Genistein对破骨细胞分化和骨吸收功能的影响及其机制探讨

Effect of genistein on osteoclastic bone resorption and its mechanism

目的 探讨三羟异黄酮类药Genistein对破骨细胞性骨吸收的作用及其机制。方法1,2 5 (OH) 2 D3 诱导小鼠骨髓细胞形成破骨样细胞 ,分别加入 0mol/L、10 -9mol/L、10 -8mol/L、10 -7mol/L、10 -6mol/L、10 -5mol/LGenistein ,于培养 3,5和 8d分别计数抗酒石酸酸性磷酸酶阳性细胞个数 ;于培养 12d利用图像分析系统对骨吸收陷窝的面积进行测量分析。另从小鼠颅盖骨分离培养获得原代成骨细胞 ,分别加入 0 ,10 -8,10 -7,10 -6和 10 -5mol/LGenistein并以 10 -9mol/L 17β 雌二醇为对照 ,采用RT PCR的方法检测Genistein对骨保护因子 (osteoprotegerin ,OPG)及破骨细胞分化因子 (receptoractivatorofNF κBligand ,RANKL)mRNA表达的影响。结果 利用 1,2 5 (OH) 2 D3 成功诱导出破骨样细胞 ;随Genistein浓度增加 ,抗酒石酸酸性磷酸酶阳性细胞 (单核、双核和多核 )个数和骨吸收陷窝面积呈剂量依赖性减少。同时 ,应用Genistein后原代成骨细胞中OPG和RANKL的mRAN表达都有增强 ,但其最终效应表现为剂量依赖性和时间依赖性地增大OPG/RANKL的浓度比。结论 Genistein通过抑制破骨前体细胞分化来抑制其体外骨吸收功能 ,其分子机制与OPG/RANKLmRNA表达比值的升高有关。

Objective To investigate the effect of genistein, a legume-derived isoflavone,on the osteoclastic bone resorption and to clarify the mechanism underlying this action. Methods Primary osteoclast-like cells (OLCs) were isolated from 3 week-old mice and induced by 1,25(OH) 2D 3. Then OLCs were exposed to genistein at various concentration of 0,10 -9,10 -8,10 -7,10 -6 and 10 -5mol/L. The number of TRAP+cells were counted as well as the surface area of bone resorption on bone slice.Additionally the osteoblasts were obtained from 5 day old mice and were exposed to 0, 10 -8,10 -7,10 -6 and 10 -5mol/L genistein and 10 -9mol/L 17β-estradiol. The mRNA levels of RANKL and OPG were assayed by RT-PCR. Results In vitro, the number of TRAP+cells decreased depending on the concentration of genistein as well as the area of bone resorption. By semiquantitative RT-PCR, genistein increased the mRNA levels of OPG and RANKL, and finally enlarged the ratio between them in dose and time-dependent mamer. Conclusion Genistein inhibits OLCs' bone resorption through suppressing the differentitation of preosteoclasts, and the ratio between OPG and RANKL may be the key factor.