Lomas-Soria Consuelo, Ramos-Gómez Minerva, Guevara-Olvera Lorenzo, Guevara-González Ramón, Torres-Pacheco Irineo, Gallegos-Corona Marco A, Reynoso-Camacho Rosalía
Research and Graduate Studies in Food Science, School of Chemistry, University of Querétaro, Cerro de las Campanas, S/N, Querétaro, Qro., 76010 Mexico.
Int J Mol Sci. 2011;12(12):8431-48. doi: 10.3390/ijms12128431. Epub 2011 Nov 29.
Diabetic nephropathy (DN) is a major complication of diabetes and is caused by an imbalance in the expression of certain genes that activate or inhibit vital cellular functions of kidney. Despite several recent advances, the pathogenesis of DN remains far from clear, suggesting the need to carry out studies identifying molecular aspects, such as gene expression, that could play a key role in the development of DN. There are several techniques to analyze transcriptome in living organisms. In this study, the suppression subtractive hybridization (SSH) method was used to generate up- and down-regulated subtracted cDNA libraries in the kidney of streptozotocin (STZ)-induced diabetic rats. Northern-blot analysis was used to confirm differential expression ratios from the obtained SSH clones to identify genes related to DN. 400 unique SSH clones were randomly chosen from the two subtraction libraries (200 of each) and verified as differentially expressed. According to blast screening and functional annotation, 20.2% and 20.9% of genes were related to metabolism proteins, 9% and 3.6% to transporters and channels, 16% and 6.3% to transcription factors, 19% and 17.2% to hypothetical proteins, and finally 24.1 and 17.2% to unknown genes, from the down- and up-regulated libraries, respectively. The down- and up-regulated cDNA libraries differentially expressed in the kidney of STZ diabetic rats have been successfully constructed and some identified genes could be highly important in DN.
糖尿病肾病（DN）是糖尿病的主要并发症，由激活或抑制肾脏重要细胞功能的某些基因表达失衡所致。尽管近年来取得了一些进展，但DN的发病机制仍远未明确，这表明需要开展研究来确定可能在DN发生发展中起关键作用的分子层面因素，如基因表达。有多种技术可用于分析生物体中的转录组。在本研究中，采用抑制性消减杂交（SSH）方法构建链脲佐菌素（STZ）诱导的糖尿病大鼠肾脏中上调和下调的消减cDNA文库。利用Northern印迹分析来确认从获得的SSH克隆中得到的差异表达率，以鉴定与DN相关的基因。从两个消减文库（各200个）中随机选取400个独特的SSH克隆，并验证其差异表达。根据Blast筛选和功能注释，下调文库和上调文库中分别有20.2%和20.9%的基因与代谢蛋白相关，9%和3.6%与转运蛋白和通道相关，16%和6.3%与转录因子相关，19%和17.2%与假定蛋白相关，最后分别有24.1%和17.2%与未知基因相关。已成功构建了在STZ糖尿病大鼠肾脏中差异表达的下调和上调cDNA文库，一些鉴定出的基因可能在DN中具有高度重要性。
