鸭油甘油二酯和维生素D<sub>3</sub>对结肠炎大鼠结肠菌群、脾脏过氧化物酶体增殖物激活受体γ mRNA表达量及结肠组织发育的影响

doi:10.3969/j.issn.1006-267x.2020.03.045

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摘要本试验旨在研究不同水平的鸭油甘油二酯和维生素D₃对结肠炎大鼠结肠菌群、脾脏过氧化物酶体增殖物激活受体γ（PPARγ）mRNA表达量及结肠组织发育的影响。选择100只大鼠，随机分为模型组和对照组，对照组每组4个重复，模型组每组16个重复，每个重复均5只大鼠；对照组大鼠自由饮用蒸馏水，模型组大鼠自由饮用5%葡聚糖硫酸钠水溶液，进行连续7 d的造模试验。造模成功后，选取10只正常大鼠作为对照组（Ⅰ组），另选择建模成功的大鼠70只，随机分成7个组，分别为模型组（Ⅱ组）和试验组（Ⅲ~Ⅷ组）；每组均2个重复，每个重复5只。采用2×3两因素交叉析因试验设计，鸭油甘油二酯添加水平分别为2.5、5.0、10.0 mL/kg，维生素D₃添加水平分别为12.5、25.0 μg/kg。试验期为3周。结果表明：1）鸭油甘油二酯与维生素D₃交互作用对Ace指数、Chao1指数影响显著（P<0.05），与模型组相比，试验组操作分类单元（OTUs）数量、Ace指数、Chao1指数均显著增加（P<0.05），其中5.0 mL/kg鸭油甘油二酯和12.5 μg/kg维生素D₃组的OTUs数量、Ace指数、Chao1指数最高，微生物的物种丰度高于其他5个试验组。2）主成分分析图显示，5 mL/kg鸭油甘油二酯和12.5 μg/kg维生素D₃组与对照组距离最近，微生物相似性最高。3）与模型组相比，试验组的厚壁菌门（Firmicutes）、变形菌门（Proteobacteria）、γ-变形菌纲（Gammaproteobacteria）、志贺氏菌属（Shigella）、普拉氏菌属（Prevotella）和支原体菌属（Allobaculum）比例显著降低（P<0.05），拟杆菌门（Bacteroidetes）、拟杆菌属（Bacteroides）比例显著增加（P<0.05），梭状芽孢杆菌纲（Clostridia）比例无显著变化（P>0.05）。4）鸭油甘油二酯和维生素D₃交互作用对绒毛高度、隐窝深度影响显著（P<0.05）；与模型组相比，试验组的结肠绒毛高度、绒毛高度/隐窝深度显著增加（P<0.05），隐窝深度显著变浅（P<0.05）。5）鸭油甘油二酯和维生素D₃交互作用对脾脏PPARγ mRNA表达量影响显著（P<0.05）；与模型组相比，试验组（除10.0 mL/kg鸭油甘油二酯和25.0 μg/kg维生素D₃组）脾脏PPARγ mRNA表达量显著增加（P<0.05），其中5.0 mL/kg鸭油甘油二酯和12.5 μg/kg维生素D₃组脾脏PPARγ mRNA表达量最高。6）脾脏PPARγ mRNA表达量与绒毛高度、绒毛高度/隐窝深度呈极显著正相关（P<0.01），与隐窝深度呈极显著负相关（P<0.01）。由此可见，不同水平的鸭油甘油二酯和维生素D₃能够改善结肠炎大鼠的结肠区系微生物结构，增加有益微生物比例，促进结肠肠道发育，上调脾脏PPARγ mRNA表达量，并且对肠道发育具有调控作用，以5.0 mL/kg鸭油甘油二酯和12.5 μg/kg维生素D₃组合为最优。

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	刘亚楠
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	孔敏
	孙京新

