肝硬化自发性腹膜炎患者肠道优势菌群分析初探

Analysis of predominant gastrointestinal microbiota in hepatic cirrhosis with spontaneous bacterial peritonitis

目的探讨肝硬化自发性腹膜炎(SBP)患者肠道优势菌群多样性变化的规律。方法采集肝硬化SBP初始诊治患者和健康受试者各30例,收集粪便样本,进行PCR-DGGE优势菌群结构分析,并应用定量PCR技术检测肠道内乳杆菌属、双歧杆菌属和肠杆菌科细菌的含量。结果肝硬化SBP患者肠道优势菌群结构复杂程度明显增多,SBP组和健康对照组患者分别能够聚类到不同的簇中,提示两组受试者肠道优势菌群结构明显不同。定量PCR检测结果发现,肠道优势细菌中双歧杆菌的含量明显下降(7.90±0.96 vs.9.15±1.21),而肠杆菌科细菌的数量明显增多(8.95±0.88 vs.7.31±0.40),两组间比较差异具有统计学意义(P<0.05),这与SBP常见的病原菌有一定的相关性。结论肝硬化SBP患者肠道优势菌群结构明显变化,双歧杆菌属细菌明显减少,肠杆菌科细菌明显增加,肠道微生态失衡可能是SBP发生的内在原因。

Objective To investigate the diversity of predominant gastrointestinal microbiota in cirrhotic patients with spontaneous bacterial peritonitis （SBP）. Methods 30 initially diagnosed cases of hepatic cirrhosis with SBP and 30 healthy control participants were included in the study. After collecting the feces from these participants, the total genomic bacterial DNA was extracted. The diversity and structure of the predominant gastrointestinal microbiota were explored using culture-independent PCR-denaturing gradient gel electrophore- sis （DGGE）, and the numbers of gastrointestinal Lactobacillus spp. , Bifidobacterium spp. and Enterobacteriaceae were determined with quantitative real-time PCR. Results The structure of the predominant gastrointestinal microbiota in cirrhotic patients with SBP was much more complex than that in healthy participants. The participants in SBP group and Control group could be divided into different clusters. Our data indicated that the predominant gastrointestinal microbiota in the two groups were significantly different. The results of quantitative real-time PCR showed that the number of Bifidobacterium spp. significantly decreased in SBP group, compared with control group （7.90 ＋-0.96 vs 9.15 ＋ 1.21 ） , while the number of Enterobacteriaceae significantly increased （8.95 -＋ 0.88 vs. 7.31 ：~ 0.40） , which showed statistically significant differences （ P 〈 0.05 ）. The change of the predominant gastrointestinal microbiota might be associated with the common SBP pathogens. Conclusion The diversity of the predominant gastrointestinal microbiota in cirrhotic patients with SBP changed significantly, while the number of Bifidobacterium spp. decreased and the numbers of Enterobacteriaceae increased obviously, which indicated that the gastrointestinal dysbiosis might be the underlying causes of SBP.