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目的 :观察大黄素对骨髓基质细胞向成骨细胞方向分化的影响 ,探讨大黄素对干细胞分化方向的调节作用。方法 :用密度梯度离心的方法和传代贴壁筛选的方法相结合 ,分离纯化骨髓基质细胞。对硝基苯磷酸盐法判断是否成功诱导骨髓基质细胞向成骨细胞方向分化 ,碱性磷酸酶染色法、放射免疫测定法观察大黄素对骨髓基质细胞向成骨细胞方向分化的影响。结果 :结合密度梯度离心的方法和传代贴壁筛选的方法 ,分离纯化后可得到成分较为均一的骨髓基质细胞 ,其间存在有间质干细胞 ,经诱导后向成骨细胞方向分化。大黄素可增加大鼠骨髓基质细胞成骨诱导后细胞碱性磷酸酶的含量 ,其中10 -6mol·L-1浓度组作用 7d时效果明显。大黄素在 2 .5× 10 -7～ 10 -5mol·L-1浓度范围内未能增加细胞上清液和细胞裂解液中骨钙素含量。结论 :大黄素能促进骨髓基质细胞向成骨细胞方向分化 ,这一作用主要体现在骨形成过程的早期 ,对矿化期没有促进作用。
Objective: To observe the effects of emodin on the differentiation of bone marrow stromal cells into osteoblasts, and to explore the regulatory effects of emodin on stem cell differentiation. METHODS: The bone marrow stromal cells were isolated and purified by the method of density gradient centrifugation and passaging method. The nitrophenyl phosphate method was used to determine whether bone marrow stromal cells were successfully induced to differentiate into osteoblasts. Alkaline phosphatase staining and radioimmunoassay were used to observe the effects of emodin on the differentiation of bone marrow stromal cells into osteoblasts. Results :Combined with the method of density gradient centrifugation and method of adherence screening, bone marrow stromal cells with homogeneous components were obtained after separation and purification. There existed mesenchymal stem cells between them and differentiated into osteoblasts after induction. Emodin can increase the alkaline phosphatase level in osteoblasts induced by osteoblasts in rats, and the concentration of 10 -6 mol·L -1 concentration group had a significant effect on 7 days. Emodin failed to increase the content of osteocalcin in the cell supernatant and cell lysate in the range of 2. 5×10 -7 to 10 -5 mol·L -1 . Conclusion: Emodin can promote the differentiation of bone marrow stromal cells into osteoblasts. This effect is mainly reflected in the early stage of bone formation and has no promoting effect on mineralization.