目的 研究淫羊藿苷基于成骨细胞和破骨细胞蛋白质组学的抗骨质疏松作用机制。方法 四甲基偶氮唑蓝(MTT)法测定细胞的增殖,ELISA法测定碱性磷酸酶(ALP)和抗酒石酸酸性磷酸酶(TRAP)的活性,茜素红染色观察成骨细胞骨矿化结节的形成,以骨片上形成的骨吸收陷窝的数目和面积表征破骨细胞的骨吸收活性,以二维凝胶电泳观察成骨及破骨细胞蛋白质组的变化,应用TOF/MS/MS对差异表达的蛋白进行鉴定。结果 淫羊藿苷显著增加成骨细胞的骨形成,抑制破骨细胞的骨吸收,上调或下调成骨和破骨细胞各9个蛋白的表达。结论 淫羊藿苷通过参与能量代谢、氧化应激、嘌呤合成和细胞生长分化的等生物学过程调控骨代谢。
Abstract
OBJECTIVE To explore the anti-osteoporotic mechanism of icariin based on osteoblast and osteoclast proteomics. METHODS The cell proliferation was determined by MTT assay. The activity of alkaline phosphatase (ALP) in osteoblasts and tartrate-resistant acid phosphatase (TRAP) in osteoclast were measured using ELISA kit. Alizarin red staining was used to observe the formation of bone mineralized nodules in osteoblasts. The number and area of bone resorption pit formed on bone slices by osteoclasts were used to characterize the activity of osteoclastic bone resorption. Two-dimensional gel electrophoresis was used to observe the changes of icariin-treated osteoblast and osteoclast proteomics. TOF/MS/MS analysis was applied to identify the differentially expressed proteins. RESULTS Icariin significantly enhanced the osteoblastic bone formation and inhibited the osteoclastic bone resorption, and up-regulated or down-regulated expression of nine proteins in osteoblast and osteoclast, respectively. CONCLUSION Icariin regulates bone metabolism through involving multiple biological process, including energy metabolism, oxidative stress, purine synthesis and cell growth and differentiation.
关键词
淫羊藿苷 /
成骨细胞 /
破骨细胞 /
蛋白质组学 /
差异蛋白
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Key words
icariin /
osteoblasts /
osteoclast /
proteomics /
differential proteins
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中图分类号：
R965
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参考文献
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脚注
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基金
国家自然科学基金项目资助(90709023)
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