从新疆哈萨克族2型糖尿病患者粪便中分离的气单胞菌对小鼠的作用研究

目的从新疆哈萨克族2型糖尿病患者粪便中分离培养气单胞菌,并回接到小鼠体内,检测其对小鼠血糖、血脂等2型糖尿病相关中间表型的作用。方法 (1)收集新鲜哈萨克族2型糖尿病患者的粪便样本,用气单胞菌专属培养基进行分离培养,微量生化反应管进行初步鉴定;提取所得菌株的DNA,使用气单胞菌属引物对此DNA进行特异性扩增,结合全自动细菌鉴定仪对菌株进行鉴定;(2)将气单胞菌菌液回接小鼠,采用羰花青素染料(CM-Dil)标记法对菌定植力情况进行检测。(3)检测气单胞菌菌液对小鼠体重、随机血糖(GLU)、空腹血糖(FBG)、总胆固醇(TC)和甘油三酯(TG)的作用,进行葡萄糖耐量(OGTT)、胰岛素耐量(ITT)试验。结果 (1)所得到的菌株经全自动细菌鉴定仪确定为气单胞菌;(2)从小鼠粪便中分离培养后的细菌能检测到荧光标记;(3)与正常饮食组小鼠相比,气单胞菌稀释液、原液、浓缩液组小鼠体重均升高;气单胞菌原液组小鼠GLU升高。小鼠FBG、TC和TG在各组间差异均无统计学意义;气单胞菌菌液组小鼠口服葡萄糖耐量试验血糖曲线下面积增加;气单胞菌原液、浓缩液组小鼠胰岛素耐量试验血糖曲线下面积增加。结论 (1)从新疆哈萨克族2型糖尿病患者粪便中分离培养得到气单胞菌;(2)从糖尿病患者粪便中分离的气单胞菌可定植于小鼠体内,并使小鼠的体重增加,GLU升高,给予气单胞菌稀释液、菌原液、菌浓缩液后第8周,小鼠出现糖耐量受损,胰岛素敏感性下降。

Influence of Aeromonas spp. isolated from the feces of Xinjiang Kazakhs T2DM patient on mice

Abstract: Objective To isolate and culture Aeromonas spp. from feces of Xinjiang Kazakhs with type 2 diabetes mellitus, and explore its influence on the related phenotypes of type 2 diabetes mellitus such as blood glucose and blood lipid in mice. Methods (1) Fresh fecal samples of Kazakhs with type 2 diabetes mellitus were collected to isolate Aeromonas spp. by using specific media; micro-biochemical reaction were adopted for preliminary identification; specific primer was used to amplify the DNA of the strains; and fully automated bacterial identification system was used for strain indentification. (2) Aeromonas spp. was inoculated into the mice, and the colonization force was detected by CM-Dil labeling method. (3) The influence of Aeromonas spp. on the body weight, glucose (GLU), fasting blood glucose (FBG), total cholesterol (TC) and triglyceride (TG) of the mice were determined. Oral glucose tolerance test (OGTT) and insulin Tolerance test (ITT) were conducted. Results (1) The isolated strain was identified as Aeromonas spp. by fully automated bacterial identification system. (2) Fluorescent markers were detected in the bacteria isolated from mouse feces. (3) Compared with the normal diet group, the body weights of mice treated with diluted solution, stock solution and concentrated solution of Aeromonas spp. increased; the random blood sugar levels of mice in the stock solution group increased. No significant differences were seen in fasting blood glucose, total cholesterol and triglyceride among groups. The AUCs of glucose in OGTT increased; the AUCs of glucose in ITT increased in the stock solution group and concentration group. Conclusion Aeromonas spp. isolated from fecal samples of Kazakhs with type 2 diabetes mellitus.(2) The colonization was successful and increased the weight of mice, GLU increased, at 8 weeks, the Aeromonas dilution, bacterial stock solution, bacterial concentration of impaired glucose tolerance, ITT decreased insulin sensitivity.