小檗碱对骨髓间充质干细胞成骨分化的影响

Effect of berberine on the osteogenic differentiation of bone marrow mesenchymal cells

目的探讨小檗碱对体外培养SD大鼠骨髓间充质干细胞成骨分化的影响。方法应用MTT法检测不同浓度小檗碱对骨髓间充质干细胞增殖情况的影响,以对硝基苯磷酸盐法检测碱性磷酸酶活性,等离子体直读光谱仪检测钙沉积,放射免疫法检测骨钙素含量,茜素红法染色观察钙化结节形成。结果小檗碱在1×10-9～1×10-5mol.L-1浓度范围对骨髓间充质干细胞增殖无明显影响。小檗碱浓度为1×10-7和1×10-6mol.L-1培养3～7d能明显提高骨髓间充质干细胞内碱性磷酸酶活性。小檗碱在1×10-7～1×10-5mol.L-1浓度范围培养22d能明显促进骨髓间充质干细胞骨钙素合成和分泌。小檗碱在浓度为1×10-6和1×10-5mol.L-1培养28d时钙盐沉积量和钙化结节形成增加。结论小檗碱可促进骨髓间充质干细胞向成骨细胞方向分化,增强骨钙化。这可能为其防治骨质疏松的机制之一。

Aim To investigate the effect of berberine on osteogenic differentiation of rat bone marrow mesenchymal stem cells（MSCs）.Methods Methylthiazolyl tetrazolium（MTT）assay was applied to determine the MSCs proliferation treated by berberine at different concentration.Alkaline phosphatase（ALP）activity was detected using p-nitrophenyl phosphate assay.Bone gla protein（BGP）was determined by radioimmunoassay.Calcium was detected via inductively coupled plasma direct reading spectrometer.Nodule formation of calcium was observed with alizarin red stain.Results Berberine at concentration from 1×10^-9 to 1×10^-5 mol·L^-1for 24 hours and 96 hours has no effect on the proliferation of MSCs.Berberine at concentration 1×10^-7 and 1×10^-6 mol·L^-1for 3 days and 7 days effectively stimulates the activity of ALP in MSCs.Berberine at concentration 1×10^-7 to 1×10^-5 mol·L^-1for 22 days effectively increases the synthesis and secretion of BGP in MSCs.Berberine at concentration from 1×10^-5 and 1×10^-6 mol·L^-1for 28 days effectively increases the calcium deposition and the nodule formation of calcium.Conclusion Berberine improves the differentiation of MSCs into osteoblasts,which may be one of its mechanism of preventment and treatment of osteoporosis.