离体培养海马神经干细胞中自噬现象的观察  

Observation of autophagy in cultured hippocampal neural stem cells

作　　者：董富兴[1] 张振中[1] 刘亚萍[2] 胡莹莹[3] 曲学彬[1] 于红丽[1] 姚瑞芹[1] 

机构地区：[1]徐州医学院神经生物学研究中心,徐州221004 [2]徐州医学院形态学实验教学中心,徐州221004 [3]南京医科大学康达学院基础医学部,连云港222000

出　　处：《神经解剖学杂志》2016年第1期56-60,共5页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(81271345;81302519);江苏省高校自然科学研究面上项目(15KJB180018)

摘　　要：目的:观察离体培养海马神经干细胞中自噬现象。方法:分离新生1 d大鼠海马组织,培养、扩增和传代,鉴定后取培养7 d的细胞,用RT-PCR法检测自噬特异性蛋白LC3和Bclin1的mRNA表达,Western Blot检测LC3和Bclin1的蛋白表达,透射电子显微镜观察神经干细胞中的自噬体的超微结构。结果:分离培养的神经干细胞呈Nestin免疫荧光阳性,且能分化为β-Ⅲtubulin^+的神经细胞和GFAP^+的胶质细胞;RT-PCR和Western Blot分别检测出神经干细胞有LC3、Bclin1的mRNA和蛋白表达;电镜下观察到神经干细胞胞质内有自噬体和自噬溶酶体。结论:正常离体培养的大鼠海马神经干细胞存在自噬现象,提示自噬可能对神经干细胞正常生理功能的维持有重要作用。Objective： To observe the autophagy in cultured hippocampal neural stem cells in vitro. Methods： After isolating hippocampus tissue from newborn rats 1 d, cells were cultured and identified. Ceus wene collected after cultured for 7 d and RT-PCR assay was used to detect the autophagy-specific protein LC3 and Belinl mRNA expression. Protein expression of LC3 and Bclinl were detected by Western Blot technology. Furthermore, we used transmission electron microscope to observe the ultrastructure of autophagosomes in neural stem cell. Results： Neural stem cells cultured for 7 d were nestin positive and could differentiate into β-Ⅲ tubulin positive neural cells and GFAP positive glial cells. LC3 and Bclinl mRNA and protein were expressed in neural stem cells detected by RT-PCR and Western Blot, respectively. Auto- phagosomes and autolysosomes were found in neural stem cells under the transmission electron microscope. Conclusion： The autophagy is present in cultured intact rat hippocampal neural stem cell, which suggests that autophagy may play an important role in maintaining normal physiological function of neural stem cells.

关 键 词：神经干细胞 自噬 LC3 BECLIN1 超微结构 大鼠 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]