Hu Y X, Yu L, Liu H J, Cheng M L
Department of Infectious Diseases, Guizhou Medical University Affiliated Hospital, Guiyang 550004, China.
Zhonghua Gan Zang Bing Za Zhi. 2017 Apr 20;25(4):291-296. doi: 10.3760/cma.j.issn.1007-3418.2017.04.011.
To investigate the changes in the composition of intestinal microbiota in mice with acute liver failure and identify characteristic bacteria, and to provide a basis for the diagnosis and treatment of acute liver failure with intestinal microbiota disorders. A total of 30 specific pathogen-free male BALB/c mice were used in this study, including 25 mice in the model group and 5 mice in the control group. An acute liver failure model was induced by D-galactosamine. Microbial DNA was extracted from intestinal contents in different segments of the lower digestive tract (ileum and colon) and feces and then were amplified using PCR. The regions of 16S V3-V4 were subjected to high-throughput sequencing, followed by bioinformatics analyses, including OTU hierarchical clustering, species annotation, alpha-diversity analysis, and LEfSe (LDA Effect Size) analysis. Comparison of continuous data was made using t-test, while comparison of categorical data was made using χ2 test. A total of 10 mice survived in the two groups, with 80% mortality rate in the model group. The alpha-diversity analysis revealed increased bacterial diversity and abundance in the ileum, increased bacterial diversity and reduced bacterial abundance in the colon, and reduced bacterial diversity and insignificantly changed bacterial abundance in feces in the model group compared with the control group. Based on the optimized classification level, significantly reduced abundance of Clostridiaceae (44.95% ± 19.28% vs 7.51% ± 16.57%, = 0.011) in the ileum, whereas significantly increased abundance of Rikenellaceae (1.08% ± 1.01% vs 4.18% ± 2.39%, = 0.028), S24-7 (4.75% ± 4.87% vs 22.77% ± 13.05%, = 0.020), and F16 (0.24% ± 0.28% vs 2.18% ± 1.61%, = 0.029) in the colon were found in model group compared with the control group. The LEfSe analysis demonstrated that there were significant differences in Staphylococcaceae and S24-7 between the two groups, and S24-7 could be defined as the characteristic bacteria. Intestinal microbiota disorders, especially the excessive growth of microbes in the ileum, are observed in mice with acute liver failure. Moreover, acute liver failure may be closely associated with the excessive growth of S24-7.
探讨急性肝衰竭小鼠肠道微生物群组成的变化，鉴定特征性细菌，为肠道微生物群紊乱所致急性肝衰竭的诊断和治疗提供依据。本研究共使用30只无特定病原体的雄性BALB/c小鼠，其中模型组25只，对照组5只。用D-半乳糖胺诱导急性肝衰竭模型。从下消化道不同节段（回肠和结肠）的肠内容物及粪便中提取微生物DNA，然后用PCR进行扩增。对16S V3-V4区域进行高通量测序，随后进行生物信息学分析，包括OTU层次聚类、物种注释、α多样性分析和LEfSe（线性判别分析效应大小）分析。连续数据比较采用t检验，分类数据比较采用χ2检验。两组共有10只小鼠存活，模型组死亡率为80%。α多样性分析显示，与对照组相比，模型组回肠细菌多样性和丰度增加，结肠细菌多样性增加但丰度降低，粪便细菌多样性降低且丰度变化不显著。基于优化的分类水平，模型组回肠中梭菌科丰度显著降低（44.95%±19.28% vs 7.51%±16.57%，P = 0.011），而结肠中理研菌科（1.08%±1.01% vs 4.18%±2.39%，P = 0.028）、S24-7（4.75%±4.87% vs 22.77%±13.05%，P = 0.020）和F16（0.24%±0.28% vs 2.18%±1.61%，P = 0.029）丰度显著增加。LEfSe分析表明，两组葡萄球菌科和S24-7存在显著差异，S24-7可被定义为特征性细菌。急性肝衰竭小鼠存在肠道微生物群紊乱，尤其是回肠微生物过度生长。此外，急性肝衰竭可能与S24-7过度生长密切相关。
