RACK1与小鼠空间学习记忆的相关性研究及自噬相关因子测定  

Correlation of RACK1 level in hippocampus and learning and memory in mice and the measurement of the autophagy related genes

作　　者：朱杰君[1] 刘立涛[1] 王文[1] 万莉红[1] 

机构地区：[1]四川大学华西基础医学与法医学院药理教研室,四川成都610041

出　　处：《四川生理科学杂志》2014年第4期147-151,共5页Sichuan Journal of Physiological Sciences

基　　金：国家自然科学基金资助(编号:81100989)

摘　　要：目的:研究RAKC1与小鼠空间学习记忆的相关性,同时通过自噬相关因子的检测探索自噬在该相关性中所起作用。方法:通过侧脑室注射Aβ25-35(10μL,1mg·mL-1)诱导小鼠学习记忆障碍,通过Morris水迷宫评价小鼠的空间学习记忆能力,采用HE染色法观察小鼠海马神经元损伤情况,同时以Realtime RT-PCR及Western blotting法分别测定小鼠海马组织中RACK1的mRNA及蛋白水平并将二者分别与小鼠空间学习记忆能力进行相关性分析。进一步采用Realtime RT-PCR法检测Beclin1的mRNA水平,采用Western blotting法检测BECLIN1及LC3II/LC3I的蛋白水平,并将两种蛋白水平分别与RACK1蛋白水平进行相关性分析。结果:侧脑室注射Aβ25-35明显导致小鼠空间学习记忆障碍(P=0.021)及海马神经元的广泛损伤(P=0.003),同时小鼠海马RACK1mRNA和蛋白水平也显著降低(P=0.00,P=0.05),且二者分别与小鼠的空间学习记忆能力正相关(P=0.05,P=0.011)。进一步研究发现,Aβ25-35导致小鼠海马BECLIN1、LC3II/LC3I蛋白水平增高(P=0.00,P=0.01)同时这两种蛋白水平分别与RACK1蛋白水平呈负相关(P=0.021,P=0.005)。结论:小鼠RACK1蛋白水平与空间学习记忆过程正相关,可能通过负性调节自噬过程介导。Objective： To investigate the correlation between the RACK1 and learning and memory in mice and to determine whether autophagy is involved in it. Methods. Male Kunming mice were injected intraventricularly with Aβ25-35 （10μL, 1 mg · mL-1 ） to establish the AD model. Morris water maze was used to assess the ability of learning and memory in mice. The damage induced by Aβ25-35 was evaluated by HE staining, mRNA levels of Rack1, Beclinl in hippocampus were determined by Realtime RT-PCR and protein levels of RACK1, BECLIN1, LC3II/LC3I were evaluated by Western blotting. Results： Compared with the NS group, Aβ25-35 administration induced a significantly impairment in learning and memory（P=0. 021） and a extend neurodemage in hippocam- pus（P=0. 003）. The expression of Rackl, both in mRNA and protein levels, were increased in the Aβ25-35 group（P=0. 00,P= 0. 05） and has shown to positively correlate with the capability of learning and memory respectively（P=0. 05, P=0. 011）. Compared with the NS group, Aβ25-35 group increase the protein expression of BECLIN1 and LC3II/LC3I（P=0. 00, P=0. 01）. And the negative correlation between BECLIN1 and RACK1, LC3II/LC3I and RACK1 were demonstrated（P=0. 021, P= 0. 005）. Conclusion. RACK1 plays an important role in learning and memory in mice by negatively regulates the process of autophagy.

关 键 词：空间学习记忆 海马 RACK1 自噬 

分 类 号：R749.16[医药卫生—神经病学与精神病学]