两种不同肝脏灌注方法对大鼠肝脏保护作用的比较

目的探讨两种不同的肝脏灌注方法对冷保存的离体大鼠肝脏保护作用的差异，为后期离体大鼠肝脏保存实验提供简单而有效的方法。方法制备两种不同的单纯冷保存的离体大鼠肝脏灌注模型，分为实验组（经门静脉和腹主动脉灌注法，10只）；对照组（经门静脉、腹主动脉及肝动脉灌注法，10只），灌注后取出肝脏置于4℃威斯康星大学保存液保存，12h后切取肝组织。行苏木素-伊红（HE）染色，观察肝组织病理损伤情况，并且用免疫组织化学染色法检测核因子（NF）-κB（SABC法）的表达情况。结果经12h冷保存后，两组大鼠肝脏组织病理示：肝脏小叶结构紊乱，肝细胞变性、肿胀及空泡形成，肝血窦和中央静脉有不同程度的淤血，部分肝窦内皮细胞坏死并脱落，但两组之间无明显差别。两组之间NF-κB阳性表达率差异无统计学意义（P〉0．05）。结论经门静脉和腹主动脉灌注法与经门静脉、腹主动脉及肝动脉灌注法，对离体大鼠肝脏的保护作用无差异，但前者操作相对简单，更容易在今后研究大鼠离体肝脏的实验中应用。

Comparison of the effect of organ protection between two different liver perfusion methods in rats

Objective To compare the difference of protective effect for cold stored isolated liver of rats between the two different liver perfusion methods, and to provide a simple and effective method for the later storage experiment of isolated liver of rats. Methods Two different liver perfusion methods were used for cold stored isolated liver of rats. Rats were divided into experiment group （ perfnsion via portal vein and abdominal aorta, n = 10） and control group （perfusion via portal vein, abdominal aorta and hepatic artery, n = 10）. After perfusion, the liver was taken out and stored in University of Wisconsin （UW） solution at 4 ℃ for 12 hours. Then the liver tissue was stained by hematoxylin-eosin （HE） and the pathological injury was observed. Expression of nuclear factor （NF） - κB was measured by immunohistochemical method （ SABC method）. Results After cold storage for 12 hours, the pathological results of liver tissue in two groups showed liver lobule structure disorder, degeneration, swelling and cavity formation in liver cell, different degrees of cogestion in hepatic sinusoids and central vein and necrosis and detachment of endothelial cell in hepatic sinus. But there was no significant difference in pathological changes between the two groups. There was also no significant difference in the positive expression of NF-κB between two groups （P 〉 0. 05）. Conclusions There is no significant difference in the isolated liver protective effect between the two methods. However, via the portal vein and abdominal aorta perfusion method is more simple and easy to apply in experiments for isolated rats＇ liver.