γ-分泌酶抑制剂DAPT对神经前体细胞系分化的作用研究  

Research on Regulation Function of γ-Secretase Inhibitor DAPT on the Differentiation of Neural Precursor Cell Line

作　　者：王琪[1] 李赫冬[1] 

机构地区：[1]四川大学华西第二医院发育与干细胞研究所,成都610041

出　　处：《生物医学工程学杂志》2015年第1期126-130,共5页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(30971633;31171045)

摘　　要：本研究目的为探究γ-分泌酶抑制剂DAPT(3,5-二氟苯乙酰-L-丙氨酰-S苯基甘氨酸t-丁酯)对神经元前体细胞系分化的影响,检验经DAPT诱导产生更多可用于移植的神经元的可能性。培养神经元前体细胞系GE6时,以培养基中加入4μmol/L DAPT的为实验组,不加DAPT的为对照组,分化4d,采用免疫荧光染色的方法分别检测两组细胞Tuj1、GFAP、O4的阳性率,采用qRT-PCR分别检测两组细胞Tuj1mRNA、GFAP mRNA相对表达量。免疫荧光染色表明实验组Tuj1阳性率较对照组增加,而GFAP和O4的阳性率较对照组减少,这些差异均有统计学意义(P<0.05)。qRT-PCR中两组Tuj1和GFAP mRNA的变化趋势与免疫荧光染色结果一致。结果提示DAPT能促进神经元前体细胞系向神经元分化,抑制其向胶质细胞分化,可以用DAPT诱导产生更多的可用于移植的神经元。This study aims to investigate the effect ofγ-Secretase Inhibitor DAPT,（N-[N-（3,5-Difluorophenacetyl）-L-alanyl]-S-phenylglycine t-butyl ester）,on the differentiation of neural precursor cells and the production of neurons in the neural precursor cell line GE6.GE6 was cultured in medium with 4μmol/L DAPT added as the experimental group and the untreated medium separately as the control group.After 4days of differentiation,we carried out the following experiments.We used immuno-fluorescent staining to observe the ratio of Tuj1,GFAP and O4 positive cells.We also used qRT-PCR to detect the effect of the DAPT on Tuj1 and GFAP mRNA transcription in the GE6.The results of immuno-fluorescent staining indicated that the Tuj1 ratio of experimental group was higher compared to that of the control group,but the GFAP and O4 ratio of experimental group was lower than that of the control group.The differences were statistically significant（P〈0.05）.The result of qRT-PCR was in accordance with immunofluorescent staining results.It was well concluded that DAPT could promote the neurogenic differentiation of neural precursor cell line rather than leading to gliogenic differentiation.More neurons could be obtained for transplantation with the addition of DAPT.

关 键 词：DAPT 神经元前体细胞系 神经元 分化 

分 类 号：R741[医药卫生—神经病学与精神病学]