冷冻保存大鼠肝脏离体再灌注时嘌呤核苷磷酸酶活性和透明质酸吸收率的变化

Rat liver ischemia and reperfusion injury in UW, HTK and Celsior

目的探讨不同保存液保存大鼠肝脏后离体再灌注时嘌呤核苷磷酸酶（PNP）活性和透明质酸吸收率的变化及意义。方法分别采用UW液、HTK液和Celsior液灌洗、冷保存Wistar大鼠肝脏16及24h，然后用37℃的Kreb-Henseleit液在常温下连续灌注90min，分别于灌注0、15、30、60和90min时，从灌注液中取样，测定嘌呤核苷磷酸酶活性和外源性透明质酸吸收率的变化，据此评价肝窦内皮细胞的状况。结果经过16h的低温保存，在再灌注60min以前，HTK液组灌注液中PNP的含量明显高于UW液组和Celsior液组（P〈0．01）；再灌注60min后，HTK液组和Celsior液组灌注液中PNP的含量明显高于UW液组（P〈0．01）。经过24h的低温保存，在再灌注15min后，HTK液组灌注液中PNP含量明显高于Celsior液组（P〈0．01），而Celsior液组又明显高于Uw液组（P〈0．01）。透明质酸的吸收率均为负值，说明内源性透明质酸的释放大于外源性透明质酸的吸收，且随着保存时间和再灌注时间的延长，这一趋势更加明显，其中HTK液组最明显，Celsior液组次之。结论随着低温保存和再灌注时间的延长，肝脏中PNP活性逐渐升高，外源性透明质酸的吸收率下降，二者可作为评价肝脏缺血一再灌注损伤的指标。

Objective To study the changes of purine nucleoside phosphorylase （PNP） and rat liver exogenous hyaluronic acid （HA） absorption after cold storage and normothermic reperfusion in University of Wisconsin （UW）, Celsior and histidine-tryptopan-ketoglutarate （HTK） solutions during ischemia and reperfusion, Methods After different periods of cold storage in different preservation solutions, the rat livers were reperfused for 90 min with 37 ℃ Krebs-Henseleit solution using isolated rat liver perfuse model. The PNP and the absorption of exogenous HA were assayed in perfusate at different time points of reperfusion, Results After cold storage for 16 h, 60 rain before reperfusion, PNP level of rat liver in HTK was obviously higher than that in UW and Celsior （both P〈0. 01） ; after 60 min, PNP in HTK and Celsior was higher than that in UW. During reperfusion the rat livers preserved in UW and Celsior had a higher absorption of exogenous HA than in HTK （P〈0. 01） ; after cold storage for 24 h, 15 min after reperfusion, PNP level in HTK was obviously higher than that in Celsior, and the PNP in Celsior was higher than that in UW. Rat livers preserved in UW had a higher absorption of exogenous HA than in Celsior and HTK （P〈0. 01）. Conclusion The PNP level in rat liver was increased and the absorption of exogenous HA was decreased with the prolongation of cold storage time and reperfusion time. Both PNP level and absorption of exogenous HA could be used as the indicators to evaluate hepatic liver ischemia-reperfusion injury.