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骨髓间充质干细胞促进大鼠80%肝切除后残肝再生及修复作用的研究*

龚慧宾,沈中阳,宋红丽,郑卫萍,董冲,张静   

  1. 天津市第一中心医院
  • 收稿日期:2010-05-04 修回日期:2010-09-18 出版日期:2011-02-15 发布日期:2011-02-15
  • 通讯作者: 宋红丽

The Effects of Bone Marrow Mesenchymal Stem Cells on Liver Regeneration and Repair after 80% Hepatectomy in Rat

  • Received:2010-05-04 Revised:2010-09-18 Published:2011-02-15 Online:2011-02-15
  • Contact: Hong-Li Song

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摘要： 摘要 目的：骨髓间充质干细胞（Bone marrow mesenchymal stem cells，BMSCs）对大鼠自体80%肝切除后残余肝脏再生及修复的作用。方法：采用贴壁筛选法分离纯化鉴定BMSCs，取6-8周和体重200-220g的大鼠,建立大鼠80%肝切除模型，分为实验组16只，经门静脉注射BMSCs，对照组20只，注射等量生理盐水，采用生化方法检测肝功能，流式细胞仪和免疫组化检测肝细胞周期和增殖细胞核抗原（proliferation cell nuclear antigens；PCNA）。结果：①体外成功分离培养、增殖BMSCs；②术后生存率：实验组术后生存率为93.7%，对照组为75%；③实验组肝组织显示肝细胞坏死及炎性细胞增生、细胞水肿、气球样变与对照组相比明显减轻；④实验组术后1d、3d、7d，肝功能ALT、AST恢复明显优于对照组，差异有统计学意义（P＜0.05）；⑤实验组肝细胞周期中S期细胞比例与对照组相比明显增高（P＜0.05）；⑥ 残肝免疫组化：实验组PCNA表达水平明显高于对照组（P＜0.05）。结论：BMSCs可以促进大鼠80%肝切除后残余肝脏细胞的再生及修复。

关键词: 骨髓间充干细胞, 肝切除, 大鼠, 再生, 修复

Abstract: Abstract Objective: To investigate the effects of rat bone marrow mesenchymal stem cells on liver regeneration and repair after 80% hepatectomy. Methods: BMSCs were isolated and purified by adherent method, identified by surface markers via flow cytometry. 36 rats model of 80% hepatectomy were randomized into 2 groups: experimental group contains 16 rats with BMSC injection via their portal veins and control group contains 20 rats with saline by the same way. Using biochemical methods to test liver function, the flow cytometry and immunohistochemistry to detect cell cycle and proliferation cell nuclear antigens. Results: ① We isolated, cultured and proliferated BMSCs successfully in vitro. ② Survival rate after operation: survival rate was 93.7% in the experimental group and 75% in control group. ③ Liver tissue display that the hepatocyte necrosis and inflammatory cells proliferation and cellular edema, ballooning degeneration markedly reduced significantly in experimental group compared with the control group. ④ 1std, 3th d, 7th d after operation, the levels of ALT and AST recover significantly in experimental group than in the control group, the difference was statistically significant (P <0.05). ⑤ The S phase of cell cycle was significantly higher in experimental group than in the control group (P <0.05). ⑥ Remnant liver immunohistochemistry showed that PCNA expression was significantly higher in the experimental group than in the control group (P <0.05). Conclusion: BMSCs can improve significantly rat liver regeneration and repairment after 80% hepatectomy.

Key words: bone marrow mesenchymal stem cells, hepatectomy, rat, regeneration, repair