大鼠肝再生过程中肝细胞质膜微区差异蛋白质鉴定与分析

动物肝是具有极强再生能力的器官,研究并阐明肝再生的机制可为肝移植等与肝损伤相关的疾病治疗提供理论依据.质膜包括"脂筏(lipid rafts)"和"质膜微囊(caveolae)"的微区,具有参与胞吞胞饮、信号转导、运输胆固醇等重要功能.肝再生过程中,肝质膜微区脂筏蛋白质受到内部调控的影响会发生改变.捕获脂筏微区信号蛋白分布的变化,对于理解和阐明肝再生过程中信号通路途径有重要意义.本研究应用成熟的大鼠2/3肝切除模型结合蔗糖密度梯度离心法,提取假手术组与肝再生组大鼠肝细胞质膜,并进一步纯化获得质膜微区蛋白质.通过SDS-PAGE分离以及ESI-QTOF质谱鉴定,对获得的质膜微区蛋白质进行差异分析.结果显示,有30个微区蛋白质差异表达,其中13个上调、17个下调.生物信息学分析表明,所鉴定到的蛋白质主要参与细胞增殖、程序性死亡、细胞凋亡等调控,同时涉及到与肝再生密切相关的血管生成等信号通路.本文为质膜微区蛋白质的研究提供了方法上的参考以及相关基础数据,为后续临床肝再生的研究奠定了一定的基础.

Proteomic Profiling of Membrane Microdomain Proteins from Hepatocytes during Rat Liver Regeneration

Liver has a remarkable capacity of regeneration after hepatectomy. The research to understand liver regeneration provides clues for the treatment of liver injury and liver transplantation. As specialized regions of plasma membrane, lipid rafts and caveolae microdomain, play important roles in endocytosis, pinocytosis, signal transduction and transportat of cholesterol. During liver regeneration, the protein components of lipid raft in the microdomains were changing in respond to internal signals. Analysis of the varied lipid raft protein composition is helpful to reveal the involved signal pathways. In this study, sucrose density centrifugation were used to purify the plasma membrane from 2/3 partial hepatectomy rat model and compare with the sham group. The membrane proteins were separated by SDS-PAGE and identified by ESI-Q-TOF MS/MS analysis. Among the differential expressed proteins, 13 were up regulated and 17 were down regulated, with functions in cell proliferation, programmed cell death, apoptosis and angiogenesis signaling pathway. Our results provided clues to identify microdomains of signaling components on liver regeneration.