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[摘要]
目的 观察淫羊藿水提取物对SD大鼠骨髓间充质干细胞(MSCs)骨向分化能力及转化生长因子β1(TGF-β1)、骨形态发生蛋白2(BMP-2)表达的影响,阐释其防治骨质疏松症的作用机制。方法 全骨髓贴壁法分离、纯化大鼠MSCs;以碱性磷酸酶(ALP)活性及ALP染色阳性率确定淫羊藿水提取物促MSCs骨向分化的最佳质量浓度,并以该质量浓度进行MSCs骨向分化实验。按是否添加经典成骨诱导液将大鼠分为对照组、成骨诱导剂组、淫羊藿水提取物(500 μg/mL)组、淫羊藿水提取物(500 μg/mL)+成骨诱导剂组,每3~4 d各组更换含相应物质培养液1次,连续干预14 d。通过检测各组ALP、I型胶原(Col I)、骨钙素(BGP)和钙化结节等骨向分化指标的变化,以评价淫羊藿水提取物对MSCs骨向分化能力的影响;通过检测TGF-β1、BMP-2的表达,研究淫羊藿水提取物促MSCs骨向分化的作用机制。结果 淫羊藿水提取物最佳促MSCs骨向分化的质量浓度为500 μg/mL;成骨诱导剂组、淫羊藿水提取物组及淫羊藿水提取物+成骨诱导剂组均能促进MSCs骨向分化及上调TGF-β1和BMP-2的表达。结论 淫羊藿促进MSCs骨向分化,上调TGF-β1、BMP-2的表达可能是其作用机制之一。
[Key word]
[Abstract]
Objective To investigate the effects of epimedium water extract (EWE) on the ability of osteogenic differentiation and the expression of TGF-β1 and BMP-2 in inducing mesenchymal stem cells (MSCs) differenting into osteoblast and to further explain its mechanism. Methods MSCs were isolated and purified by differential time adherent method; the most effective concentration of EWE on the osteogenic differentiation of MSCs was confirmed by the activity of alkaline phosphatase (ALP) and positive rates of ALP staining; according to the differently induced condition, MSCs were divided into four groups: control, classic (induced by the classic osteoblast-induced system), EWE (induced by the most effective concentration of EWE on the osteogenic differentiation), and EWE + classic (induced by the combination of classic osteoblast-induced system and the most effective concentration of EWE on osteogenic differentiation) groups. ALP, type I collagen (Col I), bone gla protein (BGP), and calcium nodes in each group were detected and compared to indicate the osteogenic differentiation of each group. TGF-β1 and BMP-2 in each group were detected by ELISA. Results The most effective concentration of the EWE on the osteogenic differentiation of MSCs was 500 μg/mL. The classic, EWE, and EWE + classic groups could promote the osteogenic differentiation of MSCs and increase the expression of TGF-β1 and BMP-2. Conclusion The EWE could promote the osteogenic differentiation of MSCs. The increase of the expression of TGF-β1 and BMP-2 in the induced groups of EWE may be the mechanism of improving the differentiation of MSCs into osteoblast.
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[基金项目]
国家自然科学基金资助项目(81173619);广东省教育厅育苗工程项目(LYM10025);广东省自然科学基金博士启动(S2012040007531)