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

1.平滑肌细胞源性迁移因子（SDMF）的纯化：从培养的大鼠主动脉平滑肌细胞（SMC）的100 L条件培养基中，通过四步柱纯化SDMF。SDMF的分子量为58 KD，等电点为10.0。SDMF对SMC和迁移具有特异性（无促有丝分裂活性）。SDMF的最大活性是PDGF的4倍。SDMF基因的克隆：纯化后的SDMF经酸裂解，产生35 KD和25 KD的片段，其中35 KD片段的N端被封闭。对25 KD片段的N端10个氨基酸进行了检测，并以此为基础合成了cDNA探针。SDMF基因克隆正在进行中。SDMF表达的调控：SDMF在合成型SMC中表达，而在收缩型SMC中不表达，在合成型SMC中，动脉粥样硬化内膜来源的SMC比正常中膜来源的SMC表达高1.5倍。在测试的各种细胞因子中，只有TGF-β抑制SDMF的表达。SDMF抑制剂：在测试的内源性细胞因子中。PDGF-AA、TGF-β和C型利钠肽以浓度依赖性方式抑制SDMF刺激的SMC迁移，在所测试的钙拮抗剂中，只有硝苯地平抑制SDMF活性。

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.