急性肝衰竭时小鼠胆汁内微生态变化的研究

Microflora Changes in Bile of Acute Hepatic Failure Mice

目的:研究急性肝衰竭对小鼠胆汁内菌群的影响。方法:将12只SPF级健康BALB/c小鼠分为肝衰竭模型组(M组)和对照组(K组),M组小鼠采用D-氨基半乳糖建腹腔注射建立肝衰竭模型,K组小鼠腹腔注射等量生理盐水作为对照,造模后24 h,采集小鼠血清行丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、总胆红素(TBi L)及胆碱酯酶(CHE)生化检测;采集小鼠胆汁,通过高通量测序技术测定细菌16Sr DNA的V3+V4区细菌丰度及多样性;HE染色观察肝脏组织的病理变化,LEf Se分析肝衰竭前后各段的差异菌。结果:血清生化指标和HE染色结果提示,M组小鼠肝衰竭模型建立成功;K、M两组小鼠的胆汁内均包含拟杆菌门、厚壁菌门、变形菌门、蓝藻菌门和放线菌门这5个优势菌门;门水平上,M组小鼠胆汁内Thermi的相对丰度高于K组,差异有统计学意义(P<0.05);科水平上,与K组相比,M组小鼠胆汁内乳杆菌科增多(P<0.05)。结论:急性肝衰竭可能会导致胆汁内乳酸杆菌科增多。

Objective：To study the influence of acute liver failure（ ALF） on the changes of bile flora in mice. Methods：Twelve mice were randomly divided into 2 groups：control group（ group K） and ALF group（ group M）. ALF in group M were induced by intraperitoneal injection of D-galactosamine（ D-Ga IN）（ 3. 0 g/kg）. Mice in group K were injected with normal saline instead. Serums were collected in 24 hours after model made to determine alanine aminotransferase（ ALT）,aspartat aminotransferase（ AST）,total bilirubin（ TBIL）,and cholinesterase（ CHE）. Mice bile was collected to determine bacterial abundance and diversity in V3 ＋ V4 region of bacterial 16 Sr DNA by using high throughout sequencing technique. Liver tissues were collected for pathological examination with HE staining. Bacterial translocation and changes associated with acute liver injury were identified by LEf Se（ LDA Effect Size） analysis. Results：The serum biochemistry indexes and HE staining check results showed that the liver failure mouse model in group M was established successfully. In bile of mice of both groups were dominant bacteria phylums of Bacteroidetes,Firmicutes,Proteobacteria,Cyanobacteria and Actinobacteria. At phylum level,for Thermi of bile,the relative abundance in group M（ 0. 2237%） were significantly higher than that in group K（ 0. 055%）. At family level,compared with group K,there were more species of Lactobacillaceae in bile of group M,the difference was significant（ P〈0. 05）. Conclusion：ALF may result in an increase of Lactobacillaceae species in the bile.