Elucidation of the mechanism of caediovascular remodeling and development of therapy by proteomic analysis

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The objective of this study is to elucidate the mechanism of cardiovascular remodeling and the development of its therapy. By introducing research proteomics, and, mainly on transcription factor KLF5 which had an important role for the cardiovascular remodeling which we identified in the past, we perform a comprehensive isolation of the co-factors of the factor which affect the pathogenic process of the cardiovascular diseases, a functional analysis by the loss of function system. This study finally elucidates transcription control mechanism by the transcription factor network in the nucleus which assumed KLF5 a model and aims for developing the prevention of the organ remodeling and a cure. PDGF-A promotes a migration/the increase of the smooth muscle cell, and it is a growth factor playing an important role for the onset of the atherosclerosis. It was found to be a target gene of KLF5 by the examination of our past, and it became clear that -71 to -55bp domains were important in cardiovascular remodeling. At first, as a concrete plan in this study, we identify protein which bind -71 to -55bp domains of PDGF-A gene. I follow screening of the KLF5 binding protein basically. To be concrete, we composed the double-stranded DNA of -71 to -55bp domains that were the KLF5 binding site of the PDGF-A promoter and biotinylated the one end. Using streptoavidin and a strong bond of the biotin, we fixed it on the metal beads surface. I reacted the nuclear extract of the cell in this, and pull down. Then, we developed SDS-PAGE and identified a specific band by in-gel trypsin digestion method. Some interesting factors were identified. Based on the biochemical characteristic of the KLF5 binding factor, I pursued the pathogenic mechanism from in the animal level to the cell level. Above all, new mechanism in the defense mechanism of the cardiovascular tissues from oxidation stress became clear.