过表达肝细胞核因子4α脐带间充质干细胞促进小鼠大部肝切后肝再生  

Overexpression of hepatocyte nuclear factor 4α in umbilical cord mesenchymal stem cells promotes liver regeneration after subtotal hepatectomy in mice

作　　者：吴宁[1] 马永[1] 翟秀宇[2] 张玉玲[3] 张勇[1] 张祺祺 杭化莲[4] 夏强[4] 卞建民[1] 

机构地区：[1]南京医科大学附属南京医院普外科,南京210006 [2]吉林大学第一医院泌尿外二科,吉林130021 [3]遵义医学院附属医院贵州省细胞工程重点实验室,遵义563003 [4]上海交通大学医学院附属仁济医院肝脏外科,上海200127

出　　处：《中华老年多器官疾病杂志》2015年第1期64-70,共7页Chinese Journal of Multiple Organ Diseases in the Elderly

基　　金：南京市科学技术委员会基金（201308014）;国家自然科学基金（81100306）;上海市科委医学引导类基金（134119a9501）

摘　　要：目的：探讨人源性脐带间充质干细胞（UC-MSC）以及过表达肝细胞核因子4α（HNF4α）的UC-MSC能否促进小鼠大部肝切后肝再生。方法体外分离、培养、鉴定人UC-MSC，慢病毒转染过表达HNF4α。体外收集细胞培养上清液，将L02与上清液共培养，通过细胞增殖实验试剂盒（CCK8）检测细胞增殖活性。体内实验建立肝大部切除模型（约70%），分别经尾静脉向肝切除小鼠移植0.9%生理盐水（NS）、MSC、MSC-HNF4α。48h后比较3组肝切后肝再生的变化，通过免疫组化来观察肝标本Ki67的表达。结果成功分离鉴定UC-MSC，并且成功建立稳定过表达HNF4α的MSC。体外实验MSC组和MSC-HNF4α组中L02的增殖能力都明显高于NS组（P＜0.01），MSC组高于MSC-HNF4α组（P＜0.05）。同样体内实验相对于NS组，经MSC或MSC-HNF4α细胞处理的肝脏，其Ki67的表达明显高于NS组（ P＜0.01），同样MSC组高于MSC-HNF4α组（P＜0.05）。结论 UC-MSC和过表达HNF4α的MSC都分泌一系列因子促进肝再生。Objective To investigate whether human umbilical cord mesenchymal stem cells （UC-MSC） and the cells with overexpression of hepatocyte nuclear factor 4α （HNF4α） can promote liver regeneration after subtotal hepatectomy in mice. Methods After human UC-MSC were isolated, cultured and identified, the cells were transfected with lentiviral vector LV-HNF4α. Then the cell supernatant was collected and cultured with L02 cells, and the L02 cell viability was detected by cell counting kit-8 （CCK8） assay. In the in vivo study, a total of 18 mice were randomly divided into 3 groups （n=6 for each group）：normal saline （NS） group, MSC group and MSC-HNF4α group, and received an injection of NS, MSC and MSC-HNF4α（5×10^6/ml）, respectively via tail vein in 2 h before the establishment of subtotal hepatectomy model （70%）. In 48 h later, the expression of Ki67 in the liver tissues was detected by immunohistochemical assay, and the results were compared among 3 groups. Results UC-MSC was successfully isolated from human umbilical cord, and the UC-MSC with stable overexpression of HNF4α were established. In vitro study indicated that the L02 cells showed stronger proliferative ability when cocultured with MSC and MSC-HNF4α than cultured alone （P〈0.01）, and those cocultured with MSC-HNF4α stronger than the cells with MSC （P〈0.05）. In vivo study showed similar findings： the expression of Ki67 was stronger in the mice treated with MSC or MSC-HNF4αthan those with NS （P〈0.01）, and also overexpression of HNF4αresulted in more significant expression of Ki67 （P〈0.05）. Conclusion Both MSC and MSC-HNF4αsecret some cytokines to promote liver regeneration.

关 键 词：小鼠 肝再生 间充质干细胞 肝细胞核因子4Α 

分 类 号：R333.4[医药卫生—人体生理学]