脉冲强磁场对新生大鼠神经干细胞β-catenin表达的影响-康复医学与理疗学专业毕业论文.docxVIP

华中科技大学硕士学位论文 华 中 科 技 大 学 硕 士 学 位 论 文 PAGE 1 PAGE 1 脉冲强磁场对新生大鼠神经干细胞 β-catenin 表达的影响 摘 要 目的： 通过检测神经干细胞 β-catenin 基因和蛋白的表达，初步探讨脉冲强磁场 （0.1Hz,4T, 8 次）促神经干细胞增殖的作用机制。 方法： 取新生 SD 大鼠双侧脑室下区神经干细胞，无血清培养两周， 然后将细胞分 成实验组和对照组，实验组给予 0.1HZ,4T 脉冲强磁场干预 8 次，对照组置相同条件 下不行干预。此后第 1、3、5、7 天，分别用 RT-PCR 和 Western-Blot 方法检测两组神 经干细胞 β-catenin 基因和蛋白的表达。 结果： 干预后第 1、3、5、7 天, RT-PCR 法显示实验组神经干细胞β-catenin 基因表 达较对照组明显增高（P＜0.05)；Western-Blot 也显示实验组神经干细胞β-catenin 蛋白 表达较对照组明显增高（P＜0.05），增高水平在干预后第 7 天最显著。 结论： 新生大鼠神经干细胞经 0.1Hz, 4T 脉冲强磁场干预 8 次后，β-catenin 基因及 蛋 白 表 达 均 增 高 ， 提 示 脉 冲 强 磁 场 促 神 经 干 细 胞 增 殖 的 机 制 可 能 是 通 过 激 活 Wnt/β-catenin 通路实现。 关键词 脉冲强磁场 神经干细胞 增殖 机制 Wnt/β-catenin The Study of High-intensity Pulsed Electromagnetic Fields on expression of β-catenin of Neural Stem Cells of Neonatal Rat in vitro Abstract Object: To study the mechanism of High-intensity Pulsed Electromagnetic Fields （0.1HZ，4T，8 pulses）on facilitating the proliferation of neural stem cells by detecting the expression of β-catenin gene and protein. Methods: Neural stem cells (NSCs) were isolated from the sub ventricular zone (SVZ) of neonatal rats, and cultured in serum-free medium supplemented for two weeks. Then NSCs were divided into two groups, NSCs of experimental group were exposed in 0.1HZ, 4T High-intensity Pulsed Electromagnetic Fields for 8 pulses，and NSCs of control group were given sham stimulation. The gene and protein expressions of β-catenin of NSCs were assayed by RT-PCR and Western-blot at the 1st, 3rd, 5th, 7th day after stimulation respectively. Results: The NSCs cloned spheres were round shaped and translucent, and showed red fluorescence after stained by anti-nestin (cy3). The RT-PCR results showed β-catenin gene were highly expressed in HIPEMF exposed NSCs group (p ＜ 0.05), and the Western-blot results showed β-catenin protein were higher in experimental group (p＜0.05), especially at the 7th day after stimulation, the differences were statistically significant. Co