不同浓度地塞米松对人骨髓间充质干细胞体外增殖及凋亡的影响

The effect of dexamethasone on the proliferation and apoptosis of human bone marrow mesenchymal stem cells in vitro

目的观察地塞米松对体外培养人骨髓间充质干细胞增殖及凋亡的影响。方法利用1.073g/mLPercoll分离液以梯度密度分离法获取人骨髓间充质干细胞(hMSCs),进行体外培养。成骨诱导组细胞用地塞米松、抗坏血酸和β-甘油磷酸钠处理,分别进行ALP染色和矿化结节染色。实验组细胞分别以10-9mol/L、10-8mol/L、10-7mol/L和10-6mol/L地塞米松加以干预,MTT法检测各组细胞的增殖率;流式细胞仪定量分析hMSCs的凋亡率。结果成骨诱导组细胞ALP染色和矿化结节染色均为阳性,对照组为阴性;低浓度(10-9mol/L)地塞米松对细胞的体外增殖无明显影响(P>0.05),10-8mol/L及以上浓度地塞米松可明显抑制细胞的增殖(P<0.01);hMSCs凋亡率随地塞米松浓度的增加而升高(P<0.01)。结论地塞米松、抗坏血酸和β-甘油磷酸钠可促使hMSCs成骨分化,10-8mol/L及以上浓度的地塞米松可明显抑制细胞的增殖,地塞米松能促进体外培养的人骨髓间充质干细胞凋亡。

Objective To observe the effect of dexamethasone on the proliferation and apoptosis of human mesenchy-mal stem cells（hMSCs） derived from bone marrow in vitro.Methods The primary human MSCs were isolated and cultured in vitro.For osteogenic induction,the hMSCs were cultured with medium containing Dexamethasone,sodium β-glycerophos-phate,and ascorbic acid.The hMSCs were examined by Alkaline Phosphatase（ALP）,Von Kossa staining.In subcultures.hM-SCs were respectively treated with different concentration dexamethasone（10^-9mol/L,10^-8mol/L,10^-7mol/L and10^-6mol/L）.The proliferation of hMSCs was evaluated using MTT method, Apoptosis was examined quantitatively with flow cytometer.Results The results of ALP,Von Kossa staining of hMSCs were positive,while those of the control group were negative.The optical density values of hMSCs treated with high concentration dexamethasone（10^-8mol/L,10^-7 mol/L and10^-6 mo1/L） for 6 days were significantly lower than those in the controls（P〈0.01）.Dexamethasone increased the apoptosis rates of hMSCs cultured in vit-ro.Conclusion The hMSCs can be differentiated to osteoblasts with osteogenic potential in vitro.The high concentration dexamethasone inhibits the proliferation of hMSCs.Dexamethasone leads to a increase of apoptosis rate.