抗生素联合粪菌移植构建人源化小鼠的可行性研究

Feasibility study on construction of humanized mice with antibiotics combined with fecal bacteria transplantation

目的:选择一种广泛使用的人源化小鼠构建方法,对其可行性进行验证,为后续粪菌移植研究提供理论依据。方法:用抗生素连续喂养SPF级小鼠3周,期间记录小鼠一般情况,3周后通过革兰氏染色评估小鼠粪便微生物的量,并观察小鼠肠道形态学变化。给予抗生素处理后小鼠移植粪菌1周,之后采用RT-PCR测定粪菌移植供体及受体粪便微生物的含量,评价粪便微生物定植效果。结果:抗生素处理的小鼠每日饮用水(P=0.0003)及饲料消耗量(P=0.0012)均明显减少;且抗生素处理可以明显消耗小鼠原始肠道微生物。粪菌移植后,接受1号人粪菌的小鼠粪菌中厚壁菌门、拟杆菌门及二者比例分别占接受2号人粪菌小鼠的15.4%、112.0%和13.7%;而1号人粪菌中厚壁菌门、拟杆菌门及二者比例分别占2号人的33.7%、104.0%和32.0%,受体的肠道微生物与人类供体趋势相同。结论:抗生素处理联合粪菌移植使SPF级小鼠人源化可行且安全。

Objective:To select a common method of humanized mice and verify its feasibility with the aim to provide a theoretical basis for subsequent fecal bacteria transplantation research.Methods:Specific pathogen free(SPF)mice were fed with antibiotics mixed with sterilized drinking water for 3 weeks,during which the general situation of mice was recorded.After 3 weeks,the number of microbes in mice's feces was evaluated by Gram staining.After antibiotic treatment,mice were transplanted fecal bacteria by gavage for a week.Then,the contents of fecal bacteria in the donor and recipient feces were determined by RT-PCR,to evaluate the colonization effect of fecal bacteria.Results:Antibiotic treatment had an effect on weight,feed and drinking water consumption of mice,and the fecal bacteria in mice decreased obviously.After fecal bacteria transplantation,the Firmicutes,Bacteroidetes and Firmicutes/Bacteroidetes of the mice receiving Donor1 fecal bacteria accounted for 15.4%,112.0%and 13.7%of the mice receiving Donor2 fecal bacteria,respectively.The Firmicutes,Bacteroidetes and Firmicutes/Bacteroidetes of Donor1 fecal bacteria accounted for 33.7%,104.0%and 32.0%of Donor2,respectively.The recipient's gut microbes had the same trend as human donors.Conclusion:These results indicate that antibiotic treatment combined with fecal bacterial transplantation is feasible and safe for humanizing SPF mice.