大黄素诱导人骨髓间充质干细胞向成骨细胞分化的研究

Emodin Induced Differentiation of Human Bone Marrow Mesenchymal Stem Cell Osteoblasts

目的:探讨大黄素对人骨髓间充质干细胞向成骨细胞方向分化的影响。方法:体外分离、培养及扩增人骨髓MSCs,用流式细胞仪检测人骨髓MSCs表面抗原的表达,用倒置光学显微镜、透射电镜、四甲基偶氮唑盐比色、碱性磷酸酶染色、碱性磷酸酶活性测定等研究人骨髓MSCs增殖和分化规律。采用不同方法诱导第3代人骨髓MSCs向成骨细胞分化。结果:人骨髓MSCs贴壁生长,呈成纤维细胞外观。流式细胞仪检测显示CD29、CD44表达为阳性,CD34、CD45、HLA-DR表达为阴性。大黄素对人骨髓MSCs增殖的影响:对照组和DXM(地塞米松)组与大黄素+DXM组比较均具有显著差异(P<0.05)。大黄素对人骨髓MSCs分化的影响:对照组和DXM组与大黄素+DXM组比较均具有显著差异(P<0.01)。结论:大黄素能促进人骨髓MSCs向成骨细胞方向分化。

Objective To investigate the effect of emodin on the differentiation of human bone marrow mesenchymal stem cells（MSCs） into osteoblast in vitro Methods MSCs were isolated and cuhured and expanded in vitro and their surface antigens were detected by flow cytometry. The proliferation and differentiation of MSCs were examined by phase - contrast microscopy, transmission electron microscopy, MTT alkaline phosphatase（ALP） staining and the determination of ALP activities. And then different methods were used to induce the third-passage MSCs to differentiate into osteoblast in vitro. Results MSCs proliferated, and adhered to the plastic surface and looked like long spindle-shape fibroblasts. CD29 and CD44 expressed positive, CD34, Cd45 and HLA-DR negative by flow cytometry. Emodin induced more proliferation of MSCs than in the emodin ＋ dexamethasone（DXM） group, with significant differences in control group and DXM group, respectively（ P 〈 0.05）. Emodin induced no more differentiation of MSCs than in emodin ＋ DXM group, while control group and DXM group had significant differences. Conclusion Emodin can induce orientation differentiation of MSCs into osteoblast.