不同胆道灌洗方法对大鼠肝内胆管冷保存再灌注损伤的影响

Effect of different methods of bile duct lavage on cold preservation-reperfusion injury of intrahepatic bile duct in rats

目的观察不同胆道灌洗方法对大鼠移植肝肝内胆管冷保存再灌注损伤的影响。方法应用大鼠原位肝移植模型,将88只SD大鼠随机分为假手术组、胆道非灌洗组、UW液胆道灌洗组、生理盐水(NS)胆道灌洗+UW液肝内胆道灌注保存组、HTK液胆道灌洗+UW液肝内胆道灌注保存组、HTK液胆道灌洗+HTK液肝内胆道灌注保存组。移植肝置于4℃林格液中保存2 h后行原位肝移植。移植肝再灌注后24 h,检测血清总胆红素(TB)、直接胆红素(DB)、碱性磷酸酶(AKP)、γ-谷酰转肽酶(GGT)及胆汁中GGT、葡萄糖(Glu)含量。在光镜及电镜下观察肝内胆管上皮细胞的形态学变化。结果与非灌洗组比较,胆道灌洗组术后各项指标明显改善(P<0.01);HTK液及NS灌洗组较UW液灌洗组术后指标改善明显(P<0.05)。病理检测发现非灌洗组胆道损伤明显,各灌洗组胆道损伤程度明显改善,HTK液灌洗+UW或HTK液灌注组对胆管上皮细胞的损伤较轻。结论移植肝冷保存前进行胆道灌洗可以明显减轻胆管上皮细胞的损伤,4℃HTK液灌洗+4℃UW或HTK液灌注保存效果比较理想。

Objective To explore the effect of different methods of bile duct lavage on the cold preservation-reperfusion injury of liver graft intrahepatic bile duct. Methods By using the model of ortho- topic liver transplantation, 88 SD rats were randomly allocated into 6 groups：hile duct non-lavage group,4 ℃ UW solution bile duct lavage group,4 ℃ normal saline bile duct lavage ＋4 ℃ UW solution bile duct perfusion group,4 ℃ HTK solution lavage ＋ 4 ℃ UW solution perfusion group,4 ℃ HTK solution lavage ＋ 4 ℃ HTK solution perfusion group, and sham operation group. The levels of serum total biliruhin （ TB）, direct biliruhin （DB）, alkaline phosphatase （AKP） and gamma glutamyl transferase （GGT） were determined, and intrahepatic bile duct epithelial cells were observed under the light microscopy and electron microscopy. Results AU the indexes were improved significantly in the lavage group as compared with the non-lavage group （P 〈0.01 ） ,while some of indexes in the HTK solution and NS lavage group better than the UW solution lavage group after liver transplantations （ P 〈 0. 05）. Morphology confirmed that the bile duct injuries in the non-lavage group were conspicuous ,which were improved significantly in all bile lavage groups. The protective effect of bile duct was most ideal in the HTK lavage group. Conclusion Bile duct lavage should be performed before liver graft preservation to avoid the remained bile injuring the epithelium of intrahepatic bile duct; The preservation effect of 4℃ HTK solution ravage ＋ 4℃ UW or HTK solution perfusion is ideal.