Dillmann W H, Mehta H B, Barrieux A, Guth B D, Neeley W E, Ross J
Circ Res. 1986 Jul;59(1):110-4. doi: 10.1161/01.res.59.1.110.
Recent evidence indicates that different forms of stress, including hypoxia, can induce specific proteins called heat-shock or stress proteins in various types of mammalian cells. These studies examined whether myocardial ischemia can result in increased levels of proteins with molecular weight and isoelectric point characteristics similar to those described for heat-shock or stress proteins. The left anterior descending coronary artery of the dog heart was completely occluded; normal and ischemic myocardial samples were obtained 6 hours after occlusion; and total cardiac proteins and RNA were prepared. Ribonucleic acid was translated in vitro in a modified rabbit reticulocyte lysate system, and [35S]-methionine-labelled translational products as well as unlabelled cardiac proteins were separated by two-dimensional gel electrophoresis. Total proteins were visualized by silver staining and in vitro translation products quantified by fluorometry. A translatable mRNA coding for a 71,000 dalton peptide with an isoelectric point of 5.8 was markedly increased in the ischemic myocardium after 6 hours of ischemia. A protein with similar migration characteristics was detected in ischemic myocardium but not in normal myocardium. These results indicate that an mRNA coding for a translational product with similar migration characteristics of heat-shock protein 71 is induced by ischemia in the dog heart.
最近的证据表明,包括缺氧在内的不同形式的应激可在各种类型的哺乳动物细胞中诱导产生称为热休克蛋白或应激蛋白的特定蛋白质。这些研究探讨了心肌缺血是否会导致分子量和等电点特征与热休克蛋白或应激蛋白相似的蛋白质水平升高。将狗心脏的左冠状动脉前降支完全闭塞;闭塞6小时后获取正常和缺血心肌样本;并制备总心脏蛋白和RNA。在改良的兔网织红细胞裂解物系统中进行RNA体外翻译,并用二维凝胶电泳分离[35S]-甲硫氨酸标记的翻译产物以及未标记的心脏蛋白。通过银染使总蛋白可视化,并用荧光法对体外翻译产物进行定量。缺血6小时后,缺血心肌中编码一种等电点为5.8、分子量为71,000道尔顿的肽的可翻译mRNA显著增加。在缺血心肌中检测到一种具有相似迁移特征的蛋白质,但在正常心肌中未检测到。这些结果表明,狗心脏缺血可诱导编码具有与热休克蛋白71相似迁移特征的翻译产物的mRNA。