新疆哈萨克族肥胖儿童菌群人源化动物模型的建立  

Establishment of Human Flora- associated Animal Model for Obese Children of Xinjiang Kazakh Nationality

作　　者：刘新彩[1] 徐佩茹[1,2] 李敏 艾比白.艾尔肯[1] 王红清 

机构地区：[1]新疆医科大学第一附属医院儿科,新疆乌鲁木齐市830054 [2]新疆维吾尔自治区循证医学研究所

出　　处：《中国全科医学》2016年第8期936-940,共5页Chinese General Practice

基　　金：国家自然科学基金资助项目(81460165);新疆维吾尔自治区自然科学基金资助项目(2014211C056)

摘　　要：目的利用无菌小鼠建立新疆哈萨克族肥胖儿童菌群人源化（HFA）动物模型,为深入研究哈萨克族儿童肥胖的病理生理机制提供新的研究方式。方法收集新疆哈萨克族7-13岁肥胖儿童新鲜粪便,男女各10例。20只3周龄无菌级C57BL/6J小鼠（雌雄各半）,按照性别进行分组并编号。小鼠灌胃接种哈萨克族肥胖儿童粪便菌悬液0.1 ml/次,隔日一次,共接种3次,且接种时遵循性别一致原则。于接种后第7天收集小鼠新鲜粪便标本,提取基因组DNA后,采用特异性引物对16S rRNA V3区进行PCR扩增,对扩增产物运用DGGE方法分离并鉴定,以了解HFA动物模型中肠道菌群的定植情况。结果接种哈萨克族肥胖儿童肠道菌群后的无菌小鼠盲肠均减小为正常小鼠大小;HFA动物模型的PCR-DGGE指纹图谱显示各观察标本均形成复杂而多样的菌属条带。结论本实验成功利用无菌C57BL/6J小鼠建立哈萨克族肥胖儿童HFA动物模型,该动物模型是探索哈萨克族儿童特征性肥胖与肠道菌群关系的新的选择方式。Objective To establish human flora associated（ HFA） animal models related with Kazakh obese children in Xinjiang based on germ- free（ GF） mice and to provide a new way for the further study of the physiopathologic mechanism of obese children of Kazakh nationality. Methods We collected fresh feces from 20 Xinjiang Kazakh obese children who were 7-13 years old; and each gender 10 cases. We selected 20 three- week- old GF C57 BL /6J mice（ half male and half female） and assigned them into two groups and numbered them. The mice received gavage inoculation of bacterium suspension once every two days,given three times totally and each time 0. 1 ml; the principle of gender consistency was followed. On the 7th day after inoculation,fresh feces samples were collected, and genome and DNA were extracted; specific primers were employed to conduct PCR amplification on V3 region of 16 S rRNA; DGGE method was adopted to separate and identify amplification products,in order to understand the engraftment of intestinal flora in HFA animal models. Results The cecums of HFA mice were reduced to the normal size after inoculation; PCR-DGGE fingerprint of HFA mice showed that complex and diverse bacterial genus stripes formed in each sample. Conclusion The establishment of HFA animals related with Kazakh obese children based on GF C57 BL /6J mice was successful. HFA animal model is a new method for the study of the relationship between Kazakh obese children and gut microbiota.

关 键 词：肥胖症 儿童 哈萨克族 模型 动物 菌群人源化动物 无菌小鼠 

分 类 号：R723.14[医药卫生—儿科]