肝动脉结扎对梗阻性黄疸大鼠残肝肝细胞凋亡及再生的影响

Influence of hepatic artery ligation on hepatocyte apoptosis and liver regeneration of rats with obstructive jaundice

目的探讨正常肝脏状态下和梗阻性黄疸状态下,肝固有动脉结扎对大鼠肝细胞凋亡和肝脏再生的影响。方法设计动物实验。80只Wistar雄性大鼠分为4组,每组大鼠20只,即:建立梗阻性黄疸模型后3 d施行70%肝切除+肝固有动脉结扎+胆肠内引流组(A组);建立梗阻性黄疸模型后3 d施行70%肝切除+胆肠内引流(B组);假手术后3 d施行70%肝切除+肝固有动脉结扎(C组)和假手术后3 d施行70%肝切除(D组)。各组分别在肝大部切除模型建立后1、2、3和6 d共4个时间点观察动物生理状态并采集血液及残肝组织标本。记录各组术后大鼠的生理状态及死亡情况;检测血清肝功能指标:胆红素(TB)、丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)及白蛋白(ALB);采用TUNEL法检测残肝组织肝细胞的凋亡;采用Brdu检测残肝再生情况,并进行统计分析。结果 A、B 2组大鼠间及C、D 2组大鼠间术后生存率比较差异无统计学意义(均P>0.05)。A组和B组大鼠的术后肝功能差异无统计学意义(P>0.05),但术后早期的白蛋白合成A组要弱于B组(术后3 d,P<0.05);C组和D组术后肝功能指标差异也无统计学意义(P>0.05),但术后早期C组的白蛋白合成要弱于D组(术后1 d和3 d均P<0.05)。A、B、C 3个组术后1 d残肝凋亡指数达到最大,而后逐渐下降;而D组残肝凋亡指数在4个时间点上均为最低。术后1 d,C、D 2组肝细胞增殖达到高峰,增殖指数大小为D组>C组>B组>A组,各组之间差异均有统计学意义(均P<0.05)。术后6 d,A、B组肝细胞增殖指数无明显增大,C、D组肝细胞增殖指数明显降低,4个组的肝细胞增殖指数均处于较低水平。结论结扎肝动脉将增加残肝肝细胞凋亡,减弱肝细胞再生。但结扎肝动脉并未增加梗阻性黄疸大鼠术后死亡风险。

Objective To assess the influence of hepatic artery ligation on survival, hepatocyte apoptosis and regeneration of rats with obstructive jaundice. Methods Eighty adult male Wistar rats were divided into four groups： group A, suffered 70% hepatectomy＋hepatic artery ligation＋biliary drainage after 3 days of establishing obstructive jaundice model; group B, suffered 70% of hepatectomy＋biliary drainage after 3 days of establishing obstructive jaundice model; group C, suffered 70% of hepatectomy＋hepatic artery ligation after 3 days of sham operation; group D, suffered 70% of hepatectomy after 3 days of sham operation. Five rats of each group were sacrificed on 1, 2, 3, and 6 days after second operation. Liver function, hepatocyte apoptosis and liver regeneration were detected. Results Postoperative survival rates were not significantly different between group A and group B, similarly between group C and group D （allP〉0.05）. There was no significantly different in liver function of group Aversus group B, and group Cversus group D （P〉0.05）, but the synthesis of album on 1 d or 3 d after operation were significant difference （group Aversus group B,P〈0.05; group Cversus group D,P〈0.05）. Both of the group A, group B and group C had the highest apoptotic index on 1 d after operation, whereas the group D had the lowest hepatic apoptotic index among four group after the surgery. The regeneration indexes were as follow： group D〉group C〉group B〉group A （allP〈0.05）. On y 6 d after operation, the regeneration indexes of group A and group B did not increase, while those of group C and group D decreased remarkably. However, the regeneration indexes of four groups were lower than the mean level. Conclusions Hepatic artery ligation will increase hepatocyte apoptosis and weaken liver regeneration. However, for rats with obstructive jaundice, hepatic artery ligation didn＇t increase the risk of postoperative mortality.