淫羊藿对骨髓间充质干细胞向成骨细胞方向分化以及FN表达的影响

Effect of Herba Epimedii on the Formations of Osteoblasts and the Expression of FN

目的研究中药淫羊藿对骨髓间充质干细胞(mesenchymal stroma cells,MSCs)向成骨细胞方向分化以及对纤维连接蛋白(FN)表达的影响,探讨其促成骨作用的分子机制。方法分别用淫羊藿含药血清和地塞米松干预诱导体外培养的骨髓间充质干细胞,即分为淫羊藿组(M组)、地塞米松组(D组)和阴性对照组(C组),分别于诱导培养的第2、5、8天对细胞进行Giemsa染色和FN免疫细胞化学染色,于诱导培养的第8天对细胞进行矿化结节染色。结果①随着培养天数的增加,M组和D组细胞中有大量的立方形和多角形细胞形成,C组细胞以梭形为主;②随着培养天数的增加,三组细胞的FN表达均呈现先下降后上升的趋势,但M组和D组FN上升的速率较C组快;③M组和D组均有大量矿化结节形成。结论淫羊藿具有促进MSCs向成骨细胞分化的能力。淫羊藿在骨髓MSCs向成骨细胞分化早期,对FN的表达具有一定的抑制作用,之后可能具有促进作用,提示FN可能是淫羊藿促成骨细胞分化作用后期的辅助因子。

Objective To study the effects of Herba Epimedii on formations of osteoblasts and the expression of FN and eplore the molecule mechanism in helping bone formation by Herba Epimedii. Methods Bone marrow mesenchymal stem cells culture systems were divided into M,D and C groups by different additions. Herba Epimedii serum which was collected from the Wistar rats fed with Herba Epimedii was added to M group;dexamethason was added to D group;and C group was added nothing as the control group. On the second,fifth and eighth days of the experiment,Giemsa staining and immunocytochemistry were used to observe the shape of the cells and the expression of FN. Alizarin red S staining was also used to detect mineralized nodes on the eighth day. Results （1） There were so many cube cells and polygon cells in M and D groups, but not in C group as culturing days going on. （2）Expression of FN was decreased from the second day to the fifth day and increased from the fifth day to eighth day in all the three groups,but the speeds of increase were quicker in M and D groups than in C group. （3）Many mineralized nodes were found in M and D groups. Conclusion Herba Epimedii will induce bone marrow mesenchymal stem cells to differentiate into osteoblasts and inhibit the expression of FN at the beginning of the osteoblasts＇ formation,demonstrating FN is a cofactor in promoting the osteoblasts growing mature by Herba Epimedii.