Effect of icariin and curculigoside on inhibition of osteoclastic formation, differentiation and bone absorption
-
摘要: 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-(OH)2Vitamin D3和地塞米松;或人巨噬细胞集落刺激因子(MCSF)和核因子κ B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。Abstract: 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.
-
Key words:
- icariin /
- curculigoside /
- osteoclasts /
- cytoskeleton /
- synergism
-
[1] 林 殷, 郭 华, 林 毅,等. 命门用药之特点初探(三)[J]. 北京中医药大学学报,2008,31(5):303. [2] 徐忠明,周志昆. 黄芪三仙汤对绝经后妇女骨密度和骨矿含量指标影响的研究[J]. 深圳中西医结合杂志,2009, 19(2):84. [3] 沈自尹. 淫羊藿总黄酮对应一个药靶的探索过程[J] 中药新药与临床药理. 2003,14(5):293. [4] Jiao L, Cao DP, Qin LP, et al. Antiosteoporotic activity of phenolic compounds from Curculigo orchioide[J]. Phytomedicine, 2009, 16:874. [5] 刘祖德,臧鸿声,欧阳跃平.新生大鼠颅骨成骨性细胞体外生长过程研究[J].解剖学报,1995, 26(2):157. [6] Li B,Yu S.Genistein prevents bone resorption diseases by inhibiting bone resorption and stimulating bone formation[J].Biol Pharm Bull,2003, 26(6):780. [7] Zhang Q,Qin L,He W,et al.Coumarins from Cnidium monnieri and their antiosteoporotic activity[J].Planta Med,2007, 73(1):13. [8] 鲍蕾蕾,秦路平,卞 俊,等. 巴戟天甲基异茜草素对破骨细胞性骨吸收的影响[J]. 解放军药学学报, 2009,25(6):508. [9] Stavros CM.Birth and death of bone cells:Basic regulatory mechanism and implications for the pathogenesis and treatment of osteoporosis[J].Endocrine Reviews,2000,21(2):115. [10] Mitra SK, Hanson DA,Schlaepfer DD.Focal adhesion kinase:in command and control of cell motility[J].Nat Rev Mol Cell Biol,2005,6:56. [11] Ory S, Brazier H, Pawlak G, et al. Rho GTPases in osteoclast:Orchestrators of podosome arrangement[J].Eur J Cell Biol,2008,87:469. [12] 姚志红. Rho GTPase在信号转导和细胞骨架中的作用[J].国际病理科学与临床杂志,2009,29(1):20. [13] Zhang Z,Neff L,Bothwell ALM,et al.Calcitonin induces dephosphorylation of pyk2 and phosphorylation of focal adhesion kinase in osteoclasts[J].Bone,2002,31(3):359. [14] Xiong WC, Feng X. Pyk2 and FAK in osteoclasts[J]. Frontiers in Bioscience, 2003, 8:1219. -
点击查看大图
计量
- 文章访问数: 3660
- HTML全文浏览量: 409
- PDF下载量: 595
- 被引次数: 0
下载:
