胶质源性神经营养因子基因修饰骨髓间充质干细胞与缺氧复氧神经细胞的共培养

背景：课题组前期研究表明，胶质源性神经营养因子基因修饰的骨髓间充质干细胞能稳定地表达胶质源性神经营养因子 mRNA，但骨髓间充质干细胞/胶质源性神经营养因子分泌的胶质源性神经营养因子蛋白对缺氧复氧损伤的神经细胞是否有积极影响，尚不清楚。目的：观察胶质源性神经营养因子基因修饰的骨髓间充质干细胞对缺氧复氧神经细胞的保护作用。方法：胶质源性神经营养因子基因修饰的骨髓间充质干细胞经诱导后与缺氧复氧神经细胞共培养，采用Hoechst33258和细胞免疫荧光染色分别检测神经细胞凋亡及生长相关蛋白43在共培养细胞中的表达。结果与结论：骨髓间充质干细胞/胶质源性神经营养因子组的神经细胞凋亡率显著低于骨髓间充质干细胞组和缺氧组(P < 0.05)，骨髓间充质干细胞/胶质源性神经营养因子组表达生长相关蛋白43的吸光度值明显高于骨髓间充质干细胞组(P < 0.05)。提示骨髓间充质干细胞/胶质源性神经营养因子通过抑制缺氧复氧神经细胞凋亡和促进共培养细胞生长相关蛋白43表达发挥神经保护作用。

Bone marrow mesenchymal stem cells modified by glial cell line-derived neural factor gene co-cultrued with neurons subjected to anoxia-reoxygenation

BACKGROUND:Preliminary studies in workgroup indicated that glial-derived neurotrophic factor (GDNF) gene modified by bone marrow mesenchymal stem cells (BMSCs) (BMSCs/GDNF) can stably express the mRNA of GDNF, but whether GDNF protein in BMSCs/GDNF affects neurons subjected to anoxia-reoxygenation positively is unclear.OBJECTIVE:To observe the protective effects of BMSCs/GDNF on neurons subjected to anoxia-reoxygenation.METHODS:BMSCs/GDNF were co-cultured with anoxia-reoxygenation nerve cells. Hoechst33258 staining was used to detect apoptosis of neurons. Immunofluorescent-chemistry method was used to detect growth-associated protein-43 (GAP-43) expression in co-cultured cells.RESULTS AND CONCLUSION:The rate of neurons apoptosis in the BMSCs/GDNF group was lower than that of the BMSCs group and anoxia group (P < 0.05). Absorbance of growth-associated protein-43 (GAP-43) expression in the BMSCs/GDNF group was higher than that in the BMSCs group (P < 0.05). It is indicated that BMSCs/GDNF have neuroprotective effects due to inhibiting apoptosis of neurons subjected to anoxia-reoxygenation and increasing GAP-43 expression in co-cultured cells.