Development of specific inhibitors smooth muscle cell-derived migration factor (SDMF) and their clinical application

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1. Purification of smooth muscle cell derived migration factor (SDMF) : SDMF was purified from 100L of conditioned media dirived from cultured rat aortic smooth muscle cells (SMC) through four steps of columns. The molecular weight and isoelectric point of isoelectric point of SDMF were 58KD and PH 10.0, respectively. SDMF was specific to SMC and migration (no mitogenic activity) . The maximal activity of SDMF was four-fold that of PDGF.2. Cloning of SDMF gene : Purified SDMF was subjected to acid break down, producing the fragments of 35KD and 25KD.N-terminus of 35KD fragment was being blocked. 10 amino acids of N-terminus of 25KD fragment were detected and cDNA probes were synthesized from this amino acid information. The SDMF gene cloning is now in progress.3. Regulation of SDMF expression : SDMF was expressed in synthetic SMC but not in contractile SMC.Of synthetic phenotype, SMC derived from atheromatous intima expressed more SDMF than SMC derived from normal media by 1.5 fold. Among various cytokines tested, only TGF-beta inhibited expression of SDMF.4. Inhibitors of SDMF : Among endogenous cytokines tested. PDGF-AA,TGF-beta, and C-type natriuretic peptide inhibited migration of SMC stimulated by SDMF in a concentration dependent manner.Among calcium antagonists tested, only nifedipine was inhibitory of SDMF activity.