【摘要】 目的观察腹腔注射纳米氧化锌的小鼠肠道菌群种类变化。方法将6只健康雄性昆明小鼠随机分为实验组和对照组,实验组给予氧化锌纳米溶液(0. 5 g/kg体质量),对照组给予PBS溶液,均每天1次,连续腹腔注射14 d,注射结束3 d后处死小鼠,解剖肠道,获得粪便标本,并提取DNA,扩增含V4 16S r DNA高变区的片段,并用MiSeq测序仪对PCR产物进行测序,进行物种丰度差异分析,然后进行Alpha、Beta多样性分析。结果 6个粪便样本中共分析出肠道菌群17门、26纲、42目、64科、115属;实验组和对照组分别存在2 936、2 981个操作分类单元(OTU),共享OTU的百分比为19. 95%;实验组厚壁菌门、拟杆菌门、变形杆菌门、柔膜菌门所占百分比分别为39%、33%、25%、3%,对照组分别为68%、19%、11%、1%;实验组小鼠肠道幽门螺杆菌属、颤杆菌属、另枝菌属所占百分比分别为46%、5%、22%,对照组分别为21%、15%、35%。物种丰度差异分析示,与对照组相比,实验组中异常球菌-栖热菌门、拟杆菌属增多,臭味杆菌属和颤杆菌属减少(P均<0. 05)。Alpha多样性分析示,与对照组比较,实验组多样性指数均下降; Beta多样性分析示PC1变化率65. 97%,PC2变化率17. 30%。结论纳米氧化锌腹腔注射后小鼠肠道菌群各种类占比在门、属水平均发生了变化,异常球菌-栖热菌门、拟杆菌属增多,臭味杆菌属和颤杆菌属减少;肠道菌群多样性减少,不均一性增加。更多还原

【Abstract】 Objective To observe the changes in the intestinal flora species in mice after intraperitoneal injection of Zinc oxide nanoparticles( Zn O NPs). Methods Six healthy male Kunming mice were randomly divided into the control group and experimental group. Mice in the control group were peritoneally injected with 0. 2 m L phosphate buffered saline( PBS),once a day,for 14 days,and experimental group with the same volume of Zn O NPs solution. Mice were sacrificed three days after drug intervention,and then we dissected gut and extracted DNA from the fecal samples. The fragments encompassing V4 16 S r DNA hypervariable regions were amplified from the fecal samples,and the PCR products of V4 were sequenced by Illumina MiSeq for species richness difference analysis,and Alpha diversity analysis and Beta diversity analysis. Results A total of 17 phyla,26 classes,42 orders,64 families,115 genera were analyzed from the 6 stool samples. Sequencing results showed 2 981 and 2 936 operational taxonomic unit( OTU) in the control group and experimental group,respectively,and the percentage of shared OTU was 19. 95%. The percentages of Firmicutes,Bacteroides,Proteobacteria,and Tenericutes were 39%,33%,25%,and 3%,respectively,in the experimental group,versus 68%,19%,11%,and 1%,respectively,in the control group. At the genus level,the percentages of Helicobacter,Oscillibacter,and Alistipes were 46%,5%,and 22% in the experimental group,and 21%,15%,and 35%,respectively in the control group. Compared with the control group,the experimental group had a lower diversity index and a higher number of Deinococcus-Thermus( at the phylum level),Bacteroides( at the genus level)( P < 0. 05),a lower number of species of Odoribacter and Oscillibacter( at the genus level)( P < 0. 05). Alpha diversity analysis showed that the diversity index of the experimental group decreased as compared with that of the control group; Beta diversity analysis showed that the percentage of PC1 was approximately 65. 97%,and the percentage of PC2 was approximately 17. 30%. Conclusions After intraperitoneal injection of Zn O NPs,the proportion of various types of intestinal flora in mice has changed at the phylum and genus levels; Deinococcus-Thermus and Bacteroides increase,Odoribacter and Oscillibacter decrease,and the diversity of intestinal flora in mice decreases,and the heterogeneity increases.更多还原