中文摘要：

目的用在线二维纳升高效液相结合串联质谱（2D nanoLC—MS／MS）技术分析体外培养大鼠原代肝星状细胞（HSC）的蛋白质表达谱。方法分离培养大鼠原代HSC，取体外培养第10天自动活化的大鼠HSC总蛋白质，通过2D nanoLC—MS／MS对酶解所获肽段进行分离和鉴定，并对鉴定的蛋白质进行细胞定位和功能分类。结果从50μg HSC总蛋白质中鉴定出1014种蛋白质，相对分子质量范围为7832～588364；主要分布于细胞核、细胞骨架、线粒体、内质网，分别占25％、17％、13％、13％。蛋白质功能主要与核酸代谢、细胞器组装、信号传导、能量代谢等生理进程有关，分别占17％、14％、10％、9％；其中细胞骨架蛋白α-平滑肌肌动蛋白、波形蛋白和结蛋白是活化大鼠HSC特异性表达的蛋白质。结论本研究获得了目前较为全面的活化大鼠原代HSC的蛋白喷表达谱，为深入研究HSC的活化机制奠定了基础。

英文摘要：

Objective To profile the protein expression in activated rat hepatic stellate cells （HSCs）. Methods Primary rat HSCs were isolated and cultured in vitro. After 10 days in vitro culture, the HSCs were activated. Total protein extracted from these activated HSCs were digested, and the obtained peptides were analyzed by using online 2D nanoLC-MS/MS. The identified proteins were classified according to their distrbutions and functions. Results 1014 proteins were identified from 50 μg HSCs protein extract, the molecular weights of these proteins ranged from 7832 Da to 588 364 Da. Most of these proteins resided in nucleus, cytoskeleton, mitochondrion and endoplasmic reticulum. And these proteins were mainly involved in nucleic acid metobolism, organelle organization, signal transduction and energy generation. Among these proteins, alpha-smooth muscle actin, vimentin and desmin were specifically expressed in activated HSCs. Conclusion To the best of our knowledge, this is the most comprehensive protein expression profile of activated rat HSCs.