丁酸钠对幼鼠回肠发育、炎症因子及物理屏障功能的调控作用

doi:10.11843/j.issn.0366-6964.2025.03.027

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (3): 1278-1289.doi: 10.11843/j.issn.0366-6964.2025.03.027

丁酸钠对幼鼠回肠发育、炎症因子及物理屏障功能的调控作用

张萱¹^,²(), 杨雪¹^,², 李新科¹^,², 郑楠¹^,², 孟璐¹^,²^,*()

1. 中国农业科学院北京畜牧兽医研究所农业农村部奶及奶制品质量安全控制重点实验室，北京 100193
2. 中国农业科学院北京畜牧兽医研究所农业农村部奶产品质量安全风险评估实验室(北京)，北京 100193

收稿日期:2024-04-16 出版日期:2025-03-23 发布日期:2025-04-02
通讯作者: 孟璐 E-mail:18647394594@163.com;menglu@caas.cn
作者简介:张萱(2000-)，女，内蒙古呼和浩特人，硕士生，主要从事奶产品质量安全风险评估和营养功能评价研究，E-mail: 18647394594@163.com
基金资助:
国家重点研发计划(2022YFD1600104)；中国农业科学院科技创新工程(ASTIP-IAS12)；国家现代农业产业技术体系(CARS36)

The Regulatory Effects of Sodium Butyrate on Ileal Development, Inflammatory Factors and Physical Barrier Function in Young Mice

ZHANG Xuan¹^,²(), YANG Xue¹^,², LI Xinke¹^,², ZHENG Nan¹^,², MENG Lu¹^,²^,*()

1. Key Laboratory of Quality & Safety Control for Dairy Products of China Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China
2. Laboratory of Quality and Safety Risk Assessment for Dairy Products of China Ministry of Agriculture and Rural Affairs (Beijing), Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received:2024-04-16 Online:2025-03-23 Published:2025-04-02
Contact: MENG Lu E-mail:18647394594@163.com;menglu@caas.cn

摘要/Abstract

摘要：

旨在利用细胞试验确定丁酸钠对人小肠上皮细胞(FH_S 74 int cells)中细胞因子和紧密连接蛋白表达调控的最适浓度，并进一步探讨在该浓度范围下丁酸钠对幼鼠回肠发育、炎症因子及物理屏障功能的影响。细胞试验选取FHs 74 int细胞，探究不同浓度丁酸钠对细胞活力、炎症因子及紧密连接蛋白表达水平的影响。动物试验选取刚出生的C57BL/6J幼鼠，共32只，随机分为4组，每组8只，分别为对照组(0.9%生理盐水)、SB20组(20 mg·kg^-1 BW丁酸钠)、SB350组(350 mg·kg^-1 BW丁酸钠)和SB1000组(1 000 mg·kg^-1 BW丁酸钠)，连续给药30 d后对幼鼠体重、血常规以及回肠组织学形态、回肠细胞因子和紧密连接蛋白基因表达水平进行检测。在细胞试验中，12.5、25 mmol·L^-1丁酸钠显著提高了细胞闭锁蛋白(occludin)的基因和蛋白表达量，25 mmol·L^-1丁酸钠显著上调了细胞GPR109A的基因表达量以及闭锁小带蛋白(ZO-1)的基因和蛋白表达量。在动物试验中各组幼鼠的体增重以及血常规指标均无显著差异。通过回肠病理学发现，350 mg·kg^-1 BW丁酸钠显著提高了幼鼠的回肠绒毛高度，降低了隐窝深度，同时提高了回肠抗炎因子IL-4、IL-10的mRNA相对表达量，降低促炎因子IL-18的mRNA相对表达量；1 000 mg·kg^-1 BW丁酸钠显著提高了回肠绒毛高度/隐窝深度的比值(P < 0.05)，降低隐窝深度和促炎因子TNF-α的mRNA相对表达量。综上，25 mmol·L^-1的丁酸钠能够促进FH_S 74 int细胞occludin和ZO-1以及GPR109A的基因表达，维护屏障功能；350 mg·kg^-1 BW的丁酸钠能够促进幼鼠回肠形态结构发育，提高回肠绒毛高度，降低隐窝深度，同时调节肠道炎症因子和紧密连接蛋白的基因表达。

关键词: 丁酸钠, FH_S 74 int细胞, 肠道屏障, 紧密连接蛋白

Abstract:

The purpose of this study was to determine the optimal concentration of sodium butyrate in regulating the expression of cytokines and tight junction proteins in human intestinal epithelial cells (FH_S 74 int cells) by cell experiments, and to further explore the effects of sodium butyrate on ileum development, inflammatory factors and physical barrier function in young rats. In cell experiments, FH_S 74 int cells were selected to explore the effects of different concentrations of sodium butyrate on cell viability, expression levels of inflammatory factors and tight junction protein. In animal experiment, 32 newborn C57BL/6J mice were randomly divided into 4 groups, each with 8 mice, namely the control group (0.9% normal saline), SB20 group (20 mg·kg^-1 BW sodium butyrate), SB350 group (350 mg·kg^-1 BW sodium butyrate), and SB1000 group (1 000 mg·kg^-1 BW sodium butyrate). After continuous administration for 30 days, body weight, blood routine, ileal histological morphology, ileal cytokine and tight junction protein gene expression levels of mice were detected. In vitro, 12.5 and 25 mmol·L^-1 sodium butyrate increased the gene and protein expression of occludin in cells, and 25 mmol·L^-1 sodium butyrate significantly increased the gene expression of GPR109A and the gene and protein expression of ZO-1. In animal experiment, there were no significant differences in body weight and blood routine indexes among the groups of mice. Through ileal pathology, it was found that 350 mg·kg^-1 BW sodium butyrate significantly increased the ileal villus height but reduce the crypt depth of young mice (P < 0.05). At the same time, it increased the relative mRNA expression of ileal anti-inflammatory factors IL-4 and IL-10, while decreasing the relative mRNA expression of pro-inflammatory factor IL-18. 1 000 mg·kg^-1 BW sodium butyrate significantly increased the ratio of villus height to crypt depth of ileum (P < 0.05), decreased the crpyt depth and the relative mRNA expression of pro-inflammatory factor TNF-α. In conclusion, 25 mmol·L^-1 sodium butyrate can promote the gene expression of occludin, ZO-1, and GPR109A in FH_S 74 int cells, maintaining barrier function. Sodium butyrate at 350 mg·kg^-1 BW can promote the development of the ileum morphological structure of mice, increase villus height, reduce crypt depth, and simultaneously regulate the gene expression of intestinal inflammatory factors and tight junction proteins.

