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Proteomics analysis of plasma membrane from liver sinusoidal endothelial cells after partial hepatectomy by an improved two-dimensional electrophoresis

机译:改进的二维电泳技术对部分肝切除术后肝窦内皮细胞质膜的蛋白质组学分析

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摘要

Liver regeneration is an angiogenesis-associated phenomenon. To identify key plasma membrane (PM) proteins of endothelial cells involved in the initiation of angiogenesis during liver regeneration, the PM of liver sinusoidal endothelial cells (LSEC) at 72 h after partial hepatectomy was enriched by an established in vivo membrane density perturbation method. The differentially expressed membrane proteins compared to those from sham operation were quantified using an improved two-dimensional 16-BAC/SDS-PAGE and identified by LC-MS/MS. Several proteins were further confirmed by cICAT labeling quantitative strategy. A total of 47 proteins were identified including known and novel proteins involved in angiogenesis or liver regeneration, such as inducible nitric oxide synthase, type IV collagen, and integrin beta3. Our results indicated that the combination of the membrane density perturbation strategy and the improved two-dimensional electrophoresis (2-DE) method are useful for investigating the endothelial dysfunctions in vivo.
机译:肝再生是与血管生成有关的现象。为了确定参与肝再生过程中血管新生的内皮细胞关键质膜(PM)蛋白,通过建立的体内膜密度扰动方法富集部分肝切除术后72 h肝窦形内皮细胞(LSEC)的PM。使用改良的二维16-BAC / SDS-PAGE定量分析与假手术相比差异表达的膜蛋白,并通过LC-MS / MS进行鉴定。 cICAT标记定量策略进一步证实了几种蛋白质。共鉴定出47种蛋白质,包括与血管生成或肝脏再生有关的已知和新型蛋白质,例如诱导型一氧化氮合酶,IV型胶原和整联蛋白beta3。我们的结果表明,膜密度摄动策略和改进的二维电泳(2-DE)方法的结合可用于研究体内内皮功能障碍。

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