Analysis of gut microbiota in obese population with type 2 diabetes mellitus

doi:10.3969/j.issn.1673-8640.2016.010.003

Abstract

Abstract:

Objective To investigate the characteristics of gut microbiota in obese population with type 2 diabetes mellitus. Methods A total of 21 obese patients with type 2 diabetes mellitus（study group） and 21 obese subjects with normal glucose（control group） were enrolled. The fecal samples were collected and determined by Illumina Miseq high-throughput sequencing for the V3 region of 16S rRNA gene. The abundance and distribution were analyzed by cluster analysis. Gut microbiota was analyzed by real-time quantitation polymerase chain reaction（PCR）.Results The analysis of DNA sequence was successfully performed. The rare faction curves showed that adequate sequencing depth was achieved,and the coverage indices were >0.99. The Ace and Chao1 indices of study group were lower than those of control group（P<0.05）. The Shannon index or Simpson index of study group was lower or higher than that of control group（P<0.05）. The results of real-time quantitation PCR showed that the quantities of Actinobacteria and Firmicutes were lower（P<0.05）,while that of Bacteroidetes was higher in study group than that in control group（P<0.05）. The quantities of Proteobacteria and Fusobacteria had no statistical significance between the 2 groups（P>0.05）. The quantities ofVeillonella,Clostridium Ⅳ and Bacteroides were higher,and the quantities of Lactobacillus,Blautia coccoides-Eubacterium rectale,Prevotella and Bifidobacterium were lower in study group compared with those in control group（P<0.05）. Conclusions The obese population with type 2 diabetes mellitus has low abundance and diversity of gut microbiota. The quantitation analysis of the composition of gut microbiota has a certain significance.

Key words: Gut microbiota, Type 2 diabetes mellitus, Obese, High-throughput sequencing

CLC Number:

R446.5

ZENG Yipeng, HU Yan, WU Ping, FENG Xin'ge, GU Chengying, GUO Yafang. Analysis of gut microbiota in obese population with type 2 diabetes mellitus[J]. Laboratory Medicine, 2016, 31(10): 848-853.

Figures/Tables 7

References 13

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菌种	引物序列	产物序列长度（bp）
拟杆菌门	F: CATGTGGTTTAATTCGATGAT	126
拟杆菌门	R: AGCTGACGACAACCATGCAG
厚壁菌门	F: ATGTGGTTTAATTCGAAGCA	126
厚壁菌门	R: AGCTGACGACAACCATGCAC
放线菌门	F: CGCGGCCTATCAGCTTGTTG	600
放线菌门	R: CCGTACTCCCCAGGCGGGG
变形杆菌门	F: CATGACGTTACCCGCAGAAGAAG	195
变形杆菌门	R: CTCTACGAGACTCAAGCTTGC
梭形杆菌门	F: CCCTTCAGTGCCGCAGT	273
梭形杆菌门	R: GTCGCAGGATGTCAAGAC
拟杆菌属	F: GAGAGGAAGGTCCCCCAC	106
拟杆菌属	R: CGCTACTTGGCTGGTTCAG
普雷沃菌属	F: GGTTCTGAGAGGAAGGTCCCC	121
普雷沃菌属	R: TCCTGCACGCTACTTGGCTG
Ⅳ型梭菌属	F: GCACAAGCAGTGGAGT	239
Ⅳ型梭菌属	R: CTTCCTCCGTTTTGTCAA
乳杆菌属	F: GAGGCAGCAGTAGGGAATCTTC	126
乳杆菌属	R: GGCCAGTTACTACCTCTATCCTTCTTC
韦荣球菌属	F: ACCAACCTGCCCTTCAGA	343
韦荣球菌属	R: CGTCCCGATTAACAGAGCTT
Blautia球菌-直肠真杆菌属	F: CGGTACCTGACTAAGAAGC	429
Blautia球菌-直肠真杆菌属	R: AGTTTCATTCTTGCGAACG
双歧杆菌属	F: CTCCTGGAAACGGGTGG	550
双歧杆菌属	R: GGTGTTCTTCCCGATATCTACA

