小剂量干扰素-γ通过P27kip1影响少突胶质细胞前体分化的机制  

Low-dose interferon-γ influences differentiation of oligodendrocyte precursor cell through P27kip1

作　　者：张敬华[1] 谢利娟[1] 朱建幸[1] 

机构地区：[1]上海交通大学附属新华医院儿内科,200092

出　　处：《中华围产医学杂志》2010年第4期318-323,共6页Chinese Journal of Perinatal Medicine

摘　　要：目的 探讨小剂量干扰素-γ（interferon-γ,INF-γ）调节少突胶质细胞前体P27kip1表达的有关信号通路.方法 新生1 d的SD大鼠无菌条件下取出大脑皮层制成混合胶质单细胞悬液,用完全培养基培养7～10 d,采用振荡结合差速贴壁方法并用无血清化学条件培养基培养获取少突胶质细胞前体.少突胶质细胞前体培养同时向各组培养细胞中分别添加IFN-γ、AG490、Fludarabine,采用逆转录-聚合酶链反应技术、Western印迹和流式细胞技术检测P27kip1 mRNA及蛋白的表达和对细胞分化的影响.结果 （1）IFN-γ组较对照组细胞P27kip1 mRNA和蛋白表达量减少（t=85.535,P〈0.05;t=12.481,P〈0.05）,IFN-γ＋AG490组和IFN-γ＋Fludarabine组较IFN-γ组细胞P27kip1 mRNA和蛋白表达量增加（P〈0.05）.（2）IFN-γ组的JAK2/STAT1磷酸化程度高于其他3组（P〈0.05）.（3）IFN-γ组髓鞘碱性蛋白阳性细胞百分比为（68.42±2.53）%,相对于对照组[（88.21±1.97）%]明显减少（t=10.682,P〈0.05）,IFN-γ＋AG490组髓鞘碱性蛋白阳性细胞百分比为（57.63±2.75）%,相对IFN-γ组进一步下降（t=5.002,P〈0.05）,而IFN-γ＋Fludarabine组髓鞘碱性蛋白阳性细胞百分比[（79.53±4.15）%]相对IFN-γ组却有所增加（t=3.957,P〈0.05）.结论小剂量INF-γ通过JAK2/STAT1信号通路调节少突胶质细胞前体P27kip1的表达.Fludarabine能有效减弱小剂量INF-γ对少突胶质细胞前体分化成熟的抑制作用.Objective To expore the mechanism of low-dose interfone-γ（IFN-γ） influences on differentiation of oligodendrocyte precursor cell. Methods The cerebral cortex samples were obtained from one day old SD rats to form mixed single cell suspensions. After culturing in full medium for 7 to 10 days, succession and differential velocity adherent technique were performed to acquire oligodendrocyte precursor cell and cultured in serum-free medium. IFN-γ, AG490 and Fludarabine were added during the culture of oligodendrocyte precursor cell and reverse transcription-polymerase chain reaction, Western blot and flow cytometry were performed to evaluate the expression of intracellular P27kip1 and its influence on the differentiation of oligodendrocyte precursor cell. Results （1）The expression of P27kip1 mRNA and protein was lower in IFN-γ group than in control group （t=85. 535, P〈0. 05;t= 12. 481, P〈0. 05）, while the expression of P27kip1 mRNA and protein in IFN-γ＋AG490 group and IFN-γ＋Fludarabine group were both higher than those in IFN-γ group （P〈0. 05）. （2） The phosphorylation levels of JAK2/STAT1 in INF-γ group were higher than that in the other three groups （P〈0. 05）. （3） The percentage of myelin basic protein positive cells was （68. 42 ± 2. 53）% in IFN-γ group, lower than that in control group [（88.21 ± 1.97）%]（t=10.682, P 〈 0.05）. Myelin basic protein positive cells in IFN-γ ＋ AG490 group were （57. 63 ±2. 75） %, lower than those in the IFN-γ group. The same figure in IFN-γ＋Fludarabine group were （79. 53±4. 15）% , higher than those in IFN-γ group （t = 3.957, P〈0.05）. Conclusions Low-dose IFN-γ can regulate the expression of intracellular P27kip1 through JAK2/STAT1 signal transduction pathway and Fludarabine may participate in this process and improve the differentiation and maturation of oligodendrocyte precursor cell.

关 键 词：干扰素Ⅱ型 酪氨酸磷酸化抑制剂 阿糖腺苷 周期素依赖激酶抑制剂P27 少突神经胶质 细胞分化 

分 类 号：R[医药卫生]