摘要
目的:采用蛋白质组学方法研究低氧条件下培养小鼠肝细胞膜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,于低氧条件下(5%CO2,4.5 mg/L O2)培养8 h后提取膜蛋白,然后采用双向电泳法分离差异表达的蛋白,凝胶银染后切取差异蛋白点进行MALDI-TOF质谱检测,对获取的数据采用Mascot软件在NCBInr数据库内检索,并采用Western blottng方法验证差异蛋白的表达。结果:发现低氧组小鼠肝细胞膜蛋白图谱中有28个蛋白点表达量与对照组有显著差异(P<0.05),根据数据库检索结果,鉴定了低氧时低表达的膜蛋白9个和低氧时高表达的膜蛋白7个。Western blotting验证结果与蛋白质组学分析一致。结论:低氧时,抑素、细胞色素b-c1复合体亚单位1和血红素结合蛋白p22HBP可能在细胞感受外界环境的氧变化及随后的细胞内低氧信号传递的过程中起到了非常重要的作用。
AIM： To identify the differential expression of mice hepatic cell membrane proteins after hypoxic exposure. METHODS： The hepatic cells of C57BL/6 mice were cultured for 8 h under hypoxic or normoxic conditions and the membrane proteins were extracted. The differentially expressed membrane proteins were separated by two - dimensional electrophoresis. The technique of matrix - assisted laser desorption/ionization mass time - of - flight spectrometry（ MALDI - TOF - MS ） was used to analyze the proteins. RESULTS ： Compared with the proteins extracted from the cells under nor- moxic condition with a threshold of 1.5 - fold, 28 differentially expressed proteins were identified in the proteins extracted from the cells under hypoxie condition according to the database NCBInr 20071130, in which 9 were down - regulated and 7 were up - regulated, including prohibitin, cytochrome b - el complex subunit 1 and p22HBP. CONCLUSION ： The resuhs suggest that prohibitin, cytoehrome b - el complex subunit 1 and p22HBP might play important roles in sensing of cel- lular oxygen and the related signal transduction pathways.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2011年第2期338-342,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.30271444)
关键词
低氧
肝细胞
膜蛋白质
蛋白质组
Hypoxia
Hepatocytes
Membrane protein
Proteome
