UCP2^(+/+)和UCP2^(-/-)小鼠的鉴定及表型初步分析

Identification and elementary phenotype analysis of UCP^(2+/+) and UCP^(2-/-) mice

目的对解耦联蛋白2(UCP2)转基因(UCP2^(+/+))和UCP2敲除(UCP2^(-/-))小鼠进行鉴定及表型初步分析。方法采用PCR法鉴定UCP2^(+/+)和野生型(C57BL/6J)小鼠UCP2基因的表达。采用Western blot法检测UCP2^(+/+)、UCP2^(-/-)和C57BL/6J小鼠肝脏UCP2蛋白的表达。绘制0~12周龄UCP2^(+/+)、UCP2^(-/-)和C57BL/6J小鼠生长曲线,并肉眼观察UCP2^(+/+)和UCP2^(-/-)小鼠在外观和行为上有无异常。分别采用高脂饮食和普通饮食饲喂UCP2^(+/+)、UCP2^(-/-)和C57BL/6J小鼠12周,检测其空腹血糖和胰岛素水平。结果UCP2^(+/+)小鼠UCP2基因片段为280bp,且肝脏表达相对分子量约为32kD的UCP2蛋白。与C57BL/6J小鼠相比,UCP2^(+/+)小鼠肝脏UCP2表达量升高(P<0.05),而UCP2^(-/-)小鼠肝脏未见UCP2蛋白表达。UCP2^(+/+)和UCP2^(-/-)小鼠与C57BL/6J小鼠相比,在外观和行为上未见异常。与C57BL/6J小鼠相比,饲喂普通饲料对UCP2^(+/+)和UCP2^(-/-)小鼠空腹血糖和胰岛素水平影响差异无统计学意义(P>0.05)。与饲喂普通饲料相比,高脂饲料能升高C57BL/6J和UCP2^(+/+)小鼠空腹血糖水平(P<0.05),提高UCP2^(-/-)小鼠胰岛素的分泌(P<0.05)。结论成功鉴定了小鼠UCP2^(+/+)和UCP2^(-/-)基因型;高脂饮食下,UCP2与小鼠血糖水平和胰岛素分泌相关。

Objective To identify UCP2（＋/＋） and UCP2（-/-） mice and analyze elementarily the phenotypes.Methods The genotypes of mice were identified by PCR and the liver expression of UCP2 protein was identified by Western blot.The growth curves of UCP2（＋/＋） ,UCP2（-/-） and C57BL/6J mice were drawn in the age of 0-12 weeks and the appearance and behavior of the mice with UCP2（＋/＋） and UCP2（-/-） were observed.The blood glucose and insulin levels of the mice with UCP2（＋/＋） ,UCP2（-/-） and C57BL/6Jwere measured after feeding with high-fat diet or normal diet for 12 weeks.Results The UCP2 gene fragment of UCP2（＋/＋） mice was 280 bp and the UCP2 protein had a molecular weight of 32kD.Compared with C57BL/6Jmice,the expression of UCP2in liver tissues of UCP2（＋/＋） mice was increased（P〈0.05） ,but there was no UCP2protein expression in liver tissues of UCP2（-/-） mice.The mice with UCP2（＋/＋） and UCP2（-/-） had no abnormality in the appearance and behavior compared with C57BL/6J mice.Compared with C57BL/6J mice,there was no significant difference in fasting blood glucose and insulin between UCP2（＋/＋） and UCP2（-/-） mice fed with normal diet（P〈0.05） .Compared with normal diet feeding,high-fat diet feeding increased the fasting blood glucose of UCP2（＋/＋） and C57BL/6 Jmice and insulin secretion of UCP2（-/-） mice（P〈0.05） .Conclusion The phenotypes of UCP2（＋/＋） and UCP2（-/-） mice have been successfully identified.UCP2 is associated with blood glucose level and insulin secretion in the mice fed with high-fat diet.