槲皮素对骨髓间充质干细胞增殖和骨向分化的影响

Effects of quercetin on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells

目的:通过观察槲皮素对骨髓间充质干细胞增殖和骨向分化的影响,探讨其调控骨髓间充质干细胞及其防治骨质疏松症的作用机制。方法:运用全骨髓贴壁法分离培养骨髓间充质干细胞,并通过形态学、流式细胞仪检测表面标志物及分化能力鉴定所取得的细胞;运用MTT法检测不同浓度槲皮素对大鼠骨髓间充质干细胞增殖的影响,以确定其最佳促增殖剂量。实验分为空白组、槲皮素组、经典诱导组(地塞米松+β-甘油磷酸钠+维生素C),检测诱导分化过程中ALP活性和钙化结节数目,RT-PCR的方法检测BMP-2 mRNA的变化。结果:经MTT法检测,浓度为5μmol/L和10μmol/L的槲皮素组OD值较空白组明显升高;与空白组相比,槲皮素组与经典诱导组ALP活性及矿化结节均显著增加,并均可上调MSCs骨向分化过程中BMP-2 mRNA的表达。结论:槲皮素能促进骨髓间充质干细胞的增殖和骨向分化,可能是其促进骨折愈合、防治骨质疏松症的细胞学机制之一。

Objective： Investigate the effects of quercetin on the bone marrow stromal cells proliferation and osteoblast-differentiation ability. To provide a probable action mechanism of traditional Chinese medicine prevent and cure OP in stem cell aspect. Methods： BMSCs was isolated and purified by adherent method,BMSCs was identified by morphologic characteristics,surface antigens detected by flow cytometer; MTT method was used to confirm the most effective concentration of quercetin and cytokines on the proliferation of BMSCs. Groups were divided：the blank group,quercetin group and classics group（ DEX ＋ β-GP ＋ Vit C）; The ALP activity and the number of mineralized nodules were measured to indicate the osteoblast function. Detect the change of BMP-2 mRNA by RT-PCR method. Results： MTT assay showed that compared with the control group,the quercetin of 5 mol / L and 10 mol / L two concentration could promote the value of OD. Compared with the control group,the ALP activity and mineralized nodules were significantly increased in the group of quercetin and the classical induction group,and both of them can up regulate the expression of BMP-2 mRNA in the osteogenic differentiation process of MSCs. Conclusion： We demonstrated that the quercetin could stimulate the proliferation and osteogenic differentiation of bone marrow mysenchymal stem cells. Perhaps the effect of Quercetin on BMSCs is one of cytology mechanisms of promoting the concrescence of bone fracture and preventing and treating osteoporosis.