2型糖尿病患者肠道拟类菌属和双歧杆菌属多样性的PCR-DGGE指纹图谱分析  

Analysis of Bacteroides and Bifidobacterium diversity in fecal samples from patients with type Ⅱdiabetes by using PCR-denaturing gradient gel electrophoresis fingerprint

作　　者：吴晓康[1] 徐纪茹[1] 张旭燕[1] 马超锋[1,2] 韩蕾[1] 申艳星[3] NAWAZ M 王娟[1] 

机构地区：[1]西安交通大学医学院免疫学与病原生物学系,环境与疾病相关基因教育部重点实验室,陕西西安710061 [2]西安市疾病预防控制中心病毒检验科,陕西西安710054 [3]陕西中医学院,陕西咸阳712046

出　　处：《西安交通大学学报（医学版）》2010年第4期475-480,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家重点基础研究发展规划(973计划)项目(No.2004CB418503)~~

摘　　要：目的分析2型糖尿病患者肠道拟类菌属和双歧杆菌属的多样性及其与健康个体的差异,探讨糖尿病与肠道优势菌属的相关性。方法收集2型糖尿病患者和健康志愿者粪样,提取样品中菌群总DNA,利用拟类菌和双歧杆菌16SrRNA种属特异性引物,进行PCR扩增,将产物用变性梯度凝胶电泳分离,获得肠道菌群分子指纹图谱,进行多样性和相似性等特征分析。结果糖尿病组和健康人组样品DGGE图谱多样性指数范围分别是:拟类菌属0.89(0.75～0.95),0.95(0.89～0.97);双歧杆菌属:0.85(0.75～0.94),0.87(0.73～0.93)。组内成对相似性系数累积曲线分析,拟类菌属:小于0.5的Cs值在糖尿病组占到总Cs值个数的85.83%,而健康人组只占到35.71%;双歧杆菌属:小于0.4的Cs值在糖尿病组占到总Cs值个数的90%,健康人组占到总Cs值个数的60.7%。结论糖尿病患者和健康人肠道拟类菌属和双歧杆菌属指纹图呈现个体特异性,糖尿病患者两种菌属多样性与健康个体无显著差异,而糖尿病患者个体之间的相似性较健康个体之间低。Objective To analyze the diversity of Bacteroides and Bifidobacterium in type 2 diabetes patients and its difference from that of healthy individuals so as to explore the relation between diabetes and gut predominant microbiota. Methods Bacterial DNAs from type 2 diabetes patients and healthy individuals were extracted from fecal samples and then characterized by PCR-denaturing gradient gel electrophoresis （DGGE） with genus-and group-specific primers targeting 16S rRNA for Bacteroides and Bifidobacterium. After DGGE profilings was obtained,the diversity and similarity analyses were carried out by the number of band,Shannon-Weaver （H＇）,cluster analysis and the cumulative distribution curve of similarity. Results The range of Shannon-Weaver （H＇） in diabetes group and healthy group was 0.89（0.75-0.95） and 0.95（0.89-0.97） for Bacteroides; 0.85（0.75-0.94） and 0.87（0.73-0.93） for Bifidobacterium. The cumulative distribution curve of intra-group similarity showed that the Dice similarity coefficient （Cs） lower than （or equal to） 0.5 took up 85.83% of the total Cs for Bacteroides in diabetes group and 35.71% in healthy group; the Cs lower than （or equal to） 0.4 constituted 90% of the total Cs for Bifidobacterium in diabetes group and 60.7% in healthy group. Conclusion DGGE profiling has shown individual specificity in diabetic group and healthy group for Bacteroides and Bifidobacterium. There is no significant distinction in diversity of Bacteroides and Bifidobacterium between the two groups. The intra-group similarity of Bacteroides and Bifidobacterium is lower in diabetic group than in healthy group.

关 键 词：2型糖尿病 肠道菌群 变性梯度凝胶电泳 指纹图谱技术 多样性 

分 类 号：R587.1[医药卫生—内分泌] Q938.1[医药卫生—内科学]