淫羊藿苷和仙茅苷协同抑制破骨细胞的形成、分化和骨吸收功能

目的 研究淫羊藿苷和仙茅苷配伍抑制破骨细胞形成、分化和骨吸收的相互作用关系.方法 用1,25-(0H)2Vitamin D3和地塞米松；或人巨噬细胞集落刺激因子(MCSF)和核因子κB受体活化因子配体(RANKL)诱导骨髓单核细胞使其分化为破骨细胞抗酒石酸酸性磷酸酶(TRAP)染色进行阳性破骨细胞鉴定；磷酸苯二钠法测定抗酒石酸酸性磷酸酶活性；将破骨细胞和骨片共同培养,以计算机图像分析测定骨吸收陷窝的面积；以Hoechst 33258和罗丹明-鬼笔环肽染色,激光共聚焦显微镜观察破骨细胞肌动蛋白环(F-actin Ring)的形态；用Western-blot分析细胞骨架相关蛋白的表达.结果 淫羊藿苷和仙茅苷在1×10-6mol/L浓度下可协同抑制破骨细胞的形成、降低抗酒石酸酸性磷酸酶的活性,减少破骨细胞在骨片上形成的骨吸收陷窝面积,抑制破骨细胞骨架F-actin环的构建及其调控因子Rho GTPases和灶性黏附激酶(focal adhesion kinase,FAK)的表达.结论 淫羊藿苷和仙茅苷协同抑制破骨细胞性的骨吸收,为药对淫羊藿仙茅的配伍提供了实验依据.

Effect of icariin and curculigoside on inhibition of osteoclastic formation, differentiation and bone absorption

Objective To investigate the interaction relationship of icariin and curculigoside on the osteoclastic formation, differentiation and bone resorbing.Methods Multinucleated osteoclasts were induced by 1, 25-dihydroxyvitamine D₃ and dexamethasone from bone marrow cells with the presence of primary osteoblastic cells, or induced with RANKL and MCSF.The activity of TRAP was measured by p-nitrophenyl sodium phosphate assay.The bone absorption pit area on the bone slices formed by osteoclast was measured by computer image processing.The F-actin ring were stained with Hoechst 33 258 and rhodamine-phalloidin, and then observed under the confocal scanning laser microscopy. The Western-blot was used to analysis the expression of protein which was related to the cytoskeleton of osteoclasts. Results Icariin and curculigoside at the concentration of 1×10^-6mol/L synergistically inhibited the osteoclastic formation, decreased the activity of TRAP, and reduced the area of bone resorption pit on bone slices, inhibited the organization of F-actin ring and the expression of FAK and Rho GTPases. Conclusions Icariin and curculigoside synergistically inhibited the osteoclastic bone resorption, which provided scientific basis for the theory of the compatibility of traditional Chinese medicine Epimedii Folium and CurculiginIs Rhizoma.