圣草酚改善5×FAD小鼠认知功能并调控Nogo-A/NgR/ROCK2信号通路  

Eriodictyol improves cognitive function of 5×FAD mice by regulating Nogo-A/NgR/ROCK2 signaling pathway

作　　者：李梦迪 李娜 郭敏芳[2] 孟涛 于婧文[2] 李雁冰 马存根 尉杰忠[1,2,3] Li Mengdi;Li Na;Guo Minfang;Meng Tao;Yu Jingwen;Li Yanbing;Ma Cungen;Yu Jiezhong(Research Center of Neurobiology/The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation for Multiple Sclerosis of State Administration of Traditional Chinese Medicine,Shanxi University of Chinese Medicine,Jinzhong 030619,Shanxi Province,China;Institute of Brain Science,Shanxi Datong University,Datong 037009,Shanxi Province,China;Datong Fourth People’s Hospital,Datong 037009,Shanxi Province,China)

机构地区：[1]山西中医药大学神经生物学研究中心/国家中医药管理局益气活血法治疗多发性硬化重点研究室,山西省晋中市030619 [2]山西大同大学脑科学研究所,山西省大同市037009 [3]大同市第四人民医院,山西省大同市037009

出　　处：《中国组织工程研究》2023年第32期5097-5102,共6页Chinese Journal of Tissue Engineering Research

基　　金：山西省2022年度“四个一批”科技兴医创新计划项目(2022XM33),项目负责人:尉杰忠;山西省基础研究计划项目(20210302123476),项目负责人:郭敏芳;山西省卫健委医学科技领军团队(2020TD05),项目负责人:马存根;国家中医药管理局多发性硬化益气活血重点研究室开放课题项目(2021-KF-21S),项目负责人:李梦迪。

摘　　要：背景:大量研究表明,神经生长抑制因子A(nerve growth inhibitory factor A,Nogo-A)可以通过与其下游分子结合激活Rho相关卷曲螺旋激酶(Rho-associated kinase,ROCK),从而对神经轴突的生长发挥抑制作用,与阿尔茨海默病中β-淀粉样蛋白的累积密切相关。实验室前期已经证明圣草酚能够改善5×FAD小鼠的认知功能障碍,但其对Nogo-A/NgR/ROCK2信号通路的调控尚未明确。目的:基于Nogo-A/NgR/ROCK2信号通路,探讨圣草酚对5×FAD小鼠认知功能的影响。方法:选取8月龄雄性C57BL/6小鼠分为野生组、野生给药组,8月龄雄性5×FAD小鼠分为模型组、圣草酚治疗组,每组8只,从小鼠33周龄开始,野生给药组和圣草酚治疗组每日腹腔注射圣草酚10μL/g,野生组和模型组注射同等体积的生理盐水,连续给药2个月,进行水迷宫、Y迷宫等行为学检测,检测结束后采用Western blot和免疫组织化学方法检测小鼠脑内β-淀粉样蛋白1-42,Nogo-A,NgR,p75NTR,Lingo1,ROCK2,p-ROCK2等相关靶蛋白的表达。结果与结论:①行为学检测结果显示,与野生组和野生给药组相比,模型组小鼠空间记忆能力与新事物探索能力均下降;与模型组相比,圣草酚治疗组空间记忆能力与新事物探索能力得到改善;②免疫组织化学与Western blot结果显示,与野生组和野生给药组相比,模型组β-淀粉样蛋白1-42、神经生长抑制因子Nogo-A及其神经生长抑制因子受体NgR,p75NTR,Lingo1的表达均升高;与模型组相比,圣草酚治疗组相关蛋白表达均有所下调;③与野生组和野生给药组相比,模型组Rho激酶相关蛋白ROCK2及p-ROCK2表达均升高;与模型组相比,圣草酚治疗组相关蛋白表达下调;④结果表明,圣草酚改善5×FAD小鼠的认知功能障碍与抑制Nogo-A/NgR/ROCK2信号通路相关。BACKGROUND:Numerous studies have shown that nerve growth inhibitory factor(Nogo-A)can activate Rho-associated coiled-coil kinase(ROCK)by binding to its downstream molecules,thereby exerting an inhibitory effect on nerve axon growth,which is closely related to the accumulation ofβ-amyloid protein in Alzheimer’s disease.Pre-laboratory work has demonstrated that eriodictyol improves cognitive dysfunction in 5×FAD mice,but its modulation of the Nogo-A/NgR/ROCK2 signaling pathway has not been clarified.OBJECTIVE:To investigate the effect of eriodictyol on cognitive function in 5×FAD mice via the Nogo-A/NgR/ROCK2 signaling pathway.METHODS:Male C57BL/6 mice,8 months old,were randomized into wild group and wild triodictyol-treated group,while 8-month-old male 5×FAD mice were divided into model group and eriodictyol-treated group,with eight animals in each group.Starting from 33 weeks of age,the wild eriodictyol-treated group and the eriodictyol-treated group were injected intraperitoneally with 10μL/g eriodictyol daily,and the wild and model groups were injected with the same volume of saline for 2 months.Behavioral assays such as Morris water maze and Y-maze tests were performed,and the expression ofβ-amyloid 1-42,Nogo-A,NgR,p75NTR,Lingo1,ROCK2,p-ROCK2 and other related target proteins in brain were detected by western blot and immunohistochemistry after testing.RESULTS AND CONCLUSION:Behavioral assay results showed that the spatial memory ability and novelty exploration ability were decreased in the model group compared with the wild group and the wild eriodictyol-treated group,while these abilities were improved in the eriodictyol-treated group compared with the model group.Immunohistochemistry and western blot results showed that the expression ofβ-amyloid 1-42,Nogo-A and its receptors such as NgR,p75NTR and Lingo-1 were increased in the model group compared with the wild group and the wild eriodictyol-treated group,and were all down-regulated in the eriodictyol-treated group compared with the model group.Th

关 键 词：阿尔茨海默病 圣草酚 5×FAD小鼠 神经保护 认知功能 

分 类 号：R459.9[医药卫生—治疗学] R332[医药卫生—临床医学] R741.02