黄芪多糖对电离辐射骨髓间充质干细胞向神经分化潜能的影响

摘要

Aim To investigate the effect of Astragalus polysaccharide(APS) on the differentiation ability of human bone marrow mesenchymal stem cells(bone marrow mesenchymal stem cells,BMSCs) into nerve cells after irradiated by X rays.Methods We used different concentrations of APS to treat BMSCs which were exposed in 2 GyX ray radiation,then we selected the proliferation ability of BMSCs by CCK-8 method.We also counted the nerve cells that were differentiated from BMSCs.Then we detected the expression of Nissl bodies by Toluidine blue staining,and we also tested the expression of Nestin and NSE by Western blot.Results Compared with the control group,the ability of cell proliferation significantly decreased after X ray radiation(P<0.05),compared with radiation group,the proliferation ability of BMSCs treated by APS significantly increased(P<0.05).Moreover, BMSCs had the best proliferation ability which was treated by APS in the concentration of 50 mg·L-1.Then we chose this dose as the best drug concentration.The number of nerve cells significantly decreased after radiation(P<0.05).Neural cells significantly increased(P<0.05) after treated by APS.We found the expression of Nissl bodies decreased after radiation(P<0.05),and increased significantly(P<0.05) after treated by APS.The expression of Nestin and NSE decreased after radiation(P<0.05), and their expression level increased after treated by APS(P<0.05).Conclusion APS has protective effects on the differentiation potential of human bone marrow mesenchymal stem cells into nerve cells exposed in ionizing radiation.%目的 研究黄芪多糖对X射线照射人骨髓间充质干细胞向神经分化的干预作用.方法 采用CCK-8 法筛选不同浓度的黄芪多糖作用于2GyX射线辐射人骨髓间充质干细胞的增殖能力;细胞形态计数X射线对骨髓间充质干细胞分化为神经细胞数量的影响;甲苯胺蓝染色检测X射线对骨髓间充质干细胞分化为神经细胞中尼氏体的表达量;Western blot检测神经巢蛋白(Nestin)及神经元特异性烯醇化酶(neuron-specific enolase,NSE)表达变化.结果 与对照组相比,X射线照射后细胞增殖水平明显降低(P<0.05),与单纯照射组比较,加药照射组骨髓间充质干细胞增殖水平显著增加(P<0.05),其中黄芪多糖浓度为50 mg·L-1时促增殖作用最强,设为最佳药物浓度;单纯辐射后骨髓间充质干细胞分化为神经细胞的数量明显降低(P<0.05),当给予黄芪多糖干预后分化为神经细胞的数量明显上升(P<0.05);辐射后骨髓间充质干细胞分化为神经细胞中尼氏体的表达量降低(P<0.05),当给予黄芪多糖干预后,尼氏体的表达量明显增多;单纯辐射后Nestin及NSE表达量降低(P<0.05),而当给予黄芪多糖干预后,Nestin和NSE表达量升高(P<0.05).结论 黄芪多糖对电离辐射人骨髓间充质干细胞向神经分化潜能的影响具有保护作用.