【摘要】 目的观察不同浓度的甲基强的松龙(甲强龙)对体外培养的大鼠骨髓间充质干细胞(MSCs)的生物学特性的影响。方法全骨髓培养法获得MSCs,通过表面抗原和多能分化能力鉴定。不同浓度的甲强龙分别处理MSCs,流式细胞仪分析各组细胞的凋亡率;MTS/PMS法检测各组细胞的增殖率;不同浓度的甲强龙加抗坏血酸和β-甘油磷酸诱导MSCs成骨分化,茜素红染色(AR-S)、ALP染色检测各组细胞的成骨分化能力。结果 10-6mol/L及以上浓度的甲强龙可明显促进MSCs的凋亡(P<0.01)并明显抑制MSCs的增殖(P<0.01),10-7mol/L及以下浓度的甲强龙对MSCs的凋亡率和增殖抑制作用的影响与对照组差别没有统计学意义(P>0.05);10-7及10-6mol/L甲强龙加抗坏血酸和β-甘油磷酸组茜素红染色和ALP染色阳性,10-8及10-5mol/L组茜素红染色和ALP染色阴性。结论高浓度的甲强龙(10-6mol/L及以上)能抑制MSCs的增殖并促进凋亡,而低浓度的与对照组差别不大。适当浓度(10-7及10-6mol/L)的甲强龙可以诱导MSCs的成骨分化,而过高或过低浓度(10-8及10-5mol/L)的甲强龙则没有这种作用。更多还原

【Abstract】 Objective To observe the effect of methylprednisolone on the biological characteristics of SD rats bone marrow mesenchymal stem cells(MSCs) in vitro.Methods The MSCs were isolated from SD rats and cultured in vitro.By detecting the surface antigen and pluripotent differentiation ability,we identified them as MSCs.In subcultures,MSCs were respectively treated with different concentrations of Methylprednisolone.The proliferation of MSCs was evaluated by MTS/PMS assay;apoptosis was examined with flow cytometer;MSCs were respectively treated with different concentrations of Methylprednisolone accompanied by ascorbic acid and β-glycerol phosphate for 21 days.We used alizarin red staining(AR-S) and the alkaline phosphatage(ALP) staining to detect its osteogenic differentiation ability.Results The optical density value of MSCs treated with high concentration of methylprednisolone was significantly lower than that of the controls(P<0.01).The apoptosis rate of MSCs increased when treated with high concentration of methylprednisolone(P<0.01).Treated with 1×10-6 and 1×10-7mol/L methylprednisolone accompanied by ascorbic acid and β-glycerol phosphate for 21 days,the group′s alizarin red staining and ALP staining were darker than the other concentration one(1×10-5 and 1×10-8).Conclusion High concentration of methylprednisolone inhibits the proliferation of MSCs and leads to an increase of its apoptosis rate,while low concentration one does not.Only the appropriate concentration of methylprednisolone can induce the differentiation of MSCs to osteoblagtic cells.The low or high one can not.更多还原