肝细胞移植治疗肝硬化合并小肝综合征大鼠模型的效果观察

Effect of hepatocyte transplantation in treatment of rats with liver cirrhosis and small-for-size syndrome

目的观察肝细胞移植对大鼠肝硬化合并小肝综合征的治疗效果。方法对40只雄性SD大鼠用四氯化碳皮下注射加乙醇饮用建立肝硬化大鼠模型。从肝硬化大鼠模型中随机抽取30只,分为A组(术前3 d进行肝细胞移植)、B组(手术时行肝细胞移植)、C组(不进行肝细胞移植处理),每组10只,进行70%肝脏切除,术后观察各组大鼠生存率、不同时点血生化以及肝脏HE染色病理切片变化情况。符合正态分布的计量资料多组间比较采用方差分析,进一步两组间比较,方差齐采用LSD-t检验,方差不齐采用Games-Howell检验。生存率比较采用log-rank检验。结果A组术后存活率(90%,9/10)明显高于B组(50%,5/10)和C组(30%,3/10)(χ2=6.440,P=0.04)。术后24 h A、B、C 3组ALT平均值分别为(860.1±39.9)U/L、(904.6±35.6)U/L、(927.4±17.0)U/L(F=6.808,P=0.007),AST平均值分别为(896.3±44.7)U/L、(923.8±31.6)U/L、(950.0±16.3)U/L(F=3.666,P=0.047),Alb平均值分别为(25.6±0.5)g/L、(24.8±0.6)g/L、(23.4±0.4)g/L(F=26.577,P<0.001),A组的恢复情况均有好于B、C两组的趋势,但差异不全有统计学意义。至术后240 h 3组ALT平均值分别为(97.8±10.0)U/L、(128.2±20.7)U/L、(150.5±14.8)U/L(F=13.816,P=0.001),AST平均值分别为(108.7±10.8)U/L、(142.0±14.1)U/L、(161.0±21.2)U/L(F=17.220,P<0.001),Alb平均值分别为(29.1±0.6)g/L、(28.0±0.2)g/L、(27.0±0.2)g/L(F=15.629,P=0.001)。术后72 h肝组织病理学检查结果显示,C组大鼠肝索结构消失,大块肝细胞坏死及炎性细胞浸润,A组仅见肝细胞气球样病变,点状细胞坏死,少量炎性细胞浸润。结论肝细胞移植可有效促进肝硬化大鼠术后小肝综合征肝功能恢复及提高生存率,且治疗效果可能与细胞移植的时间点有关。

Objective To investigate the effect of hepatocyte transplantation(HCT)in the treatment of rats with liver cirrhosis and small-for-size syndrome(SFSS).Methods A total of 40 male Sprague-Dawley rats were given subcutaneous injection of carbon tetrachloride and oral administration of alcohol to establish a rat model of liver cirrhosis,among which 30 were randomly selected and divided into group A(hepatocyte transplantation on day 3 before surgery),group B(hepatocyte transplantation during surgery),and group C(no hepatocyte transplantation),with 10 rats in each group.All rats were given the resection of 70%of the liver,and the groups were compared in terms of survival rate,blood biochemistry at different time points,and change in liver pathological section based on HE staining.An analysis of variance was used for comparison of normally distributed continuous data between multiple groups,and the least significant difference t-test(for data with homogeneity of variance)or the Games-Howell test(for data with heterogeneity of variance)was used for further comparison between two groups;the log-rank test was used for comparison of survival rate.Results Group A had a significantly higher postoperative survival rate than groups B and C[90%(9/10)vs 50%(5/10)and 30%(3/10),χ2=6.440,P=0.04].For groups A,B,and C at 24 hours after surgery,the mean level of alanine aminotransferase(ALT)was 860.1±39.9 U/L,904.6±35.6 U/L,and 927.4±17.0 U/L,respectively(F=6.808,P=0.007),the meal level of aspartate aminotransferase(AST)was 896.3±44.7 U/L,923.8±31.6 U/L,and 950.0±16.3 U/L,respectively(F=3.666,P=0.047),and the mean level of albumin(Alb)was 25.6±0.5 g/L,24.8±0.6 g/L,and 23.4±0.4 g/L,respectively(F=26.577,P<0.001);group A tended to have better recovery than groups B and C,while some differences had no statistical significance.For groups A,B,and C at 240 hours after surgery,the mean level of ALT was 97.8±10.0 U/L,128.2±20.7 U/L,and 150.5±14.8 U/L,respectively(F=13.816,P=0.001),the mean level of AST was 108.7±10.8 U/L,142.0±14.1 U/L,and 161.0±21.2 U/L,respectively(F=17.220,P<0.001),and the mean level of Alb was 29.1±0.6 g/L,28.0±0.2 g/L,and 27.0±0.2g/L,respectively(F=15.629,P=0.001),with significant differences between the three groups.At 72 hours after surgery,liver histopathological examination showed disappearance of hepatic cord,massive hepatocyte necrosis,and inflammatory cell infiltration in group C,while group A only had balloon-like lesions,punctate cell necrosis,and a small amount of inflammatory cell infiltration.Conclusion Hepatocyte transplantation can effectively promote liver function recovery and improve survival rate of rats with liver cirrhosis and SFSS,and its therapeutic effect is associated with the time point of transplantation.