摘要
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.
出处
《中国骨质疏松杂志》
CAS
CSCD
2004年第3期272-276,共5页
Chinese Journal of Osteoporosis
基金
国家自然科学基金资助项目 ( 3 0 2 714 12 )
北京大学"985"专项基金资助项目
