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首页> 外文期刊>Brain research. Brain research protocols >Analysis of gene expression of aldose reductase and sorbitol dehydrogenase in rat Schwann cells by competitive RT-PCR method using non-homologous DNA standards.
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Analysis of gene expression of aldose reductase and sorbitol dehydrogenase in rat Schwann cells by competitive RT-PCR method using non-homologous DNA standards.

机译：使用非同源DNA标准品通过竞争性RT-PCR方法分析大鼠雪旺细胞中醛糖还原酶和山梨糖醇脱氢酶的基因表达。

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摘要

Aldose reductase (AR) and sorbitol dehydrogenase (SDH) are the enzymes constituting the polyol pathway, an alternate route of glucose metabolism. A wealth of experimental data has indicated the involvement of the polyol pathway in the pathogenesis of diabetic complications. However, there has been surprisingly little research on the relative abundance of SDH to AR in the tissues affected in diabetes. We therefore developed a competitive RT-PCR system to simultaneously determine the mRNA levels of these two enzymes in small amounts of samples, and studied their expression in Schwann cells isolated from adult rat sciatic nerves. Although both AR and SDH mRNA were expressed in the Schwann cells, the levels of SDH cDNA were much lower than those of AR cDNA. The induction of AR mRNA expression in the Schwann cells under hyperosmotic conditions was similarly detected by Northern blot analysis and our competitive RT-PCR method. The RT-PCR system developed in this study may be a useful tool in ascertaining the relative contributions of AR and SDH to the metabolic derangements resulting from the acceleration of polyol pathway activity in the target organ of diabetic complications.
机译：醛糖还原酶（AR）和山梨糖醇脱氢酶（SDH）是构成多元醇途径（葡萄糖代谢的另一种途径）的酶。大量的实验数据表明，多元醇途径参与了糖尿病并发症的发病机制。然而，在受糖尿病影响的组织中，关于SDH与AR相对丰度的研究很少，令人惊讶。因此，我们开发了一种竞争性RT-PCR系统，可同时测定少量样品中这两种酶的mRNA水平，并研究了它们在从成年大鼠坐骨神经中分离出的雪旺细胞中的表达。尽管AR和SDH mRNA均在雪旺氏细胞中表达，但SDH cDNA的水平远低于AR cDNA。通过Northern印迹分析和我们的竞争性RT-PCR方法相似地检测了高渗条件下雪旺细胞中AR mRNA表达的诱导。在这项研究中开发的RT-PCR系统可能是确定AR和SDH对由糖尿病并发症目标器官中多元醇途径活性加速导致的代谢紊乱的相对贡献的有用工具。

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