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

目的：报道肝缺血再灌注后热休克蛋白水平升高，热缺血前诱导热休克蛋白产生缺血耐受。我们假设热休克蛋白72 （HSP72）作为供肝活力的指标，评估了HSP72的量与基因表达水平和不同缺血时间之间的相关性。方法：采用大鼠肝局部缺血模型制备肝局部缺血模型。分别在热缺血0、15、30、45、60和90分钟后，采用逆转录聚合酶链反应（RT-PCR）技术和Western blot检测HSP72和HSP72的基因表达（mRAM）。结果：缺血30min后，观察HSP72的表达。与缺血30分钟相比，缺血90分钟后HSP72水平无明显变化。mRNA水平在缺血30分钟后升高，但缺血45分钟后没有升高。在缺血45分钟及更长时间后观察HSP72和hspmrna水平的分离。这些数据提示，HSP72和HSP72 mRNA水平的检测可作为肝缺血再灌注移植肝细胞生存能力的指标。

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.