骨唾液酸蛋白对破骨细胞前体RAW264．7增殖、分化作用的信号通路

摘要

Objective To explore the key signaling pathways and molecules of bone sialoprotein (BSP) to promote proliferation and differentiation of preosteoclasts RAW264.7. Methods RAW264.7 cells were treated with BSP and the inhibitors of signaling pathway molecules. MTS and TRAP staining kits were used to evaluate the effects of proliferation and differentiation. The activity assay kits of Calcineurin, AKT, JNK, ERK and p38 were used to detect their activities. Results Inhibiting ERK and p38 can inhibit cell proliferation induced by BSP significantly, while inhibiting AKT, Calcineurin and PI3K can reduce the role of promoting the differentiation by BSP. With increasing treatment time of BSP, the activity of Calcineurin in RAW264.7 cells increased. Furthermore, the activity of AKT was maximum at 48 h after treated with BSP, while the activity of JNK, ERK and p38 were maximum at 24 h after treated with BSP. Conclusions BSP mainly regulates the ERK and p38 of MAPK pathway to promote RAW264.7 cell proliferation, and regulates AKT, PI3K and Calcineurin pathways to promote RAW264.7 cell differentiation.%目的：探讨骨唾液酸蛋白（BSP）促进破骨细胞前体RAW264．7增殖和分化的关键信号通路与分子。方法：培养RAW264．7细胞，加BSP及信号通路相应抑制剂处理后，用MTS检测细胞的增殖，用TRAP染色试剂盒检测细胞的分化，用Calcineurin、AKT、JNK、ERK和p38活性检测试剂盒检测其活性。结果：抑制ERK和p38分子可显著抑制BSP促细胞增殖作用；抑制AKT、PI3K、Calcineurin分子可显著降低BSP的促分化作用；随着BSP处理时间的增加，RAW264．7细胞内Calcineurin活性增加；AKT活性在BSP作用48 h时最大，JNK、p38和ERK活性则在BSP作用24 h时最大。结论：BSP主要通过调节细胞内MAPK通路中的ERK和p38促进RAW264．7细胞的增殖，通过调节AKT、PI3K、Calcineurin通路促进RAW264．7细胞的分化。