杜仲含药血清诱导骨髓间充质干细胞定向分化蛋白质组学研究_药学论文.docVIP

杜仲含药血清诱导骨髓间充质干细胞定向分化蛋白质组学研究_药学论文 杜仲含药血清诱导骨髓间充质干细胞定向分化蛋白质组学研究_药学论文 作者：曾建春 樊粤光 刘建仁 易春智 阎亮 【摘要】 　　目的研究杜仲含药血清诱导骨髓间充质干细胞 (Mesenchymal stem cells,MSCs) 向成骨细胞定向分化过程的蛋白表达差异，分析杜仲诱导MSCs向成骨细胞分化的可能调控机制。方法对杜仲含药血清干预前、干预后第2天、第7天，第14天的细胞提取总蛋白，通过双向电泳分离、PDQuest-7.3.0胶图分析软件进行分析找出差异蛋白，并进行质谱分析、数据库查询建立功能蛋白质组。结果双向电泳分离得到641个蛋白点，选取其中75个点进行质谱分析，经数据库查询得到6个功能蛋白。结论实验认为补肾中药杜仲可能通过上调波形蛋白、核纤层蛋白A的表达促进细胞分化，下调钙网织蛋白表达，释放细胞内钙，参与钙结节的形成，与体内骨矿化过程有关。过氧化物歧化酶、膜联蛋白A6与细胞的活力有关，调节着细胞的分裂增殖与分化，但是否与定向成骨细胞分化有着密切关系尚有待进一步研究。 【关键词】 杜仲　骨髓间充质干细胞　蛋白质组学　双向电泳 　　Abstract:ObjectiveTo study the protein change in the process of MSCs differentiation to osteoblas guiding by blood serum containing Duzhong（Cortex Eucommiae）, and its possible regulation mechanisms.MethodsTotal protein of cells interfered by blood serum containing Duzhong（Cortex Eucommiae）for 0day, 2days, 7days,14days, was extracted, runouted by two-dimensional electrophoresis and searched for different protein spot by PDQuest-7.3.0 software for analyzing gel picture.Functional proteome was established by mass chromatographic analysis and data base querying. Results641 protein spots were obtained, chose 75 spots to receive mass chromatographic analysis, and 6 functional protein were obtained by data base querying.ConclusionChinese drug Duzhong（Cortex Eucommiae）may up-regulate express of vimentin and lamin A to promote cell differentiation, down-regulate express of calreticulin to release intracellular Ca2+ which is related to Ca- nodus and bone mineralization in bodies. SOD and annexin A6 are concerned with corpuscular vigor, regulate corpuscular division growth and differentiation, but whether they have intimate relation with directional differentiation from MSCs to osteoblast is unknown, waiting for further research. 　　Key words:Duzhong（Cortex Eucommiae）； MSCs； Proteome； Two-dimensional electrophoresis 骨髓间充质干细胞（MSCs）是一种多功能干细胞，在不同条件下可向成骨细胞、软骨细胞、心肌细胞等分化。中医学认为，肾藏精，主骨，生髓，经前期研究发现，杜仲含药血清能够诱导MSCs向成骨细胞分化并促进细胞增殖，但对于MSCs向成骨细胞分化这一复杂过程的机制缺乏相关研究。 　　蛋白质组学研究是后基因组学时代又一重要研究工具，已经