Estrogenic effect of total flavonoids from Herba Epimedii on osteogenic differentiation
(2.Jiujiang University, Jiujiang 332000)
【Abstract】The purpose of this study was to investigate the estrogic effect of total flavonoids of Herba Epimedii (EF) on the osteogenesis of human bone marrow mesenchymal stem cells (hBMSCs). The first generation of hBMSCs were cultured, subcultured, and purified, and the fourth passage of hBMSCs were used to detect the cell surface markers CD90 and CD45 by flow cytometry. In the drug experiment, the effect of different concentrations of EF on the proliferation of hBMSCs was detected by CCK-8 assay. Three groups were designed: blank, EF, and EF + ICI182780. Alizarin red staining was used to observe the formation of calcium nodules and RT-PCR was used to detect the expression of OPN and ALP. The results indicated that the CD90 expressed and CD45 did not express on the surface of purified hBMSCs. Low concentration of EF (2.5 and 5 μg/mL) promoted the proliferation of hBMSCs, while high concentration of EF (10, 20, and 40 μg/mL) inhibited the proliferation of hBMSCs. Cells cultured with EF presented osteoblast-like morphology under the microscope. Alizarin red staining showed that EF can promote the formation of calcium nodules, while estrogen receptor (ER) inhibitor ICI182780 can antagonize this effect of EF. EF can up-regulate the expression of osteoblast-associated genes OPN and ALP (P < 0.05), and ICI182780 can inhibit the effect of EF (P < 0.05). In conclusion, EF can promote the osteogenic differentiation of hBMSC by activating ER.
【Keywords】 total flavonoids of Herba Epimedii; hBMSCs; osteogenic differentiation; estrogen;
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