[关键词]
[摘要]
目的 研究淫羊藿低糖苷组分(LGFEH)对UMR-106细胞和波尼松龙诱导的骨质疏松斑马鱼的作用。方法 将不同质量浓度的LGFEH与UMR-106细胞共同培养后,MTT法测定LGFEH对UMR-106细胞增殖的影响,通过测定细胞内外碱性磷酸酶(ALP)活性评价LGFEH对细胞分化的影响。将受精后3 d的斑马鱼幼鱼分为空白培养基组、0.5% DMSO溶媒对照组、泼尼松龙组、依替膦酸二钠组、LGFEH(1、2、4、8、16 μg/mL)组。每天换液至受精后9 d,处死,采用茜素红对各组斑马鱼幼鱼骨骼染色,并以显微检测、数码成像方法定量分析骨骼染色区域。结果 LGFEH不仅可以促进成骨样细胞UMR-106细胞的增殖,促进其合成和分泌ALP,而且能够显著增加斑马鱼头部骨骼染色面积(矿化面积)和染色累积吸光度值(骨密度)。结论 LGFEH体内及体外具有较好的抗骨质疏松作用。
[Key word]
[Abstract]
Objective To assess the potential effect of the low glycosides fraction from Epimedii Herba (LGFEH) on UMR-106 cells and zebrafish model with osteoporosis induced by Prednisolone. Methods The MTT assay was used to detect the effect of LGFEH on the proliferation of UMR-106 cells, and at the same time, the influence of LGFEH to the UMR-106 cells differentiation was observed through testing the ALP activity of UMR-106 cells using biochemical assays. Zebrafish larvae at 3 d post fertilization were divided into blank control group, 0.5% DMSO group, Prednisolone group, etidronate disodium group, and LGFEH (1, 2, 4, 8, and 16 μg/mL) groups. All groups were incubated in 24-well plates for 9 d until execution, and then zebrafish skeleton was anesthetized and fixed for staining with alizarin red. Quantitative analysis of the stained area was performed by microscopic inspection and digital imaging methods to reflect the amount of bone mineralization. Results The LGFEH significantly increased both proliferation and ALP activities of UMR-106 cells. Furthermore, compared with model group, head skeleton mineral area and IOD of the LGFEH groups were significantly increased. Conclusion Our results indicate that the LGFEH might be beneficial for treating osteoporosis.
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[基金项目]
国家自然科学基金资助项目(81303275,81573620);2013苏州市医疗器械与新医药专项(ZXY2013022)