热休克对心肌细胞中αB-晶状体蛋白移位及表达的影响

摘要

The effects of heat shock pretreatment on translocation andexpression of αB-crystallin were evaluated in neonatal rat cardiomyocytes. Results: ① Western blot analysis demonstrated that cytosolic soluble αB-crystallin rapidly translocated to insoluble intracellular structure at 10 or 30 minutes and returned to cytosolic soluble compartment at about 1～2h after heat shock. ② Double staining of immunofluorescence showed that αB-crystallin redistributed in cytosol after heat shock, while HSC70 translocated from cytosol to nucleus. ③ Quantitative analysis with immunoelectronic microscopy demonstrated αB-crystallin translocated to cytoskeletal structure such as Z-line and nuclear envelope after heat shock. ④ Northern-blot showed that heat shock induced αB-crystallin mRNA expression. The rapid translocation and increasing expression of αB-crystallin after heat shock suggests that this small heat shock protein may play an important protective roles during the early and late phases of myocardial ischemic preconditioning.%采用热休克对新生大鼠心肌细胞进行处理，观察αB-晶状体蛋白在细胞内的分布及其表达变化。结果：①Western-blot显示热休克后胞浆中可溶性αB-晶状体蛋白迅速移位至细胞内的不溶性结构中，2h左右基本复位；②免疫双荧光细胞化学显示热休克导致αB-晶状体蛋白在细胞浆内发生分布变化，而HSC70从胞浆向胞核移位；③免疫电镜定量分析进一步证实αB-晶状体蛋白在热休克时主要移向Z线及核膜等细胞骨架部位；④Northern-blot显示热休克诱导心肌细胞αB-晶状体蛋白mRNA的表达增加。热休克处理使αB-晶状体蛋白迅速移位至细胞骨架，表明其在心肌预适应早期相中可能发挥保护作用。而αB-晶状体蛋白的诱导表达，可能是心肌预适应延迟相保护的重要机制之一。