中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (32): 5097-5102.doi: 10.12307/2023.589

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

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

李梦迪1，2，李  娜 1，2，郭敏芳2，孟  涛2，于婧文2，李雁冰1，2，马存根1，2，尉杰忠1，2，3   

  1. 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
  • 收稿日期:2022-08-30 接受日期:2022-10-19 出版日期:2023-11-18 发布日期:2023-03-22
  • 通讯作者: 尉杰忠，博士，教授，博士生导师，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；山西大同大学脑科学研究所，山西省大同市 037009；大同市第四人民医院，山西省大同市 037009 马存根，博士，教授，博士生导师，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；山西大同大学脑科学研究所，山西省大同市 037009
  • 作者简介:李梦迪，女，黑龙江省哈尔滨市人，山西中医药大学在读硕士，主要从事神经退行性变疾病的基础和应用研究。 李娜，女，陕西省榆林市人，山西中医药大学在读硕士，主要从事神经退行性变疾病的基础和应用研究。
  • 基金资助:
    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

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

Li Mengdi1, 2, Li Na1, 2, Guo Minfang2, Meng Tao2, Yu Jingwen2, Li Yanbing1, 2, Ma Cungen1, 2, Yu Jiezhong1, 2, 3   

  1. 1Research 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; 2Institute of Brain Science, Shanxi Datong University, Datong 037009, Shanxi Province, China; 3Datong Fourth People’s Hospital, Datong 037009, Shanxi Province, China
  • Received:2022-08-30 Accepted:2022-10-19 Online:2023-11-18 Published:2023-03-22
  • Contact: Yu Jiezhong, MD, Professor, Doctoral supervisor, 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 Ma Cungen, MD, Professor, Doctoral supervisor, 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
  • About author:Li Mengdi, Master candidate, 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 Li Na, Master candidate, 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
  • Supported by:
    Shanxi Province "Four Batches" Science and Technology for Medical Innovation Program in 2022, No. 2022XM33 (to YJZ); Shanxi Province Basic Research Program, No. 20210302123476 (to GMF); Medical Science and Technology Leading Team Project of Shanxi Provincial Health Care Commission, No. 2020TD05 (to MCG); Open Project of the Key Research Laboratory of Benefiting Qi for Acting Blood Circulation for Multiple Sclerosis of State Administration of Traditional Chinese Medicine, No. 2021-KF-21S (to JMD)

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摘要：

文题释义：

圣草酚：是一种天然二氢黄酮类化合物，广泛存在于水果、蔬菜、中药中，过去人们曾利用富含圣草酚的植物治疗过敏性鼻炎、风湿等疾病，随后展开的研究证明了圣草酚有抗氧化应激、抗炎等作用，并对神经退行性疾病有一定的治疗效果。
5×FAD小鼠：为携带5个突变基因的APP/PS1小鼠，5个突变基因分别为APP中的瑞典型(K670N/M671L)、佛罗里达型(I716V)和伦敦型(V717I)突变，以及PS1中的M146L和L286V突变，是目前应用较为广泛的阿尔茨海默病模型之一。
Nogo-A/NgR/ROCK2信号通路：Nogo-A能够与下游相应受体结合，最终激活下游的ROCK2信号，参与细胞的各种生理活动，包括抑制神经轴突的生长，从而影响神经元的再生。

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10 μL/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。

https://orcid.org/0000-0002-6106-9146(李梦迪)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 阿尔茨海默病, 圣草酚, 5×FAD小鼠, 神经保护, 认知功能

Abstract: 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 of β-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 10 μL/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 of β-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 of β-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. The expression of Rho kinase-related proteins ROCK2 and p-ROCK2 was increased in the model group compared with the wild group and wild eriodictyol-treated group, but was down-regulated in the eriodictyol-treated group compared with the model group. These findings suggest that eriodictyol treatment ameliorates cognitive dysfunction in 5×FAD mice, which is associated with inhibition of the Nogo-A/NgR/ROCK2 signaling pathway.  

Key words: Alzheimer’s disease, eriodictyol, 5×FAD moues, neuroprotection, cognitive function

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