猪骨髓间充质干细胞与肝细胞共培养的比较蛋白质组学研究  

Comparative proteomic analysis of primary hepatocytes co-cultured with bone marrow mesenchymal stem cells in vitro

作　　者：顾劲扬[1] 施晓雷[1] 袁献温[1] 张悦[1] 肖江强[1] 丁义涛[1] 

机构地区：[1]南京大学医学院附属鼓楼医院肝胆研究所,210008

出　　处：《中华实验外科杂志》2014年第10期2131-2134,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81170417、81300337）;江苏省自然科学基金资助项目（BK2012081）;江苏省卫生厅青年科研课题（Q201307）;中国博士后基金资助项目（2011M500904）;江苏省博士后科研资助计划项目（1102149C）;南京市卫生青年人才培养工程（qrx11168）

摘　　要：目的 比较猪骨髓间充质干细胞（MSCS）与原代肝细胞构建共培养体系前后肝细胞内蛋白质表达谱的差异.方法 体外建立肝细胞与MSCS最适共培养体系.流式细胞仪分选共培养肝细胞）,并设立肝细胞、MSCS单纯培养组作为对照.采用双向凝胶电泳分离后,将差异蛋白点进行胶内原位切割、酶解后,获得肽质指纹谱.并通过Western blot技术进一步验证.结果 经质谱鉴定出34个蛋白点,Western blot检测证实囊性纤维化跨膜传导调节因子（CFTR）、白细胞介素（IL）-6、人高迁移率族蛋白（HMG）-1变化水平与蛋白质组结果基本一致.结论 这些功能蛋白的差异表达可能与MSCS和原代肝细胞体外共培养下肝细胞特异性功能改善、细胞生存时间延长、免疫调节存在必然联系.Objective To identify the differentially expressed proteins in hepatocytes within the optimal co-cultivation system with bone marrow mesenchymal stem cells （MSCS）.Methods The optimal co-cultivation of MSCS and primary hepatocytes was generated in vitro.CD44-CD45-cell population （hepatocytes） was sorted from co-culture system by flow cytometry.Protein samples were isolated by twodimensional （2D） gel electrophoresis.Bioinformatics was employed to analyze the relevance between the altered proteins.The protein expression levels of interest were confirmed by Western blotting.Results Thirty-four altered protein spots were unambiguously identified with high confidence,which are likely associated with hepatic metablic functions,cell cycle,cellular proliferation and apoptosis,as well as immune modulation.Conclusion Several differentially expressed functional proteins within co-cultured hepatocytes have been revealed by comparative proteomics,which may be associated with enhanced hepatic functionality,prolonged cellular survival,and immune modulation.

关 键 词：猪肝细胞 骨髓间充质干细胞 共培养 蛋白质组 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]