ASSESSMENT OF EARLY GRAFT FUNCTION BY EXPRESSION LEVELS OF HEAT SHOCK PROTEIN AND ITS mRNAs IN CANINE LIVER ISCHEMIAREPERFUSION

目的：据报道，肝脏缺血再灌注后热休克蛋白水平升高，在热缺血前诱导热休克蛋白表达可导致缺血耐受。我们假设热休克蛋白72(HSP72)作为供体肝脏存活的一个指标，评估了HSP72的数量和基因表达水平以及不同的缺血时间之间的相关性。方法：采用大鼠部分肝缺血模型。分别于0为对照组、热缺血15、30、45、60、90min后，用逆转录聚合酶链式反应(RT-PCR)和Western印迹分析检测HSP72和HSP72的基因表达。结果：缺血30min后，观察到HSP72的表达。与缺血30min相比，缺血90min后HSP72表达水平无明显变化。缺血30min后，mRNA表达水平升高，45min后无明显变化。缺血45min及更长时间后，HSP72和HSP基因表达水平呈解离状态。提示HSP72和HSP72mRNA的检测可作为肝移植缺血再灌流后存活的指标。

Purpose : It is reported that the level of heat shock protein elevated after ischemia-reperfused liver, and induction of it before warm ischemia resulted in ischemic tolerance. We hypothesized that the heat shock protein 72 (HSP72) as a index of viability in donor liver, evaluated the correlation between the amount of HSP72 and the gene expression levels, and various ischemic times. Method : Partial liver ischemia model was prepared using rat models. The gene expression (mRAM) of HSP72 and HSP72 were detected by reverse transcription polymerase chain reaction (RT-PCR) technique and Western blot analysis after 0 as control, 15, 30, 45, 60 and 90 minutes of warm ischemic times, respectively. Results : After 30 minutes ischemia, expression of HSP72 was observed. The level of HSP72 did not change after 90 minutes ischemia compared with that of 30 minutes ischemia. The level of mRNA elevated by 30 minutes ischemia, but did not increase after 45 minutes ischemia. Dissociation between HSP72 and HSP mRNA level was observed after 45 minutes and more longer ischemia models. Those data suggested that the detection of the levels of HSP72 and HSP72 mRNA would become as a index of viability in ischemia-reperfusion liver graft.