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

发布时间：2018-07-13 13:28

【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1d,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(P0.05)。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(P0.01),存活率增加(P0.01);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2d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。
[Abstract]:Aim: to observe the liver repair of acute hepatic failure mice induced by carbon tetrachloride (CCl4) at different concentrations after transplantation of hepatic progenitor cells (HPCs), and to evaluate the repair ability of HPCs on liver injury, so as to establish an ideal liver failure model for evaluating the effect of HPCs transplantation. Methods Ninety-six ICR mice were randomly divided into control group, negative control group (0.1l4 group, 0.5l4 group, 2.0l4 group, 10.0l4 group) and experimental group (0.1l4 HPCs group, 0.5l4 HPCs group, 2.0l4 HPCs group and 10.0l4 HPCs group). Mice in blank control group were given normal saline by stomach, mice in negative control group and experimental group were given different doses of CCL _ 4 by intragastric perfusion for 1 day. Mice in experimental group and negative control group were treated with routine anesthesia and were treated intraperitoneally. Mice in the experimental group were injected with HPCs through spleen, and mice in the negative control group were injected with the same dose of aseptic saline. The vital signs and survival rate of mice were observed dynamically, and the activities of aspartate aminotransferase (AST) and alanine aminotransferase (alt) in serum were measured. The hepatic histopathological changes of mice in each group were observed by HE staining. HPCs were labeled with Hoechst33342. The distribution of transplanted HPCs in the liver and the expression of cytokeratin 18 (CK18) and cytokeratin 18 (CK18) were detected by confocal laser microscopy. The expression of albumin (ALB). Results: compared with the blank control group, the survival rate, liver index and alt activity of mice in 0.1% and 0.5l4 groups were not significantly changed (P0.05). Pathological examination showed that the effect of HPCs transplantation was not obvious. Compared with 2.0l4 group, the activity of AST and alt and liver index in 2.0l4 HPCs group were decreased (P0.01), the survival rate was increased (P0.01), and histopathological examination showed that hepatocyte regeneration and the expression of CK18 and ALB in a large number of exogenous hepatocytes were similar to those of endogenous hepatocytes. The mice in 10.0l4 group and 10.0l4 HPCs group died within 2 days. The therapeutic effect of HPCs could not be evaluated. Conclusion the transplantation of 2.0l4 can improve the survival rate of mice with acute liver failure, improve the activities of AST and alt, and can repair the liver injury, and the model of acute liver failure induced by 2.0l4 can effectively reflect the effect of transplantation.
【作者单位】：重庆医科大学附属儿童医院儿科研究所儿童发育疾病研究教育部重点实验室儿科学重庆市重点实验室重庆市儿童发育重大疾病诊治与预防国际科技合作基地;重庆医科大学附属儿童医院干细胞研究室;
【基金】：国家自然科学基金青年科学基金资助课题(81100309)
【分类号】：R285.5

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