肝素寡糖通过抑制PKC-α影响血管平滑肌细胞增殖的作用(英文)

Effect of heparin-derived oligosaccharide on vascular smooth muscle cell proliferation through inhibition of PKC-α expression

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。

In this study,the effect of heparin-derived oligosaccharide（HDO） on bovine vascular smooth muscle cell（VSMC） proliferation and signal transduction mechanism involved were investigated.The levels of PKC-α protein and mRNA were determined by cell-based ELISA,RT-PCR,Western blotting and immunocytochemical methods.Meanwhile,mRNA levels of c-jun,c-myc and c-fos were assayed by RT-PCR method.The results showed that HDO inhibited newborn calf serum（NCS）-induced expression of PKC-α and proto-oncogenes,which may be one of the mechanisms for the inhibition of VSMC proliferation by HDO.Flow cytometry analysis indicated that HDO blocked NCS-induced cell cycle progression by arresting cells at G0/G1 phase.The results imply that HDO inhibits VSMC proliferation by moderating the gene level of PKC-α,eventually inhibiting proto-oncogene mRNA expression and blocking G1/S transition.