肠道菌群与维吾尔族2型糖尿病的关联研究
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摘要
目的:比较分析新疆地区维吾尔族2型糖尿病个体与正常血糖个体肠道菌群结构的差异。方法:采集30例维吾尔族2型糖尿病个体与33例维吾尔族正常血糖个体的粪便样品,提取粪便样品中细菌的混合DNA,利用16S rDNA的一对细菌通用引物扩增粪便样品中菌群的基因,最后对PCR产物进行纯化,克隆,测序,分析肠道菌群的组成情况及其差异。结果:(1)比较维吾尔族2型糖尿病组与正常血糖组肠道菌群基因的PCR扩增产物,两者琼脂糖凝胶电泳时均在200bp～1500bp之间有条带出现,比较出现的条带数目,两组出现的平均条带数目有统计学差异,正常血糖组平均条带数目多于2型糖尿病组。(2)维吾尔族2型糖尿病个体与正常血糖个体肠道菌群基因在Genebank序列数据库上利用blast程序进行同源性比对,二者碱基序列有差别。(3)维吾尔族2型糖尿病个体与正常血糖个体肠道菌群的基因序列在Genebank数据库上用blast程序分别与引物相对应的细菌基因序列进行同源性比对,正常血糖个体肠道菌群的基因序列与芽胞杆菌(Uncultured Bacillus sp.AY242538)和乳酸球菌(Lactococcus lactis AM849109)的同源性比对率为100%,2型糖尿病个体肠道菌群的基因序列与这两个细菌的同源性比对率分别为90%和93%,可以确定,正常血糖个体肠道菌群中存在芽胞杆菌和乳酸球菌,2型糖尿病个体肠道菌群中缺失两细菌。结论:维吾尔族2型糖尿病个体与正常血糖个体相比,肠道菌群有差异,2型糖尿病个体的部分肠道细菌缺失。
Objective:To study difference of intestinal microflora between type 2 diabetes mellitus and euglycemia samples in xinjiang uygur.Methods:30 patients with type 2 diabetes Mellitus(T2DM) were selected and there were 33 subjects with euglycemia as the controls in xinjiang uygur.we collected their samples of feceals and extracted composite DNA of bacterium in these samples.Primers were used to amplify V6 to V8 variable regions of the bacterial 16S rDNA.Eventually,cloning and sequencing the purified PCR products,we analysed the composition of the microbial population.Results:(1) Some bands were appeared between 200bp and 1500bp when performing in agarose gel electrophoresis.There were significant differences on the bands in PCR products of intestinal microflora between T2DM groups and control groups(P＜0.05).(2) There were different gene orders in the intestinal microflora of the two groups with blast soft ware analysised on Genebank.(3) The obtained sequences were compared with sequence of known bacteria listed in the genebank nucleotide sequence database by using blast soft ware,euglycemia sample's similarity were 100%compared with Uncultured Bacillus sp.(AY242538)and Lactococcus lactis(AM849109),T2DM sample's similarity were 90% and 93%.Therefore,The bacteria identified included members of Uncultured Bacillus sp. and Lactococcus lactis in euglycemia sample.Conclusions:Compared with the samples, there were different intestinal microfloras with T2DM and euglycemia samples.T2DM sample lacked two bacterias in intestinal microfloras.
Objective:To study difference of intestinal microflora between type 2 diabetes mellitus and euglycemia samples in xinjiang uygur.Methods:30 patients with type 2 diabetes Mellitus(T2DM) were selected and there were 33 subjects with euglycemia as the controls in xinjiang uygur.we collected their samples of feceals and extracted composite DNA of bacterium in these samples.Primers were used to amplify V6 to V8 variable regions of the bacterial 16S rDNA.Eventually,cloning and sequencing the purified PCR products,we analysed the composition of the microbial population.Results:(1) Some bands were appeared between 200bp and 1500bp when performing in agarose gel electrophoresis.There were significant differences on the bands in PCR products of intestinal microflora between T2DM groups and control groups(P＜0.05).(2) There were different gene orders in the intestinal microflora of the two groups with blast soft ware analysised on Genebank.(3) The obtained sequences were compared with sequence of known bacteria listed in the genebank nucleotide sequence database by using blast soft ware,euglycemia sample's similarity were 100%compared with Uncultured Bacillus sp.(AY242538)and Lactococcus lactis(AM849109),T2DM sample's similarity were 90% and 93%.Therefore,The bacteria identified included members of Uncultured Bacillus sp. and Lactococcus lactis in euglycemia sample.Conclusions:Compared with the samples, there were different intestinal microfloras with T2DM and euglycemia samples.T2DM sample lacked two bacterias in intestinal microfloras.
引文
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[14]傅晓英,周焰,阚金娥等.2型糖尿病大血管病变相关因素多元回归分析.中国糖尿病杂志,2002,10(2):101-103.
[15]张菱,李德霞,傅晓英等.223例糖尿病人膳食现状调查.中国糖尿病杂志,2001,9(4):218-220.
[16]买苏木.马合木提,程祖亨,热娜等.自然长寿者的生活环境.新疆医科大学学报,2001,24(增刊)123-124.
[17]张菱,李德霞,傅晓英等.223例糖尿病人膳食现状调查.中国糖尿病杂志,2001,9(4):218-220.
[18]蒙昌金,莫炳强,田明照,冯桂湘等.小鼠限量对小鼠免疫功能、肠道菌群及寿命的影响.中国微生态学杂志,1999,(11)3:56-59.
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[20]Yan WL,Yang XY,Zheng Y J,et al.The Metabolic Syndrome in Uygur and Kazak Population.Diabetes Care[M].Alexandria.2005;28:(25)54-2556
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