
- trefoil factor 3 /
- goblet cells /
- rats, Sprague-Dawley

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