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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: To observe the effect of osteoprotegerin on endothelial cell number and endothelial nitric oxide synthase (eNOS) and its effect on endothelial cells. Methods: Immortalized human endothelial cells were cultured routinely. After treated with different concentrations of osteoprotegerin (0, 0.5, 2.0, 4.0, 6.0 and 8.0 μg / L) for 48 h, The number of cells was detected by cell counting kit (CCK-8). The expression of eNOS protein was detected by immunocytochemistry. Quantitative expression of eNOS gene was detected by real-time quantitative PCR. Results: Compared with the control group (0.936 ± 0.146), the proliferation of endothelial cells was significantly increased in the 0.8 μg / L group (1.291 ± 0.200; P <0.05); the expression of eNOS protein (P <0.05), 6.0 μg / L and 8.0 μg / L (P <0.01), respectively, while it was significantly increased in the group of 2.0 μg / L, 4.0 μg / L and 4.0 μg / L , 6 · 0μg / L, 8 · 0μg / L group eNOS mRNA were significantly higher than the control group (P <0.01). CONCLUSION: Osteoprotegerin can promote the proliferation of endothelial cells in the range of human body concentration fluctuation. It may promote the repair of endothelial cells during the process of anti-atherosclerosis, and has a positive correlation with concentration gradient.