唑来膦酸对共培育体系中atp6v0d2基因表达及破 - 第三军医大学学报.doc

唑来膦酸对破骨细胞生成及Syk基因表达的影响 王宏利，李 鹏，刘 强，董 伟，戚孟春，李金源 （063000 唐山，河北联合大学口腔医学院口腔颌面外科教研室） [摘要] 目的唑来膦酸 ZOL)在共培养体系中对破骨细胞(osteoclastOC)生成的影响。方法小鼠颅骨成骨细胞与单核巨噬细胞RAW264.7共培5×10-7mol/L ZOL处理24 h组（ZOL组）。应用TRAP染色、牙本质吸收陷窝检测OC生成及骨吸收情况；Real-time PCR、Western blot、免疫荧光化学检测Syk基因表达。结果 细胞均有TRAP阳性多核OC生成，并在牙本质磨片上形成吸收陷窝；。结论OC生成和骨吸收功能，表达[关键词] 唑来膦酸Spleen tyrosine kinase in a osteoclast-osteoblast co-culture system in vitro Wamg Hongli, Li peng, Liu Qiang, Dong Wei, Qi Mengchun, Li Jinyuan (College of Stomatology, Hebei United University, Tangshan, Hebei Province ,063000,China) [Abstract] Objective To explore the effect of zoledronate acid (ZOL) on gene expression of spleen tyrosine kinase (Syk) and osteoclastogenesis in an osteoclast-osteoblast co-culture system. Methods Mice calvarial osteoblasts and RAW264.7 cells were co-cultured and the cells were divided into 2 groups. Group A served as control and group B received ZOL treatment (5×10-7mol/L) for 24h. TRAP staining and dentin resorption lacunae examination were used to detect osteoclast formation and resorption function, while Real-time PCR, Western blot and immunofluorescent chemistry were applied to examine Syk gene expression. Results TRAP positive cells and dentin resorption lacunae were observed in both groups. However, group B showed less TRAP positive multi-nucleated OC and resorption lacunae when compared with group A (p0.01). Syk mRNA and protein were also significantly down-regulated in group B (p0.01). Immunofluorescent examination showed that Syk was strongly expressed in cytoplasm of group A and formed actin-like rings at cell periphery; however, it was only weakly expressed in group B and no actin-like rings formed. Conclusion In the co-culture system, ZOL could significantly inhibit formation and resorption function of multi-nucleated osteoclasts and this may be attributed, at least partly, to its inhibition on gene expression of Syk. [Key words] Zoledronate; osteoclastogenesis; Spleen tyrosine kinase, tartrate-resistant