Comparative Proteomic Study Of The Proteins Related To Hcy Induced Proliferation Of Vascular Smooth Muscle Cells

Object:Comparative proteomic analysis method was used to explore the differential expressed protein in vascular smooth muscle cells (VSMC) induced by Homocysteine (Hcy), for clarifying the mechanism and the protein basis of proliferation. Methods: FACS and MTT were used to observe the effect of Hcy on the proliferation of VSMC. For exploring VSMC proteins correlated with Hcy-induced proliferation, two-dimensional electrophoresis (2-DE) and MALDI-TOF mass spectrometry were applied, obtained piptide finger print were queried in NCBInr database. The function of some differential expressed proteins, such as aldose reductase and pyruvate kinase were conformed their activity by biochemical test. Results: Hcy induces the proliferation of VSMC in a dose and time dependent manner. There are total 1019+ 52 protein spots detected on the 2-DE images of VSMC, the match rate of all images were 84.29%. After comparing the protein profiles of VSMC between Hcy induced group and control group, 11 protein spots were found altered stabilized and distinctly, 9 of them has been identified by MALDI-TOF mass spectrometry,. These identifid proteins were laminC LIM and SH3 protein 1,similar to Wdr1 protein, Aldose reductase,Pyruvate kinase M2,Tpil protein ,calreticulin and two of them were Vimentin. Biochemical tests for glycolytic metabolism in mitochondria also supported the results from proteomic analysis. Conclusion: Among these altered proteins,glycolytic metabolism proteins, cytoskeleton proteins, structural domain proteins and calreticulin are involved. These findings provided clues for understanding the mechanism of proliferation effect on VSMC.