杜仲诱导骨髓间充质干细胞成骨分化的有效部位

背景：体外细胞培养实验发现，杜仲能够诱导骨髓间充质干细胞成骨分化。目的：分离确认杜仲诱导骨髓间充质干细胞成骨分化的有效部位。方法：体外培养SD大鼠骨髓间充质干细胞。提取分离杜仲有效部位，杜仲粉末经体积分数60%乙醇提取，再用半制备高效液相色谱仪法收集6个组分，编号为B2.1.1、B2.1.2、B2.1.3、B2.1.4、B2.1.5、B2.1.6。取第3代骨髓间充质干细胞，加入不同浓度的杜仲分离物，诱导培养6 d，同时设非诱导对照。荧光定量 PCR法测定成骨分化标记物碱性磷酸酶的表达变化。结果与结论：在高效液相色谱仪分离的6个组分中，B2.1.1和B2.1.3两个组分具有显著刺激骨髓间充质干细胞成骨分化标志基因碱性磷酸酶表达的作用，与非诱导对照相比，其表达分别达到3.73和4.74倍。实验初步分离了杜仲诱导骨髓间充质干细胞成骨分化有效部位，为最终分离和确认有效物质的化学成分提供了资料。

The active fractions of eucommia bark for osteogenic induction of bone marrow mesenchymal stem cells

BACKGROUND:In vitro cell culture experiments have demonstrated that eucommia bark can induce the osteogenic differentiation of bone marrow mesenchymal stem cells.OBJECTIVE:To determine the active fractions of eucommia bark in osteogenic differentiation of bone marrow mesenchymal stem cells.METHODS:Sprague-Dawley rat bone marrow mesenchymal stem cells were in vitro cultured. The active fractions of eucommia bark were extracted. The eucommia bark power was extracted by 60% ethanol, and then six ingredients were collected using high performance liquid chromatography method and numbered B2.1.1, B2.1.2, B2.1.3, B2.1.4, B2.1.5, B2.1.6. Passage 3 bone marrow mesenchymal stem cells were treated with different concentrations of eucommia bark for 6 days. At the same time, a control group was set. The expression level of alkaline phosphatase, an osteogenic differentiation marker, was determined by fluorescent quantitative PCR method.RESULTS AND CONCLUSION:Among the six active fractions isolated by high performance liquid chromatography, B2.1.1 and B2.1.3 significantly stimulated the expression of alkaline phosphatase in bone marrow mesenchymal stem cells. The expression level of alkaline phosphatase was 3.73 and 4.74 times higher respectively compared to the control group. The active fractions for inducing osteogenic differentiation of bone marrow mesenchymal stem cells were preliminarily isolated, which provides data for finally isolating and confirming the chemical ingredients of the active fractions of eucommia bark.