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  本文關鍵詞： 腸道菌群 慢性乙型肝炎 Miseq測序　出處：《廣西醫(yī)科大學》2015年碩士論文　論文類型：學位論文

【摘要】：目的：比較慢性乙型肝炎患者及乙肝相關性肝硬化患者與健康人群的腸道菌群多樣性和結構差異,探討慢性肝炎與腸道菌群之間的相互關系,為慢乙肝及其相關性肝硬化的治療提供新思路。方法：按照2010年中華醫(yī)學會肝病學分會和感染病學分會修訂的《慢性乙型肝炎防治指南》診斷標準,選擇廣西醫(yī)科大學第一附屬醫(yī)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統(tǒng)疾病者39例,其中慢性乙型肝炎組(CHB組)19例,乙肝后肝硬化20例(LC組,均為失代償期)。健康對照組9例。所有研究對象近1個月內均未使用抗生素及益生菌制劑。收集所有48例研究對象糞便樣本,提取細菌組DNA,送深圳華大基因科技服務公司進行Illumina Miseq測序,分析各樣本物種組成和豐度,并對各組內和組間物種多樣性差異和菌群結構差異進行分析。結果：(1)在所研究健康組及病例組中,擬桿菌門和厚壁菌門總比例達腸道菌群的80%以上；(2)健康組shannon指數(shù)大于肝硬化組,差異有統(tǒng)計學意義(P0.05)；健康組simpson指數(shù)小于慢乙肝組,差異有統(tǒng)計學意義(P0.05); CHB組Observed species指數(shù)大于肝硬化組,差異有統(tǒng)計學意義(P0.05)。表明健康組、慢乙肝組、肝硬化組腸道菌群物種多樣性呈逐漸減少趨勢；(3)健康組、慢乙肝組及肝硬化組菌群結構呈現(xiàn)出較大差異,在聚類分析中各組互有交叉,但總體上表現(xiàn)出聚類趨勢,可見健康組、慢乙肝組與肝硬化組各組內均有其特征性菌群結構；(4)在門分類水平進行組間顯著性差異分析,可看出與健康組比較,慢乙肝組擬桿菌門豐度增加(p0.05)；肝硬化相較健康組無明顯差異；肝硬化組與慢乙肝組比較,肝硬化組擬桿菌門豐度減低,變形菌門豐度增加(p0.05)。而在屬分類水平,共有23種占比在0.5%以上的菌屬鑒定出有組間差異,其中慢乙肝及肝硬化組擬桿菌門下的機會致病菌普雷沃氏菌屬、TM7菌門下的blautia菌屬豐度均較健康組增加(p0.05)；肝硬化組韋榮氏球菌屬豐度增高(p0.05)。結論：1、各組中擬桿菌門和厚壁菌門均為腸道兩大優(yōu)勢菌群；2、隨慢乙肝病情進展,腸道菌群多樣性減少,而腸道菌群高多樣性與健康狀態(tài)有關；3、慢乙肝疾病不同階段,腸道菌群結構有顯著差異；4、 慢乙肝及肝硬化組主要表現(xiàn)為擬桿菌門及厚壁菌門下菌屬豐度改變。
[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.
【學位授予單位】：廣西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R512.62

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本文編號：1528466