基于16S rDNA测序分析酒精性脂肪肝病进展中小鼠肠道菌群变化

Analysis of gut microbiota changes in alcoholic fatty liver disease in progression based on 16S rDNA sequencing technology

  • 摘要:
    目的 采用16S rDNA测序技术分析酒精性脂肪肝病(AFLD)发展过程中小鼠肠道菌群的变化,并探讨小鼠AFLD进展的可能机制。
    方法 将60只雄性C57BL/6小鼠随机分为对照组(35只)和模型组(25只)。适应性喂养5 d后,对照组小鼠每天喂食Lieber-DeCarli对照流质饲料(TP4030C),模型组小鼠每天喂食含有4%乙醇的Lieber-DeCarli流质饲料(TP4030B),连续30 d后,采用苏木精—伊红(HE)染色法观察肝脏病理变化;16S rDNA测序法分析肠道菌群组成结构。
    结果 与对照组相比,AFLD模型组小鼠肠道菌群的α多样性结果显示其物种丰富度降低(P<0.05)。Beta多样性结果表明两组小鼠肠道菌群结构组成和丰度存在显著差异(P<0.05)。与对照组相比,模型组粪杆菌属(Faecalibaculum)、杜博菌属(Dubosiella)、异杆菌属(Allobaculum)、瘤胃球菌属UCG-013(Ruminococcaceae UCG-013)等菌属相对丰度升高,乳酸杆菌属(Lactobacillus)、毛螺菌属(Lachnospiraceae NK4A136 group)等菌属相对丰度降低。模型组小鼠肠道菌群在丙氨酸转移酶、谷胱甘肽水解酶和组蛋白乙酰转移酶通路活性显著增加(P<0.05)。
    结论 含4%乙醇的Lieber-DeCarli流质饲料成功构建了AFLD C57BL/6小鼠模型,AFLD小鼠肠道菌群的结构和组成均发生变化,酒精可能通过破坏肠道微生物稳态,增加丙氨酸转移酶、谷胱甘肽水解酶和组蛋白乙酰转移酶通路活性加快AFLD的进展。

     

    Abstract:
    Objective  To analyze the changes of gut microbiota in the progression of alcoholic fatty liver disease(AFLD) using 16S rDNA sequencing technology, and to explore the possible mechanism of AFLD progression in mice.
    Methods  A total of 60 male C57BL/6 mice were randomly divided into control group (n=35) and model group (n=25). After adaptive feeding for 5 days, mice in the control group were fed a control Lieber-DeCarli fluid feeds (TP4030C) daily, and mice in the model group were fed Lieber-DeCarli fluid feeds (TP4030B) containing 4% ethanol daily. After 30 consecutive days, hematoxylin-eosin (HE) staining was used to observe the pathological changes of the liver. 16S rDNA sequencing was used to analyze the composition and structure of gut microbiota.
    Results  Compared with the control group, Alpha diversity results of the gut microbiota of mice in the AFLD model group showed reduced species richness (P < 0.05). Beta diversity results indicated significant differences in the structural composition and abundance of the gut microbiota of the two groups of mice (P < 0.05). Compared with the control group, the relative abundance of Faecalibaculum, Dubosiella, Allobaculum, Ruminococcaceae UCG-013 were higher in the model group, while the relative abundance of Lactobacillus and Lachnospiraceae NK4A136 group were lower in the model group. The activity of gut microbiota in alanyltransferase, glutathione hydrolase and histone acetyltransferase pathways was significantly increased in the model group of mice (P < 0.05).
    Conclusion  AFLD C57BL/6 mouse model is successfully constructed on Lieber-DeCarli fluid feeds containing 4% ethanol, and both the structure and composition of the gut microbiota of AFLD mice are altered. Alcohol may accelerate the progression of AFLD by disrupting gut microbial homeostasis and increasing the activities of alanyltransferase, glutathione hydrolase, and histone acetyltransferase pathways.

     

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