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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 investigate the effect of hepatic artery ligation on hepatocyte apoptosis and liver regeneration in normal liver and obstructive jaundice. Methods Design animal experiments. Eighty Wistar male rats were divided into 4 groups, 20 rats in each group, namely: 70% hepatectomy + hepatic artery ligation + cholangio-intestine drainage group (group A) 3 days after establishment of obstructive jaundice model; The model of obstructive jaundice was treated with 70% hepatectomy and cholangio-intestine drainage (B group) on the third day after operation; 70% of hepatectomy + hepatic artery ligation (C group) and 3 days after sham operation Liver resection (group D). Each group were observed at 4, 1, 2, 3, and 6 days after the establishment of the subhepatic partial hepatectomy model, and blood and residual liver tissue samples were collected. The physiological status and death of the rats in each group were recorded. The indexes of liver function such as serum bilirubin (TB), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and albumin (ALB) TUNEL method was used to detect the apoptosis of liver cells in residual liver tissue. Brdu was used to detect the regeneration of residual liver tissue and statistical analysis was made. Results There was no significant difference in postoperative survival between A, B 2 rats and C and D 2 rats (all P> 0.05). There was no significant difference in postoperative liver function between group A and group B (P> 0.05), but albumin synthesis in group A was weaker than group B after operation (P <0.05 on day 3); C There was no significant difference in postoperative liver function between group D and group D (P> 0.05). However, the albumin synthesis of group C was weaker than that of group D after operation (P <0.05 on both 1 and 3 days after operation). The apoptotic index of residual liver reached the maximum at 1 d after operation in group A, B and C, and then gradually decreased. The apoptotic index of residual liver in group D was the lowest at 4 time points. At 1 day after operation, the proliferation of hepatocytes in groups C and D 2 peaked. The proliferation index was in group D> C group> B group> A group. The difference was statistically significant (all P <0.05). On the 6th day after operation, the proliferation index of hepatocytes in group A and group B had no obvious increase, and the index of proliferation of hepatocytes in groups C and D decreased obviously. The proliferation index of hepatocytes in group A and B were all at a low level. Conclusion Ligation of hepatic artery will increase the apoptosis of liver cells and reduce the regeneration of hepatocytes. However, ligation of the hepatic artery did not increase the risk of postoperative death in obstructive jaundice rats.