不同氧浓度培养小鼠肝细胞线粒体蛋白的蛋白质组学研究  

Proteomic Study of Mitochondrial Proteins in Mouse Hepatocytes Exposed to Different Concentrations of Oxygen

作　　者：孙鲁宁[1] 孙娇[1] 张宁[1] 张静萍[2] 张海鹏[1] 

机构地区：[1]中国医科大学病理生理学教研室,沈阳110001 [2]中国医科大学附属盛京医院手术室,沈阳110004

出　　处：《中国医科大学学报》2011年第3期196-199,203,共5页Journal of China Medical University

基　　金：国家自然科学基金资助项目(30900567)

摘　　要：目的采用蛋白质组学方法研究低氧和高氧条件下培养小鼠肝细胞线粒体蛋白表达的变化情况并探讨其病理生理学意义。方法直接消化法分离C57BL/6小鼠肝细胞,分别于低氧、高氧或常氧条件下培养8 h后提取线粒体蛋白,然后采用双向电泳法(2-DE)分离差异表达的蛋白,凝胶银染后切取差异蛋白点进行基质辅助激光解析电离飞行时间(MALTI-TOF)质谱检测,对获取的数据采用Mascot软件在NCBInr数据库内检索。结果共检测出19种差异表达的线粒体蛋白。低氧组小鼠肝细胞线粒体蛋白图谱中有16个蛋白点表达量与常氧组有统计学差异(P<0.05);高氧组小鼠有12个蛋白点表达量与常氧组有统计学差异(P<0.05),其中低氧组和高氧组均与常氧组有统计学差异的蛋白点有9个(P<0.05)。结论高氧时线粒体复合体Ⅲ高表达可能是造成细胞氧化损伤的基本病理基础,而超氧化物歧化酶2表达上调可以清除过多的活性氧;与此同时,参与损伤基因修复的与残基X相连的二磷酸核苷表达增多可以减轻细胞内的氧化损伤。Objective To identify the expression of mitochondrial proteins in mouse hepatocytes exposed to low or high concentration of oxygen by using proteomic approach.Methods C57BL/6 mouse hepatocytes were isolated and cultured for 8 hours under hypoxic,hyperoxic,or normoxic condition respectively,and then the mitochondrial proteins were extracted.The differentially expressed mitochondrial proteins were separated with two dimensional electrophoresis（2-DE） and analyzed by matrix assisted laser desorption ionization/time-of-flight mass spectrometry（MALDI-TOF-MS）.Results Totally 19 differentially expressed mitochondrial proteins were found.There were significant differences in the expression levels of 16 proteins between hypoxic and normoxic groups（P 0.05）,12 proteins between hyperoxic and normoxic groups（P 0.05）,and 9 proteins between hypoxic and hyperoxic groups（P 0.05）.Conclusion The high expression of cytochrome b-c1 complex subunit 1 under hyperoxic condition might be invovled in the pathological basis of cellular oxidative damage,and superoxide dismutase 2 could scavenge excessive reactive oxygen species during oxidative stress.The nucleoside diphosphate linked moiety X might play an important role in reducing the oxidative damage.

关 键 词：低氧 高氧 肝细胞 线粒体蛋白 蛋白质组 

分 类 号：R363.2[医药卫生—病理学]