Cuprizone 对少突胶质细胞相关蛋白的影响

目的: 研究双环己酮草酰双腙（cuprizone）对少突胶质细胞（oligodendrocytes，OLs）相关蛋白的影响。方法: 新生24 h Sprague-Dawley（SD）大鼠的大脑皮层胶质细胞混合培养7 天后，采用恒温摇床振荡分离与差异贴壁方法并结合条件培养基纯化培养细胞，纯化培养3 d 后加入含有25 nmol·L-1 cuprizone 培养基继续培养3 d。光学显微镜观察细胞形态；免疫荧光染色鉴定细胞类型。Western Blot 检测MBP、Fyn、Erk1/2 等蛋白的含量。结果：获得高纯度成熟的少突胶质细胞，荧光MBP 染色阳性。相较于正常细胞组蛋白，Cuprizone 组细胞MBP，Erk1/2 及磷酸化Fyn（416 位点）蛋白含量明显降低。结论：Cuprizone 对髓鞘形成细胞- 少突胶质细胞相关蛋白的表达成抑制作用。

Effect of Cuprizone on Oligodendrocyte-associated Proteins

Objective：To investigate the effect of cuprizone on the oligodendrocyte （OLs）related proteins. Methods: Mixed glial cells of cortices from neonatal rats（24 h after born） wereprimarily cultured for 7 d，and cells were purified by horizontal orbital shaking，differential adhesion and defined culture media. After three days，OLs were exposed to 25 nmol·L-1 cuprizone and further cultured for 3 days. Cells were observed by optical microscope and identified by immumofluorescence staining with MBP and DAPI. MBP，Fyn，Erk1/2 and other proteins were tested by Western Blot. Results：High purity mature OLs （specific marker MBP） were obtained. Compared to normal cells，the contents of MBP，Erk1/2 and p-Fyn （416 point） were significantly reduced in the cells exposed to Cuprizone. Conclusion: Cuprizone inhibits the expression of oligodendrocyte-associated proteins.