Maekawa K, Tanimoto T, Okada S, Suzuki T, Suzuki T, Yabe-Nishimura C
Division of Drug Evaluation, National Institute of Health Sciences, 1-1-43 Hoenzaka, Chuo-ku, Osaka 540-0006, Japan.
Brain Res Mol Brain Res. 2001 Mar 5;87(2):251-6. doi: 10.1016/s0169-328x(01)00009-2.
To investigate the polyol pathway activity in Schwann cells, we determined the mRNA levels of aldose reductase (AR) and sorbitol dehydrogenase (SDH) in cultured cells under hyperglycemic or hyperosmotic conditions using competitive RT-PCR technique. The expressions of AR and SDH mRNAs in Schwann cells were unaltered by high (30 mM) glucose content in the medium. On the other hand, osmotic stress elicited significant increases in AR mRNA without any effect on SDH mRNA expression. The levels of AR mRNA determined by this RT-PCR system were significantly correlated with AR activity, as well as the levels of sorbitol accumulated in Schwann cells cultured under hyperosmotic conditions. These findings suggest that in contrast to the induction of AR expression by osmotic stress, high glucose per se does not up-regulate expression of the enzymes constituting the polyol pathway in Schwann cells. 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 leading to diabetic complications.
为研究雪旺细胞中的多元醇途径活性,我们使用竞争性逆转录聚合酶链反应(RT-PCR)技术,测定了在高血糖或高渗条件下培养细胞中醛糖还原酶(AR)和山梨醇脱氢酶(SDH)的mRNA水平。培养基中高浓度(30 mM)葡萄糖含量并未改变雪旺细胞中AR和SDH mRNA的表达。另一方面,渗透压应激导致AR mRNA显著增加,而对SDH mRNA表达无任何影响。通过该RT-PCR系统测定的AR mRNA水平与AR活性以及在高渗条件下培养的雪旺细胞中积累的山梨醇水平显著相关。这些发现表明,与渗透压应激诱导AR表达相反,高糖本身并不会上调雪旺细胞中构成多元醇途径的酶的表达。本研究中开发的RT-PCR系统可能是确定AR和SDH对导致糖尿病并发症的代谢紊乱相对贡献的有用工具。