| Surgical resection of hepatic 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 of physiological angiogenesis and the closest approximation of pure liver regeneration unaccompanied by cellular injury. Because the liver is composed of multiple cell populations, rebuilding of the liver after 70% partial hepatectomy (PH) requires the synergistical actions of various liver cell types. It is well documented that liver sinusoidal endothelial cells (LSECs) are the largest number of liver non-parenchymal cells and the second largest number among all resident liver cells next to hepatocytes, and play a pivotal role in liver regeneration (LR), especially in the angiogenesis of regeneration. After resection of the liver, formed the liver cells Island without blood vessels firstly, which posses the normal organizational structure only after the formation of the liver sinusoidal endothelial cells. It is difficult to illustrate the whole variation of liver regeneration in gene level, as the variation of gene expression is mainly in the quality rather than the emergency of new gene expression. The study of proteomics posses the characteristics of large-scale, high-throughput, high sensitivity, which is appropriate to the whole study of liver regeneration. Given the important role of liver sinusoidal endothelial cells in liver regeneration as well as the advantages of the subcellular proteomics, the study is going to explain the mechanism of liver regeneration and angiogenesis via the comparison of proteomics between the group of surgical (PHx) and sham operation (SHam). Were use the combined means of Cationic colloidal silicon perfusion and sucrose density gradient centrifugation to isolate plasma membrane of the liver sinusoidal endothelial cells (LSECs). Then the plasma membrane protein were separated by SDS-PAGE and identified by tandem mass spectrometry(LC-MS/MS). We identified 74 differentially expressed proteins from the two models. Compared to the sham operation control group,47 proteins were up-regulated and 27 proteins were down-regulated at 72 h after 70% PHx, which were categorized into functional groups including angiogenesis, cell division, apoptosis, cell cycle control, cell movement, angiogenesis, cell differentiation, metabolic and inflammatory reaction.Glycosylation is one of the most important post-translational modifications in protein, which plays an extremely important role in vivo. The change in the quality of glycoproteins and glycosylation sites and even the structure of glycopeptides, which may affect the activities of cell directly or indirectly. Therefore, in the study of glycosylation and its changes about the protein of liver regeneration has a great significance. We pay close attention on the membrane of liver sinusoidal endothelial cells (LSECs) in the group of surgical (PHx) and sham operation (SHam),through trypsin digestion respectively under the same conditions and lectin affinity chromatography(LAC), identified by tandem mass spectrometry(LC-MS/MS) lastly. In the group of surgical (PHx),40 glycopeptides with 42 glycosylation sites were identified, which resulted in the identification of 30 membrane glycoproteins; In the group of sham operation (SHam),35 glycopeptides with 35 glycosylation sites were identified, which resulted in the identification of 27 membrane glycoproteins. And we found five peptides have been identified in the two groups of PHx and Sham, but their corresponding glycosylation has been changed. I believe that this significant discovery will help us to explore and study the liver regeneration,and afford important theoretical basis to us. |
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