维吾尔族新发2型糖尿病患者肠道乳杆菌属和多形拟杆菌的定量研究
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摘要
目的定量研究维吾尔族新发2型糖尿病(T2DM)患者和糖耐量正常(NGT)人群肠道中乳杆菌属(Lactobacillus genus)和多形拟杆菌(Bacteroides thetaiotaomicron)的相对水平。方法严格按照纳入标准、排除标准收集维吾尔族新发T2DM患者96例,NGT 98例。提取所有研究对象的粪便细菌总DNA后,采用16SrDNA基因实时荧光定量PCR(Real-time PCR)对乳杆菌属和多形拟杆菌的水平进行定量检测;运用Pearson相关性分析乳杆菌属与研究对象的BMI、腰围、臀围、收缩压(SBP)、舒张压(DBP)、空腹血糖(FBG)、甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白(HDL-C)和低密度脂蛋白(LDL-C)的相关性。结果 16SrDNA基因Real-time PCR结果显示:(1)与新疆维吾尔族NGT组相比,乳杆菌属水平在新发T2DM中较低,差异有统计学意义(t=8.557,P=0.000)。但是多形拟杆菌在两组差异无统计学意义(t=0.524,P=0.601);(2)新疆维吾尔族上述人群肠道中乳杆菌属水平与FBG呈负相关(r=-0.334,P=0.000),腰围呈负相关(r=-0.170,P=0.018),TC呈负相关(r=-0.178,P=0.013),TG呈负相关(r=-0.157,P=0.030),收缩压呈负相关(r=-0.255,P=0.000)。结论 Lactobacillus genus水平在肠道中降低可能与2型糖尿病的发病,血糖和血脂代谢有关,其机制需进一步研究探讨。
Objective To analyze quantitatively the Lactobacillus genus and Bacteroides thetaiotaomicron in the gut microbiota of Xinjiang Uyghurs with new-onset T2 DM.Methods 96 cases of new-onset T2 DM and98 cases of normal glucose tolerance(NGT)were selected according to inclusion criteria.The total DNAs were extracted from the fecal microbiota,and the amounts of Lactobacillus genus and Bacteroides thetaiotaomicronin the gut microbiota were determined with 16 SrDNA gene Real-time fluorescence quantitative PCR amplification.The correlations of Lactobacillus genus and Bacteroides thetaiotaomicron with BMI,waist circumference,hip circumference,systolic blood pressure(SBP),diastolic blood pressure(DBP),fasting blood glucose(FBG),triglycerides(TG),total cholesterol(TC),high-density lipoprotein(HDL-C)and low-density lipoprotein(LDL-C)were analyzed by using Pearson analysis.Results(1)Compared with the NGT group,the abundance of Lactobacillus genus was significantly lower in the new-onset T2 DM group(t=8.557,P=0.000).There was no significant difference in the abundance of Bacteroides thetaiotaomicron between two groups(t=0.524,P=0.601).(2)The abundances of Lactobacillus genus in the two group were negatively correlated with the level of FBG(r=-0.334,P=0.000),waist circumference(r=-0.170,P=0.018),TC(r=-0.178,P=0.013),TG(r=-0.157,P=0.030)and SBP(r=-0.255,P=0.000).Conclusion The decreased abundance of Lactobacillus genus may be related to the pathogenesis of new-onset T2 DM,the mechanism of which needs to be further studied.
Objective To analyze quantitatively the Lactobacillus genus and Bacteroides thetaiotaomicron in the gut microbiota of Xinjiang Uyghurs with new-onset T2 DM.Methods 96 cases of new-onset T2 DM and98 cases of normal glucose tolerance(NGT)were selected according to inclusion criteria.The total DNAs were extracted from the fecal microbiota,and the amounts of Lactobacillus genus and Bacteroides thetaiotaomicronin the gut microbiota were determined with 16 SrDNA gene Real-time fluorescence quantitative PCR amplification.The correlations of Lactobacillus genus and Bacteroides thetaiotaomicron with BMI,waist circumference,hip circumference,systolic blood pressure(SBP),diastolic blood pressure(DBP),fasting blood glucose(FBG),triglycerides(TG),total cholesterol(TC),high-density lipoprotein(HDL-C)and low-density lipoprotein(LDL-C)were analyzed by using Pearson analysis.Results(1)Compared with the NGT group,the abundance of Lactobacillus genus was significantly lower in the new-onset T2 DM group(t=8.557,P=0.000).There was no significant difference in the abundance of Bacteroides thetaiotaomicron between two groups(t=0.524,P=0.601).(2)The abundances of Lactobacillus genus in the two group were negatively correlated with the level of FBG(r=-0.334,P=0.000),waist circumference(r=-0.170,P=0.018),TC(r=-0.178,P=0.013),TG(r=-0.157,P=0.030)and SBP(r=-0.255,P=0.000).Conclusion The decreased abundance of Lactobacillus genus may be related to the pathogenesis of new-onset T2 DM,the mechanism of which needs to be further studied.
引文
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[19]LU Jia,FANG Xiucai.Mechanism of Lactobacillus in the treatment of irritable bowel syndrome[J].Clin J Gastroenterol Hepatol,2017,26(2):218-221.(in Chinese)陆佳,方秀才.乳杆菌属治疗肠易激综合征的机制[J].胃肠病学和肝病学杂志,2017,26(2):218-221.
[20]Wang G,Li X,Zhao J,et al.Lactobacillus casei CCFM419attenuates type 2diabetes via a gut microbiota dependent mechanism[J].Food Funct,2017,8(9):3155-3164.
[21]TANG Ying.Influence of Bacteroides thetaiotaomicron on blood glucose in rats and relationship between hyperglycemia and atherosclerosis disease in diabetic rats[D].Nanchang:Medical College of Nanchang University,2014.(in Chinese)唐英.多形拟杆菌对大鼠血糖的影响及糖尿病大鼠诱发动脉粥样硬化的实验研究[D].南昌:南昌大学医学院,2014.
[22]Ravcheev DA,Godzik A,Osterman AL,et al.Polysaccharides utilization in human gut bacterium Bacteroides thetaiotaomicron:Comparative genomics reconstruction of metabolic and regulatory networks[J].BMC Genomics,2013,14:873.
[23]Wrzosek L,Miquel S,Noordine ML,et al.Bacteroides thetaiotaomicron and Faecalibacterium prausnitzii influence the production of mucus glycans and the development of goblet cells in the colonic epithelium of a gnotobiotic model rodent[J].BMC Biol,2013,11:61.
[24]YI Jinyang.Correlation between Bacteroides thetaiotaomicron,Methanobrebibacter smithii and glucagon-like peptide-1in intestinal in type 2diabetes mellitus db/db mice[D].Urumqi:Xinjiang Medical University,2012.(in Chinese)易金阳.db/db 2型糖尿病模型小鼠肠道多形拟杆菌,产甲烷短杆菌与GLP-1的相关性研究[D].乌鲁木齐:新疆医科大学,2012.
[25]XIE Jingwen,WANG Ye,ZHU Ming,et al.The effects of Bacteroides thetaiotaomicron on diabetic mouse models[J].Chin J Microecol,2013,25(8):869-881.(in Chinese)谢婧雯,王烨,朱明,等.多形拟杆菌对糖尿病模型小鼠的影响[J].中国微生态学杂志,2013,25(8):869-881.
[26]Peterson DA,Planer JD,Guruge JL,et al.Characterizing the interactions between a naturally primed immunoglobulin A and its conserved Bacteroides thetaiotaomicron species-specific epitope in gnotobiotic mice[J].J Biol Chem,2015,290(20):12630-12649.
[2]Semenkovich CF,Danska J,Darsow T,et al.American diabetes association and JDRF research symposium:Diabetes and the microbiome[J].Diabetes,2015,64(12):3967-3977.
[3]CENG Yipeng,HU Linghong,FENG Xinge.Review of specific biomarker of gut microbiota with type 2diabetes[J].Int JLab Med,2016,7(15):2142-2144.(in Chinese)曾艺鹏,胡凌红,冯新格.2型糖尿病患者肠道菌群特异性生物标记物研究进展[J].国际检验医学杂志,2016,7(15):2142-2144.
[4]Pollak M.The effects of metformin on gut microbiota and the immune system as research frontiers[J].Diabetologia,2017,60(9):1662-1667.
[5]Forslund K,Hildebrand F,Nielsen T,et al.Disentangling type 2diabetes and metformin treatment signatures in the human gut microbiota[J].Nature,2015,528(7581):262-266.
[6]Byun R,Nadkarni MA,Chhour KL,et al.Quantitative analysis of diverse Lactobacillus species present in advanced dental caries[J].J Clin Microbiol,2004,42(7):3128-3136.
[7]YI Jinyang,WANG Ye,ZHU Ming,et al.Comparison of standard sample preparation methods with real time fluorescence quantitative PCR of Bacteroides thetaiotaomicronin intestinal from db/db type 2diabetic mellitus mice[J].J Xinjiang Med Univ,2012,35(6):716-720.(in Chinese)易金阳,王烨,朱明,等.db/db2型糖尿病模型小鼠肠道多形拟杆菌实时荧光定量PCR标准品制备方法的比较研究[J].新疆医科大学学报,2012,35(6):716-720.
[8]Blandino G,Inturri R,Lazzara F,et al.Impact of gut microbiota on diabetes mellitus[J].Diabetes Metab,2016,42(5):303-315.
[9]Goldstein EJ,Tyrrell KL,Citron DM.Lactobacillus species:taxonomic complexity and controversial susceptibilities[J].Clin Infect Dis,2015,15(60Suppl 2):S98-107.
[10]Yakovlieva M,Tacheva T,Mihaylova S,et al.Influence of Lactobacillus brevis 15 and Lactobacillus plantarum 13 on blood glucose and body weight in rats after high-fructose diet[J].Benef Microbes,2015,6(4):505-512.
