炎症性肠病肠道黏膜细菌谱的研究

Mucosal bacteria in Inflammatory Bowel Diseases patients revealed by 16S rDNA gene cloning and sequencing analysis

目的以16S-rDNA基因序列测定方法研究日本炎症性肠病（inflammatoryboweldisease，1BD）患者与正常对照肠镜标本中细菌构成谱。方法纳入2010年11月-2011年6月在日本名古屋大学附属病院接受肠镜检查患者14例，其中4例活动期溃疡性结肠炎（ulcerativecolitis，UC）患者（8个标本），3例缓解期UC（5个标本），2例活动期克罗恩病（Crohn’Sdisease，CD）（4个标本），和正常对照5例（10个标本），共收集肠镜27个标本。应用16S-rDNA基因序列测定方法检测肠道黏膜细菌谱。比较不同组间、不同部位标本的细菌构成差异。结果共取得796个细菌16S-rDNA基因序列。这些序列分属于5个门（phylum）：厚壁菌（Firmicutes）、拟杆菌（Bacteroidetes）、变形杆菌（Proteobacteria）、梭杆菌（Fusobacteria）和未分类细菌（unclassifiedBacteriao活动期UC组和活动期CD组中厚壁菌门细菌构成比低于缓解期UC组和正常对照组。CD组中细菌种类减少，CD组中变形杆菌门细菌高于UC组和对照组。结论本研究证实了IBD组肠道黏膜细菌构成谱不同于正常对照组，且有细菌种类减少趋势。

Objectives We conducted a study on comparing mucosa-associated bacterial communities between IBD pa- tients and non-IBD controls by using 16S rDNA cloning and sequencing method. Methods Fourteen patients undergoing colonoscopy at Nagoya University Hospital from November 2010 to June 2011 were included in the study, including four active ulcerative colitis patients （n=8）, three remission UC （n=5）, two active CD （n=4） and five non-IBD patients （n = 10）. 16S rDNA sequencing was used to identify mucosal-associated bacterial communities. Results 796 clones of bac- terial 16S rDNA gene sequences were gathered from intestinal biopsies. Preliminary results revealed that there were five phyla, Firmicutes, Bacteroidetes, Proteobacteria, Fusobacteria and unclassified bacteria within the mucosal bacterial communities. Compared with non-IBD and UC in remission, the portion of Firmicutes reduced in active UC and CD, more Proteobacteria in CD than the others. And bacterial diversity was reduced in IBD, particularly in CD at phylum level. Conclusion The results may support the dysbiosis hypothesis observed in inflammatory bowel disease patients relative to non- IBD controls.