大鼠肝移植重建肝动脉对胆管超微结构和并发症的影响

Effects of hepatic artery reconstruction in rat with orthotopic liver transplantation on ultrastructure of intrahepatic biliary epithelial cells and biliary complications

目的观察大鼠原位肝移植重建肝动脉对肝内胆管上皮细胞缺血再灌注损伤后超微结构及术后胆道并发症的影响。方法228只SD大鼠分为假手术组（8只）、肝移植重建肝动脉组（55对）和未重建肝动脉组（55对）。重建肝动脉组和未重建肝动脉组分别于肝脏复流后0．5、3、6、12、24、36、48h取材，用透射电镜观察肝内胆管上皮细胞的超微结构，通过计算机图像分析系统对线粒体形态计量分析；观察术后胆道并发症。结果两组肝内胆管上皮细胞损伤均有加重，表现为线粒体肿胀、嵴模糊或消失、微绒毛减少等超微结构改变，至24h达高峰，以后逐渐恢复。术后两组线粒体平均面积和周径随时间的延长逐渐增大，线粒体数密度随时间延长而减少。在24h，两组缺血再灌注损伤最显著，之后均开始缓解。在24、36、48h，两组线粒体平均面积、平均周径比较，差异均有统计学意义（t=-3．566，-7．780，-4．730，-4．610，-2．599，-5．370，P〈0．05）；在36、48h，两组线粒体平均数密度比较，差异有统计学意义（t=4．619，4．000，P〈0．05）。重建肝动脉组的胆道并发症发生率低于未重建肝动脉组（x^2=4．286，P〈0.05）。结论大鼠肝移植重建肝动脉对肝内胆管上皮细胞缺血再灌注损伤后的超微结构具有保护作用，有利于术后恢复和减少胆道并发症的发生。

Objective To observe the effects of hepatic artery reconstruction in rat with orthotopic liver transplantation （OLT） on uhrastructure changes of intrahepatic biliary epithelial cells after ischemia reperfusion injury and postoperative biliary complications. Methods Male SD rats were divided into sham operation group （SO）, OLT with hepatic artery reconstruction group （HA） and OLT without hepatic artery reconstruction group （NA）. The liver tissue samples were collected at 0.5, 3, 6, 12, 24, 36, 48 hours after ischemia reperfusion in both HA group and NA group. The ultrastructure of intrahepatic biliary epithelial cells was observed by transmission electron microscope, and the morphometric analysis by the computer image analysis system. The postoperative biliary complications were also observed. Results In HA group and NA group, the injury of intrahepatic biliary epithelial cells aggravated gradually along with the reperfusion. The ultrastructure changes, such as enlarged mitochondria, blurred or disappeared cristae and decreased number of microvilli, were most significant at 24 hours and then recovered gradually. The mitochondrial average area and average perimeter were increased and the mitochondrial numerical density was decreased gradually along with the reperfusion. At 24, 36, 48 hours, the mitochondrial average area and average perimeter in NA group were significantly greater than those in HA group （t = - 3. 566, -7. 780, -4.730, - 4.610, - 2. 599, - 5. 730, P 〈 0.05）. The average numerial density of mitochondia in NA group was significantly less than that in HA group at 36, 48 hours（ t = -4. 619, 4. 000, P 〈 0.05）. The incidence rate of biliary complications in HA group is significantly lower than that in NA group （ X^2 = 4. 286, P 〈 0.05 ）. Conclusions Hepatic artery reconstruction in rat with OLT has an protective effect on the ultrastructure of intrahepatic biliary epithelial cells after ischemia reperfusion. It is beneficial to the recovery of intrahepatic bile duct epithelial cells and can reduce the incidence rate of biliary complications.