氯化钴作用人脐带间充质干细胞的差异蛋白质组学分析

Differential proteomic analysis in human umbilical cord mesenchymal stem cells induced by cobalt chloride

目的用蛋白质二维凝胶电泳图谱，结合质谱技术鉴定、分析化学低氧模拟剂氯化钴（CoCl2）作用人脐带间充质干细胞（MSC）前后蛋白质组的差异表达。方法CoCl2作用人脐带MSC，应用双向凝胶电泳技术（2-DE）分离细胞总蛋白。ImageMaster 2D Platinum软件分析差异蛋白质点，用基质辅助激光解析串联飞行时间质谱对差异表达蛋白做质谱鉴定，对差异蛋白进行功能分类。结果建立了CoCl2作用人脐带MSC的蛋白质组图谱，鉴定出26个差异表达蛋白，其中11个表达上调，15个表达下调。其生物学功能涉及糖代谢、蛋白质代谢和修饰、脂类代谢、辅酶与辅基、细胞周期、免疫和防御、细胞结构和运动、信号转导、蛋白靶向和定位、神经细胞的活动、肌肉收缩等。其中过氧化物还原酶-1（Prdx1）表达下调，α-烯醇化酶（ENO1）、单胺囊泡转运蛋白1（VAT1）表达上调。结论低氧对人脐带MSC的影响涉及多蛋白及多个功能通路。

Objective To analyze the differential proteomics in human umbilical cord mesenchymal stem cells （MSC） induced by chemical hypoxia-mimetic agent cobalt chloride （ CoCl2 ） by two-dimensional gel electrophoresis （2-DE） and mass-spectrometry. Methods 2-DE was performed to separate proteins from treated and untreated human umbilical cord MSC with CoCl2. 2-DE images were analyzed by ImageMaster 2D Platinum software 6.0. The differential expressed proteins was identified by MALDI-TOF-MS. The differential proteins were classified based on their functions. Results 2-DE reference patterns of CoCl2 treated human umbilical cord MSC were established. A total of twenty-six differential proteins were identified, of them eleven proteins were up-regulated and fifteen down-regulated. Their biological functions involved in carbohydrate metabolism, protein metabolism and modification, lipid metabolism, coenzyme and prosthetic group metabolism, cell cycle, immunity and defense, cell structure and motility, signal transduction, protein targeting and localization, neuronal activities, muscle contraction, etc. Peroxiredoxinl （Prdx） was down-regulated, whereas al- pha-enolase （ENO1） and vesicle amine transport protein 1 homolog （VAT1） up-regulated. Conclusion The effects of hypoxia on human umbilical cord MSC were participated by multiple proteins and involved in multiple functional pathways.