损伤脊髓匀浆上清对骨髓间充质干细胞分泌髓鞘前脂蛋白、脑源性神经营养因子的影响

背景：损伤脊髓匀浆上清成分复杂,其中不仅存在多种化学物质,而且也存在着多种细胞因子,这些物质能否影响骨髓间充质干细胞的增殖分化和分泌功能还不清楚。目的：探讨损伤脊髓匀浆上清成分对骨髓间充质干细胞分泌的脑源性神经营养因子和髓鞘前脂蛋白的影响。方法：贴壁法分离纯化Wistar大鼠骨髓间充质干细胞,稳定传到第3代后,分别用正常和损伤的Wistar大鼠脊髓匀浆上清诱导培养20d。免疫荧光染色检测神经元特异性烯醇化酶阳性细胞,ELISA法检测培养液内髓鞘前脂蛋白、脑源性神经营养因子的含量,即时定量-PCR检测髓鞘前脂蛋白mRNA、脑源性神经营养因子mRNA水平。结果与结论：损伤脊髓匀浆上清液诱导培养骨髓间充质干细胞后,神经元特异性烯醇化酶阳性细胞率和培养液内脑源性神经营养因子、髓鞘前脂蛋白的含量在各个时间点均较正常脊髓匀浆上清液对骨髓间充质干细胞培养组高。提示,损伤的脊髓匀浆上清液能够诱导骨髓间充质干细胞分泌脑源性神经营养因子、髓鞘前脂蛋白,有利于向神经细胞方向分化。

Effects of injured spinal cord extracts on brain-derived neurotropic factor and myelin proteolipid protein in bone marrow mesenchymal stem cells

BACKGROUND：The component of injured spinal cord extracts is complex and includes some chemical substances or cytokines. It remains unclear if these factors may influence cellular proliferation and secretion of bone marrow mesenchymal stem cells （BMSCs） or not. OBJECTIVE：To investigate the effect of injured spinal cord extracts on secretion of brain-derived neurotropic factor （BDNF） or myelin proteolipid protein （PLP） in BMSCs. METHODS：BMSCs were obtained from Wistar rats. After 3 generation of cell proliferation,BMSCs were induced with normal or injured spinal cord extracts for 20 days. Neuron specific enolase （NSE） positive cells were detected by immunofluorescence staining. Levels of BDNF and PLP in BMSCs cultured supernatant were evaluated using enzyme linked immunosorbent assay （ELISA） and BDNF and PLP mRNA was determined by real time polymerase chain reaction assay. RESULTS AND CONCLUSION：The rate of NSE positive cells and levels of BDNF and PLP that were treated with injured spinal cord extracts were greater significantly at different cultured time points than normal spinal cord extracts. These results showed that the injured spinal cord extracts could induce secretion of BDNF and PLP in BMSCs that is to the benefit of cellular differentiation into neuronal cells.