目的:观察骨保护素对内皮细胞数量变化及内皮型一氧化氮合酶(eNOS)的影响,探讨其对内皮细胞的作用。方法:常规培养永生化的人内皮细胞,用不同浓度(0,0.5,2.0,4.0,6.0,8.0μg/L)的骨保护素作用细胞48h后,用细胞计数试剂盒(cellcount kit,CCK-8)来检测细胞的数量;免疫细胞化学检测eNOS蛋白的表达水平,实时定量PCR检测eNOS基因的定量表达。结果:与对照组(0.936±0.146)相比,在8.0μg/L组的内皮细胞增殖量明显增加(1.291±0.200;P〈0.05);eNOS蛋白表达在2.0μg/L、4.0μg/L组(P〈0.05)和6.0μg/L、8.0μg/L组(P〈0.01)均显著增加;而4.0μg/L、6.0μg/L、8.0μg/L组的eNOS mRNA较对照组均呈显著增高(P〈0.01)。结论:骨保护素在人体的浓度波动范围内具有促进内皮细胞增殖效应,在抗动脉粥样硬化过程中,可能具有促内皮细胞修复作用,且与浓度梯度有明显的正相关性。
Objective:Observe the effects of osteoprotegerin (OPG) on the changes of endothelial cell numbers and nitric oxide synthase (eNOS), in order to investigate the influence of OPG on endothelial cells.Methods:The conventional cultures of immortalized human endothelial cells, with different concentration of OPG (0, 0.5, 2.0, 4.0, 6.0, 8.0 μg/L) cultivating for 48 h, were detected the number of cells with the cell count kit (CCK-8), eNOS protein expression with immunocytochemical method, eNOS mRNA levels with real-time quantitative PCR.Results:compared to the control group (0.936±0.146), the cell number of 8.0 μg/L group (1.291±0.200) was evidently elevated (P〈0.05); compared to the control group, eNOS protein expression in 2.0 μg/L, 4.0 μg/L group (P〈0.05) increased significantly, and increased more remarkably in 6.0 μg/L, 8.0 μg/L groups (P〈0.01); in 4.0 μg/L, 6.0 μg/L, 8.0 μg/L groups eNOS mRNA were significantly increased (P〈0.01) compared to the control group as well. Conclusion:the concentrations of OPG in human ranges promoted endothelial cell proliferation. As the anti-atherosclerotic factor, OPG may promote endothelial cells' repairation, and had evidently positive correlation with the concentration gradient.