关键词 ：鸭油甘油二酯, 维生素D₃, 结肠炎, 大鼠, 结肠菌群, 肠道发育, PPAR&gamma

Abstract：This study aimed to explore the effects of different levels of duck oil diglyceride and vitamin D₃ on colonic flora, spleen peroxisome proliferators activated receptor γ (PPARγ) mRNA expression and colonic tissue development of colitis rats. A total of 100 rats were randomly divided into model group and control group, each control group contained 4 replicates, each model group contained 16 replicates, and each replicate contained 5 rats; rats in control group free drank distilled water, rats in model group free drank 5% dextran sulfate sodium, and the modeling test for consecutive lasted for 7 days. After the successful molding, selected 10 normal rats as a control group (group Ⅰ), then selected 70 rats of successful molding randomly divided into 7 group, which were model group (group Ⅱ) and experimental groups (groups Ⅲ to Ⅷ); each group contained 2 replicates and each replicate contained 5 rats. A 2×3 cross factor analysis experiment design was adopted, the addition levels of duck oil diglyceride were 2.5,5.0 and 10.0 mL/kg, respectively, and the addition levels of vitamin D₃ was 12.5 and 25.0 μg/kg, respectively. The experiment lasted for 3 weeks. The results showed as follows:1) the interaction of duck oil diglyceride and vitamin D₃ had significant effects on Ace index and Chao1 index (P<0.05), compared with the model group, the operation classification unit (OTUs) number, Ace index and Chao1 index of experimental groups were significantly increased (P<0.05), the OTUs number, Ace index and Chao1 index of 5.0 mL/kg duck oil diglyceride and 12.5 μg/kg vitamin D₃ group were the highest, and the microbial species abundance was higher than other five experimental groups. 2) The principal component analysis figure showed that the 5.0 mL/kg duck oil diglyceride and 12.5 μg/kg vitamin D₃ group was closest to the control group, with the highest microbial similarity. 3) Compared with the model group, the proportions of Firmicutes, Proteobacteria, Gammaproteobacteria, Shigella, Prevotella and Allobaculum of experimental groups were significantly decreased (P<0.05), the proportions of Bacteroidetes and Bacteroides were significantly increased (P<0.05), and the Clostridia proportion had no significant change (P<0.05). 4) The interaction of duck oil diglyceride and vitamin D₃ had significant effects on villus height and crypt depth (P<0.05); compared with the model group, the villus height and villus height/crypt depth of experimental groups were significantly increased (P<0.05), and the crypt depth was significantly decreased (P<0.05). 5) The interaction of duck oil diglyceride and vitamin D₃ had significant effects on spleen PPARγ mRNA expression (P<0.05); compared with the model group, the spleen PPARγ mRNA expression of experimental groups (except 10.0 mL/kg duck oil diglyceride and 25.0 μg/kg vitamin D₃ group) was significantly increased (P<0.05), and the spleen PPARγ mRNA expression of 5.0 mL/kg duck oil diglyceride and 12.5 μg/kg vitamin D₃ group was the highest. 6) The spleen PPARγ mRNA expression was significantly positively correlated with villus height and illus height/crypt depth (P<0.01), and negatively correlated with crypt depth (P<0.01). In conclusion, different levels of duck oil diglyceride and vitamin D₃ can improve the microbial structure of colon flora of colitis rats, increase the proportion of beneficial microorganisms, promote the development of colon intestinal tract, up-regulate the spleen PPARγ mRNA expression, and have regulatory effect on intestinal development. The combination proportion of 5.0 mL/kg duck oil diglyceride and 12.5 μg/kg vitamin D₃ is the optimal combination.

Key words： duck oil diglyceride vitamin D₃ colitis rats colon flora intestinal development PPARγ

收稿日期: 2019-09-07

PACS:

S816.7

基金资助:国家水禽产业技术体系专项基金（CARS-43-11）；山东省2018年度农业重大应用技术创新项目

通讯作者: 王宝维,教授,硕士生导师,E-mail:wangbw@qau.edu.cn E-mail: wangbw@qau.edu.cn

作者简介: 刘亚楠(1995-),女,山东荣成人,硕士研究生,研究方向为营养与保健。E-mail:657347874@qq.com

引用本文:

刘亚楠, 王宝维, 葛文华, 张名爱, 谢玉娥, 孔敏, 孙京新. 鸭油甘油二酯和维生素D₃对结肠炎大鼠结肠菌群、脾脏过氧化物酶体增殖物激活受体γ mRNA表达量及结肠组织发育的影响[J]. 动物营养学报, 2020, 32(3): 1382-1395.
LIU Yanan, WANG Baowei, GE Wenhua, ZHANG Ming'ai, XIE Yu'e, KONG Min, SUN Jingxin. Effects of Duck Oil Diglyceride and Vitamin D₃ on Colonic Flora, Spleen Peroxisome Proliferators Activated Receptor γ mRNA Expression and Colonic Tissue Development of Colitis Rats. Chinese Journal of Animal Nutrition, 2020, 32(3): 1382-1395.

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http://www.chinajan.com/CN/10.3969/j.issn.1006-267x.2020.03.045 或 http://www.chinajan.com/CN/Y2020/V32/I3/1382

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