骨桥蛋白反义核苷酸对大鼠血管平滑肌细胞增殖的影响

Effect of osteopontin antisense nucleotide on proliferation of rat vascular smooth muscle cells

背景:骨桥蛋白在血管损伤后新生内膜中的表达明显上调,而且能促进血管平滑肌细胞和外膜细胞的增殖、迁移。目的:探索骨桥蛋白反义核苷酸对大鼠血管平滑肌细胞增殖的影响。方法:培养大鼠A10主动脉血管平滑肌细胞,通过MTT比色法测定骨桥蛋白反义核苷酸对平滑肌细胞增殖的影响,流式细胞仪测定骨桥蛋白反义核苷酸对平滑肌细胞周期分布的影响;通过RT-PCR方法检测血管平滑肌细胞转染骨桥蛋白反义核苷酸对增殖细胞核抗原表达的影响。结果与结论:骨桥蛋白反义核苷酸对血管平滑肌细胞的增殖有明显的抑制作用,随时间延长对细胞增殖抑制率下降。骨桥蛋白反义核苷酸主要作用于G1/S限制点,能阻止血管平滑肌细胞由G0/G1期向S期推进,从而将血管平滑肌细胞阻滞于G0/G1期,抑制血管平滑肌细胞的增殖。血管平滑肌细胞转染骨桥蛋白反义核苷酸后,增殖细胞核抗原的表达水平均较对照组降低(P<0.05)?因此,得出骨桥蛋白反义核苷酸通过阻止血管平滑肌细胞由G0/G1期向S期推进,从而抑制血管平滑肌细胞的增殖,进而抑制血管内膜增生。

BACKGROUND： The expression of osteopontin （OPN） is obviously up-regulated in neointimal formation after vessel damage and promotes the proliferation and migration of vascular smooth muscle cells （VSMCs） and adventitial cells. OBJECTIVE： To explore the effect of OPN antisense nucleotide on proliferation of rat vascular smooth muscle cells. METHODS： A10 aortic VSMCs of rat were cultured. The effects of OPN antisense nucleotide on proliferation of VSMCs were observed by MTT colorimetric assay. Flow cytometry was performed to track cell cycle progression. RT-PCR was done to detect the effect of VSMCs transfected OPN antisense nucleotide on the antigen expression of proliferated cells. RESULTS AND CONCLUSION： OPN antisense nucleotide significantly inhibited VSMCs proliferation, but the inhibitory rate was decreased with time prolonged. OPN antisense nucleotide blocked VSMCs cycle in G0/G1 phase, and inhibited the proliferation of VSMCs. Compared with the control group, the antigen expression of proliferated cells were decreased obviously in the OPN antisense nucleotide group （P 0.05）. Accordingly, OPN antisense nucleotide can block VSMCs cycle in G0/G1 phase, thus, inhibits the proliferation of VSMCs and suppresses the vascular intima hyperplasia.