NO供体对大鼠成骨细胞增殖分化及BMP-2表达的影响

摘要

Objective:To study the effects of nitric oxide( NO) on the proliferation differentiation and expression of bone morphogenetic protein ? (BMP -2) in rat osteoblasts. Methods-. Osteoblasts were isolated from calvarial bone of SD rats by combined means of enzymatic digestion and fragments. Then the cultured cells were added in NOC - 18 of concentration in 0, 1, 10, 100 umol,and the cell proliferation was measured by thiazoly blue (MTT) assay. The protein expression of BMP -2 was determined by immunocytochemisty. Results; NOC ?18 had significant proliferation effect on osteoblasts and all the expression of osteoblasts BMP -2 were increased. Among the 3 groups,the group with NOC - 18 10 umol had the strongest osteoblast proliferation. Conclusion; NO had dual effects on proliferaton and differentiation in rat osteoblasts. Moderate low concentration of NO could stimulate proliferation and differentiation of osteoblast and increase bone formation; high concentration of NO had weak effects on stimulating proliferation and differentiation of osteoblast, even inhibiting osteoblasts differentiation. The results showed that the effects of NO in osteoblast was correlated with the expression of BMP -2.%目的:研究NO对大鼠成骨细胞的增殖分化、碱性磷酸酶(ALP)活性及对BMP-2表达的影响.方法:体外分离培养新生SD大鼠颅骨成骨细胞,加入不同浓度的NO供体(NOC-18),不同时间段采用噻唑蓝(MTT)比色法检测成骨细胞增殖,测定细胞ALP活性；应用免疫细胞化学法检测BMP-2的表达.结果:3个浓度组NO对成骨细胞均有显著的增殖作用,3组成骨细胞BMP-2表达均增加,其中以NOC-18 10 μmol组对成骨细胞增殖的作用最强.结论:NO对大鼠成骨细胞增殖和分化有双重调节作用,适量低浓度的NO能够刺激成骨细胞增殖分化,增强细胞的骨形成作用,提高成骨细胞ALP活性；高浓度NO对成骨细胞的增殖分化作用弱,NO对成骨细胞的作用与调节和细胞BMP-2的表达有关.