补骨脂素对大鼠骨髓间充质干细胞成骨分化的影响

目的研究不同浓度的补骨脂素对大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs)成骨分化的影响,为临床应用补骨脂素治疗骨质疏松症提供理论依据。方法选取8周龄健康雄性SD大鼠,取双侧股骨、胫骨,对大鼠的BMMSCs进行分离、原代培养和细胞表型鉴定。取第3代大鼠BMMSCs,体外利用成骨诱导培养基进行不同浓度补骨脂素的药物诱导,MTT法检测不同浓度的补骨脂素对大鼠BMMSCs生长的影响;通过碱性磷酸酶(ALP)活性测定、Western blot法和茜素红染色方法评价不同浓度补骨脂素对大鼠BMMSCs成骨分化能力。结果第3代大鼠BMMSCs表面抗原符合干细胞鉴定标准,成骨诱导后给予15μmol/L补骨脂素对大鼠BMMSCs的增殖作用最佳,ALP活性、Runx-2表达和钙化结节数目均明显高于经典成骨诱导组、5μmol/L和10μmol/L和20μmol/L补骨脂素组。结论15μmol/L补骨脂素成骨诱导促进BMMSCs向成骨分化,从而起到防治骨质疏松的作用。

The effect of psoralen on the osteogenic differentiation of rat bone marrow mesenchymal stem cells

Objective To investigate the effect of different concentrations of psoralen on the osteogenic differentiation of rat bone marrow mesenchymal stem cells(BMMSCs),and to provide theoretical basis for clinical treatment of osteoporosis with psoralen.Methods BMMSCs were were isolated from bilateral femur and tibia and cultured,and cell phenotype was identified in eight-week-old healthy male SD rats.The effect of psoralen on the growth of BMMSCs were detected by MTT method,the osteogenic differentiation ability of BMMSCs was evaluated by Western blot,alkaline phosphatase(ALP)activity and alizarin red staining.Results The surface antigens of rat BMMSCs meets the criteria for stem cell identification at the third generation.The proliferation rates of BMMSCs were the highest at 15μmol/L psoralen after osteogenesis induction(compared to 5μmol/L,10μmol/L and 20μmol/L).The activity of ALP,the expression level of Runx-2 and the number of calcified nodules were significantly increased in 15μmol/L psoralen group than in classical osteogenesis induction group,and other concentrations psoralen groups(5μmol/L,10μmol/L and 20μmol/L).Conclusion Psoralen promotes BMMSCs to differentiate into osteoblasts,and would be useful in preventing and treating osteoporosis.