玛咖醇提取物促进大鼠骨髓间充质干细胞成骨分化的作用

Effect of Maca Alcohol Extract Promotes Osteogenic Differentiation of Rat Bone Marrow Mesenchymal Stem Cells

摘要: 目的：初步探讨玛咖醇提取物(MAE)对大鼠骨髓间充质干细胞(BMSCs)向成骨细胞分化的影响。方法：黑玛咖用乙醇浸泡后旋转蒸发制备玛咖醇提取物浸膏。分离SD大鼠的股骨和胫骨利用全骨髓贴壁方法在体外培养BMSCs，通过细胞表面标志鉴定细胞；细胞传代培养至第3代，将浸膏用成骨诱导培养基稀释为不同浓度(0、1.25、2.5、5、10 mg/ml)作用于BMSCs，以不含药物的培养基作为空白对照，采用CCK-8法检测MAE对BMSCs增殖与毒性，利用茜素红染色观察钙化结节形成的数量，评价MAE对BMSCs成骨分化的影响。结果：MAE浸膏1 g相当于原材料5 g；大鼠BMSCs流式鉴定结果为CD45：4.53%，CD44：71.4%，CD90+：85.6%，细胞纯度较好；MAE对BMSCs的IC₅₀是8.445 mg/ml，且MAE呈剂量依赖性抑制细胞增殖，药物浓度为2.5 mg/ml时细胞增殖最为明显，此时茜素红染色强度最强。结论：MAE能够诱导大鼠BMSCs向成骨细胞分化，且最佳作用浓度为2.5 mg/ml。

Abstract: Objective: To investigate the effect of maca alcohol extract on osteogenic differentiation of rat bone marrow mesenchymal stem cells. Methods: Black maca was soaked in ethanol and then rotary evaporation was used to prepare maca alcohol extract (MAE). The bone marrow mesenchymal stem cells (BMSCs) were isolated from femur and tibia of SD rats. BMSCs were cultured in vitro by whole bone marrow adherent method and identified by cell surface markers. The cells were subcultured to the third generation, and MAE was diluted into different concentrations (0, 1.25, 2.5, 5, 10 mg/mL) in osteogenic induction medium to treat BMSCs. The drug-free medium was used as blank control, and the proliferation and toxicity of MAE on BMSCs were detected by CCK-8 method. Alizarin red staining was used to observe the number of calcified nodules and evaluate the effect of MAE on osteogenic differentiation of BMSCs. Results: 1 g MAE was equivalent to 5 g raw material; flow cytometric identification results were CD45: 4.53%, CD44: 71.4%, CD90+: 85.6%, the cell purity was good; the IC₅₀ of stem cells was 8.445 mg/ml, and the MAE inhibits cell proliferation in a dose-dependent manner. The cell proliferation was most obvious when the drug concentration was 2.5 mg/ml. At this time, the intensity of Alizarin Red staining was the strongest. Conclusion: MAE can promote BMSCs to differentiate into osteoblasts, and the optimal concentration is 2.5 mg/ml.