高通量测序分析老年血液透析患者肠道菌群的改变
Evaluation of Intestinal Microflora by High-throughput Gene Sequencing in Elderly HD Patients
胡建广 1钟小仕 1闫静 1覃丹平 1郑媛媛 1谭荣韶 1刘岩1
作者信息
- 1. 广东省广州市红十字会医院/暨南大学医学院附属广州红十字会医院肾内科,广州临床病态营养研究所 广州 510220
- 折叠
摘要
目的:了解老年血液透析(hemodialysis,HD)患者肠道菌群分布和丰度的变化规律.方法:采集51例HD患者作为疾病(Case)组,53例血肌酐正常的健康志愿者作为对照(Control)组;采集研究对象晨起新鲜粪便样品,抽提其中微生物总基因组DNA,采用Ⅱ代基因测序技术对粪便样品16S rDNA V3高变区进行扩增测序;划分可操作分类单元(operational taxonomic unit,OTU),使用QIIME分析平台对所有OTU进行物种注释、多样性分析、主坐标分析及肠型分析;采用PICRUSt法预测肠道微生物基因功能及其在KEGG通路上的丰度差异,运用R Studio软件对数据进行统计学分析.结果:两组样品检测出肠道菌群分属14个门和252个属;厚壁菌门和拟杆菌门在Case组和Control组均为最优势菌门,丰度和分别为88.50(±13)%和93.55(±8.8)%,差异无统计学意义;Case组厚壁菌门的丰度下降显著(P<0.01),拟杆菌门的丰度升高显著(P<0.01),厚壁/拟杆比值明显减低;Case组有27个菌属的丰度与Control组相比差异有统计学意义(P<0.01),分别属于厚壁菌门(21个)、拟杆菌门(3个)以及变形菌门(3个),其中10个菌属的丰度升高,以拟杆菌属升高最为显著,17个菌属的丰度降低,以柔嫩梭菌、罗斯菌属、巨单胞菌属等肠道益生菌减少为主;Case组肠型分析为拟杆菌肠型,三大营养物质分解代谢相关的基因表达明显增强,与Control组的柔嫩梭菌肠型差异有统计学意义(P<0.01).结论:老年HD患者肠道菌群呈门结构组成和菌属丰度紊乱,提示血肌酐升高和血液透析可能影响肠道菌群的生存环境,主要表现为厚壁菌门细菌减少,拟杆菌门细菌适应性繁殖,引起拟杆菌属丰度增加以及糖代谢、脂肪代谢和氨基酸代谢相关的基因表达增强,机体能量缺乏超出肠道菌群的代偿能力导致能量代谢失衡,最终导致PEW(protein-energy wasting,PEW)的发生、发展.
Abstract
Objective:To investigate distribution and abundance of intestinal flora in elderly HD patients. Methods:51 eld-erly patients receiving maintenance hemodialysis as case group and 53 healthy volunteers with normal serum creatinine as control group were recruited in this study;Total microbial genomic DNA was extracted from fresh fecal samples which were collected in both groups early morning;V3 hypervariable region of 16S rDNA has been amplified and sequenced by II-generation gene sequencing technolo-gy;All OTUs have been noted and analyzed using QIIME after OUT was classified. We have predicted intestinal microbial gene func-tion and its abundance difference on KEGG pathway with PICRUSt method. All statistical analysis were performed using R Studio a-nalysis software. Results:Intestinal flora detected in both groups belong to 14 phylum and 252 genera; Firmicutes and Bacteroides were the most predominant bacteria phylum in both groups, sum of abundance were 88. 50 ( ± 13)% and 93. 55 ( ± 8. 8)%, but the difference was not statistically significant;Firmicutes decreased and Bacteroides significantly increased in case group, F/B significant-ly reduced (P<0. 01);At genus level, we found that there were 27 significant differences in case group compared with control group (P<0. 01), respectively belonging to Firmicutes(21), Bacteroides(3) and Proteobacteria(3);Relative abundance of 10 genera has increased, especially Bacteroides; Relative abundance of 17 genera has decreased, such as Faecallbacterium, Loseburia, Mega-monas, and et al;Three major nutrients catabolic gene expression was significantly enhanced in case group with Bacteroides-domina-ted, compared to the control group with Faecallbacterium-dominated(P<0. 01). Conclusion:The pioneering work of this study has showed disorders in structural composition and species abundance in elderly HD patients, suggesting that elevated serum creatinine and hemodialysis may affect the living environment of intestinal microflora, which leads to reduction of Firmicutes and reproduction of Bacteroides, and gene expressions in glucose metabolism, fat metabolism and amino acid metabolism have been enhanced. But the lack of energy beyond compensatory ability of intestinal flora has contributed to occurrence and development of energy metabolic imbal-ance and PEW ( protein-energy wasting, PEW) .
关键词
血液透析/肠道菌群/蛋白质-能量消耗/16SrDNAKey words
Hemodialysis/Microbial flora/Protein-Energy/Wasting/16S rDNA引用本文复制引用
基金项目
广东省省级科技计划项目(2016A020215023)
广州市临床医学研究与转化中心基金资助项目(2060404)
出版年
2017
NETL
NSTL
维普
万方数据