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目的以新生大鼠颅盖骨成骨细胞和由骨髓单核细胞诱导的破骨细胞为模型,观察仙茅代表性酚苷类成分仙茅苷、仙茅素A、苔黑酚葡萄糖苷和苔黑酚龙胆二糖苷的抗骨质疏松作用。方法 MTT法测定成骨细胞的增殖;磷酸苯二钠法测定成骨细胞碱性磷酸酶(alkaline phosphatase,ALP)和破骨细胞抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)的活性;茜素红染色观察成骨细胞骨矿化结节的形成;TRAP染色测定破骨细胞的数目;罗丹明-鬼笔环肽染色和激光共聚焦显微镜观察成骨细胞细胞骨架和破骨细胞肌动蛋白环的结构和形态;将破骨细胞与骨片共同培养,计算机图像处理测定破骨细胞在骨片上形成的骨吸收陷窝的面积。结果仙茅苷在10-9和10-8 mol/L的浓度下可促进成骨细胞的增殖(P<0.05),10-7~10-5 mol/L浓度时抑制破骨细胞TRAP的活性(P<0.05)。仙茅苷、苔黑酚葡萄糖苷和苔黑酚龙胆二糖苷可减少破骨细胞的数目,抑制破骨细胞的形成(P<0.05)。仙茅苷在10-10 mol/L,仙茅素A、苔黑酚葡萄糖苷和苔黑酚龙胆二糖苷在10-9 mol/L浓度时均可增加成骨细胞ALP的活性和骨矿化结节的形成(P<0.01),在一定程度上使1,25-二羟维生素D3损伤的成骨细胞细胞骨架的结构得以恢复;减少破骨细胞在骨片上形成的骨吸收陷窝面积,破坏破骨细胞伪足和F-actin的结构。结论仙茅酚苷类成分仙茅苷、仙茅素A、苔黑酚葡萄糖苷和苔黑酚龙胆二糖苷均可促进成骨细胞的骨形成,抑制破骨细胞的骨吸收,具有显著的抗骨质疏松作用。
OBJECTIVE: To observe the effects of curculigo, curculigin A, cethenol glucoside and moss Anti-osteoporosis effect of hematoporphyrins glycosides. Methods The proliferation of osteoblasts was determined by MTT assay. The activity of alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP) Alizarin red staining was used to observe the formation of osteoblasts in osteoblasts. The numbers of osteoclasts were determined by TRAP staining. The osteoblasts and osteoclasts were observed by rhodamine-phalloidin staining and confocal laser scanning microscope The structure and morphology of the protein ring; co-culture of osteoclasts and bone slices, and the area of bone resorption lacuna formed on osteoclasts by computer image processing. Results Curculigin promoted the proliferation of osteoblasts at the concentrations of 10-9 and 10-8 mol / L (P <0.05), and inhibited the TRAP activity of osteoclasts at the concentration of 10-7-10-5 mol / L (P <0.05). Curculigin, cethenol glucosidase and oroghenol gentiobioside reduced the number of osteoclasts and inhibited the formation of osteoclasts (P <0.05). Curcumin increased the activity of ALP in osteoblasts at 10-10 mol / L, curculigin A, orcinol and tocopherol gentiobioside at the concentration of 10-9 mol / L and bone mineral (P <0.01). The structure of osteoblast cytoskeleton injured by 1,25-dihydroxyvitamin D3 was restored to a certain extent and the bone resorption lacunae formed on osteoclasts were reduced , Disrupts the osteoclast pseudopodia and the structure of F-actin. Conclusions Curculigoside, curculin A, orcinoliglucoside, and tighenol gentiobioside all promote osteoblast formation and inhibit bone resorption of osteoclasts, which is significant Anti-osteoporosis effect.