基于16S rRNA基因测序技术对成人隐匿性自身免疫糖尿病患者肠道菌群特征的分析  

Analysis characterization of gut microbiome in latent autoimmune diabetes in adults based on 16S rRNA gene sequencing

作　　者：潘念思 刘净文 孙衍[2] 金苗苗[2] 刘金霞[3] 申一凡 杨石美 黄林青 牛晓红[2] Pan Niansi;Liu Jingwen;Sun Yan;Jin Miaomiao;Liu Jinxia;Shen Yifan;Yang Shimei;Huang Linqing;Niu Xiaohong(Graduate School of Changzhi Medical College,Changzhi 046000,China;Department of Endocrinology,Heji Hospital Affiliated to Changzhi Medical College,Changzhi 046000,China;Department of Medical Clinical Laboratory,Heji Hospital Affiliated to Changzhi Medical College,Changzhi 046000,China.)

机构地区：[1]长治医学院研究生院,长治046000 [2]长治医学院附属和济医院内分泌科,长治046000 [3]长治医学院附属和济医院医学检验科,长治046000

出　　处：《中华糖尿病杂志》2022年第8期809-816,共8页CHINESE JOURNAL OF DIABETES MELLITUS

基　　金：中南大学糖尿病免疫学教育部重点实验室开放课题研究项目(502401004);山西省“四个一批”科技兴医创新计划项目(2020XM25)。

摘　　要：目的基于16S rRNA基因测序技术分析成人隐匿性自身免疫糖尿病(LADA)患者肠道菌群的特征。方法本研究为病例对照研究。选取2019年9月到2020年10月就诊于山西省长治医学院附属和济医院内分泌科的LADA及2型糖尿病(T2DM)患者作为研究对象,并从该院体检中心同期体检的人群中选取健康人群作为正常对照(NCP)者。收集研究对象谷氨酸脱羧酶抗体(GADA)、蛋白酪氨酸磷酸酶2抗体(IA-2A)等,收集研究对象粪便16S rRNA基因序列。使用微生物组分析软件QIIME(version 1.8.0)分析反映肠道菌群丰富性和多样性的alpha多样性指标和反映组间结构相似性的Beta多样性指标,采用t检验或单因素方差分析、Mann‐Whitney U检验或Kruskal-Wallis H检验、χ^(2)检验进行组间比较,基于PICRUST分析微生物功能基因和代谢预测途径的变化特征,采用Spearman相关性分析法分析肠道菌群与各生化及免疫指标的相关性。结果共纳入42例研究对象,NCP组14名,T2DM组14例,LADA组14例。3组间的alpha多样性指数差异均无统计学意义(P>0.05),Beta多样性差异有统计学意义(P<0.05)。与NCP组相比,LADA组的短链脂肪酸产生菌属如毛螺菌属(Lac_Lachnospira)、罗氏菌属(Lac_Roseburia)、瘤胃球菌属(毛螺菌科)[Lac(Ruminococcus)]均降低(P<0.05),与T2DM组相比,LADA组的瘤胃球菌属(毛螺菌科)降低(P<0.05)。与NCP组相比,LADA组的苯丙氨酸合成、初级胆汁酸生物合成、次级胆汁酸生物合成等代谢预测功能途径降低(P<0.05)。Spearman相关性分析结果显示,GADA抗体滴度与瘤胃球菌属(毛螺菌科)呈负相关(r=-0.44),与普雷沃菌属(r=0.38)、小类杆菌属(r=0.34)、瘤胃球菌属(瘤胃球菌科)(r=0.53)均呈正相关(均P<0.05);IA-2A抗体滴度与巨球菌属呈正相关(r=0.13,P<0.05)。结论LADA患者存在较为特异的肠道微生物群结构,并伴随着短链脂肪酸产生菌属如毛螺菌属、罗氏菌属和瘤胃球菌属(毛螺菌科)的Objective To explore the characterization of gut microbiota in latent autoimmune diabetes in adults(LADA)based on 16S rRNA gene sequencing.Methods This was a case-control study.Patients with LADA and type 2 diabetes mellitus(T2DM)were selected from the Department of Endocrinology,Heji Hospital Affiliated to Changzhi Medical College from September 2019 to October 2020,and healthy people were selected as normal control people(NCP)from the physical examination center of this hospital.The data of glutamic acid decarboxylase antibody(GADA),protein tyrosine phosphatase 2 antibody(IA-2A),etc.,and the 16S rRNA gene sequence of the gut microbiota were collected.The analysis software QIIME(version 1.8.0)was used to analyze the alpha diversity index reflecting the diversity and richness of gut microbiota and the Beta diversity index reflecting the structural similarity between groups.The t-test or analysis of one-way variance,Mann-Whitney U test,Kruskal-Wallis H test or χ^(2) test were adopted for comparison among the groups.The changes of microbial functions were predicted based on the PICRUST.Spearman correlation was used to analyze the correlation between gut microbiota and biochemical data.Results A total of 42 subjects were included,including 14 in NCP group,14 in T2DM group and 14 in LADA group.There was no significant difference in alpha diversity index(P<0.05),but there was significant difference in Beta diversity(P<0.05)among the three groups of gut microbiota.Compared with the NCP group,the short-chain fatty acid producing bacteria such as Lachnospira,Roseburia and Lac_(Ruminococcus)genera significantly decreased(P<0.05),and the Lac_(Ruminococcus)genera significantly decreased in LADA group compared with the T2DM as well(P<0.05).Compared with NCP group,the metabolic prediction function pathways such as phenylalanine synthesis,primary bile acid biosynthesis and secondary bile acid biosynthesis in LADA group decreased(P<0.05).Spearman correlation analysis showed that the GADA antibody titer was negatively correlate

关 键 词：糖尿病 2型 成人隐匿性自身免疫糖尿病 肠道菌群 

分 类 号：R587.1[医药卫生—内分泌]