S期激酶相关蛋白2在急性肝功能衰竭大鼠肝组织中的表达

目的 探讨急性肝功能衰竭(ALF)大鼠S期激酶相关蛋白2(Skp2)表达变化及意义.方法 雄性SD大鼠256只,其中240只用D-氨基半乳糖溶液制备ALF模型.将大鼠分为ALF模型组、裸肝细胞移植组和微囊化肝细胞移植组,经腹腔分别注射RPMI 1640培养液、裸肝细胞悬液、微囊化肝细胞悬液各2 mL.另外6只作为健康对照组,另10只用于肝细胞分离.免疫组织化学法检测不同时间点大鼠肝细胞中Skp2蛋白的表达,全自动生化分析仪测定各组ALT、AST、TBil值,并观察各组生存率,多组样本间比较采用单因素方差分析.结果微囊化肝细胞腹腔移植可较裸肝细胞更好地降低ALF大鼠ALT、AST、TBil水平(P<0.05).造模36 h时,ALF模型组、裸肝细胞移植组和微囊化肝细胞移植组肝中Skp2-标记指数分别为(28.2±6.1)%、(41.4±10.5)%和(68.0±10.8)%(F=29.08,P<0.05),三组各15只大鼠168 h时各有4、6和11只存活.结论 动态观察Skp2表达可较好地判断ALF肝细胞的再生情况.

Expression of S-phase kinase associated protein 2 in liver tissues of rats with acute liver failure

Objective To investigate the expression of S-phase kinase associated protein 2 (Skp2) in rats with acute liver failure (ALF) and its significance. Methods There were 256 male SD rats used in this study, among which 240 were injected with D-galactosamine (D-GaIN) to set up ALF model. The rats were divided into 3 groups: ALF model group, free hepatocellular transplantation group, microencapsulated hepatocyte transplantation group, which were intraperitoneally injected with 2 mL of RPMI 1640 culture medium, free hepatocellular suspension and microencapsulated hepatocyte suspension, respectively. The other 6 rats were in control group and the rest 10 rats were used for hepatocyte isolation. Expressions of Skp2 protein in hepatocytes of rats at different time points were detected by immunohistochemical technique. Levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBil) were detected by automatic biochemistry analyzer. The survival rate in each group was observed. Comparisons among groups were done using one-factor analysis of variance. Results Levels of ALT, AST and TBil decreased more significantly by intraperitoneal transplantation of microencapsulated hepatocytes than those by intraperitoneal transplantation of free hepatocytes (P<0. 05). Skp2 labeling indices after 36 h of injection in ALF model group, free hepatocellular transplantation group and microencapsulated hepatocyte transolantation grouo were (28. 2±6.1) %, (41.4± 10. 5) % and (68. 0±10.8) %, respectively (F=29. 08 , P<0. 05). There were 4, 6 and 11 out of 15 rats survived in the 3 groups, respectively. Conclusion The dynamic observation of Skp2 expression could be used to judge the regeneration of hepatocytes.