肝动脉桥式置管转流对供肝及胆道组织细胞凋亡的保护作用

Protection value of transfering the blood by a bridging duct of hepatic artery on apoptosis in orthotopic liver transplantation

目的验证肝动脉桥式置管转流这一方法是否能够减轻肝动脉缺血(hepatic artery ischemia,HAI)引起的肝胆细胞凋亡。方法应用简易犬肝自体原位移植模型,将24只杂交犬随机分为肝动脉缺血组(HAI组,8只)、肝动脉桥式置管转流组(TBB组,8只)及对照组(8只)。在制模后,3组动物均于冷灌注后不同时点切取肝脏、胆道组织,分别用戊二醛及40 g/L多聚甲醛溶液固定,进行电镜观察和TUNEL染色,观察肝胆组织形态学改变及细胞凋亡情况,并计算两组的凋亡指数。结果在冷灌注后2 h,HAI组电镜下出现较明显的肝胆细胞凋亡现象;肝动脉桥式置管转流组肝胆细胞凋亡现象少见;对照组难以找到凋亡肝胆细胞。TUNEL法染色切片显示,冷灌注后3组肝胆组织内均有少量凋亡细胞,其凋亡指数相差不大(P>0.05);随冷灌注后时间的延长,3组凋亡细胞的数量均有所增加,但HAI组凋亡指数的升高更为显著,肝动脉桥式置管转流组次之,对照组变化不大,3组间差别有统计学意义(P<0.01)。结论肝动脉桥式置管转流对肝移植时肝动脉缺血所导致的肝胆细胞凋亡具有显著保护作用,应在继续完善改进的基础上实施于临床。

Objective To validate whether the method of transfering the blood by a bridging duct between the recipient＇s spleen artery and the donor＇s gastroduodenal artery can lessen the apoptosis of liver and bile duct caused by hepatic artery ischemia or not. Methods Altogether 24 cross-fertilized dogs were divided into HAl group, transfering the blood by a bridging duct group （TBB group） and control group. After the models were induced, we cut the samples of liver and bile duct in all groups at different time points to fix in glutaraldehyde and 4% citromint and observed the apoptosis variety of liver and gall and calculated the apoptotic index. Results Two hours after cold perfusion, under electron microscope, apoptotic phenomenon could be easily observed in HAl group, singularly in TBB group, while almost none in control group. Slice staining by TUNEL showed that after cold perfusion each group had a few apoptotic cells, but there was no distinct difference in apoptotic index among the three groups. However, with the time prolonging after cold perfusion, although the-quantities of the apoptotic cells increased in all groups, the index increased most distinct in HAl group, least in the control group, the TBB group took the middle, and apoptotic index differed notably （P〈0.01）. Conclusion Transfering the blood by a bridging duct of hepatic artery can lessen the apoptosis of liver and bile duct caused by HAl significantly. This method should be put in practice in clinic based on constant improvement.