【摘要】 目的:研究肝素十二糖(dp12)对血管平滑肌细胞(vascular smooth muscle cells,VSMCs)增殖作用的影响,并分析其分子水平的作用机制。方法:通过以10%新生牛血清诱导牛胸主动脉血管平滑肌细胞增殖建立模型,然后考察实验室精制的不同浓度的肝素寡糖对血管平滑肌细胞增殖作用的影响;四甲基偶氮唑盐法(MTT)检测dp12对VSMCs增殖的影响;流式细胞仪检测细胞周期分布;RT-PCR法检测ERK1/2转录变化情况;Western Blotting法检测ERK1/2和p-ERK1/2的表达。结果:MTT结果显示不同浓度的dp12可以明显抑制由10%新生牛血清诱导的血管平滑肌细胞的增殖;细胞周期实验揭示dp12抑制血清诱导的VSMCs从G1期进入S期,影响细胞周期;dp12通过下调ERK基因的转录进而下调ERK1/2的表达,此外dp12抑制ERK1/2的磷酸化进而影响细胞增殖。结论:dp12使VSMCs在G1期/S期阻滞,通过抑制ERK基因的转录和ERK蛋白的磷酸化抑制VSMCs增殖,可能是dp12抗VSMCs增殖的机制之一。更多还原

【Abstract】 AIM: To investigate the effects of 12-degree of polymerization heparin oigosaccharid(dp12) on proliferation in vascular smooth muscle cells(VSMCs) and to analyze their activated signal transduction pathways. METHODS: In this study,the proliferation of VSMCs of bovine thoracic aorta was induced by 10% NCS and the effects of dp12 on VMSCs proliferation followed upon different concentrations.MTT assay was evaluated to indicate the proliferation of VSMCs;flow cytometry was used to detect the cell cycle;the ERK mRNA and phosphor protein of ERK were detected by reverse transcription polymerase chain reaction(RT-PCR) and western-blotting respectively.RESULTS: MTT assay showed the inhibitory effects of dp12 on VSMCs induced by 10% NCS;flow cytometry revealed that the inhibited proliferation was a result of the interference of VSMCs entering S phase from G1 phase;western-blotting and RT-PCR profiles proved that dp12 inhibited the transcription and phosphorylation of ERK1/2.CONCLUSION: 12-Degree of polymerization heparin oigosaccharid inhibits the VSMCs entering S phase from G1 phase,it also inhibits the transcripttion and phosphorylation of ERK1/2,which might be the putative anti-proliferation mechanism of VSMCs.更多还原