新生大鼠肝细胞脾内移植联合鼠肝再生增强因子治疗大鼠急性肝衰竭的疗效与免疫学机制  

Intrasplenic transplantation of newborn rat hepatocytes with rALR for treating rats with acute hepatic failure

作　　者：陈曜[1] 袁刚[1] 孙航[1] 刘杞[1] 

机构地区：[1]重庆医科大学病毒性肝炎研究所、教育部感染性疾病分子生物学重点实验室,400010

出　　处：《中华肝脏病杂志》2007年第5期378-382,共5页Chinese Journal of Hepatology

基　　金：国家自然科学基金（30170843、30500457）;重庆市科委重点项目（[2001]38、CSTC2006bb5276）

摘　　要：目的探讨新生大鼠肝细胞脾内移植联合重组鼠肝再生增强因子（rALR）治疗大鼠急性肝衰竭的疗效与免疫学机制。方法采用D-氨基半乳糖（1．2 g／kg）诱导并建立大鼠急性肝衰竭模型,36 h后随机分为6组：Ⅰ组：模型对照组;Ⅱ组：经脾脏注射等渗盐水1 ml,腹腔注射等渗盐水1 ml／d;Ⅲ组：经脾脏注射rALR 1 ml（50μg／kg）,腹腔注射rALR 1 ml（50μg·kg^-1·d^-1）;Ⅳ：经脾脏移植2×10^7同种异体新生大鼠肝细胞,腹腔注射等渗盐水1 ml／d;Ⅴ组：经脾脏移植2×10^7同种异体新生大鼠肝细胞,腹腔注射rALR 1 ml（50μg·kg^-1·d^-1）;Ⅵ组：经脾脏移植2×10^7同种异体新生大鼠肝细胞,腹腔注射环孢菌素1 ml（10 mg·kg^-1·d^-1）。观察大鼠每天存活率;术后第1、5天及2周处死大鼠以获取肝、脾组织,观察脾内移植肝细胞的组织学改变;在术前、术后第1、2、5、12天采集静脉血,采用放射免疫法测定血清IL-1β和TNFα的浓度。结果Ⅰ、Ⅱ和Ⅲ组大鼠存活时间差异无统计学意义,Ⅳ组有部分大鼠长期存活,2周存活率为33％,Ⅴ组大鼠2周存活率显著高于Ⅳ组,而与Ⅵ组大鼠2周存活率比较差异无统计学意义。Ⅳ、Ⅵ组脾内移植的肝细胞可存活5～7 d,Ⅴ组脾内移植肝细胞可存活至2周以上。术后第1天,Ⅳ组血清IL-1β水平显著高于Ⅴ组和Ⅵ组（P〈0．05）;术后第5天,Ⅳ组血清IL-1β水平仅与Ⅵ组差异有统计学意义（P〈0．05）。术后第1天,TNFα的浓度在Ⅱ组显著高于Ⅳ、Ⅴ、Ⅵ组（P〈0．05）。结论新生大鼠肝细胞脾内移植与rALR联合治疗大鼠急性肝衰竭有一定疗效,可提高D-氨基半乳糖诱导肝衰竭大鼠的存活率;移植肝细胞在脾脏可存活2周以上,其机制可能与促进肝细胞再生、预防移植肝细胞凋亡及轻微抑制细胞免疫有关。Objective To evaluate the effects of newborn rat hepatocyte intrasplenic transplantation combined with rat augmenter of liver regeneration （rALR） injection in treating rats with acute hepatic failure. Methods Acute hepatic failure （AHF） was induced in rats using D-gal （1.2 g/kg）. The rats were then randomly divided into 6 groups. Group Ⅰ received no further treatment and served as blank controls; group Ⅱ received 1ml buffered saline once through intrasplenic injection; group Ⅲ received 1 ml rALR; group Ⅳ received 2×10^7/ml hepatocytes; group V received 2×10^7 hepatocytes suspended in 1 ml rALR （50 μg/kg） and group Ⅵ received 2×10^7 hepatocytes in 1ml cyclosporine A （10 mg/kg）. The rats of the various treated groups were sacrificed at day 1, 5 and at week 2 and their livers and spleens were examined histopathologically. Blood samples of the rats were also obtained to determine the levels of TNF α and IL- 1β. Results There were no significant differences in survival between group Ⅰ, Ⅱ and Ⅲ rats. 33.3% of the group Ⅳ rats survived for 2 weeks. At week 2, the survival rate of group Ⅴ rats was significantly higher than that of group Ⅳ, but there was of no statistical significant increase when compared to that of group Ⅵ rats. Hepatocytes transplanted into spleens survived for 5-7 days in the spleens of group Ⅳ and Ⅵ rats, but they survived at least 2 weeks in group V. The average serum TNF α level in group Ⅳ was significantly higher than that in groups Ⅴ and Ⅵ on the first postoperative day, but after four days, only the difference between group Ⅳ and group Ⅵ was significant （P 〈 0.05）. The average serum level of TNF α in group Ⅱ was higher than that in groups Ⅳ, Ⅴ and Ⅵ on the first postoperative day （P 〈 0.05）, but there were no sigificant differences between those in groups Ⅳ,Ⅴ and Ⅵ on the 1st and the 5th postoperative days. Conelusion Newborn rat hepatocyte intrasplenic transplantation combined with rALR is effective in t

关 键 词：大鼠 肝功能衰竭 急性 肝细胞 肝再生增强因子 

分 类 号：R686[医药卫生—骨科学]