克罗恩病患者肠道菌群多样性变化研究初探  

Alterations of bacterial diversity of intestinal microbiota in patients with Crohn′s diseases

作　　者：沈海燕[1] 阮水良[1] 徐水凌[2] 陆其明[1] 杨志宏[1] 罗鼎天[1] 陈春晓[3] 

机构地区：[1]嘉兴学院医学院附属第二医院消化内科,浙江嘉兴314000 [2]嘉兴学院医学院,浙江嘉兴314000 [3]浙江大学医学院附属第一医院消化内科,浙江杭州310003

出　　处：《中国微生态学杂志》2016年第6期662-666,共5页Chinese Journal of Microecology

基　　金：嘉兴市科技局科技研究计划资助项目(2012A1071-1)

摘　　要：目的探讨克罗恩病(CD)患者肠道菌群多样性变化。方法收集8例确诊CD患者的粪便标本(CD组),以同期8例健康人群粪便作为对照组(CN组)。收集新鲜粪便样本提取DNA,并将样本进行PCR扩增,最终进行16SrRNA数据分析。结果测序共获得990 890条高质量序列,CD组和CN组多样性指数Shannon和Simpson比较差异有统计学意义(t=-3.802和t=-2.886,P<0.05)。OTUs(可操作分类学单位)比较韦恩图显示CD组共有7 437株菌种,而CN组共有7 744株菌种,其中3 438株菌种重叠。LDA差异贡献分析图提示CD患者肠道菌群以芽胞杆菌、变形菌、乳杆菌目、放线菌目、异常球菌纲、栖热菌目、假单胞菌目和交替单胞菌目等为主要特征差异细菌菌类,而CN组则以双歧杆菌目及双歧杆菌科,Odoribacteraceae、Christensenellaceae、弧菌目及弧菌科等为主要特征差异细菌菌类。据物种进化树的样本菌落分布图显示:两组样本厚壁菌门、放线菌门、拟杆菌门、变形菌门、疣微菌门、酸杆菌门和螺旋菌门占主要门类分布。CD组及CN组细菌科类分析结果:毛螺菌科、韦荣球菌科、瘤胃菌科、丹毒丝菌科、红蝽菌科、紫单胞菌科、双歧杆菌科、普雷沃菌科、拟杆菌科、产碱菌科和链球菌科在两组样本中共同存在。同时从该进化树可发现,CD组中红蝽菌科、产碱菌科、丹毒丝菌科和链球菌科相对于CN组优势丰度分布,而普雷沃菌科、拟杆菌科和双歧杆菌科比CN组丰度明显减少。此外,Dethiosulfovibrionaceae、丙酸杆菌科、盐单胞菌科、希万菌科等为CD组特有菌种。结论 CD组的菌群多样性明显少于CN组,CD组有其特有优势菌群。Objective To explore the changes in the diversity of intestinal microbiota in patients with Crohn′s diseases（CD）.Methods 8patients with Crohn′s disease were enrolled in this study,while 8healthy participants were used as the controls.DNAs were extracted from fresh fecal specimens and amplified by PCR.The resulting data of 16 SrRNA gene were analyzed.Results 990,890high-quality reads were obtained and used for downstream analysis.Indicators of the alpha diversity,such as the Shannon and the Simpson indices,demonstrated that the level of diversity of the intestinal microbiota in group CD was significantly lower than that of the healthy controls（t=-3.802 and t=-2.886,P〈0.05）.Operational taxonomic units（OTUs）Venn diagram showed 7,437 strains of bacteria in CD group and 7,744 strains in control group,among which 3,438 strains overlapped.LDA difference-contribution analysis showed that the main bacteria were Bacilli,Gammaproteobacteria,Lactobacillales,Actinomycetales,Deinocicci,Thermales,Pseudomonadales and Alteromonadales in CD group,and Bifidobacteriales,Bifidobacteriaceae,Odoribacteraceae,Christensenellaceae,Vibrionales and Vibrionaceae in control group,respectively.According to evolutionary tree of colony,Actinobacteria,Bacteroidetes,Proteobacteria,Verrucomicrobia,Acidobacteria and Spirochaetes were prominant in both groups.Lachnospiraceae,Veillonellaceae,Ruminococcaceae,Erysipelotrichaceae,Corobacteriaceae,Porphyromonadaceae,Bifidobacteriaceae,Prevotellaceae,Bacteroidaceae,Alcaligenaceae and Streptococcaceae existed in both groups.Compared with the control group,Corobacteriaceae,Alcaligenaceae,Erysipelotrichaceae and Streptococcaceae appeared dominant abundance distribution in CD group,while Prevotellaceae,Bacteroidaceae and Bifidobacteriales were on the contrary.In addition,Dethiosulfovibrionaceae,Propionibacteriaceae,Halomonadaceae and Shewanellaceae were special microbiota in CD group.Conclusion Bacterial diversity in CD group was obviously decreased compared with control group,and CD group ha

关 键 词：克罗恩病 肠道菌群 16SrRNA高通量测序 多样性 

分 类 号：R574.62[医药卫生—消化系统]