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

目的：有研究表明，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（HSP 72）作为一个指标的活力在供体肝脏，评估HSP 72的数量和基因表达水平之间的相关性，和不同的缺血时间。方法：采用大鼠模型制备部分肝缺血模型。采用逆转录-聚合酶链反应（RT-PCR）和Western blot方法检测热缺血0、15、30、45、60和90 min后HSP 72和HSP 72的mRNA表达。结果：缺血30 min后，可见HSP 72的表达。缺血90 min与缺血30 min相比，HSP 72水平无明显变化。缺血30分钟时mRNA水平升高，缺血45分钟后mRNA水平不再升高。缺血45分钟及更长时间后，HSP 72和HSP mRNA表达水平均出现分离。提示HSP 72及其mRNA水平的检测可作为缺血再灌注供肝存活的一个指标。

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.