基于16SrDNA高通量测序技术对肾移植术前后肠道菌群变化的研究  

Preliminary study of gut microbiota changes before and after kidney transplantation based on 16S

作　　者：李钢 贺腾辉 罗用文 唐倩[2] 解俊杰 李超 药晨 孙忠蔚 李昆 柏宏伟 王洪阳 陈荣[4] 刘双[5] 刘兆宇 肖征[5] 张晓[5] 石炳毅 钱叶勇 宋阳[4,5] Li Gang;He Tenghui;Luo Yongwen;Tang Qian;Xie Junjie;Li Chao;Yao Chen;Sun Zhongwei;Li Kun;Bai Hongwei;Wang Hongyang;Chert Rong;Liu Shuang;Liu Zhaoyu;Xiao Zheng;Zhang Xiao;Shi Bingyi;Qian Yeyong;Song Yang(Institute of Organ Transplantation of Chinese PLA,No.309 Hospital of PLA and Beijing Key Laboratory of Organ Transplantation and Immunoregulation,Beijing 100091,China;Department of Clinical Laboratory, No.309 Hospital of PLA,Beijing 100091,China;Department of Kidney Transplantation,Affiliated Hospital of Qingdao University,Qingdao 266071,China;Department of Clinical Laboratory,PLA General Hospital,Beijing 100853,China;Department of Clinical Laboratory,Hainan Branch of PLA General Hospital,Sanya 572013,China)

机构地区：[1]解放军第三0九医院全军器官移植研究所暨北京市器官移植与免疫调节重点实验室,北京100091 [2]解放军第三0九医院检验科,北京100091 [3]青岛大学附属医院肾移植科,266071 [4]解放军总医院检验医学中心,北京100853 [5]解放军总医院海南分院检验科,三亚572013

出　　处：《中华实验外科杂志》2019年第1期150-153,共4页Chinese Journal of Experimental Surgery

基　　金：海南省医药卫生科研项目(14A210195);海南省三亚市医疗卫生科技创新项目(2014YW42);中国博士后科学基金面上项目(2015M582828);中国人体器官捐献专项研究课题(CODMF2017011).

摘　　要：目的探讨肾移植患者手术前、后肠道菌群的变化。方法根据纳入排除标准选取在解放军第309医院全军器官移植研究所行肾移植手术的7例患者作为研究对象,收集术前3 d内及术后第21天共计14份粪便标本,提取标本中原核生物全基因组后采用16S rDNA高通量测序方法分析菌群结构。结果在门分类水平上,肾移植术后较术前,变形菌门相对丰度显著升高(P<0.01),厚壁菌门相对丰度下降(P>0.05);在纲、目、科、属分类水平上,相对丰度增加的菌属包括隶属于厚壁菌门的芽孢杆菌纲(P<0.05)及其所属的乳杆菌目(P<0.05)和肉杆菌科、颗粒链菌属;减少的菌属包括隶属于厚壁菌门梭菌纲梭菌目的毛螺旋菌科、胃瘤球菌科、真细菌属、粪球菌属以及隶属于放线菌门的红蝽菌目。术后组Shannon指数较术前呈降低趋势(P>0.05,Wilcoxon符号秩检验)。主坐标分析(PCoA)显示术前组的各标本间距离小于术后组各标本间距离,且术后组各标本在坐标中呈散在分布。结论肾移植术后肠道菌群的改变主要包括:多样性较术前呈减少趋势;变形菌门相对丰度升高,厚壁菌门相对丰度降低。由于肠道菌群结构具有高度的个体差异,不太可能以某种特定细菌类别作为统一适用于移植术后并发症的诊断标志物。但是,纵向来看,每个移植患者肠道菌群变化所遵循的规律有可能预示移植术后患者的健康或疾病状态,并指导移植术后的治疗策略。Objective To investigate the changes of intestinal microflora before and after kidney transplantation.Methods According to the inclusion and exclusion criteria, 7 patients who had undergone kidney transplantation in the 309th Hospital of PLA were selected as study subjects. A total of 14 stool specimens were collected within 3 days before and 21 days after the operation. Prokaryotic genome was analyzed by 16S rDNA high-flux sequencing after extraction from specimens.Results In the classification level of Phylum, Proteobacteria relative abundance increased significantly (P<0.01) and Firmicutes relative abundance decreased (P>0.05) after operation as compared with preoperation. In the classification level of class, order, family and genus, the relative abundance of Bacillus (P<0.05), Lactobacillales (P<0.05), Carnobacteriaceae and Granulicatella increased. The relative abundance of Lachnospiraceae, Ruminococcaceae, Eubacterium, Coprococcus and Coriobacteriales decreased. The Shannon index in postoperative group was significantly lower than that in preoperative group (P>0.05, Wilcoxon rank sum test). The principal coordinate analysis (PCoA) showed that the distance among the specimens in the preoperative group was shorter than that in the postoperative group, and the specimens in postoperative group were scattered in the coordinates.Conclusion The changes of gut microbiota after kidney transplantation mainly includes reduced diversity, increased relative abundance of Proteobacteria, and decreased relative abundance of Firmicutes. Because of high degree of individual variation in gut microbiota, it is unlikely that a specific bacterial category could be used as a diagnostic marker for complications after transplantation. However, in the vertical view, the pattern of changes in intestinal flora in each transplant patient may predict the health or disease status after transplantation, and even guide the treatment strategy.

关 键 词：肠道菌群 肾移植 16S RDNA 

分 类 号：R699.2[医药卫生—泌尿科学]