Shi S, Unakar N J, Wen Y, Tsui J, Bekhor I
Laboratory for Molecular Genetics, Doheny Eye Institute, Los Angeles, California.
Mol Cell Biochem. 1992 Sep 22;115(1):27-34. doi: 10.1007/BF00229092.
Aldose reductase (AR), a major enzyme in the polyol pathway, is thought to be responsible for accumulation of polyols in lenses exposed to high doses of galactose or glucose, and it may be linked to some of the complications found in diabetes. In this report we examined the level of expression of AR mRNA in lens epithelia undergoing development of galactose cataracts in vivo. The AR mRNA was quantitated by Northern blot hybridization with a [35S]-RNA transcript from a previously described AR cDNA clone. This was done on normal lens epithelia and on epithelia from lens of rats fed a diet of Purina Chow containing 50% galactose for periods of from 6 hr to 20 days. We found AR mRNA to elevate to about 5-fold the control levels by 12-24 hr on galactose, then decrease to the control levels by day 4. The increase in AR mRNA appears to be transitory. The high abundance in AR mRNA by 24 hr on galactose was confirmed by in situ hybridization. At later periods, from 8 to 20 days on galactose, a slow increase in AR mRNA took effect, as we have previously reported. Changes in the levels of galactose and dulcitol between 0 and 96 hr were also quantitated by gas chromatography, showing that there was a significant increase in both galactose and dulcitol occurring throughout the experimental period.(ABSTRACT TRUNCATED AT 250 WORDS)
醛糖还原酶(AR)是多元醇途径中的一种主要酶,被认为是导致暴露于高剂量半乳糖或葡萄糖的晶状体中多元醇积累的原因,并且它可能与糖尿病中发现的一些并发症有关。在本报告中,我们检测了体内正在发生半乳糖性白内障的晶状体上皮细胞中AR mRNA的表达水平。通过与先前描述的AR cDNA克隆的[35S]-RNA转录本进行Northern印迹杂交来定量AR mRNA。对正常晶状体上皮细胞以及喂食含50%半乳糖的普瑞纳饲料6小时至20天的大鼠晶状体上皮细胞进行了此项检测。我们发现,在半乳糖作用下,AR mRNA在12 - 24小时内升高至对照水平的约5倍,然后在第4天降至对照水平。AR mRNA的增加似乎是短暂的。半乳糖作用24小时时AR mRNA的高丰度通过原位杂交得到证实。如我们之前所报道,在半乳糖作用后期,即8至20天,AR mRNA缓慢增加。还通过气相色谱法定量了0至96小时之间半乳糖和木糖醇水平的变化,结果表明在整个实验期间半乳糖和木糖醇均显著增加。(摘要截短至250字)