MicroRNA及其靶基因在糖尿病肾病小鼠肾脏的表达

 验证与糖尿病肾病小鼠肾脏相关的microRNAs的表达并运用实时荧光定量PCR分析靶基因与糖尿病肾病的关系。以db/db小鼠为模型组（DN组），db/m小鼠为正常组（NC组），定期测量小鼠的体重、血糖、甘油三酯、总胆固醇及24 h尿蛋白排泄率。留取DN小鼠与NC小鼠肾脏组织，检测肾脏组织形态学染色及实时荧光定量PCR（qRT-PCR）。qRT-PCR验证差异表达的microRNAs及其靶基因的mRNA表达水平。血糖、24 h尿蛋白排泄率结果表明糖尿病肾病动物模型构建成功。与NC小鼠相比，DN小鼠肾脏miR-196a、miR-21、miR-200b表达明显升高，且差异有统计学意义（P<0.05）。miR-196a、miR-200b、miR-21的表达水平与血糖、甘油三酯、总胆固醇、24 h尿蛋白排泄率存在正相关关系（P<0.05）。利用miRNAs数据库预测miR-196a的靶基因有ANX1、HOXB7、PTEN、FOXO1、HOXB8、HOXA5等。与NC组比较，DN组ANX1、FOXO1的mRNA表达水平降低，且差异有统计学意义（P<0.05）。同时ANX1、FOXO1与24 h尿蛋白排泄率存在正相关（P<0.05）。MiR-196a可能通过调节ANX1、FOXO1的表达水平来参与糖尿病肾病的发生发展。

Expression of microRNA and target gene in kidney of diabetic nephropathy mice

In this paper, the expression of microRNAs in diabetic nephropathy is verified and the relationship between the target genes and the diabetic nephropathy is analyzed by qRT-PCR. With db/db mice as the model group (DN group) and db/m mice as normal group (NC group), we measure the weight, the blood glucose, the triglyceride, the total cholesterol and the 24 h urinary albumin excretion rate in every mice. The renal tissue from the kidney of the DN and NC mice is taken for pathological examination and qRT-PCR. The difference of the expression levels of microRNAs is confirmed by qRT-PCR, and then the mRNA expression of the target genes is verified. The DN animal model is successfully constructed according to the blood glucose and the 24 h urinary albumin excretion rate results. The expression levels of miR-196a, miR-21 and miR-200b are significantly increased in the DN mice compared with the NC mice, and the difference is statistically significant(P<0.05).The expression levels of miR-196a, miR-200b and miR-21 are positively correlated with the level of the blood glucose, the triglyceride, the total cholesterol and the 24h urinary protein excretion rate. The microRNAs database is used to predict the target genes of miR-196a, including ANX1, HOXB7, PTEN, FOXO1, HOXB8 and HOXA5. The expression levels of the ANX1 and the FOXO1 are significantly lower in the DN mice compared with the NC group, and the difference is statistically significant(P<0.05). There is a positive correlation between the ANX1, the FOXO1 and the 24 h urinary protein excretion rate. It is concluded that miR-196a may be involved in the development of the diabetic nephropathy to regulate the expressions of the ANX1 and the FOXO1.