【作者】 孙玉霞； 赵善良； 刘加洪； 张健稳； 张超； 马爱国；

【Author】 Yuxia Sun;Shanliang Zhao;Jiahong Liu;Jianwen Zhang;Chao Zhang;Aiguo Ma;School of Public Health/Institute of Nutrition and Health,Qingdao University;Linyi People’s Hospital of Shandong Province;Lung Disease Institute of Qingdao Central Hospital;

【机构】 青岛大学公共卫生学院/营养与健康研究院； 临沂市人民医院； 青岛市中心医院肺病研究所；

【摘要】 分析强化四周抗结核药物治疗对结核病患者肠道微生态的影响,收集青岛市某医院10名强化四周抗结核治疗结核病患者及10名健康对照组的粪便标本,提取细菌基因组DNA,PCR扩增16SrDNA的V3～V4区特定序列,用Illumina HiSeq平台测序。对序列进行操作分类单元(operational taxo-nomic unit,OTU)聚类和物种注释;通过Alpha多样性反映单个样品物种丰度及物种多样性;通过主坐标分析(principal co-ordinates analysis,PCoA)观察两组之间菌群结构的差异程度;通过比较门和属水平各细菌的相对丰度来描述菌群结构的差异并进一步采用线性判别分析效应量(line discriminant a-nalysis effect size,LEfSe)找出两组的差异菌。采用MannWhitney U检验对两组微生物多样性指数进行比较。四周抗结核治疗的Alpha多样性指标Chao1指数和Ace指数分别为(140.13±34.73)和(145.18±30.74),显著低于对照组的(196.56±37.12)和(197.62±31.76)(分别U=13.000,P=0.004;U=11.000,P=0.002);抗结核治疗组的Shannon指数为(2.31±0.57),与对照组(2.89±0.46)相比略有降低,但差异未见统计学意义(U=30.000,P=0.143)。基于binary jaccard的PCoA分析显示两组整体菌群结构有显著差异。抗结核治疗组与对照组相比肠道菌群组成发生改变,其中厚壁菌门、毛螺菌科的相对丰度明显低于对照组(46.4%vs 72.4%,χ~2=5.491,P=0.019;72.4%vs 43.1%,χ~2=5.851,P=0.016);而拟杆菌门、拟杆菌属的丰度明显高于对照组(39.0%vs18.8%,χ~2=4.166,P=0.041;31.4%vs 10.3%,χ~2=7.406,P=0.007)。强化四周抗结核药物治疗可使肠道菌群多样性下降,肠道菌群结构失衡;肠道毛螺旋菌科等丰度减少,拟杆菌属等丰度增加。更多还原

【Abstract】 To analyze the effect of four week intensive anti-tuberculosis drug treatment on intestinal microecology of tuberculosis patients.Methods We collected stool samples from 10 patients with four week intensive anti-tuberculosis treatment and 10 healthy controls in a hospital in Qingdao city.The total bacterial genomic DNA was extracted,and theV3-V4 hy-pervariable region in 16 S RDNA gene sequence was amplified by PCR.High throughput sequencing was performed on Illumina Miseq platform.The sequences were clustered into Operational Taxonomic Unit(OTU) and annotated with species.Alpha diversity was analyzed to describe richness and species diversity of the samples.The difference of bacterial flora structure between the two groups was observed by principal coordinate analysis.The difference of microbial community structure was described by comparing the relative abundance of species at phylum level and genus level.In addition,the bacteria with significant difference between the two groups were identified by line discriminant analysis effect size.The microbial diversity index of the two groups was compared by Mann Whitney U test.Results Chaol index and Ace index of Alpha diversity in the four week anti-tuberculosis treatmentgroup were(140.13±34.73) and(145.18±30.74)respectively,which were significantly lower than those of the control group((196.56±37.12),(197.62±31.76);U=13.000,P=0.004;U=11.000,P=0.002).The Shannon index of the anti-tuberculosis treatment group was(2.31±0.57),which was slightly lower than that of the control group(2.89±0.46),but no statistically significant difference was observed(U=30.000,P=0.143).PCoA analysis based on binary jaccard showed significant differences in the overall flora structure between the two groups.The composition of intestinal flora changed in the four-week anti-tuberculosis treatment group compared with the control group.The relative abundance of Frimicutes and Lachnospiraceae was significantly lower than that in the control group(460.4% vs 72.4%,χ~2=5.491,P=0.019;72.4% vs 43.1%,χ~2=5.851,P=0.016),while the relative abundance of Bacteroidetes and Bacteroides was significantly higher than that in the control group(39.0%vs 18.8%,χ~2=4.16,P=0.041;31.4% vs 10.3%,χ~2=7.406,P=0.007).Conclusions The diversity of intestinal flora was decreased and the structure of intestinal flora was obviously unbalanced with four week intensive anti-tuberculosis drug therapy.The abundance of Lachnospiraceae was decreased,while genus Bacteroides was increased.更多还原