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为考察肝素寡糖对血管平滑肌细胞增殖的抑制作用,并探索其相关细胞内信号转导机制,采用cell-based ELISA法、RT-PCR法、Western blotting法和免疫细胞化学法检测细胞内PKC-α蛋白水平和mRNA水平;采用RT-PCR法检测c-jun、c-myc和c-fos等3种原癌基因的mRNA水平。结果显示,肝素寡糖可以抑制新生牛血清诱导的PKC-α和原癌基因的表达,这可能是肝素寡糖抑制血管平滑肌细胞增殖的机制之一;流式细胞实验结果显示,肝素寡糖能将细胞阻滞于G0/G1期,从而阻断细胞周期进程。综上所述,肝素寡糖可能通过抑制PKC-α表达,进而抑制原癌基因的表达,阻断细胞G1/S期转换,从而抑制血管平滑肌细胞的增殖。
To investigate the inhibitory effect of heparin oligosaccharides on the proliferation of vascular smooth muscle cells and to explore the relevant intracellular signal transduction mechanism, cell-based ELISA, RT-PCR, Western blotting and immunocytochemistry were used to detect the intracellular PKC- α protein levels and mRNA levels were detected by RT-PCR. The mRNA levels of c-jun, c-myc and c-fos were detected by RT-PCR. The results showed that heparin oligosaccharides could inhibit the expression of PKC-α and proto-oncogenes induced by newborn calf serum, which may be one of the mechanisms of heparin oligosaccharides inhibiting the proliferation of vascular smooth muscle cells. Flow cytometry results showed that heparin oligosaccharide The cells are arrested in the G0 / G1 phase, thereby blocking the cell cycle progression. In summary, heparin oligosaccharide may inhibit the proliferation of vascular smooth muscle cells by inhibiting the expression of PKC-α, thereby inhibiting the expression of proto-oncogene, blocking the G1 / S phase transition.