脊髓法猴体神经毒力试验对恒河猴肠道菌群的影响研究

摘要

目的 探讨脊髓注射法猴体神经毒力试验对肠道菌群的影响.方法 用Ⅲ型脊髓灰质炎减毒活疫苗进行脊髓法猴体神经毒力试验,收集3只神经毒力试验恒河猴及3只健康恒河猴粪便样本,提取细菌总DNA,采用新一代高通量测序技术对16S rRNA基因的V3～V4高变区测序并进行物种分类、丰度及多样性分析.结果 神经毒力试验对动物脊髓造成不同程度的病理损伤;试验组粪便微生物多样性降低,门水平上两组菌群无差异,优势菌群主要为硬壁菌门、变形菌门及拟杆菌门,占比总和超过95％;属水平上有普雷沃菌属(Prevotella)、芽殖菌属(Gemmiger)等11种菌属存在显著差异(P＜0.05).结论 脊髓注射法猴体神经毒力试验对动物造成脊髓损伤并引起粪便内乳酸菌属(Lactobacillus)、普雷沃菌属(Prevotella)及芽殖菌属(Gemmiger)等菌属组成变化,为建立非人灵长类动物脊髓损伤模型提供理论依据.%Objective To investigate the effect of spinal cord injection on the intestinal microflora in rhesus monkeys.Methods The monkeys were injected with type Ⅲ live attenuated poliomyelitis vaccine into intraspinal.Fecal samples from 3 rhesus monkeys of neurovirulence test and 3 healthy rhesus monkeys were collected.Total bacterial DNAs were extracted and submitted high throughout gequencing on the V3-V4 viable region of 16S rRNA.Tags and operational taxonomic units (OTU) were then obtained and analysis of taxonomy,abundance and alpha diversity were performed.Results The pathological changes of various degree of spinal cord injuries caused by the intraspinal neurovirulence test.Microbial diversity decreased in experimental group.There was no difference between the two groups on the phylum level,and the dominant microbial community in fecal was Firmicutes,Proteobacteria and Bacteroidetes,accounting for more than 95％ of the total;Prevotella,Gemmiger and other 11 kinds of bacteria had significant difference on the genus level (P ＜ 0.05).Conclusion Neurovirulence test by in intraspinal injection caused spinal cord injury and caused composition changes of Lactobacillus,Prevotella and Gemmiger,The purpose of this study was to provide theoretical basis for the establishment of non-human primate spinal cord injury model.