基于16S rRNA高通量测序技术分析小鼠实验性结肠炎肠道菌群结构特征

doi:10.16150/j.1671-2870.2019.03.005

摘要/Abstract

摘要：

目的： 明确溃疡性结肠炎（ulcerative colitis,UC）与肠道菌群间的相互关系,为寻找简单、安全的UC治疗方法提供实验基础。方法： 通过3%硫酸葡聚糖钠（dextran sulfate sodium,DSS）诱导建立小鼠溃疡性结肠炎模型,观察检测体重变化、肠道病理改变,采用16S rRNA高通量测序技术,检测小鼠粪便中的肠道菌群,并分析比较结肠炎小鼠与正常小鼠间肠道菌群的多样性及丰度。同时,利用已有数据库,预估肠道菌群内的功能基因构成,分析2组间的菌群基因功能差异。结果： 溃疡性结肠炎小鼠体重明显减轻,且肠道上皮完整性被破坏,同时肠道菌群多样性明显降低,溃疡性肠炎组小鼠肠道内双歧杆菌属降至0.2%明显低于正常组小鼠（P<0.05）,而乳酸杆菌属平均丰度也显著降低至2.9%（P<0.05）,提示可能是通过影响机体代谢从而造成溃疡性结肠炎的发生、发展。结论： 溃疡性结肠炎小鼠的肠道菌群多样性降低,菌群分布均发生显著改变。

关键词: 溃疡性结肠炎, 肠道菌群, 16S rRNA高通量测序

Abstract:

Objective: To clarify the interrelationship between ulcerative colitis (UC ) and gut microbiome for providing an experimental basis to find a simple and safe treatment for patients with ulcerative colitis. Methods: A model of ulcerative colitis in mice was established by 3% DSS (dextran sulfate sodium), and the body weight and intestinal pathological changes of the model mice were detected. The diversity and differences of gut microbiome in feces of ulcerative colitis mice and normal mice were identified by 16S rRNA high-throughput sequencing technology. The existing gene database was used to estimate the functional gene composition of the intestinal flora and the functional differences between the two groups. Results: The results of comparative analysis showed that the weight of ulcerative colitis mice was significantly reduced, the integrity of intestinal epithelium was destroyed, and the diversity of gut microbiome was significantly reduced. Bifidobacterium in intestinal tract of ulcerative colitis mice decreased to 0.2%, which was significantly lower than that of mice in control group (P<0.05); and the relative abundance of Lactobacillus also decreased to 2.9% (P<0.05), which denoted that ulcerative colitis might be caused by its influence on metabolism of the body. Conclusions: The diversity and distribution of gut microbiome in ulcerative colitis mice have changed significantly.

Key words: Inflammatory bowel disease, Gut microbiome, 16S rRNA high-throughput sequencing

中图分类号:

R574.62

汪婷婷, 郑乃盛, 袁向亮, 沈立松. 基于16S rRNA高通量测序技术分析小鼠实验性结肠炎肠道菌群结构特征[J]. 诊断学理论与实践, 2019, 18(03): 263-270.

WANG Tingting, ZHENG Naisheng, YUAN Xiangliang, SHEN Lisong. Analysis of structural characteristics of gut microbiome in colitis mice based on 16S rRNA high-throughput sequencing[J]. Journal of Diagnostics Concepts & Practice, 2019, 18(03): 263-270.

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指标	评分（分）
炎症程度无轻度重度	0 1 2
炎症累及范围 1%~25% 26%~50% 51%~75% 75%~100%	1 2 3 4
增生程度无轻度重度	0 1 2
病变深度无黏膜下层肌层浆膜层	0 1 2 3
结肠溃疡无小(≤3 mm) 大(>3 mm)	0 1 2

差异菌属	3%DSS组平均丰度（%）	对照组平均丰度（%）	均值差异	P值	95.0%可信区间下限	95.0%可信区间上限	效应值
双歧杆菌属	0.235 67	1.799 96	-1.564 29	0.040 65	-3.031 42	-0.097 16	0.424 62
肠球菌属	3.468 21	0.976 05	2.492 16	0.042 03	0.123 36	4.860 95	0.395 84
乳酸杆菌属	2.947 98	12.120 7	-9.172 76	0.044 44	-18.017 8	-0.327 69	0.406 37
韦荣球菌属	0.902 27	0.133 94	0.7683 29	0.045 54	0.021 73	1.514 93	0.399 46