肝祖细胞移植对四氯化碳诱导急性肝衰竭小鼠模型肝损伤的修复作用

doi:10.13481/j.1671-587x.20150306

摘要/Abstract

摘要：

目的:观察肝祖细胞(HPCs)移植后不同浓度四氯化碳(CCl₄)诱导的急性肝衰竭小鼠模型肝脏修复情况,评价HPCs对肝损伤的修复能力,为评估HPCs移植疗效建立理想的肝衰竭模型。方法:96只ICR小鼠随机分为空白对照组、阴性对照组(0.1% CCl₄组、0.5% CCl₄组、2.0% CCl₄组、10.0% CCl₄组)和实验组(0.1% CCl₄+HPCs组、0.5% CCl₄+HPCs组、2.0% CCl₄+HPCs组和10.0% CCl₄+HPCs组)。空白对照组小鼠灌胃给予生理盐水,阴性对照组和实验组小鼠灌胃给予不同剂量CCl₄。CCl₄造模后1 d,将实验组和阴性对照组小鼠常规麻醉后切开腹腔,实验组小鼠经脾脏注射HPCs,阴性对照组小鼠注射等剂量无菌生理盐水。动态观察小鼠生命体征,监测其存活率;采血检测小鼠血清天门冬氨酸氨基转移酶(AST)和丙氨酸转氨酶(ALT)活性;取肝脏计算肝脏指数;HE染色观察各组小鼠肝脏组织病理学变化;Hoechst33342标记外源性HPCs,激光共聚焦显微镜检测移植HPCs在肝脏中的分布情况及肝脏标志蛋白细胞角蛋白18(CK18)和白蛋白(ALB)的表达。结果:与空白对照组比较,0.1%和0.5% CCl₄组小鼠存活率、肝脏指数、AST和ALT活性无明显变化(P>0.05)。病理学检测,肝细胞有自我修复作用,HPCs移植治疗的效果不明显。与2.0 %CCl₄组比较,2.0%CCl₄+HPCs组小鼠AST和ALT活性及肝脏指数降低(P<0.01),存活率增加(P<0.01);病理学检测可见肝细胞再生和大量外源性肝细胞,CK18和ALB表达与内源性肝细胞相当,HPCs移植治疗有效。10.0%CCl₄组和10.0% CCl₄+HPCs组小鼠2 d内全部死亡,无法评估HPCs的治疗效果。结论:HPCs移植可提高急性肝衰竭模型小鼠存活率,改善AST和ALT活性,对肝损伤具有较强的修复能力;且2.0%CCl₄诱导的ICR小鼠急性肝衰竭模型能有效反映HPCs移植治疗的效果。

关键词: 四氯化碳, 急性肝衰竭, 肝祖细胞, 移植

Abstract:

Objective To obseve the rapair effect of hepatic progenitor cells (HPCs) transplantation on the liver of mouse models of acute liver failure induced with different concentrations of carbon tetrachloride (CCl₄), and to construct one ideal mouse model of acute liver failure for evaluation on the efficacy of HPCs transplantation. Methods 96 ICR mice were randomly divided into nine groups: blank control group, negative control groups (0.1% CCl₄ group, 0.5% CCl₄ group, 2.0% CCl₄ group and 10.0% CCl₄ group) and experimental groups (0.1% CCl₄ + HPCs group, 0.5% CCl₄ + HPCs group, 2.0% CCl₄ + HPCs group and 10.0% CCl₄ + HPCs group).Normal saline was given to the mice in blank control group by intragastric administration, and the mice in negative control groups and experimental groups were administrated with different concentrations of CCl₄.One day later, the mice in experimental groups were injected with HPCs from spleen, and the mice in negative control groups were given the same dose of normal saline.14 d after the injection of HPCs, the survival rate, liver index, the activities of serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST), the histopathological changes, the distribution of exogenous hepatocytes and the expression levels of cytokeratin 18(CK18) and albumin (ALB) were detected. Results Compared with blank control group, the survival rates, liver indexes and the activities of serum ALT and AST in 0.1% CCl₄ group and 0.5% CCl₄ group had no significant changes(P>0.05);the effective treatment of HPCs transplantation was not observed because of self-repairing.Compared with 2.0% CCl₄ model group, the activities of serum ALT and AST and liver index in 2.0% CCl₄ + HPCs group were significantly decreased(P<0.01), and the survival rate was increased(P<0.01);the liver cell regeneration and a large number of exogenous hepatocytes were found by pathological detection in 2% CCl₄ + HPCs group;the therapy of HPCs transplantation was effective.All mice died in 2 d in 10.0% CCl₄ group and 10.0% CCl₄ + HPCs group, and the therapeutic efficacy of HPCs was not observed. Conclusion HPCs transplantation can increase the survival rate of the model mice of acute liver failure, improve the activities of ALT, and AST and repair the hepatic injury;HPCs transplantation treatment is effective on the mouse model of 2.0% CCl₄-induced acute liver failure.

Key words: carbon tetrachloride, acute liver failure, hepatic progenitor cells, transplantation

中图分类号:

R575.3

赵丽, 黄道超, 龚梦嘉, 李娅莎, 毕杨. 肝祖细胞移植对四氯化碳诱导急性肝衰竭小鼠模型肝损伤的修复作用[J]. 吉林大学学报(医学版), 2015, 41(03): 464-469.

ZHAO Li, HUANG Daochao, GONG Mengjia, LI Yasha, BI Yang. Repair effect of hepatic progenitor cell transplantation on acute liver failure mouse model induced by carbon tetrachloride[J]. Journal of Jilin University Medicine Edition, 2015, 41(03): 464-469.

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