双龙方干预骨髓间充质干细胞分化过程的蛋白表达  

Protein expression of differentiation of bone marrow mesenchymal stem cells induced by Shuanglong Formula

作　　者：叶能胜[1] 赵艳峰[1] 张荣利[1] 冯雪[1] 王义明[1] 罗国安[1] 

机构地区：[1]清华大学生命科学与医学研究院,北京100084

出　　处：《中成药》2007年第1期24-29,共6页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金(30271663);科技部十五公关项目(2004BA721A13);国家973中医理论专项(2005CB523503)资助

摘　　要：目的:研究中药复方双龙方(人参,丹参)干预骨髓间充质干细胞(MSCs)定向分化过程的蛋白表达;方法:采用双龙方含药血清对中国小型猪MSCs定向诱导分化为心肌样细胞的过程进行人工干预。对分化前后细胞蛋白进行二维电泳分离,PDQuest软件分析蛋白点的表达情况;对差异蛋白进行质谱分析并加以鉴定。结果:成功鉴定16个蛋白,通过对成功鉴定的蛋白功能进行数据库检索,表明这些蛋白的差异表达可能与中药干预过程有关;结论:双龙方含药血清对MSCs体外定向分化过程有一定的辅助作用;同时表明蛋白质组学技术可以应用于细胞增殖、分化和凋亡等过程的机理分析。AIM ： To investigate the protein patterns of bone marrow mesenchymal stem cells （MSCs） induced by 5-azacytidine and Shuanglong Formula（ Radix et Rhizoma Ginseng, Radix et Rhizoma Salviae Mihiorrhizae） and molecular mechanism of induced differentiation was discussed at the level of proteome. METHODS ： MSCs extracted from Chinese miniswine were induced by 5-azacytidine and Shuanglong Formula. Proteins were separated by two-dimensional gel electrophoresis, and were detected by silver staining method. The image was analyzed by PDQuest software, and the intensities of spots in the gels were used to quantify the expressions of proteins. Proteins of interest were chosen for in-gel digestion and MALDI-TOF-MS analysis, and then further identified by peptide mass fingerprinting （PMF）. RESULTS： 16 altered proteins were identified by PMF and their regulation might be caused by the addition of medicate sera containing the active components of Shuanglong Formula. The effects of medicated sera containing Shuanglong Formula on the induced-differentiation could be discussed from the regulated expression of functional proteins; CONCLUSION： Shuanglong Formula may promote the 5-azacytidine-induced differentiation of bone marrow MSCs in vitro, and our study indicate that proteomics can be used for the analysis of molecular mechanism for different cell process.

关 键 词：骨髓间充质千细胞 诱导分化 双龙方(人参 丹参) 含药血清 蛋白质组学 

分 类 号：R285.6[医药卫生—中药学]