慢性乙型病毒性肝炎患者肠道微生态的研究

发布时间：2018-02-24 02:08

本文关键词： 肠道菌群慢性乙型肝炎 Miseq测序　出处：《广西医科大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的：比较慢性乙型肝炎患者及乙肝相关性肝硬化患者与健康人群的肠道菌群多样性和结构差异,探讨慢性肝炎与肠道菌群之间的相互关系,为慢乙肝及其相关性肝硬化的治疗提供新思路。方法：按照2010年中华医学会肝病学分会和感染病学分会修订的《慢性乙型肝炎防治指南》诊断标准,选择广西医科大学第一附属医院感染性疾病科收治的慢性乙型肝炎病毒感染者,并排除合并其他嗜肝病毒肝炎及伴有其它系统疾病者39例,其中慢性乙型肝炎组(CHB组)19例,乙肝后肝硬化20例(LC组,均为失代偿期)。健康对照组9例。所有研究对象近1个月内均未使用抗生素及益生菌制剂。收集所有48例研究对象粪便样本,提取细菌组DNA,送深圳华大基因科技服务公司进行Illumina Miseq测序,分析各样本物种组成和丰度,并对各组内和组间物种多样性差异和菌群结构差异进行分析。结果：(1)在所研究健康组及病例组中,拟杆菌门和厚壁菌门总比例达肠道菌群的80%以上；(2)健康组shannon指数大于肝硬化组,差异有统计学意义(P0.05)；健康组simpson指数小于慢乙肝组,差异有统计学意义(P0.05); CHB组Observed species指数大于肝硬化组,差异有统计学意义(P0.05)。表明健康组、慢乙肝组、肝硬化组肠道菌群物种多样性呈逐渐减少趋势；(3)健康组、慢乙肝组及肝硬化组菌群结构呈现出较大差异,在聚类分析中各组互有交叉,但总体上表现出聚类趋势,可见健康组、慢乙肝组与肝硬化组各组内均有其特征性菌群结构；(4)在门分类水平进行组间显著性差异分析,可看出与健康组比较,慢乙肝组拟杆菌门丰度增加(p0.05)；肝硬化相较健康组无明显差异；肝硬化组与慢乙肝组比较,肝硬化组拟杆菌门丰度减低,变形菌门丰度增加(p0.05)。而在属分类水平,共有23种占比在0.5%以上的菌属鉴定出有组间差异,其中慢乙肝及肝硬化组拟杆菌门下的机会致病菌普雷沃氏菌属、TM7菌门下的blautia菌属丰度均较健康组增加(p0.05)；肝硬化组韦荣氏球菌属丰度增高(p0.05)。结论：1、各组中拟杆菌门和厚壁菌门均为肠道两大优势菌群；2、随慢乙肝病情进展,肠道菌群多样性减少,而肠道菌群高多样性与健康状态有关；3、慢乙肝疾病不同阶段,肠道菌群结构有显著差异；4、慢乙肝及肝硬化组主要表现为拟杆菌门及厚壁菌门下菌属丰度改变。
[Abstract]:Objective: to compare the diversity and structure of intestinal flora in chronic hepatitis B patients, hepatitis B related cirrhosis patients and healthy people, and to explore the relationship between chronic hepatitis B and intestinal flora. Methods: according to the revised guidelines for the Prevention and treatment of chronic Hepatitis B (CHB) in 2010, the Chinese Society of Hepatology and the Society of Infectious Diseases revised the diagnostic criteria for the treatment of chronic hepatitis B and its associated cirrhosis. We selected 39 patients with chronic hepatitis B virus infection admitted to the Department of Infectious Diseases, first affiliated Hospital of Guangxi Medical University, and excluded 39 patients with other hepatitis B and other systemic diseases, including 19 patients with chronic hepatitis B in CHB group. Twenty patients with posthepatitic cirrhosis in LC group were all in decompensated period and 9 in healthy control group. No antibiotics and probiotics were used in all subjects in the past one month. All the fecal samples of 48 subjects were collected. Bacterial DNA was extracted and sent to Shenzhen Huada Gene Technology Service Company for Illumina Miseq sequencing. The species composition and abundance of each sample were analyzed. The differences of species diversity and flora structure within and between groups were analyzed. The shannon index of healthy group was higher than that of cirrhosis group (P 0.05), the simpson index of healthy group was lower than that of chronic hepatitis B group, the simpson index of healthy group was lower than that of chronic hepatitis B group. The Observed species index in CHB group was higher than that in cirrhosis group, and the difference was statistically significant (P 0.05). The results showed that the species diversity of intestinal flora in healthy group, chronic hepatitis B group and cirrhosis group was decreasing gradually. The microflora structure of chronic hepatitis B group and liver cirrhosis group showed great difference. In cluster analysis, each group intersected, but overall showed a clustering trend, it can be seen that the healthy group, The characteristic microflora structure of chronic hepatitis B group and liver cirrhosis group were analyzed at the portal classification level, and it was found that there was a significant difference between the two groups in comparison with the healthy group. In chronic hepatitis B group, Bacteroides bacillus abundance increased p0.05, cirrhosis group had no significant difference compared with healthy group, liver cirrhosis group and chronic hepatitis B group had lower Bacteroidoid door abundance and higher Proteus abundance, but at the taxonomic level, there was no significant difference between cirrhosis group and chronic hepatitis B group. A total of 23 species of bacteria with a proportion of more than 0.5% were identified with different groups. Among them, the abundance of blautia in the subphylum of TM7 in patients with chronic hepatitis B and cirrhosis was higher than that in the healthy group (P 0.05), and the abundance of Veronella in cirrhotic patients was higher than that in the healthy group. Conclusion: 1: 1, Bacteroides in each group is more abundant than that in the healthy group (P < 0.05). And phylum thuringiensis were two dominant bacteria groups in the intestinal tract, and with the progress of chronic hepatitis B, The diversity of intestinal flora decreased, and the high diversity of intestinal flora was related to health status. There was significant difference in intestinal microflora structure. The changes of abundance of Bacteroides and subphylum in patients with chronic hepatitis B and liver cirrhosis were observed.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R512.62

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