[11]HU Fang,WEI Tingting,DU Caihe,et al.Variation analysis of bacterial communities in esophagus of BKS-DB mice[J].Chin JBiochem&Mol Biol,2011,27(10):972-979.(in Chinese)胡芳,魏婷婷,杜彩贺,等.2型糖尿病小鼠食道菌群的结构分析[J].中国生物化学与分子生物学报,2011,27(10):972-979.
[12]Ahn HY,Kim M,Ahn YT,et al.The triglyceride-lowering effect of supplementation with dual probiotic strains,Lactobacillus curvatus HY7601and Lactobacillus plantarum KY1032:Reduction of fasting plasma lysophosphatidylcholines in nondiabetic and hypertriglyceridemic subjects[J].Nutr Metab Cardiovasc Dis,2015,25(8):724-33.
[13]Wu Y,Zhang Q,Ren Y,et al.Effect of probiotic Lactobacillus on lipid profile:A systematic review and meta-analysis of randomized,controlled trials[J].PLoS One,2017,12(6):e0178868.
[14]Naito E,Yoshida Y,Kunihiro S,et al.Effect of Lactobacillus casei strain Shirota-fermented milk on metabolic abnormalities in obese prediabetic Japanese men:A randomised,doubleblind,placebo-controlled trial[J].Biosci Microbiota Food Health,2018,37(1):9-18.
[15]LIU Tingting,CAI Dehong.Real-time quantitative PCR analysis of intestinal Lactobacillus species in type 2diabetic patients[J].World Chin J Digestol,2012,20(15):1359-1365.(in Chinese)刘婷婷,蔡德鸿.实时定量PCR法分析2型糖尿病患者肠道乳酸杆菌菌种数量的改变[J].世界华人消化杂志,2012,20(15):1359-1365.
[16]Hove KD,Brons C,Faerch K,et al.Effects of 12weeks of treatment with fermented milk on blood pressure,glucose metabolism and markers of cardiovascular risk in patients with type 2diabetes:A randomised double-blind placebo-controlled study[J].Eur J Endocrinol,2015,172(1):11-20.
[17]Ivey KL,Hodgson JM,Kerr DA,et al.The effect of yoghurt and its probiotics on blood pressure and serum lipid profile:Arandomised controlled trial[J].Nutr Metab Cardiovasc Dis,2015,25(1):46-51.
[18]Crovesy L,Ostrowski M,Ferreira DMTP,et al.Effect of Lactobacillus on body weight and body fat in overweight subjects:A systematic review of randomized controlled clinical trials[J].Int J Obes(Lond),2017,41(11):1607-1614.
[19]LU Jia,FANG Xiucai.Mechanism of Lactobacillus in the treatment of irritable bowel syndrome[J].Clin J Gastroenterol Hepatol,2017,26(2):218-221.(in Chinese)陆佳,方秀才.乳杆菌属治疗肠易激综合征的机制[J].胃肠病学和肝病学杂志,2017,26(2):218-221.
[20]Wang G,Li X,Zhao J,et al.Lactobacillus casei CCFM419attenuates type 2diabetes via a gut microbiota dependent mechanism[J].Food Funct,2017,8(9):3155-3164.
[21]TANG Ying.Influence of Bacteroides thetaiotaomicron on blood glucose in rats and relationship between hyperglycemia and atherosclerosis disease in diabetic rats[D].Nanchang:Medical College of Nanchang University,2014.(in Chinese)唐英.多形拟杆菌对大鼠血糖的影响及糖尿病大鼠诱发动脉粥样硬化的实验研究[D].南昌:南昌大学医学院,2014.
[22]Ravcheev DA,Godzik A,Osterman AL,et al.Polysaccharides utilization in human gut bacterium Bacteroides thetaiotaomicron:Comparative genomics reconstruction of metabolic and regulatory networks[J].BMC Genomics,2013,14:873.
[23]Wrzosek L,Miquel S,Noordine ML,et al.Bacteroides thetaiotaomicron and Faecalibacterium prausnitzii influence the production of mucus glycans and the development of goblet cells in the colonic epithelium of a gnotobiotic model rodent[J].BMC Biol,2013,11:61.
[24]YI Jinyang.Correlation between Bacteroides thetaiotaomicron,Methanobrebibacter smithii and glucagon-like peptide-1in intestinal in type 2diabetes mellitus db/db mice[D].Urumqi:Xinjiang Medical University,2012.(in Chinese)易金阳.db/db 2型糖尿病模型小鼠肠道多形拟杆菌,产甲烷短杆菌与GLP-1的相关性研究[D].乌鲁木齐:新疆医科大学,2012.
[25]XIE Jingwen,WANG Ye,ZHU Ming,et al.The effects of Bacteroides thetaiotaomicron on diabetic mouse models[J].Chin J Microecol,2013,25(8):869-881.(in Chinese)谢婧雯,王烨,朱明,等.多形拟杆菌对糖尿病模型小鼠的影响[J].中国微生态学杂志,2013,25(8):869-881.
[26]Peterson DA,Planer JD,Guruge JL,et al.Characterizing the interactions between a naturally primed immunoglobulin A and its conserved Bacteroides thetaiotaomicron species-specific epitope in gnotobiotic mice[J].J Biol Chem,2015,290(20):12630-12649.