影响实验小鼠肠道菌群的多因素比较研究

目的比较研究影响实验小鼠肠道菌群的六种主要因素,为动物实验的设计和实验结果的解释提供参考,为肠道微生态菌群结构的干预与调控提供借鉴。方法采用珠磨法和酚-氯仿-异戊醇法提取肠道内容物中细菌基因组DNA,然后用16S rDNA V3区通用引物进行PCR扩增,扩增产物经DGGE电泳后用电泳条带数字化软件Quantity One和统计学软件SPSS对影响肠道菌群的六种因素(年龄、品种、饲养环境、饲料、个体差异和性别差异)进行比较研究。结果 Dice相关系数和主成分分析显示,年龄因素在早期(7和14日龄)对肠道菌群影响较大,到28日龄后,菌群结构趋向稳定,饲料和饲养环境成为主要的影响因素。品种和个体差异对肠道菌群也存在较大的影响,但性别差异影响相对较小,存在一定的随机性。结论以上六种因素均能在一定程度上影响肠道菌群,但其影响程度存在差异,在动物实验设计和实验结果解释时应充分考虑避免或利用这些影响因素。

Comparative Study for Factors that Affect Microbiota Colonization in Experimental Mice

In order to better understand the relationship among different affect factors and provide better instructions for regulating the gut microbiota,polymerase chain reaction-denaturing gradient gel electrophoresis（PCR-DGGE） was selected to investigate the effects of the six factors on gut microbiota.The Dice coefficience and principal component analysis（PCA） results were used to evaluate their effects.The results showed that age,raise environment,diet,mouse strains,individual variation and gender can affect the gut bacteria community.And samples can cluster together according to each factor except gender.However,their affect ability is different.Age is the most important factor at the early stage,and after day 14,raise environment and diet become more important.Although,the variation caused by individual is not obvious,the similar Dice coefficience is only 50~70%.These results showed that at the early stage,the effects of age and raise environment are more important and the colonized bacteria can affect the following colonization.Along with the increased of age,diet was fed to mouse,and diet become the most important factor.And diet has the ability to change the history colonized bacteria community.These results give us some reminds that we should consider these factors when design experiments and analyze results.More importantly,the results will help and lead us to know how to regulate the gut microbiota efficiently.