| 摘要: |
| 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10-8mol/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10-10mol/L,仙茅素A、苔黑酚葡萄糖苷和苔黑酚龙胆二糖苷在10-9mol/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。 |
| 关键词: 仙茅 酚苷类化合物 成骨细胞 破骨细胞 |
| DOI：10.16781/j.0258-879x.2016.05.0562 |
| 投稿时间：2015-11-18修订日期：2016-02-22 |
| 基金项目:国家自然科学基金(81274152),上海市科委科研计划项目(13401900102). |
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| Phenolic glycosides in Curculigo orchioides promotes osteoblastic bone formation and inhibits osteoclastic bone resorption |
| ZHANG Nai-dan△,JIANG Yi-ping△,XUE Li-ming,HAN Ting,ZHANG Qiao-yan,XIN Hai-liang* |
(Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, Shanghai 200433, China
△Co-first authors
*Corresponding author) |
| Abstract: |
| Objective To observe the anti-osteoporotic activity of representative phenolic glycosides (curculigoside, curculigine A, orcinol glucoside and orcinol rhamnoside) in Curculigo orchioides on osteoblasts in neonatal rat calvaria and on osteoclasts induced by myelomonocytes. Methods The proliferation of osteoblasts and osteoclasts were determined by MTT assay. The alkaline phosphatase (ALP) activity of osteoblasts and tartrate-resistant acid phosphatase (TRAP) of osteoclasts were detected by using p-PNPP-Na method. The mineralized nodule formation was determined by alizarin red-s (AR-S) staining. TRAP-positive multinucleated cells were counted through TRAP staining. The cytoskeleton of osteoblasts and F-actin ring of osteoclasts were labeled with rhodamine-conjugated phalloidin, and observed under confocal laser scanning microscope. The areas of osteoclastic bone resorption pit in co-culture of osteoblasts and osteoclasts were determined using computer image process. Results Curculigoside (10-9 mol/L and 10-8mol/L) significantly promoted the proliferation of osteoblasts and inhibited the TRAP activity of osteoclasts at the concentration of 10-7-10-5 mol/L (P<0.05). Curculigoside, orcinol glucoside and orcinol rhamnoside significantly decreased the number of osteoclasts (P<0.05). Curculigoside (10-10mol/L), curculigine A (10-9mol/L), orcinol glucoside (10-9mol/L) and orcinol rhamnoside (10-9mol/L) significantly increased the ALP activity and bone mineralized nodule formation of osteoblasts (P<0.01), reduced the injury of cytoskeleton of osteoblast induced by 1, 25-(OH)2-VD3, decreased the pit area formed by osteoclasts, and destroyed F-actin cytoskeleton structure and pseudopodia of osteoclasts. Conclusion Curculigoside, curculigine A, orcinol glucoside and orcinol rhamnoside, phenolic glycosides extracted from Curculigo orchioides, can promote osteoblastic bone formation and inhibit osteoclastic bone resorption, demonstrating a significant anti-osteoporotic activity. |
| Key words: Curculigo orchioides phenolic glycosides osteoblasts osteoclasts |
