db/db小鼠糖尿病肾病相关基因的分析和克隆

Analyzing and Cloning of Diabetic Nephropathy Relative Gene From db/db Mice

用GM U74A基因芯片分别检测了正常对照组 (db/m小鼠 )、糖尿病肾病组 (db/db小鼠 )、大黄酸治疗组 (大黄酸 15 0mg/kg治疗 12周 )肾脏基因表达谱 .发现在 12 43 7个基因 (包括表达序列标签 )中 ,与正常对照组相比 ,糖尿病肾病组有 10 85个基因表达下调 ,3 7个基因表达上调 ,其中变化幅度大于 2倍 ,表达下调的有 166个和表达上调的有 2 9个 .与糖尿病肾病组相比 ,大黄酸治疗组有 3 84个基因表达下调 ,15 5个表达上调 ,其中变化幅度大于 2倍 ,表达下调的有 47个和表达上调的有 3 0个 .在此基础上 ,对其中的一个差异表达的表达序列标签 (EST)进行了详细的生物信息学分析 ,发现它是一个未知功能基因———“REKENcDNA0 610 0 0 6H10”基因的一部分 .在用RT PCR进一步验证了其与糖尿病肾病的相关性后 ,对“REKENcDNA0 610 0 0 6H10”

Eight weeks old db/db mice were treated with or without rhein at a dosage of 150 mg/kg for 12 weeks. Kidney's total RNA was extracted and renal gene expression profiles were tested using the gene chip GM-U74A from Affymetrix company. Among the total 12 437 tested genes, 1 085 genes were down-regulated and 37 genes were up-regulated in untreated db/db mice when compared with normal control. Among them, 166 genes were down-regulated and 29 genes up-regulated more than 2 fold. It was found that 384 genes were down-regulated and 155 genes up-regulated in rhein treated db/db mice as compared with untreated db/db mice. About 47 genes were down-regulated and 30 genes up-regulated more than 2 fold. Then, an EST, which was down-regulated by about 2 fold in db/db mice and recover to normal level in rhein treated mice, was further analyzed by bioinformatical method and proved to be part of 'REKEN cDNA 0610006H10' gene, which function is unknown. After its expression level was further proved by RT-PCR, diabetic nephropathy relative gene, 'REKEN cDNA 0610006H10' gene, was cloned for further study.