ERIC-PCR和PCR-DGGE技术分析壳聚糖对抗生素引起肠道菌群失调小鼠的影响  

The effects of chitosan on antibiotic-induced intestinal microflora imbalance in mice by ERIC-PCR and PCR-DGGE

作　　者：赵鑫[1] 王佳[1] 邢宇[1] 张迪[1] 李新莉[1] 

机构地区：[1]大连医科大学生物技术系,辽宁大连116044

出　　处：《中国微生态学杂志》2017年第12期1378-1381,1386,共5页Chinese Journal of Microecology

基　　金：辽宁省教育厅科学研究项目一般项目(L2016004)

摘　　要：目的应用肠杆菌基因间重复共有序列基因扩增(ERIC-PCR)和聚合酶链式反应——变性梯度凝胶电泳(PCR-DGGE)分析壳聚糖对抗生素致肠道菌群失调小鼠的影响。方法 SPF级昆明小鼠24只,每组8只,依次分为模型组(N)、壳聚糖高(CS-H)和低浓度组(CS-L),盐酸左氧氟沙星灌胃6d后使用相应药物灌胃24d,收集鼠便,提取粪便细菌DNA,利用ERIC-PCR和PCR-DGGE电泳获得肠道菌群指纹图谱,主成分分析(PCA)和聚类分析(UPGMA)研究肠道菌群整体差异,并鉴定优势条带序列。结果 ERIC-PCR表明菌群失调组小鼠肠道中细菌条带减少明显,以300bp左右的条带为特征条带,PCR-DGGE显示屎肠球菌为优势菌型;壳聚糖灌胃小鼠肠道菌群结构组成发生改变,乳酸菌成为优势菌型。结论壳聚糖扶持乳酸菌等益生菌、抑制肠球菌等病原菌的增殖,起到益生元的作用进而调节肠道微生态均衡。Objective To observe the effects of chitosan on the intestinal microflora by using enterobacterial repetitive intergenic consensus-polymerase chain reaction（ERIC-PCR）and polymerase chain reaction-denaturing gradient gel electrophoresis（PCR-DGGE）techniques.Methods Twenty-four SPF KM mice were treated with Levofloxacin hydrochloride for 6 days and then randomly divided into the normal control group,low concentration group or high concentration group respectively（n=8）to receive the corresponding drugs for 24 days.The feces were collected for extraction of bacterial genomic DNA.PCA,UPGMA and sequence analysis were carried out by using ERIC-PCR and PCR-DGGE profilings.Results ERIC-PCR analysis showed that the bacterial bands reduced obviously in mice with microflora imbalance,and the band of 300 bp became the characteristic bands.PCR-DGGE showed that Enterococcus faecium was the dominant bacteria.After treatment with chitosan,the structure of intestinal microflora was changed,and Lactobacilli became dominant.Conclusion Chitosan can promote Lactobacilli and inhibit Enterococci,which has the effect of prebiotics to regulate intestinal microflora balance.

关 键 词：壳聚糖 肠道菌群 ERIC-PCR PCR-DGGE 

分 类 号：R965.2[医药卫生—药理学]