贵州医科大学学报

2017, v.42;No.200(05) 528-531

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急性肝损伤小鼠口腔菌群的变化
Oral Microecological Changes in Mice with Acute Liver Injury

刘华娟;余蕾;胡亚欣;程明亮;
LIU Huajuan;YU Lei;HU Yaxin;CHENG Mingliang;Guizhou Medical University;The Affiliated Hospital of Guizhou Medical University;

摘要(Abstract):

目的:研究急性肝损伤时小鼠口腔菌群的变化。方法:将健康BALB/c小鼠随机分为正常对照组(K组)和肝损伤模型组(M组),M组以腹腔注射3.0 g/kg D-氨基半乳糖建立急性肝损伤模型,K组在相同时间点腹腔注射无菌生理盐水2 m L,后继续正常饲喂24 h,禁食12 h;无菌条件下取两组小鼠眼球血血清检测丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)和血清白蛋白(ALB)等肝功能指标,取肝脏右叶做病理HE染色,通过Chao1、香农指数分析小鼠口腔盥洗液中菌群丰度及多样性检测,LDA Effect Size(LEf Se)分析和聚类树寻找差异菌群。结果:与K组相比,M组小鼠血清ALT、AST升高,ALB降低(P<0.05),小鼠肝脏HE染色示M组有部分肝细胞发生轻至中度病变,可见细胞坏死和炎症浸润,提示小鼠急性肝损伤模型复制成功;与K组相比,M组小鼠口腔菌群Chao1、香农指数降低,差异具有统计学意义(P<0.05);K、M两组小鼠的口腔内相对含量>1%的共同优势菌群有拟杆菌门、厚壁菌门、变形菌门、蓝藻菌门和放线菌门,M组小鼠口腔TM7的相对丰度明显高于K组(P<0.05);由聚类树和LDA分布柱状图可知,两组小鼠的口腔微生物具有明显差异的菌群聚类为厚壁菌门的芽孢杆菌纲的乳杆菌目。结论:急性肝损伤时小鼠口腔菌群会发生改变。
Objective: To study the changes of oral cavity flora associated with acute liver injury(ALI) in mice. Methods: Mice were randomly divided into control group(group K) and ALI group(group M). Mice in group M were given 3. 0 g/kg D-galactosamine intraperitoneal injection to edtablish ALI model,and mice in group K were given injection of 2 m L of normal saline instead. Then,all of the mice were reared normally for 24 h before fasting of 12 h. Serum ALT,AST and ALB of mice were tested,the right liver lob were taken to make HE slide for pathological observation,and the abundance and diversity of bacteria in oral washing lotion were detected and analyzed with Chao 1 and Shannon index. The bacterial translocation and changes associated with ALI were identified by LEf Se(LDA Effect Size) analysis. Results: The serum levels of ALT,AST were higher and the serum ALB level was lower in group M than those in group K(P < 0. 05). Pathological observation showed light to middle degrees of lesion in part of liver in group M including cell necrosis and inflammatory infiltration,which suggested the success of ALI model formation. The diversity and relative abundance of bacteria in oral cavity in group M decreased significantly than those in group K(P < 0. 05). The dominant bacteria phyla of both groups were Bacteroidetes,Firmicutes,Proteobacteria,Cyanobacteria and Actinobacteria. The relative abundance of oral cavity TM7 was obviously higher in group M(1.4766%) than that in group K(0. 0509%)(P < 0. 05). Clustering tree analysis showed that the markedly different flora cluster of the 2 groups was Order Lactobacillales of Family Bacillibacteria in Phylum Firmicutes. Conclusion: Bacteria flora in oral cavity change during ALI.

关键词(KeyWords): 急性肝损伤;高通量核苷酸测序;微生态;生物多样性;口腔
acute liver injury;high-throughput sequencing;micro-ecology;biological diversity;oral cavity

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基金项目(Foundation): 贵州省科学技术基金[黔科合LH字(2016)7243号]

作者(Author): 刘华娟;余蕾;胡亚欣;程明亮;
LIU Huajuan;YU Lei;HU Yaxin;CHENG Mingliang;Guizhou Medical University;The Affiliated Hospital of Guizhou Medical University;

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