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浙江大学学报（农业与生命科学版）
浙江大学学报（农业与生命科学版）  2019, Vol. 45 Issue (5): 626-634    DOI: 10.3785/j.issn.1008-9209.2018.09.261
动物科学与动物医学     
表没食子儿茶素没食子酸酯对葡聚糖硫酸钠诱导小鼠炎症性肠病的保护作用
赵悦伶1(),丁健1,何佳2,孔德栋1,徐平1(),王岳飞1
1. 浙江大学农业与生物技术学院茶叶研究所，杭州 310058
2. 浙江大学医学院免疫研究所，杭州 310058
Protective effect of epigallocatechin gallate on inflammatory bowel disease induced by dextran sulfate sodium in mice
Yueling ZHAO1(),Jian DING1,Jia HE2,Dedong KONG1,Ping XU1(),Yuefei WANG1
1. Department of Tea Science, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China
2. Institute of Immunology, Zhejiang University School of Medicine, Hangzhou 310058, China
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摘要：

通过设立空白组、对照组、表没食子儿茶素没食子酸酯（epigallocatechin gallate, EGCG）处理组，探究EGCG对葡聚糖硫酸钠（dextran sulfate sodium, DSS）诱导小鼠炎症性肠病的影响。结果发现，EGCG处理组的小鼠脾明显比对照组小，并且EGCG能够降低小鼠肠道固有层细胞和肠道组织上清液中白介素-6（interleukin-6, IL-6）、肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）的表达及分泌（P＜0.01），而对IL-10的表达与分泌无明显作用 （P＞0.05）。EGCG对小鼠肠上皮细胞紧密连接蛋白claudin-1、occludin的表达无显著影响，小鼠肠道苏木精-伊红（hematoxylin-eosin, HE）染色结果也显示肠道黏膜和上皮屏障损伤无明显变化。综上所述，EGCG能够抑制炎症因子分泌，减少由肠道炎症引起的小鼠全身性炎症反应，对DSS诱导的小鼠炎症性肠病起到一定的保护作用。
关键词： 表没食子儿茶素没食子酸酯;  葡聚糖硫酸钠;  炎症性肠病;  肠道固有层细胞    
Abstract:

We established three groups, blank, control and epigallocatechin gallate (EGCG) treatment groups, to explore the effect of EGCG on inflammatory bowel disease (IBD) induced by dextran sulfate sodium (DSS) in mice. The result showed that the spleen size of EGCG treatment group was significantly smaller than that of the control group. The detection of inflammatory factors in intestinal lamina propria cells and colon tissue supernatant in mice showed that EGCG could reduce the expression and secretion of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) (P＜0.01), and had no significant effect on interleukin-10 (IL-10) (P＞0.05). In addition, EGCG had no obvious effect on the expression of tight junction protein claudin-1 and occludin in intestinal epithelial cells. The hematoxylin-eosin (HE) staining results also showed that there was no significant alteration to the intestinal mucosa and epithelial barrier damage with EGCG. To sum up, EGCG could inhibit inflammatory factor secretion, reduce systemic inflammatory response in mice caused by colon inflammation, and play a protective role in DSS-induced inflammatory bowel disease in mice.
Key words: epigallocatechin gallate    dextran sulfate sodium    inflammatory bowel disease    intestinal lamina propria cell
收稿日期: 2018-09-26 出版日期: 2019-12-05
CLC:  R 93  
基金资助: 广东省产学研合作项目(2016B090918118)
通讯作者: 徐平     E-mail: 11616050@zju.edu.cn;zdxp@zju.edu.cn
作者简介: 赵悦伶（https://orcid.org/0000-0002-0626-7692），E-mail：11616050@zju.edu.cn
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赵悦伶
丁健
何佳
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引用本文:

赵悦伶,丁健,何佳,孔德栋,徐平,王岳飞. 表没食子儿茶素没食子酸酯对葡聚糖硫酸钠诱导小鼠炎症性肠病的保护作用[J]. 浙江大学学报（农业与生命科学版）, 2019, 45(5): 626-634.

Yueling ZHAO,Jian DING,Jia HE,Dedong KONG,Ping XU,Yuefei WANG. Protective effect of epigallocatechin gallate on inflammatory bowel disease induced by dextran sulfate sodium in mice. Journal of Zhejiang University (Agriculture and Life Sciences), 2019, 45(5): 626-634.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2018.09.261    或    http://www.zjujournals.com/agr/CN/Y2019/V45/I5/626

处理

Treatment

第1—7天

Days 1-7

第8—11天

Days 8-11

空白组

Blank

饮用水饮用水

对照组

Control

2% DSS溶液饮用水

EGCG组

EGCG

2% DSS溶液0.5% EGCG溶液
表1  模型建立和分组

基因

Gene

正向引物（5′→3′）

Forward primer (5′→3′)

反向引物（5′→3′）

Reverse primer (5′→3′)

IL-6GAAACCGCTATGAAGTTCCTCTCTGTGTTGGGAGTGGTATCCTCTGTGA
IL-10GGTTGCCAAGCCTTATCGGAACCTGCTCCACTGCCTTGCT
TNF-αCCCTCACACTCAGATCATCTTCTGCTACGACGTGGGCTACAG
occludinTTGAAAGTCCACCTCCTTACAGACCGGATAAAAAGAGTACGCTGG
claudin-1GGGGACAACATCGTGACCGAGGAGTCGAAGACTTTGCACT
表2  引物序列

情况描述

Description

得分

Score

与空白组相似的正常粪便0
轻微，肉眼可见血丝1
粪便有可见血痕2
大便带血3
脱肛和直肠出血4
表3  便血评分标准[19]

情况描述

Description

得分

Score

与空白相似的正常粪便0
粪便较黄1
粪便呈柔软状2
粪便没有具体形态3
脱肛，身体蜷缩4
表4  腹泻评分标准[19]
图1  EGCG对小鼠体质量变化率和便血、腹泻情况的影响
图2  各组小鼠肠组织长度(A)及脾大小(B)比较
图3  EGCG对小鼠肠道固有层细胞中IL-6、IL-10和TNF-α表达的影响**表示在P＜0.01水平差异有高度统计学意义。
图4  EGCG对小鼠结肠组织上清液中IL-6、IL-10和TNF-α表达水平的影响**表示在P＜0.01水平差异有高度统计学意义。
图5  各组小鼠肠道HE染色
图6  EGCG对小鼠肠上皮细胞紧密连接蛋白occludin和claudin-1 mRNA表达的影响
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