| 李俊杰
1
尹 文
1
洪 楠
2
赵 威
1△.重组腺病毒介导 HGF 修饰 ADSCs 对大鼠肝损伤模型的治疗作用[J].,2014,14(6):1043-1047 |
| 重组腺病毒介导 HGF 修饰 ADSCs 对大鼠肝损伤模型的治疗作用 |
| The Cellular Therapeutic Effect of Adipose-derived Stem Cell-expressedHGF Transplantation on Rat Liver Injury Model* |
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| DOI： |
| 中文关键词: 脂肪源性干细胞 肝细胞生长因子 肝损伤 细胞治疗 |
| 英文关键词: ADSCs HGF Liver injury Cell therapy |
| 基金项目:陕西省社会发展攻关项目（ 2011K12-48 ） |
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| 中文摘要: |
| 摘要 目的：建立重组腺病毒介导肝细胞生长因子 HGF 促 ADSCs 定向分化肝细胞的方法，并对其参与肝损伤修复能力进行验
证， 为作为治疗肝损伤细胞来源提供参考。 方法： 采用消化培养的方法， 分离 SD 大鼠腹股沟脂肪组织 ADSCs 细胞， 连续传代 3 次
对其进行纯化培养， 利用形态学鉴定、 流式细胞术检测 ADSCs 表面标志物方法对其间充质干细胞样特征进行鉴定， 加入成脂肪
细胞诱导液观察其分化成脂肪细胞的能力； 构建腺病毒表达 HGF 载体 Adeno-HGF-EGFP， 并转染 ADSCs 细胞， 利用免疫细胞化
学染色方法检测肝细胞标志分子表达水平； 最后建立大鼠肝损伤动物型， 观察 Adeno-HGF-EGFP 转染的 ADSCs 细胞参与肝损
伤修复能力情况。结果： 分离的 ADSCs 细胞形态较为一致， 绝大多数呈梭形， 排列不规则。流式细胞术结果显示， 该细胞表达
CD29、 CD90、 CD106 等间充质干细胞细胞表面标记物， 低表达造血干细胞细胞表面标记物 CD34、 CD45， 同时， 分离的 ADSCs 细
胞具有诱导分化成脂肪细胞能力； Adeno-HGF-EGFP 转染 ADSCs 后， AFP、 ALB、 CK18 等肝细胞特异性分子表达水平升高；经尾
静脉注射 ADSCs 细胞后， 肝损伤大鼠的 AST、 ALT、 TBIL 等分子表达水平恢复正常。结论： 建立了重组腺病毒介导肝细胞生长因
子 HGF 促 ADSCs 定向分化肝细胞的方法， 并且表达 HGF 的 ADSCs 细胞具有修复大鼠肝损伤模型能力， 这为通过细胞治疗肝损
伤提供了新的细胞来源。 |
| 英文摘要: |
| ABSTRACT Objective：A method was established to identify the hepatogenic transdifferentiation of adipose tissue-derived stem
cells (ADSCs) by recombinant adenovirus-mediated expression of hepatocyte growth factor (HGF) and the differentiated ADSCs'
cellular therapeutic effect was verified on rat liver injury model.Methods: SD rat inguinal adipose tissue was harvested by enzyme digestion. And the third passage of cells were collected to detect the markers of the mesenchymal stem-like cells by flow cytometry. The third
passage of isolated cells was treated with adipogenic nutrient media to verify their ability of differentiation into adipocytes. Recombinant
adenovirus Adeno-HGF-EGFP was constructed, and characterized ADSCs were transfected by recombinant adenovirus, then hepatocyte-specific markers were detected by immunocytochemical staining. Finally animal models of liver injury were used to verify the ability
of the liver to repair after damage by tail vein injection of transfected ADSCs. After being subcultured, the sizes of isolated cells
were highly homogeneous, and the majority of them were spindle-shaped with disorderly arrangement. The flow cytometry results
showed that the isolated cells expressed CD29, CD90 and CD106, but not CD34 and CD45. Results:After being cultured in inducing media, the
isolated cells could differentiate into adipocytes; the expression of the hepatic markers, AFP, ALB and CK18 increased in differentiated
ADSCs; also, levels of AST, ALT and TBIL in sera collected from the acute liver injury rats became normal after ADSCs transplantation.
Conclusion；Recombinant adenovirus-expressed hepatocyte growth factor could induce ADSCs directly differentiate into hepatocytes,
and differentiated ADSCs transplantation could bea new strategy of cell therapy to liver injury. |
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