菌种	引物序列	产物序列长度（bp）
拟杆菌门	F: CATGTGGTTTAATTCGATGAT	126
拟杆菌门	R: AGCTGACGACAACCATGCAG
厚壁菌门	F: ATGTGGTTTAATTCGAAGCA	126
厚壁菌门	R: AGCTGACGACAACCATGCAC
放线菌门	F: CGCGGCCTATCAGCTTGTTG	600
放线菌门	R: CCGTACTCCCCAGGCGGGG
变形杆菌门	F: CATGACGTTACCCGCAGAAGAAG	195
变形杆菌门	R: CTCTACGAGACTCAAGCTTGC
梭形杆菌门	F: CCCTTCAGTGCCGCAGT	273
梭形杆菌门	R: GTCGCAGGATGTCAAGAC
拟杆菌属	F: GAGAGGAAGGTCCCCCAC	106
拟杆菌属	R: CGCTACTTGGCTGGTTCAG
普雷沃菌属	F: GGTTCTGAGAGGAAGGTCCCC	121
普雷沃菌属	R: TCCTGCACGCTACTTGGCTG
Ⅳ型梭菌属	F: GCACAAGCAGTGGAGT	239
Ⅳ型梭菌属	R: CTTCCTCCGTTTTGTCAA
乳杆菌属	F: GAGGCAGCAGTAGGGAATCTTC	126
乳杆菌属	R: GGCCAGTTACTACCTCTATCCTTCTTC
韦荣球菌属	F: ACCAACCTGCCCTTCAGA	343
韦荣球菌属	R: CGTCCCGATTAACAGAGCTT
Blautia球菌-直肠真杆菌属	F: CGGTACCTGACTAAGAAGC	429
Blautia球菌-直肠真杆菌属	R: AGTTTCATTCTTGCGAACG
双歧杆菌属	F: CTCCTGGAAACGGGTGG	550
双歧杆菌属	R: GGTGTTCTTCCCGATATCTACA

组别	年龄（岁）	性别（男/女）	BMI （kg/m²）	空腹血糖(mmol/L)	糖化血红蛋白（%）	空腹胰岛素（mU/L)	甘油三酯(mmol/L)	总胆固醇(mmol/L)
对照组	52.11±8.56	11/10	30.95±2.77	12.16±3.13	10.71±2.31	15.26±6.17	2.13±1.32	5.39±1.32
研究组	48.12±9.33	13/8	31.23±2.91	5.29±1.19	4.22±1.23	9.35±3.09	2.09±1.38	5.53±1.21
P值	0.16	0.49	0.75	0.00	0.00	0.00	0.92	0.73

组别	年龄（岁）	性别（男/女）	BMI （kg/m²）	空腹血糖(mmol/L)	糖化血红蛋白（%）	空腹胰岛素（mU/L)	甘油三酯(mmol/L)	总胆固醇(mmol/L)
对照组	52.11±8.56	11/10	30.95±2.77	12.16±3.13	10.71±2.31	15.26±6.17	2.13±1.32	5.39±1.32
研究组	48.12±9.33	13/8	31.23±2.91	5.29±1.19	4.22±1.23	9.35±3.09	2.09±1.38	5.53±1.21
P值	0.16	0.49	0.75	0.00	0.00	0.00	0.92	0.73

组别	Reads指数	Coverage指数	OTU	Ace	Chao1	Shannon	Simpson
对照组	23 454.33±2 224.89	0.998 1±0.002 1	188.12±19.16	209.77±32.21	225.54±43.55	3.23±0.89	0.088 7±0.050 1
研究组	22 921.45±1 258.76	0.997 2±0.001 8	113.44±17.25	119.09±24.91	187.49±36.42	2.39±0.76	0.128 7±0.071 2
P值	0.057 6	0.143 8	0.000 0	0.000 0	0.003 8	0.002 1	0.041 5