大鼠肝部分切除后4小时差异蛋白及蛋白复合体研究
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摘要
肝脏在受到损伤或切除时能够增生的现象叫作肝再生。大鼠肝2/3切除是最接近生理状况下肝再生的动物模型,也是一种细胞周期调控研究的很好模型,被广泛应用。肝再生是一个连续的过程,可分为引发期、进展期、细胞周期阶段和终止期。在肝再生的所有阶段当中,引发期是尤为关键的一个阶段,正是在这段时间里肝细胞由静止期迅速进入细胞周期。已有的研究提示,血液中的一些因子和肝细胞中的一些因子都有可能在肝再生的引发过程中起到关键作用。本课题利用蛋白质组学技术平台,以大鼠肝2/3切除为动物模型,以肝再生引发期的手术后4小时为研究的主要时间点,对肝再生过程中的血浆、肝细胞全蛋白和肝细胞细胞质基质进行研究,试图找到一些引发期的重要变化。本课题利用双向电泳(2DE)技术对正常组、手术组和假手术组肝细胞全蛋白进行差异比较研究,找到并鉴定出19个差异蛋白;利用2DE对各组别的血浆样本进行差异比较,找到并鉴定出7个差异蛋白。本课题利用蓝色温和胶双向电泳(2D-BN)技术对各组别的肝细胞细胞质基质进行差异蛋白复合体比较研究,找到9个差异复合体,鉴定出41个复合体蛋白组分;利用2D-BN对各组别的血浆样本进行差异蛋白复合体比较研究,找到1个差异复合体,鉴定出其中的11个蛋白组分。结果反映出在肝切除后4小时时间点肝细胞的代谢、物质运输和信号转导等方面的复杂变化,并提示一些蛋白如ROCK-1、M4蛋白等可能更多的分泌入血浆中并参与肝再生的引发。这些结果构成了迄今为止单次研究得到的最大的肝再生中差异蛋白质组数据库,为肝再生的引发机制研究提供了线索和依据。
Surgical resection of hepatic lobes or chemical injury can trigger hepatocyte replication and enlarge liver mass, this phenomenon was named for liver regeneration. The model of 2/3 hepatectomy in rat is the most widely used model and the closest approximation of pure liver regeneration unaccompanied by cellular injury. Liver regeneration is a pathophysiological process can be divided into priming stage, progression stage, cell cycle stage and termination stage. Among these, the priming stage is the most impotant and special stage. Hepatocytes can quickly change their state from G0 to G1 during this short stage. The mechanism of the priming of liver regeneration is still not very clear until now. Evident shows that plasma and intracellular factors both can play an important role during this stage. In the current research, in order to reveal the priming mechanism of liver regeneration, samples (plasma, hepatocyte total protein, hepatocyte cytosol protein) of the rats at 4 h after 2/3 partial hepatectomy (PHx) was studied by two-dimensional gel electrophoresis (2DE) and two-dimensional blue native gel electrophoresis (2D-BN). Normal and Sham-operation control samples were also studied. In the 2DE studies, 19 differential proteins were found and identified in hepatocyte total protein samples and 7 differential proteins were found and identified in plasma samples. In the 2D-BN studies, 9 differential protein complexes contains 41 protein components were found and identified in hepatocyte cytosol samples and 1 protein complex contains 11 protein components were found and identified in plasma samples. The results reflect the complex changes of metabolism, substance transportation and signal transduction inside the hepatocytes at 4h after partial hepatectomy. Some of the results suggest that some protein such as ROCK-1 and M4 protein might be increasingly secreted into plasma and involved in the priming of liver regeneration. To date, the results constitute the largest database of proteome changes during liver regeneration from a single study. These finding give us the important clues for the study of the priming of liver regeneration.
Surgical resection of hepatic lobes or chemical injury can trigger hepatocyte replication and enlarge liver mass, this phenomenon was named for liver regeneration. The model of 2/3 hepatectomy in rat is the most widely used model and the closest approximation of pure liver regeneration unaccompanied by cellular injury. Liver regeneration is a pathophysiological process can be divided into priming stage, progression stage, cell cycle stage and termination stage. Among these, the priming stage is the most impotant and special stage. Hepatocytes can quickly change their state from G0 to G1 during this short stage. The mechanism of the priming of liver regeneration is still not very clear until now. Evident shows that plasma and intracellular factors both can play an important role during this stage. In the current research, in order to reveal the priming mechanism of liver regeneration, samples (plasma, hepatocyte total protein, hepatocyte cytosol protein) of the rats at 4 h after 2/3 partial hepatectomy (PHx) was studied by two-dimensional gel electrophoresis (2DE) and two-dimensional blue native gel electrophoresis (2D-BN). Normal and Sham-operation control samples were also studied. In the 2DE studies, 19 differential proteins were found and identified in hepatocyte total protein samples and 7 differential proteins were found and identified in plasma samples. In the 2D-BN studies, 9 differential protein complexes contains 41 protein components were found and identified in hepatocyte cytosol samples and 1 protein complex contains 11 protein components were found and identified in plasma samples. The results reflect the complex changes of metabolism, substance transportation and signal transduction inside the hepatocytes at 4h after partial hepatectomy. Some of the results suggest that some protein such as ROCK-1 and M4 protein might be increasingly secreted into plasma and involved in the priming of liver regeneration. To date, the results constitute the largest database of proteome changes during liver regeneration from a single study. These finding give us the important clues for the study of the priming of liver regeneration.
引文
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