共培养体系中阿仑膦酸钠对破骨细胞相关基因表达的影响

The influence of alendronate on the expression of osteoclast associated genes in an osteoblast-osteoclast co-culture system

目的:研究阿仑膦酸盐(ALN)对成骨细胞(OB)-破骨细胞(OC)共培养体系中破骨细胞生成、功能及相关基因表达的影响。方法:胰酶消化法培养鼠颅骨成骨细胞,并与骨髓细胞建立OB-OC共培养体系;体系中加入10-6mol/L PGE2、10-8mol/L VitD3诱导破骨细胞分化生成。实验分对照组和ALN(10-6mol/L)处理组。采用TRAP染色及扫描电镜检测破骨细胞生成及牙本质磨片吸收陷窝情况,并应用Real-time PCR检测破骨细胞相关基因NFATc1、c-Fos、RANKL、OPG的基因表达。结果:各组细胞均有TRAP阳性多核OC生成,并在牙本质磨片上形成吸收陷窝;但对照组所获TRAP阳性多核细胞数目、吸收陷窝数目及面积均显著大于ALN组。Real-time PCR检测结果显示对照组NFATc1、c-Fos、RANKL、OPG的基因表达均显著高于ALN组。结论:ALN在OB-OC共培养体系中可抑制OC的生成及其骨吸收功,并下调OC相关基因NFATc1、c-Fos、RANKL、OPG的基因表达。

Objective： To study the influence of alendronate on osteoclastogenesis, bone resorption and expression of osteoclast-as- sociated genes in an osteblast-osteoclast eo-euhure system. Methods： Mice calvarial osteoblasts were harvested by trypsin digestion and bone marrow cells were obtained. The cells were co-cultured in ct-MEM, PGEa （10-6 M） and VitD3 （10-s M） were added in the cultures to induce osteoclastogenesis. The cells treated by ALN（ 10-6 mol/L） were in ALN group, and those without ALN in control group. Osteoclastogenesis and their resorption function were examined by TRAP staining and scanning electron microscope （ SEM ） observation of dentin resorption lacunaes. Gene expressions of NFATcl, c-Fos, RANKL and OPG were detected by Real-time PCR. Results： TRAP positive multinuclear cells and resorption lacunaes were observed in both groups. However, control group showed more TRAP positive multinuclear cells and more resorption lacunaes than ALN group. Real-time PCR detection showed that gene ex- pressions of NFATcl, c-Fos, RANKL and OPG were higher in control group than in ALN group. Conclusion： ALN can inhibit oste- oclastogenesis, resorption function, and gene expression of osteclast-associated genes in the osteoblast-osteoclast co-culture system.