16S rRNA高通量测序分析肠道菌群对小于胎龄大鼠生长追赶的影响及其可能的机制

doi:10.16150/j.1671-2870.2020.04.010

摘要/Abstract

摘要：

目的： 探讨肠道菌群在小于胎龄（small for gestational age, SGA）大鼠生长追赶中的作用,并进一步研究肠道菌群介导SGA生长追赶（catch-up growth,CUG）的可能机制。方法： 用孕鼠全程半限制饮食法诱导建立4周龄的SGA大鼠模型,共收集54只大鼠幼鼠,满4周龄时,其中30只（55.6%）实现CUG,作为有CUG的SGA大鼠幼鼠组（CUG-SGA组）,另24只（44.4%）无生长追赶（non-catch-up growth,NCUG）的SGA大鼠幼鼠作为NCUG-SGA组。用酶联免疫吸附试验（enzyme-linked immunosorbent assay, ELISA）法检测其血清胰岛素样生长因子1（insulin-like growth factor 1,IGF-1）水平;用16S rRNA高通量测序分析幼鼠粪便中的肠道菌群;用气相色谱-质谱法检测其肠道粪便中的短链脂肪酸（short chain fatty acid,SCFA）,分析比较2组间的差异。结果： CUG-SGA组幼鼠的血IGF-1水平为（250.41±85.66） μg/L,显著高于NCUG-SGA组的（112.29±32.26） μg/L（P<0.05）;CUG-SGA组幼鼠的肠道菌群多样性Simpson指数（0.99）显著高于NCUG-SGA组（0.97）（P<0.05）;CUG-SGA组幼鼠肠道粪便中的SCFA各组分水平均高于NCUG组（P<0.05）。SGA幼鼠肠道粪便中的SCFA水平与11种差异表达的菌属相对丰度间存在相关性（|r|均>0.6,P<0.05）,其中丙酸、丁酸、异丁酸、戊酸浓度与幼鼠的体重、身长及其血IGF-1水平间均呈显著正相关（r均>0.6,P<0.05）。结论： 肠道菌群在SGA大鼠的CUG中发挥一定作用,这种作用可能是通过菌群代谢产物SCFA调节机体能量代谢和吸收来实现的。

关键词: 小于胎龄, 肠道菌群, 生长追赶, 大鼠

Abstract:

Objective: To explore the effect of intestinal flora on catch-up growth in the rats of small for gestational age (SGA) and study the possible mechanism on catch-up growth (CUG) mediated by intestinal flora. Methods: SGA model was established by half dietary restriction during the pregnancy in female rats. A total of 54 baby rats were collected and were divided into two groups at the 4th week according to whether the rats achieved CUG or not: non-catch-up growth SGA (NCUG-SGA) group and CUG-SGA group. The serum insulin-like growth factor-1(IGF-1) in baby rats were measured with enzyme-linked immunosorbent assay (ELISA), the fecal intestinal flora was investigated by Illumina sequencing of 16S rRNA gene amplicons, and the level of fecal short-chain fatty acid (SCFA) and was tested by gas chromatography-mass spectrometer（GC-MS）, respectively. The difference in composition of the intestinal flora, the level of fecal SCFA and serum IGF-1 were compared between 2 groups. Results: The level of IGF-1 in CUG-SGA group was significantly higher than the NCUG-SGA group[（250.41±85.66） μg/L vs（112.29±32.26） μg/L, P<0.05] in baby rats. Simpson's diversity index for intestinal flora in CUG-SGA group was markedly higher than NCUG-SGA group. The level of fecal SCFA in CUG group was higher than NCUG group（P<0.05). SCFAs were correlated with relative abundance of 11 types of differentially expressed bacteria genus in CUG-SGA group（|r|>0.6, P<0.05）. Meanwhile,the levels of propionate, butyrate,isobutyrate and valerate were positively correlated with body weight and length as well as the serum level of IGF-1 in CUG-SGA rats. Conclusions: Intestinal flora exerts an effect on CUG in SGA rats. It may be achieved through regulating energy metabolism and absorption by SCFA,which produced by intestinal flora metabolism in the body.

Key words: Small for gestational age, Intestinal flora, Catch-up growth, Rat

中图分类号:

R725.7

安静静, 王俊祺, 肖园, 陆文丽, 李林, 王伟, 董治亚. 16S rRNA高通量测序分析肠道菌群对小于胎龄大鼠生长追赶的影响及其可能的机制[J]. 诊断学理论与实践, 2020, 19(04): 375-380.

AN Jingjing, WANG Junqi, XIAO Yuan, LU Wenli, LI Lin, WANG Wei, DONG Zhiya. The effect of intestinal flora on catch-up growth of small for gestational age detected by high throughput sequencing of 16S rRNA gene in rats and its possible mechanism[J]. Journal of Diagnostics Concepts & Practice, 2020, 19(04): 375-380.

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组别	出生时		4周龄
组别	身长（cm）	体重（g）	身长（cm）	体重（g）
CUG-SGA组（n=30）	4.92±0.38	4.65±0.16	12.35±0.50	73.41±7.23
NCUG-SGA组（n=24）	4.65±0.51	4.54±0.18	11.61±0.27	55.87±2.67
P值	0.150	0.120	<0.001	<0.001

差异菌属	相对丰度		P值
差异菌属	NCUG-SGA组	CUG-SGA组	P值
乳酸杆菌属	25.42%	16.50%	0.028
颤螺菌属	3.29%	1.12%	0.009
未分类_瘤胃菌科	2.76%	0.19%	0.009
未分类_脱硫弧菌科	0.86%	0.45%	0.047
未分类_毛螺菌科	0.79%	0.22%	0.028
梭菌属	0.20%	0.27%	0.028
Odoribacter^*	0.14%	0.01%	0.007
厌氧棍状菌属	0.11%	0.00%	0.019
脱硫弧菌属	0.17%	0.06%	0.028
未分类_克里斯滕森菌科	0.10%	0.03%	0.009
Dehalobacterium^*	0.10%	0.03%	0.009
链球菌属	0.10%	0.31%	0.028
未分类_消化球菌科	0.06%	0.01%	0.009
嗜胆菌属	0.06%	<0.01%	0.005
Dorea^*	0.06%	<0.01%	0.005
罗斯伯里菌属	0.04%	0.17%	0.016
萨特菌属	0.03%	0.10%	0.047
未分类_链球菌科	0.01%	0.02%	0.044
未明确_细菌	0.01%	0.04%	0.028
未分类_梭菌科	<0.01%	0.02%	0.055
丁酸弧菌	<0.01%	0.04%	0.019
未明确_RF39^*	<0.01%	0.0%	0.019

组别（n）	乙酸	丙酸	丁酸	异丁酸	戊酸	异戊酸	己酸
NCUG-SGA组	3.49±0.33	1.71±0.07	1.20±0.25	0.01±0.00	0.07±0.03	0.08±0.03	0.03±0.00
CUG-SGA组	7.02±2.59	3.44±1.39	2.91±0.38	0.02±0.00	0.14±0.05	0.20±0.08	0.04±0.01
P值	0.017	0.025	<0.001	0.001	0.022	0.012	0.003