淫羊藿苷对人牙髓干细胞神经向分化作用的初步研究

Preliminary study of Icariin on neural differentiation of human dental pulp stem cells

摘要:

目的 探讨淫羊藿苷（ICA）对人牙髓干细胞（DPSC）神经向分化的作用。方法 分离培养DPSCs后，采用不同浓度ICA（0.01 μmol、0.10 μmol、1.00 μmol、10.00 μmol）处理DPSCs。CCK-8法检测ICA对细胞增殖活力的影响，确定其最佳促DPSCs增殖浓度。分别进行细胞形态学观察和Nestin细胞免疫荧光检测。Western blotting检测DPSCs神经向分化相关蛋白Nestin、βⅢ-tubulin及NSE的表达。结果 不同浓度ICA细胞相对活力值比较，差异有统计学意义（P <0.05），其中以0.10 μmol ICA细胞相对活力最高。与对照组比较，ICA组神经球的直径增加（P <0.05），Nestin荧光强度增强（P <0.05），Nestin、βⅢ-tubulin、NSE蛋白相对表达量升高（P <0.05）。结论 ICA能够有效促进DPSCs增殖，提高DPSCs神经向分化能力。

Abstract:

Objective To investigate the effects of Icariin (ICA) on the neural differentiation of human dental pulp stem cells (DPSCs).Methods After isolating and culturing DPSCs through primary culture, ICA with different concentrations (0.01 μmol, 0.10 μmol, 1.00 μmol, 10.00 μmol) was added into DPSCs. The effects of ICA in different concentrations on proliferation of DPSCs were assessed by CCK-8 assay to select the optimal concentration. Then, the effects of ICA on neural differentiation of DPSCs were observed. The cell morphologies were observed by microscope. Immunofluorescence staining was used to observe the expression of Nestin. Western blotting was used to detect the expressions of Nestin, βⅢ-tubulin and neuron-specific enolase (NSE).Results The results of variance analysis showed that there were differences in the relative vitality values of different concentrations of ICA groups (P < 0.05), and the relative vitality of 0.1 μmol ICA group was the highest. More neuroshpere formation was noted and the upregulation of Nestin was observed in the ICA group. And the diameters of neurospheres in the ICA group were significantly increased compared that of the control group (P < 0.05). Compared with the control group, the fluorescence expression of Nestin in the ICA group was increased (P < 0.05). Western blotting results showed that expressions of neural differentiation-related protein Nestin, β Ⅲ-tubulin, NSE increased significantly in the ICA group (P < 0.05).Conclusion ICA can effectively promote the proliferation and neuronal differentiation of DPSCs.