淫羊藿苷对破骨细胞活性的影响

目的:观察淫羊藿苷对破骨细胞骨吸收及凋亡的影响,探讨淫羊藿苷的抗骨质疏松作用机制。方法:体外分离、培养兔破骨细胞,与玻片及骨磨片共同培养,用10-7、10-6、5×10-6、10-5mol/L浓度的淫羊藿苷刺激破骨细胞,倒置相差显微镜下观察活体细胞、HE染色、TRAP染色及骨吸收陷窝甲苯胺蓝染色,鉴定破骨细胞,并进行骨吸收陷窝计数和面积测量,吖啶橙染色观察凋亡破骨细胞所占的比例。结果:与空白对照组比较,10-6、5×10-6、10-5mol/L浓度的淫羊藿苷组破骨细胞凋亡率均明显增高,骨吸收陷窝数目、面积明显减少,随浓度增加抑制作用增强,差异有显著性意义(P<0.05)。结论:淫羊藿苷可诱导破骨细胞凋亡,抑制骨吸收,并随浓度增加抑制作用增强。

Effects of Icariin on osteoclastic bone resorption and apoptosis in vitro

Objective:To observe the effect of Icariin on osteoclastic bone resorption and apoptosis in vitro and explore the mechanism of Icariin in preventing osteoporosis.Methods: Osteoclasts were seperated from long-limb bones of new born rabbits,cultured with glass slices and bone slices,and treated by 10^-7,10^-6,5×10^-6,10^-5 mol/L Icariin.Osteoclasts were identified by observing live cells with phase contrast microscope,HE staining,TRAP staining and toluidine blue staining of bone resorption pits.The number of bone resorption pits were counted as well as the surface area of bone resorption on bone slice.Osteoclasts were stained with acridine orange.The rates of apoptotic osteoclasts were observed under fluorescence microscope.Results:Comparing with the blank control group,the rates of apoptotic osteoclasts increased obviously and the number and area of bone resorption pits decreased evidently with increasing dose in those groups treated by 10^-6,5×10^-6,10^-5 mol/L Icariin.The differences between control group and those groups treated by 10^-6,5×10^-6,10^-5 mol/L Icariin were significant if P<0.05.Conclusion:Icariin can induce osteoclast apoptosis and inhibit bone resorption in dose-dependent manner.