麝香酮对大鼠骨髓间充质干细胞成骨能力的影响分析

Effect analysis of muscone on the osteogenic potential of rat bone marrow derived mesenchymal stem cells

目的探索麝香酮对大鼠骨髓间充质干细胞成骨能力的影响,为骨髓间充质干细胞在骨损伤修复方面的应用开发新的方法。方法采用密度梯度离心法,从大鼠骨髓中分离骨髓间充质干细胞,通过CCK-8方法对细胞的增殖能力进行评价,并进一步通过流式细胞术鉴定细胞表面的特异标记;用麝香酮(1.0mg/L)对细胞进行处理,分析处理后的骨髓间充质干细胞在成骨基因表达以及碱性磷酸酶活性方面的变化。结果从大鼠骨髓中分离间充质干细胞可在体外条件下正常生长增殖,细胞CD29、CD44、CD90均呈阳性表达,CD45呈阴性表达,与正常间充质干细胞的表型相符合;同时,实时定量PCR结果显示,麝香酮处理后的细胞中,成骨基因ALP、Collagen I、Runx2和Osterix都有一定的上调趋势,同时,细胞中碱性磷酸酶的活性也有一定程度的升高。结论麝香酮对骨髓间充质干细胞的成骨分化能力有一定的促进作用,在利用骨髓间充质干细胞进行骨损伤修复的过程中具有一定的应用价值。

Objective To explore the effect of muscone on rat bone marrow-derived mesenchymal stem cells（BMSCs）and develop a novel strategy for the application of BMSCs on the restoration of bone injury.Methods BMSCs were isolated from rat bone marrow with density gradient centrifugation and the proliferation ability and special makers on cell surface were evaluated with CCK-8methods and FACS respectively.After being treated with muscone（1.0mg/L）,the expression of osteogenic genes and the activity of alkaline phosphatase in BMSCs were further analyzed.Results The rat BMSCs could proliferate in vitro normally.The cells showed a positive expression of CD29,CD44 and CD90,as well as a negative expression of CD45,which coincided with the normal phenotype.Besides,the results of real-time quantitative PCR indicated that the expression of some osteogenic genes,ALP,Collagen I,Runx2 and Osterix,were upregulated in some degree,and the activity of alkaline phosphatase in BMSCs was also enhanced with the treatment of muscone.Conclusion In summary,muscone holds the promoter action on the osteogenic potential of rat BMSCs and application value in the restoration of bone injury with BMSCs.