通过粪菌移植建立慢性阻塞性肺疾病肠道菌群研究模型及其效果评价  

Establishment of COPD gut microbiota model with fecal microbiota transplantation and its evaluation

作　　者：李乃健[1] 戴周丽 陈炽勇 张佳欢 何芳 李靖[1] 周玉民[1] 李冰[2] 冉丕鑫[1] LI Naijian;DAI Zhouli;CHEN Chiyong;ZHANG Jiahuan;HE Fang;LI Jing;ZHOU Yumin;LI Bing;RAN Pixin(The First Affiliated Hospital of Guangzhou Medical University,State Key Laboratory of Respiratory Disease,National Center for Respiratory Medicine,Guangzhou,Guangdong 510120,P.R.China;The GMU-GIBH Joint School of Life Sciences,Guangzhou Medical University,Guangzhou,Guangdong 510120,P.R.China;Basic Medical School,Guangzhou Medical University,Guangzhou,Guangdong 510120,P.R.China)

机构地区：[1]广州医科大学附属第一医院,呼吸疾病国家重点实验室,国家呼吸医学中心,广东广州510120 [2]广州医科大学,中科学院广州生物医药与健康研究院联合生命科学学院,广东广州510120 [3]广州医科大学基础医学院,广东广州510120 [4]丽水学院医学与健康学院,浙江丽水323000

出　　处：《中国呼吸与危重监护杂志》2021年第7期465-471,共7页Chinese Journal of Respiratory and Critical Care Medicine

基　　金：国家自然科学基金(81900030、81970045);广东省“珠江人才计划”(2017BT01S155);广东省医学科研基金(A2018272);呼吸疾病国家重点实验室课题(SKLRD-QN-201908、SKLRD-OP-2020040)。

摘　　要：目的通过粪菌移植(FMT)方法在小鼠上建立慢性阻塞性肺疾病(简称慢阻肺)的肠道菌群研究模型并对其移植效果进行初步评价。方法入组健康对照、慢阻肺Ⅰ-Ⅱ级和慢阻肺Ⅲ-Ⅳ级患者各30例并收集其粪便制作粪菌混悬液,通过灌胃方法对小鼠进行FMT。移植后第29天采集全血行单个核细胞分析,对小鼠移植前后的肠道菌群进行16S rRNA测序分析,对模型小鼠进行初步疾病模型评价。结果抗生素预处理后小鼠肠道的OTU、Chao 1和Shannon指数均显著下降(P<0.001),接受健康对照和慢阻肺患者FMT后的受体小鼠其OTU、Chao 1和Shannon指数显著升高(P<0.01或P<0.05)。健康对照FMT组、慢阻肺Ⅰ-ⅡFMT组和慢阻肺Ⅲ-ⅣFMT组小鼠菌群丰度在厚壁菌门、变形杆菌门和放线门中均与磷酸盐缓冲液FMT组小鼠存在显著性差异(P<0.05或P<0.01)。慢阻肺Ⅰ-ⅡFMT组和慢阻肺Ⅲ-ⅣFMT组小鼠外周血CD3+、CD3+CD4+和CD3+CD8+细胞百分比升高(P<0.05或P<0.01),而CD3–CD19+细胞百分比下降(P<0.05)。结论通过FMT可在小鼠上成功建立慢阻肺肠道菌群研究模型。Objective To establisht a gut microbiota mice model for chronic obstructive pulmonary disease(COPD) with fecal microbiota transplantation(FMT) and its evaluation.Methods The mice received FMT from healthy individuals,COPD Ⅰ-Ⅱ subjects,or COPD Ⅲ–Ⅳ subjects.After microbiota depletion,the FMT was performed by a single oral administration of 100 μL per mouse every other day,for a total of 14 times in 28 days.On the 29 th day,the peripheral blood mononuclear cells were analyzed,the gut microbiota of mice before and after FMT was analyzed by 16 S rRNA sequencing,and the mice model were evaluated.Results The operational taxonomic units,Chao 1 and Shannon indexes of mice all decreased significantly after antibiotic treatment(P<0.001),but increased significantly after FMT from healthy individuals,COPD Ⅰ-Ⅱ subjects,or COPD Ⅲ–Ⅳ subjects(P<0.05 or P<0.01).The abundance of Firmicutes,Proteobacteria and Actinobacteria in the guts of the mice in the healthy human FMT group,COPD Ⅰ-Ⅱ FMT group and COPD Ⅲ-Ⅳ FMT group were significantly different from those of the control group who only received phosphate buffer saline instead of FMT(P<0.05 or P<0.01).The auxiliary T lymphocytes and cytotoxic T lymphocytes were higher,but B lymphocytes decreased in the peripheral blood of the mice in the COPD Ⅰ-Ⅱ FMT group and COPD Ⅲ-Ⅳ FMT group(P<0.05 or P<0.01).Conclusion FMT can successfully establish a COPD gut microbiota research model.

关 键 词：慢性阻塞性肺疾病 粪菌移植 肠道菌群 小鼠 

分 类 号：R563.9[医药卫生—呼吸系统]