Preliminary Study Of Differential Membrane Proteomics On Sinusoidal Surface During Rat Liver Regeneration, Implication For Angiogenesis

Background: Liver regeneration is a complicated processes with multiple stages, and regulated by several signaling pathways and multi-factors. Angiogenesis is the prerequisite for the generation of all the organs including liver. Hepatic sinusoid, as specialized capillary network, its reconstruction in the live generation process involves in the cell division of vessel endothelial cell and other cell types. The proliferation, migration, adherence and alignment until eventually the formation of the whole hepatic sinusoid must be related to the changes of the plasma membrane proteins on the hepatic sinusoid endothelial cell. Membrane proteomics is a very useful way to reveal the mechanism by detecting and identifying the proteins related to liver regeneration and angiogenesis.Aim: Proteomics methods have been used to study the dynamic changes of the plasma membrane proteins of the hepatic sinusoid endothelial cells. Through the further screening and data analysis, important plasma membrane proteins related to the hepatic sinusoid reconstruction were found and therefore provide the basis for the further study on the mechanism of angiogenesis.Method: Liver regeneration model was made by 2/3 hepatectomy on rat. 3 days after surgery, in vivo silica perfusion was performed both on the hepatectomy group and sham group. Plasma membranes proteins were extracted and separated on 16-BAC/SDS-PAGE electrophoresis. Different expressed protein spots were subjected to in gel digestion, and further identified by HCT Mass Spectrometry.Results: The observation under electro microscope with high resolution confirmed that the silica perfusion method is very effective way for the purification of hepatic sinusoid plasma membrane. MS data also suggested the successful separation of membrane proteins on 16-BAC/SDS-PAGE system. After MS identification and database searching, 11 up-regulated proteins and 8 down-regulated proteins related to hepatectomy had been identified. The up-regulation and down-regulation of these proteins was in response to the change in hepatic sinusoid.Conclusion: Our data showed that there were several endothelial cell plasma membrane proteins or membrane-related protein involved in the reconstruction process of hepatic sinusoid. It suggested that differential proteomic study was a useful way to reveal the mechanism of comprehensive physiological process.