摘要
目的研究RNA结合蛋白HuD在AD细胞和动物模型中的表达水平,探讨HuD与AD的关系;明确PI3K/AKT通路对神经细胞HuD表达的调控作用。方法免疫荧光检测HuD在C57BL/6小鼠脑组织中的表达水平;构建过表达APP的N2a APP细胞模型,Western blot检测HuD的表达情况;购买APP/PS1转基因小鼠,Western blot检测HuD在脑组织中的表达情况;应用LY294002处理3种神经细胞(N2a、SH-SY5Y和HT22),阻断PI3K/AKT通路,观察该通路对HuD的调控作用。结果 HuD在整个C57BL/6小鼠脑组织中均有表达,且在海马体的颗粒细胞尤为明显;在N2a APP细胞,HuD的表达水平明显低于对照组;APP/PS1转基因小鼠脑组织中的HuD水平也低于野生对照鼠;LY294002处理下调了HuD在神经细胞中的表达水平。结论 HuD在N2a APP细胞和APP/PS1转基因小鼠中均呈低表达;HuD在神经细胞中受PI3K/AKT通路的调控,为研究HuD与AD的关系及寻找新的AD诊断和治疗靶点提供了理论基础。
Objective To investigate the expression of HuD in AD cells and animal models to explore the relation- ship between HuD and AD and to clarify the role of PI3K/AKT pathway in the regulation of HuD expression in neurons. Methods The expression of HuD in brain tissue of C57BL/6 mice was detected by immunofluorescence. Western blot was used to detect the expression of HuD in N2aAPP cell model which was established by overexpression of APP. APP/PS 1 trans- genic mice were purchased and Western blot was used to detect the expression of HuD in brain tissue. N2a, SH-SY5Y and HT22 were treated with LY294002 to block the PI3K/AKT pathway and to observe the regulatory effect of this pathway on HuD. Results HuD was expressed in the whole C57BL/6 mouse brain tissue, especially in granulosa cells of hippoeampus. In N2aAPP cells, the expression level of HuD was significantly lower than that in control group; HuD in APP/PS1 transgenic mice brain LY294002 also down-regulated the expression of HuD in neurons. Conclusion HnD showed low expression in N2aAPP cells and APP/PS1 transgenie mice; HuD was regulated by PI3K/AKT pathway in neural cells, which provided a the- oretical basis for studying the relationship between HuD and AD and finding new targets for diagnosis and treatment of AD basis.
作者
欧阳婷
申秀银
王芬
王胜
罗涛
汪华侨
OUYANG Ting, SHEN Xiu-yin, WANG Fen, WANG Sheng, LUO Tao, WANG Hua-qiao(Department of Anatomy, Zhongshan School of Medicine, Sun Yat-sen Universi- ty, Guangzhou, Guangdong, 510080, Chin)
出处
《解剖学研究》
CAS
2018年第1期27-32,共6页
Anatomy Research
基金
国家自然科学基金(81501093)
广东省自然科学基金(2015A030313066
2015A030310251)
广东省科技计划项目(2014A020212622)
