小鼠部分肝切除术后肝功能指标及干细胞再生研究  

Study on liver function and stem cell regeneration after partial hepatectomy in mice

作　　者：刘洋[1] 柯镜 朱可心 邓浩君 饶慧玲 于娟 国宏莉[1] 李珊[1] LIU Yang;KE Jing;ZHU Ke-xin;DENG Hao-jun;RAO Hui-ling;YU Juan;GUO Hong-li;LI Shan(Institute of Basic Medical Science,Hubei University of Medicine,Shiyan 422000,Hubei,China)

机构地区：[1]湖北医药学院基础医学研究所,十堰422000

出　　处：《医学研究生学报》2021年第9期926-930,共5页Journal of Medical Postgraduates

基　　金：湖北医学院研究生科技创新项目(JC2020003);湖北省大学生创新创业训练计划项目(S202010929019,S202110929011)。

摘　　要：目的肝再生是多种效应细胞高度协调的增殖反应,文章旨在揭示小鼠部分肝切除术后肝再生过程中肝功能指标和干细胞增殖进程,为探究肝再生机制提供理论基础。方法对C57BL/6小鼠行肝左前叶切除术,建模成功后分别于术后第2、4、8天收集血清和肝组织标本。将小鼠分为假手术组、术后2天、术后4天和术后8天组,每组3只。通过血清生化检测、HE染色评估小鼠肝部分切除术后肝功能和肝组织变化。利用免疫组织化学检测Ki67,SOX2 and S100B在肝再生不同阶段的表达特征。结果PHx后2、4、8天肝比重均未达到假手术组水平(P<0.05)。手术后2~4 d为肝比重上升的高峰期,术后4~8 d肝比重不再明显增加。与假手术组相比,术后2天组ALT水平显著升高(P<0.05),血清AST水平呈现出类似改变,均为术后第2天急速升高,术后4、8天较术后2天有明显下降(P<0.01);与假手术组比较,术后2天组ALT/AST升高、AST/ALT降低(P<0.05);与术后2天组比较,术后4、8天组ALT/AST降低、AST/ALT升高(P<0.05)。IHC结果显示,假手术组小鼠肝正常增殖细胞(Ki67+)数为38.00±6.24,呈点状分布。PHx后第2天,小鼠肝组织内Ki67+细胞数为143.00±14.18,增殖程度明显强假手术组(P<0.01),增殖的细胞多散在分布于肝实质细胞中。术后第4天,除肝实质细胞外,增生的纤维组织细胞和浸润的炎细胞Ki67+(345.00±50.02),呈现出围绕血管分布的特性,且与术后2天组相比有显著增加(P<0.01)。术后第8天Ki67+细胞数(42.00±3.51)较术后2天组显著下降(P<0.01)。与假手术组比较,术后2天组SOX2、S100B表达增加(P<0.05);与术后2天组比较,术后4天组SOX2、S100B表达增加(P<0.05),术后8天组S100B表达减少(P<0.05)。结论AST/ALT可以作为评价肝损伤再生的指标。肝细胞和非实质细胞共同参与并加速肝再生的发生,SOX2和S100B阳性细胞在肝再生过程中发挥着重要的作用。Objective Liver regeneration was a proliferative response process involving highly coordinated cells.This research was focus on the process of liver function and stem cell proliferation in the process of liver regeneration after partial hepatectomy in mice,provided a theoretical foundation for exploring the mechanism of liver regeneration.Methods C57BL/6 mice were treated with left anterior lobectomy.After the model was successfully established,serum and liver tissue samples were collected on the 2nd,4th,and 8th days after the operation.The changes in liver function and liver tissue after partial hepatectomy were evaluated by serum biochemical test and H&E staining.Immunohistochemistry(IHC)was used to detect the expression of Ki67,SOX2,and S100B in different stages of liver regeneration.Results The liver/body ratio approached the peak level after 2-4 days of operation.In the early stage of liver regeneration,the serum ALT and AST levels increased sharply,and the level of ALT and AST decreased to normal level in the late stage.H&E and IHC results showed that hepatocellular proliferation was dominant in the early stage of hepatectomy,while nonparenchymal cells were involved in the late stage.The peak of cell proliferation was 4 days after the operation.SOX2,a marker of hepatic stem cells,showed a strong positive expression in the portal area and gradually decreased to the surrounding hepatocytes.At the same time,S100B expression in hepatocytes and bile duct epithelial cells was significantly increased,showing a diffuse positive expression.Conclusion AST/ALT can be used as an indicator to evaluate the regeneration of liver injury.Liver cells and nonparenchymal cells were involved in the liver regeneration.SOX2 and S100B positive cells play important roles in the process of liver regeneration.

关 键 词：肝部分切除术 肝再生 SOX2 S100B 

分 类 号：R657.3[医药卫生—外科学]