Knowlton A A, Brecher P, Apstein C S
Whitaker Cardiovascular Institute, Boston University School of Medicine, Massachusetts 02118.
J Clin Invest. 1991 Jan;87(1):139-47. doi: 10.1172/JCI114963.
The effect of brief myocardial ischemia on the expression of heat shock protein (HSP 70) was examined in an in vivo rabbit model of myocardial ischemia using Northern blotting. Functional studies were carried out in the open-chested anesthetized rabbit. The large marginal branch of the left circumflex was occluded four times for 5 min. Using piezoelectric crystals implanted midwall in the ischemic zone, end-diastolic length, end-systolic length, and percent segmental shortening were assessed. Expression of HSP 70 was measured by Northern blotting. A single 5-min coronary occlusion doubled the expression of HSP 70 whereas four cycles of 5 min of ischemia/5 min of reperfusion resulted in a threefold increase in HSP 70 mRNA (P less than 0.001). Measurements with the piezoelectric crystals showed mild myocardial dysfunction concomitant with the increase in HSP 70. This increase in HSP 70 mRNA after repetitive brief ischemia was transient, occurring as early as 1 h and returning to baseline by 24 h after ischemia. Western blot analysis with a monoclonal antibody to HSP 70 was used to compare sham and postischemic myocardial HSP 70 levels. Changes in the amount of HSP 70 were evident as early as 2 h and were even more striking at 24 h.
使用Northern印迹法在兔心肌缺血体内模型中检测短暂心肌缺血对热休克蛋白(HSP 70)表达的影响。在开胸麻醉的兔身上进行功能研究。左旋支的大边缘支闭塞4次,每次5分钟。使用植入缺血区心肌中层的压电晶体,评估舒张末期长度、收缩末期长度和节段缩短百分比。通过Northern印迹法测量HSP 70的表达。单次5分钟冠状动脉闭塞使HSP 70的表达增加一倍,而4个周期的5分钟缺血/5分钟再灌注导致HSP 70 mRNA增加三倍(P<0.001)。压电晶体测量显示,随着HSP 70增加出现轻度心肌功能障碍。反复短暂缺血后HSP 70 mRNA的这种增加是短暂的,最早在缺血后1小时出现,到24小时恢复到基线水平。使用抗HSP 70单克隆抗体进行Western印迹分析,比较假手术组和缺血后心肌HSP 70水平。HSP 70量的变化最早在2小时明显,在24小时更显著。