摘要
背景:肝脏自身具有强大的再生能力,临床资料显示大部分肝癌患者合并有肝硬化,切除肿瘤后肝脏质量可逐渐恢复,提示硬化的肝脏仍具备修复潜能。目的:尝试用肝部分切除方法诱导小鼠硬化肝组织内具再生能力的细胞增殖,观察再生肝脏的结构变化。方法:使用CCl4皮下注射方法制备小鼠肝硬化模型。对肝硬化小鼠行肝部分切除,于切除后1,3,5,7,10,14及21 d取小鼠肝组织并称质量。观察比较肝部分切除后不同时间小鼠再生肝组织的结构变化;评估肝再生率;免疫组化方法分析肝星形细胞标志分子结蛋白及肝星形细胞活化标志分子α-平滑肌肌动蛋白的表达变化;5-溴脱氧尿核苷标记技术对增殖细胞进行定性、定位。结果与结论:肝硬化小鼠肝部分切除后第1,3,5,7,14,21天肝再生率分别为6.58%,22.03%,21.39%,29.05%,45.22%,50.98%。苏木精-伊红染色和Masson染色显示CCl4注射8周后呈早期肝硬化病理改变,肝部分切除后肝组织结构逐步改善。免疫组化结果显示肝部分切除后第1天再生肝组织内结蛋白表达减少,但第3天至第14天呈上升趋势,21 d则明显减少;肝再生过程中各时间点α-平滑肌肌动蛋白表达依时递减。5-溴脱氧尿核苷阳性细胞在肝部分切除后1-7 d主要为肝细胞,肝部分切除后14-21 d阳性细胞定位于肝血窦内皮及窦腔内。结果表明早期肝硬化小鼠大部肝切除后3 d出现明显的再生反应,并逐步恢复正常的肝组织结构。
BACKGROUND： Liver possesses a powerful regenerative capacity. It is reported that the most patients suffering from hepatocelluler carcinoma are accompanied with liver cirrhosis. The liver weight can be gradually restored after tumor resection, which indicated a regenerative potential of cirrhosis liver.OBJECTIVE：To try to use partial hepatectomy to induce proliferation of cells with regeneration capacity in cirrhotic liver of mice, and to observe the structural changes of regenerated liver.METHODS： The liver cirrhosis model of mouse was produced by injecting CCl4 liquid subcutaneously. The mice with liver cirrhosis were subjected to partial hepatectomy. The residual liver was dissected and weighted at 1, 3, 5, 7, 10, 14 and 21 days after partial hepatectomy. The structural changes of regenerating livers were compared after partial hepatectomy at various time points. Liver regeneration rate was evaluated. Immunohistochemical method was used to analyze desmin and alpha-smooth muscle actin expression in the hepatic satellite cells. The 5-bromo-2-deoxyuridine incorporation labeling method was used to exhibit and localize the regenerating cells.
RESULTS AND CONCLUSION：The liver regeneration rates of the mice with liver cirrhosis at the 1, 3, 5, 7, 14, 21 days after the partial hepatectomy were 6.58%, 22.03%, 21.39%, 29.05%, 45.22%, and 50.98% respectively. The results of hematoxylin-eosin staining as well as Masson staining showed the pathological early cirrhosis features of the liver after CCl4 treatment for 8 weeks and the structure of residual livers improved gradually after partial hepatectomy. Immunohistochemical results demonstrated that the expression of desmin in regenerating liver was decreased at 1 day after partial hepatectomy, but showed an uptrend from 3 to 14 days and decreased at 21 days. The expression of alpha-smooth muscle actin decreased gradually at every time point after partialhepatectomy. The 5-bromo-2- deoxyuridine-positive nucleus localized in hepatocytes at 1-7 days and mainly localized in hepatic sinusoid endothelium and intracavity at 14-21 days after partial hepatectomy. The regenerative reaction of mouse cirrhotic liver appeared at 3 days after partial hepatectomy and the structure of normal regenerating liver will be improved and restored gradually.
出处
《中国组织工程研究》
CAS
CSCD
2013年第53期9196-9202,共7页
Chinese Journal of Tissue Engineering Research
基金
广东省2011年大学生创新实验项目(2011)~~
