门静脉栓塞及结扎联合肝细胞生长因子对肝纤维化大鼠肝再生的研究

Portal vein embolization and ligation combined with hepatocyte growth factor for liver regeneration in rats with hepatic fibrosis

目的探讨门静脉栓塞及结扎联合肝细胞生长因子(hepatocyte growth factor,HGF)对肝纤维化大鼠肝再生的作用。方法制备大鼠肝纤维化模型,将100只肝纤维化大鼠随机分为5组,术前对照组、门静脉栓塞组(A1组)、门静脉结扎组(A2组)、门静脉栓塞+肝细胞生长因子组(B1组)和门静脉结扎+肝细胞生长因子组(B2组),每组20只。除术前对照组外以上各组栓塞或结扎大鼠门静脉右支。每组于术前及术后1、3、7及14 d分批处死大鼠,每次5只,检测各组时间点大鼠外周血清中丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天冬氨酸氨基转移酶(aspartateaminotransferase,AST)的含量;称取各叶肝脏及全肝重量,计算术后不同时间点非栓塞及非结扎肝叶占整个肝脏的百分比;免疫组化检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)与Ki-67的表达。结果术后各组ALT、AST呈一过性升高改变,表现为术后第1天达到高峰,B1与B2组术后第1、3、7天ALT及AST峰值分别较A1与A2组低(P<0.05)。各组大鼠非栓塞及非结扎侧肝重/全肝重百分比均随时间增加而增加,术后第7、14天B1组非栓塞叶肝重/全肝重较A1组高,B2组非结扎叶肝重/全肝重较A2组高(P<0.05)。免疫组织化学改变:A1与A2组、B1与B2组术后非栓塞及非结扎侧肝叶PCNA与Ki-67的表达较术前明显增加并于第3天达高峰(P<0.05),然后缓慢下降,第14天A1与A2组恢复至术前水平(P>0.05),而B1与B2组较术前仍明显升高(P<0.05)。结论肝纤维化大鼠选择性门静脉栓塞、门静脉结扎后对侧肝脏均有再生能力;外源性HGF能明显提高肝再生能力。

Objective To investigate the portal vein embolization and portal vein ligation combined with hepatocytegrowth factor（HGF） in liver regeneration in hepatic fibrosis. Methods The rat liver fibrosis model was prepared. 100 hepaticfibrosis rats were randomly divided into 5 groups： pre operation control group, portal vein embolization group（A1）, portal veinligation group（A2）, portal vein embolization ＋ hepatocyte growth factor group（B1） and portal vein ligation ＋ hepatocyte growthfactor group（B2）, 20 rats in each group.Except for the control group, all the above groups were embolized or ligated to the rightbranch of portal vein. The rats of each group were killed（n=5） at 1, 3, 7 and 14 d after operation and five fibrosis rats werekilled before operation. The blood samples obtained from the inferior vena cava at different observation time points pre-operation and post-operation, the level of alanine aminotransferase（ALT） and aspartate aminotransferase（AST） were measured,The weight of the liver and the whole liver was calculated, and the percentage of the whole liver was calculated withoutembolized or not ligated at different time points after operation. The liver sections were immunostained for proliferating cellnuclear antigen（PCNA）and Ki-67. All data were statistically analyzed. Results The levels of serum ALT and AST raised inthe first day after operation were more than that of pre-operation（P〈0.05）. The peak values of ALT and AST in group B1 andB2 were respectively lower than those in group A1 and A2 in the first, third, seventh day（P〈0.05）. The percentage of thenonembolized and nonligated liver weight/whole liver weight after postoperative increased, the ratio of group B1 was higher thanthat of group A1, similarly, the ratio of group B2 was higher than that of group A2 in the seventh and fourteenth day（P〈0.05）.The PCNA and Ki-67 were positive in hepatocytes increased after operation, and reached peak at the third day（P〈0.05）,restored to the normal level at the fourteenth day post-operation in the group A1 and A2, however, group B1 and B2 was stillsignificantly higher than before surgery（P〉0.05）. Conclusion Fibrosis rats have the ability of regeneration in thecontralateral part of the liver after selective PVE and PVL, exogenous HGF can significantly improve liver regeneration.