维甲酸诱导骨质疏松模型大鼠骨髓间充质干细胞的特征

Characteristics of bone marrow mesenchymal stem cells in a rat model of retinoic acid-induced osteoporosis

背景:应用维甲酸直接体外干预骨髓间充质干细胞的研究较多,而维甲酸诱导骨质疏松模型大鼠骨髓间充质干细胞的相关研究尚未见报道。目的:建立维甲酸诱导的大鼠骨质疏松模型,观察模型大鼠骨髓间充质干细胞的体外特征。方法:将SD大鼠随机分为2组,实验组用维甲酸70mg/(kg·d)连续灌胃2周快速建立骨质疏松模型,对照组灌胃等体积生理盐水。采用全骨髓贴壁法体外培养两组大鼠骨髓间充质干细胞,观察两组细胞体外特征,包括细胞形态、生长曲线、成骨诱导后体外矿化能力和碱性磷酸酶分泌能力、成脂诱导后向脂肪细胞分化能力。结果与结论:①两组骨髓间充质干细胞各时期形态基本一致,第9天时实验组增殖能力强于对照组;②实验组碱性磷酸酶活力弱于对照组(P <0.05),茜素红染色、碱性磷酸酶染色亦支持碱性磷酸酶活力检测结果;③实验组脂滴数量和油红O吸光度值高于对照组(P <0.05);④结果表明,维甲酸灌胃法成功构建大鼠骨质疏松模型,其骨髓间充质干细胞成骨能力降低,成脂能力增强。

BACKGROUND:Retinoic acid has been directly used to intervene with bone marrow mesenchymal stem cells in vitro;however,there is still a lack of research on in vitro characteristics of bone marrow mesenchymal stem cells in an osteoporosis rat model induced by retinoic acid.OBJECTIVE:To establish the osteoporosis rat model induced by retinoic acid and to observe the characteristics of bone marrow mesenchymal stem cells from the rat model in vitro.METHODS:Sprague-Dawley rats were randomly divided into two groups.The experimental group was treated with retinoic acid,70 mg/kg per day for 2 weeks,in order to quickly establish osteoporosis models,while the control group was given an equal volume of normal saline.Bone marrow mesenchymal stem cells were cultured using the whole bone marrow culture method in vitro and their characteristics(cell morphology,growth curve and surface antigens)were observed.After osteogenic induction,the in vitro mineralization capacity and alkaline phosphatase secretion capacity of the cells were measured.After adipogenic induction,the adipogenic ability of cells was detected.RESULTS AND CONCLUSION:(1)Morphology of bone marrow mesenchymal stem cells was basically the same in the two groups.The proliferation capacity of the cells in the experimental group was higher than that in the control group at 9 days after culture.(2)After osteogenic induction,the alkaline phosphatase activity in the experimental group was lower than that in the control group(P<0.05),which was consistent with the results of alizarin red staining and alkaline phosphatase staining.(3)After adipogenic induction,larger number of lipid droplets and higher absorbance value for oil red O could be observed in the experimental group than the control group(P<0.05).To conclude,bone marrow mesenchymal stem cells from the osteoporosis rat model induced by retinoic acid have lower osteogenic capacity but higher adipogenic capacity than the normal cells.