解剖学报

解剖学报 ›› 2022, Vol. 53 ›› Issue (4): 432-439.doi: 10.16098/j.issn.0529-1356.2022.04.005

• 神经生物学 • 上一篇    下一篇

鹿茸多肽在阿尔茨海默病模型小鼠中抗氧化应激的作用机制

郎尉雅1 张春梅2 张羽镝2 刘忠锦2 衣同辉1 张可爽1 张海燕1*   

  1. 1.齐齐哈尔医学院基础医学院组织学胚胎学教研室, 黑龙江 齐齐哈尔 161006;2.齐齐哈尔医学院附属第一医院神经内科, 黑龙江 齐齐哈尔 161000
  • 收稿日期:2021-05-10 修回日期:2021-07-06 出版日期:2022-08-06 发布日期:2022-09-11
  • 通讯作者: 张海燕 E-mail:langweiya128@126.com
  • 基金资助:
    黑龙江省教育厅基本科研业务费基础研究项目

Anti-oxidative stress mechanism of velvet antler polypeptide in Alzheimer’s disease model mice

LANG  Wei-ya1  ZHANG  Chun-mei2 ZHANG  Yu-di2  LIU  Zhong-jin2  YI  Tong-hui1 ZHANG  Ke-shuang1  ZHANG  Hai-yan1*   

  1. 1.Department of Histology and Embryology, Basic Medical College of Qiqihar Medical University, Heilongjiang Qiqihar 161006,China; 2.Department of Neurology, the First Hospital Affiliated to Qiqihar Medical College, Heilongjiang Qiqihar 161000,China
  • Received:2021-05-10 Revised:2021-07-06 Online:2022-08-06 Published:2022-09-11
  • Contact: ZHANG Hai-yan E-mail:langweiya128@126.com

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摘要：

目的  探讨鹿茸多肽（VAP）在阿尔茨海默病（AD）模型中抗氧化应激的作用机制。   方法  选用 8 月龄雄性淀粉样前体蛋白（APP）/早老素1（PS1）双转基因小鼠随机分为模型组、VAP组，另以同窝阴性小鼠设立为对照组，每组12只。连续灌胃给药6个月后进行行为学、形态学和氧化应激相关指标检测。Western blotting 检测小鼠海马组织中APP及β-分泌酶 1（BACE1）蛋白的表达水平。另外以β-淀粉样蛋（Aβ）25～35（25 μmol/L）诱导 SH-SY5Y 细胞损伤建立AD细胞模型组，VAP组（VAP 500 mg/L+ Aβ25～35 25 μmol/L）和对照组。检测细胞凋亡、膜电位、活性氧簇（ROS）水平和氧化应激相关指标。   结果  动物实验发现，与模型组比较，VAP组小鼠逃避潜伏期缩短（P＜0.05）。模型组小鼠海马 CA1 区神经元减少且排列紊乱，VAP组排列较规则，形态有明显改善。与模型组比较，VAP组老年斑数量减少；与模型组比较，VAP组丙二醛（MDA）含量下降，超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶 （GSH-Px）含量上升，差异有统计学意义（P＜0.05）。与对照组比较，模型组APP 和BACE1含量上升，与模型组比较，VAP组APP和BACE1含量下降，差异有统计学意义（P＜0.05）。细胞培养实验发现，VAP组细胞凋亡率下降。与模型组比较，VAP组线粒体膜电位上升，ROS含量明显下降，MDA含量下降，SOD和GSH-Px含量上升，差异有统计学意义（P＜0.05）。   结论  VAP对AD模型动物和细胞的氧化应激损伤具有保护作用，可能是通过降低ROS产生，且提高抗氧化酶系活性，减少Aβ的沉积而实现的。

关键词: 鹿茸多肽, 阿尔茨海默病, 抗氧化, β-淀粉样蛋白, 线粒体, 免疫印迹法, 小鼠

Abstract:

Objective  To study the effect of velvet antler polypeptides (VAP) on antioxidant in Alzheimer’s disease model mice.    Methods  Eight months old male amyloid precursor protein (APP)/presenilin-1 (PS1) double transgenic mice were selected as Alzheimer’s disease (AD) model and divided into the model group and the VAP intervention group，12 in each group．Besides，normal mice of the same brood (with no transgene) were recruited as a control group (n=12) ．After 6 months of intragastric administration, behavior, morphology and oxidative stress related indicators were detected.SH-SY5 cells were used to establish AD model of damaged by Aβ25-35. The expression levels of APP and β-secreatase-1(BACE1) protein in mouse hippocampus were detected by Western blotting. VAP intervention group SH-SY5Y cells was cultured with VAP (500g/L) and amyloid  β(Aβ)25-35(25 μmol/L) for 24 hours. Control group cells were normally cultured by DMEM medium. Cell apoptosis, membrane potential, reactive oxygen species (ROS) levels and oxidative stress related indexes were detected.    Results  In animal models, compared with the model group, the escape latency of mice in the VAP intervention group was shortened (P＜0.05). The neuronal cells in the CA1 region of the hippocampus of the model group were reduced and arranged disorderly. The arrangement of the VAP intervention group was relatively regular, and the morphology was significantly improved. Compared with the model group, senile plaques were decreased in the VAP intervention group. Compared with the model group, the malondialdehyde (MDA) content of the VAP intervention group increased, and the superoxide dismutase（SOD）and glutathione peroxidase (GSH-Px) content increased, the difference was statistically significant. Compared with the control group, the APP and BACE1 content in the model group increased. Compared with the model group, the contents of APP and BACE1 in the VAP intervention group decreased, and the difference was statistically significant (P＜0.05). In the cell model, the apoptosis rates of the VAP intervention group decreased. Compared with the model group, the mitochondrial membrane potential of the VAP intervention group increased, the content of ROS decreased, the content of MDA decreased, and the content of SOD and GSH-Px increased. The difference were statistically significant (P＜0.05).     Conclusion  VAP has a protective effect on oxidative stress damage caused by Alzheimer’s disease model animals and cells, which may be achieved by reducing ROS production and increasing the activity of antioxidant enzymes to reduce Aβ deposition. 

Key words: Velvet antler polypeptide, Alzheimer’s disease, Antioxidant, β-amyloid, Mitochondria, Western blotting, Mouse

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