van Wagensveld B A, Reinders M E, van Gulik T M, Gelderblom H C, Frederiks W M, Wanders R J, Obertop H
Department of Surgery, University of Amsterdam, The Netherlands.
Transpl Int. 1998;11(1):38-45. doi: 10.1007/s001470050100.
Pretransplant rinse solutions have been shown to reduce reperfusion injury in cold-stored liver grafts, especially at the nonparenchymal level in sinusoidal endothelial cells (SEC). In this study, different rinse temperatures were tested in a rat liver preservation model. Livers were washed out in situ via the portal vein with cold (4 degrees C) University of Wisconsin (UW) solution, and after hepatectomy (t0), were stored for 8, 16, or 24 h of cold ischemia time (CIT). After storage, livers were flushed with UW solution at either 4 degrees C, 20 degrees C, or 37 degrees C and reperfused for 90 min (37 degrees C). Control livers were reperfused at t0 without preflush. Levels of hyaluronic acid (HA), purine nucleoside phosphorylase (PNP), AST, and LDH were measured in the reperfusion medium. Bile production was monitored during reperfusion. At the end of reperfusion, liver biopsies were taken for enzyme hystochemistry (5'-nucleotidase and LDH). After 8-h CIT and a flush at 4 degrees C, a release of endogenous HA (-7%) was observed, whereas uptake of exogenous HA occurred after the 20 degrees C flush (2%, P = NS) and after the 37 degrees C flush (24%, p < 0.001). HA release occurred at all three preflush temperatures after the 16-h CIT but was significantly lower when flushed at 37 degrees C (-10%) that at 4 degrees C and 20 degrees C (-64% and -17%, respectively, p = 0.05). After the 24-h CIT, the release of endogenous HA increased in the 4 degrees C and 20 degrees C preflush groups, but not in the 37 degrees C group. Levels of PNP and AST increased until the 24-h CIT in all groups but were significantly lower after preflush at 37 degrees C. Release of LDH did not increase with increasing periods of cold storage in any of the flush series. Compared to control livers, mean bile production during reperfusion was significantly decreased following preflush at 4 degrees C or 37 degrees C after all periods of CIT. No differences in mean bile production could be demonstrated in the three preflush groups after any period of CIT. LDH activity in liver tissue was best preserved after the 8 and 16-h CIT in combination with the 37 degrees C preflush, indicating less hepatocellular damage. In conclusion, in cold stored rat livers flushed at 37 degrees C before reperfusion, SEC and hepatocellular damage is attenuated.
移植前冲洗液已被证明可减少冷保存肝移植中的再灌注损伤,尤其是在肝血窦内皮细胞(SEC)的非实质水平。在本研究中,在大鼠肝保存模型中测试了不同的冲洗温度。通过门静脉用冷(4℃)的威斯康星大学(UW)溶液原位冲洗肝脏,肝切除术后(t0),进行8、16或24小时的冷缺血时间(CIT)保存。保存后,肝脏分别用4℃、20℃或37℃的UW溶液冲洗,并在37℃再灌注90分钟。对照肝脏在t0时不进行预冲洗直接再灌注。测量再灌注介质中透明质酸(HA)、嘌呤核苷磷酸化酶(PNP)、天冬氨酸转氨酶(AST)和乳酸脱氢酶(LDH)的水平。在再灌注期间监测胆汁分泌。再灌注结束时,取肝组织活检进行酶组织化学检测(5'-核苷酸酶和LDH)。在8小时CIT和4℃冲洗后,观察到内源性HA释放(-7%),而在20℃冲洗后(2%,P=无显著性差异)和37℃冲洗后(24%,p<0.001)出现外源性HA摄取。在16小时CIT后,在所有三个预冲洗温度下均出现HA释放,但在37℃冲洗时(-10%)显著低于4℃和20℃冲洗时(分别为-64%和-17%,p=0.05)。在24小时CIT后,4℃和20℃预冲洗组内源性HA释放增加,而37℃组未增加。在所有组中,PNP和AST水平在24小时CIT前均升高,但在37℃预冲洗后显著降低。在任何冲洗系列中,LDH的释放均未随冷保存时间的延长而增加。与对照肝脏相比,在所有CIT时间后,4℃或37℃预冲洗后再灌注期间的平均胆汁分泌显著减少。在任何CIT时间后,三个预冲洗组之间的平均胆汁分泌均无差异。在8小时和16小时CIT结合37℃预冲洗后,肝组织中的LDH活性保存最佳,表明肝细胞损伤较小。总之,在再灌注前用37℃冲洗的冷保存大鼠肝脏中,SEC和肝细胞损伤减轻。