骨形态发生蛋白2对成牙骨质细胞中硬化蛋白表达调控机理的研究

目的 探索成牙骨质细胞OCCM-30中骨形态发生蛋白2（BMP2）对硬化蛋白（SOST）表达的调控机制。方法 用2种质量浓度的BMP2（50、100 ng·mL^-1）处理成牙骨质OCCM-30细胞3、5、7 d，相同体积的PBS液为对照组，采用实时荧光定量聚合酶链反应（RT-PCR）、免疫印迹法检测SOST mRNA和蛋白的表达情况。将OCCM-30细胞分为5组：空白对照组、BMP2组、BMP2+dorsomorphin组、BMP2+SB202190组、BMP2+PD98059组，根据分组分别加入100 ng·mL^-1的BMP2和相应的试剂共培养，于3、5 d时检测SOST mRNA和蛋白的表达情况。结果 100 ng·mL^-1 BMP2对SOST表达的上调作用强于50 ng·mL^-1 BMP2，且有时间依赖性（P<0.05）。BMP2+dorsomorphin组、BMP2+ SB202190组、BMP2+ PD98059组的SOST mRNA水平和蛋白质水平均降低，其中BMP2+dorsomorphin组降低最明显（P<0.05）。结论 成牙骨质细胞中BMP2主要是通过Smad信号通路介导上调SOST的表达。

Preliminary research on the expression of sclerostin mediated by bone morphogenetic protein 2 in cementoblast

Objective This research explores the regulatory role of bone morphogenetic protein 2 (BMP2) in the expression of sclerostin in OCCM-30 cementoblast. Methods OCCM-30 cementoblasts were treated with 50 and 100 ng·mL^-1 BMP2 for 3, 5, and 7 days. SOST mRNA was detected by real-time quantitative polymerase chain reaction (RT-PCR). Western blot analysis was employed to detect the sclerostin levels in the nucleus. Five groups were prepared for the experiments: control, BMP2, BMP2+dorsomorphin, BMP2+SB202190, and BMP2+PD98059. OCCM-30 was pretreated with BMP2 for 3 and 5 days, and then the sclerostin and SOST mRNA levels were measured. Results RT-PCR and Western blot analyses showed that BMP2 upregulated the expression of SOST in a concentration-dependent manner. SOST expression increased with time (P<0.05). Moreover, sclerostin levels of BMP2+dorsomorphin, BMP2+SB202190, and BMP2+PD98059 groups were lower than that of the BMP2 group, and the sclerostin level in BMP2+dorsomorphin group was lowest (P<0.05). Conclusion The upregulation of SOST by BMP2 in OCCM-30 is mainly mediated by the BMP2/Smad signal pathway.