Key words: sodium butyrate, FH_S 74 int cells, intestinal barrier, tight junction protein

中图分类号:

S815.7

张萱, 杨雪, 李新科, 郑楠, 孟璐. 丁酸钠对幼鼠回肠发育、炎症因子及物理屏障功能的调控作用[J]. 畜牧兽医学报, 2025, 56(3): 1278-1289.

ZHANG Xuan, YANG Xue, LI Xinke, ZHENG Nan, MENG Lu. The Regulatory Effects of Sodium Butyrate on Ileal Development, Inflammatory Factors and Physical Barrier Function in Young Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(3): 1278-1289.

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基因名称 Gene name	参考序列登录号 Reference GenBank ID	引物序列(5′→3′) Primer sequences(5′→3′)	产物长度/bp Product length
GAPDH	NM_008084	F: AGGTCGGTGTGAACGGATTTG	123
		R: TGTAGACCATGTAGTTGAGGTCA
IL-10	NM_010548	F: GCTCTTACTGACTGGCATGAG	105
		R: CGCAGCTCTAGGAGCATGTG
IL-18	NM_008360	F: GACTCTTGCGTCAACTTCAAGG	169
		R: CAGGCTGTCTTTTGTCAACGA
IL-4	NM_021283	F: GGTCTCAACCCCCAGCTAGT	102
		R: GCCGATGATCTCTCTCAAGTGAT
TNF-α	NM_013693	F: CCCTCACACTCAGATCATCTTCT	61
		R: GCTACGACGTGGGCTACAG
β-actin	NM_001101	F: TTTTAGGATGGCAAGGGACTT	235
		R: GATGAGATTGGCATGGCTTTA
GPR109A	NM_177551	F: ATGTTGGCTATGAACCGCCAG	119
		R: GCTGCTGTCCGATTGGAGA
Human-IL-10	NM_000572	F: TCAAGGCGCATGTGAACTCC	176
		R: GATGTCAAACTCACTCATGGCT
Human-IL-18	NM_001562	F: TCTTCATTGACCAAGGAAATCGG	75
		R: TCCGGGGTGCATTATCTCTAC
Human-occludin	NM_001205254	F: ACAAGCGGTTTTATCCAGAGTC	89
		R: GTCATCCACAGGCGAAGTTAAT
Human-ZO-1	NM_003257	F: CAACATACAGTGACGCTTCACA	105
		R: CACTATTGACGTTTCCCCACTC

项目 Item	对照组 Control group	SB20组 SB20 group	SB350组 SB350 group	SB1000组 SB1000 group	P值 P-value
初始体重/g Initial body weight	1.45±0.15	1.53±0.08	1.45±0.10	1.50±0.14	0.582
终体重/g Final body weight	12.16±1.44	13.29±1.63	12.55±1.07	13.03±0.93	0.393
体增重/g Weight gain	10.71±1.50	11.76±1.62	11.10±1.13	11.53±0.96	0.477

项目 Item	对照组 Control group	SB20组 SB20 group	SB350组 SB350 group	SB1000组 SB1000 group	P值 P-value
白细胞数/(10⁹·L^－1) Leukocyte count	3.43±0.50	3.51±0.29	3.45±0.20	3.95±0.21	0.695
淋巴细胞数/(10⁹·L^－1) Leukocyte count	3.25±0.26	2.14±0.26	3.24±0.54	2.68±0.30	0.348
单核细胞数/(10⁹·L^－1) Mononuclear cells count	1.01±0.31	1.06±0.35	1.31±0.50	1.40±0.52	0.745
粒细胞数/(10⁹·L^－1) Granulocyte count	1.00±0.32	1.14±0.34	1.51±0.59	1.29±0.50	0.783
血红蛋白/(g·L^－1) Hemoglobin	162.22±5.15	159.04±3.76	161.16±2.45	151.00±4.86	0.262
平均血红蛋白含量/pg Mean corpuscular hemoglobin	22.38±0.36	22.56±0.46	21.82±0.39	22.54±0.68	0.689
红细胞数/(10¹²·L^－1) Erythrocyte count	7.25±0.30	7.06±0.30	7.58±0.29	6.64±0.20	0.154
血小板数/(10⁹·L^－1) Platelet count	359.40±27.57	418.40±38.76	366.00±16.12	395.20±38.60	0.542

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丁酸钠对幼鼠回肠发育、炎症因子及物理屏障功能的调控作用

The Regulatory Effects of Sodium Butyrate on Ileal Development, Inflammatory Factors and Physical Barrier Function in Young Mice

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