大鼠2/3肝切除72 h后再生肝脏质膜比较蛋白质组学研究

大鼠2/3肝切除模型为研究肝细胞增殖和生理性血管生成提供了一个很好的活体内模型.为了揭示肝再生过程中与肝细胞增殖终止相关及与血管生成启动相关的质膜蛋白质，本研究对大鼠肝2/3部分切除72 h后的肝脏质膜进行了研究：利用两步蔗糖密度梯度离心法对切除组和假手术组的肝脏质膜进行纯化；然后通过双向电泳和质谱技术对肝切除样品进行了比较分析并对几个关键蛋白程序性凋亡相关蛋白-6和丝蛋白-A进行了免疫印迹验证.相对于假手术对照组(Sham组)，21种蛋白质在切除后72 h的肝脏中上调，15种蛋白质下调.所鉴定的差异表达蛋白参与了血管生成、细胞分裂增殖和凋亡、细胞分化调控、肝脏组织重新构建、代谢及应急反应.本研究为肝脏再生及其血管生成的研究提供了理论依据.

Differential Proteomic Analysis of Plasma Membrane in Rat Liver at 72 Hours after 2/3 Partial Hepatectomy

The 2/3 partial hepatectomy(PHx) rat liver model provides an effective medium to study the regulation of hepatocyte proliferation and physiological angiogenesis.To identify key plasma membrane(PM) proteins involved in the termination mechanism of hepatocytes and the initiation of angiogenesis during liver regeneration,the differentially expressed proteins in rat liver at 72 hours after 2/3 partial hepatectomy(PHx) and sham operation(Sham) were analyzed using 2-DE combined with ESI-Q-TOF.We used a two-step-sucrose density gradient centrifugation strategy to isolate liver plasma membrane of PHx and Sham.The isolated plasma membrane proteins were separated by two-dimensional gel electrophoresis and identified by tandem mass spectrometry(LC-MS/MS).Compared to the sham operation control group,21 proteins were up-regulated and 15 proteins were down-regulated after 72 hours in 2/3 PHx group.Western blot analysis was conducted to measure the expression of programmed cell death 6-interacting protein and filamin-A.The identified differentially expressed proteins were involved in angiogenesis,cell division,apoptosis,cell differentiation,remodeling of liver tissue,metabolic and energy-related protein.The article has provided some clues to liver regeneration and development of angiogenesis.