天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (10): 1057-1062.doi: 10.11958/20210798

• 实验研究 • 上一篇    下一篇

褪黑素对年龄相关性黄斑变性模型小鼠视网膜氧化损伤的保护机制研究

董伟华，魏抗抗，帖红艳，何章彪，赵琳   

  1. 1商丘医学高等专科学校五官科教研室（邮编476006）；2郑州大学基础医学院人体解剖学教研室
  • 收稿日期:2021-04-06 修回日期:2021-05-20 出版日期:2021-10-15 发布日期:2021-10-15
  • 通讯作者: 董伟华 E-mail:dongwhlucky@163.com
  • 基金资助:
    2016年度河南省医学科技攻关计划项目

The protective effect and mechanism of melatonin on retinal oxidative damage in age-related macular degeneration model mice

DONG Wei-hua, WEI Kang-kang, TIE Hong-yan, HE Zhang-biao, ZHAO Lin #br#   

  1. 1 Department of Otolaryngology, Shangqiu Medical College, Shangqiu 476006, China; 2 Department of Human Anatomy,
    School of Basic Medicine, Zhengzhou University
  • Received:2021-04-06 Revised:2021-05-20 Published:2021-10-15 Online:2021-10-15

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摘要： 目的 探讨褪黑素（MEL）对年龄相关性黄斑变性（AMD）模型小鼠视网膜氧化损伤的保护作用及对沉默 信息调节因子1/叉头状转录因子1（SIRT1/FOXO1）通路的影响。方法 90只小鼠按随机数字表法分为正常组，模型 组，MEL低、中、高剂量组，每组18只。除正常组外，其余各组尾静脉注射25 µL/g NaIO3制备AMD小鼠模型，MEL低、 中、高剂量组分别以10、20、40 mg/kg MEL灌胃，正常组、模型组灌胃等体积生理盐水，1次/d，连续1周。眼底荧光造 影仪进行荧光素眼底血管造影（FFA），光学相干断层扫描（OCT）检测视网膜厚度；HE染色检测视网膜形态；试剂盒 检测眼眶静脉血血清中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）、过氧化氢酶（CAT）活性；Western blot法检测小鼠视网膜中SIRT1、FOXO1、Ac-FOXO1蛋白水平。结果 模型组视盘消失，血管出现收缩现象，视网膜 变白，血管出现破裂，视网膜各层细胞排列紊乱，外核层排列松散，细胞嵌入内节段/外节段，内核层细胞变大且松散 排列；随MEL剂量的升高，上述症状均逐渐改善。与正常组相比，模型组视网膜厚度，眼眶静脉血中SOD、GSH-Px、 CAT活性，视网膜中SIRT1、Ac-FOXO1/FOXO1蛋白水平降低（P＜0.05）；与模型组相比，MEL各剂量组视网膜厚度， SOD、GSH-Px、CAT活性，SIRT1蛋白水平均升高，MEL中、高剂量组视网膜中Ac-FOXO1/FOXO1蛋白水平升高（P＜ 0.05）。结论 MEL可能通过激活SIRT1/FOXO1通路实现对AMD小鼠视网膜氧化损伤的保护。

关键词: 褪黑激素, 黄斑变性, 年龄因素, 氧化性应激, 视网膜氧化损伤, SIRT1/FOXO1通路

Abstract: Objective To investigate the protective effect of melatonin (MEL) on retinal oxidative damage in agerelated macular degeneration (AMD) model mice and its effect on silent mating type information regulation 2 homolog 1 (SIRT1)/forkhead box transcription factor O1 (FOXO1) pathway. Methods Ninety mice were divided into normal group, model group, MEL low-dose, medium-dose and high-dose groups by random number table method, with 18 mice in each group. Except for the normal group, mice in the other groups were injected with 25 µL/g NaIO3 through tail vein to prepare AMD mouse model. Mice in the low-dose, medium-dose and high-dose MEL groups were gavaged with 10, 20 and 40 mg/kg MEL, respectively. Mice in the normal group and model group were gavaged with the same volume of normal saline, once a day, for a consecutive week. Fundus fluorescein angiography (FFA) was performed by fundus fluorescein angiography, and retinal thickness was detected by optical coherence tomography (OCT). The morphology of retina was detected by HE staining. The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) in serum of orbital venous blood were detected by kit. The protein levels of SIRT1, FOXO1 and Ac-FOXO1 were detected by Western blot assay. Results In the model group, the optic disc disappeared, the blood vessels contracted, the retina became white, the blood vessels ruptured, the cells in each layer of retina arranged disorderly, the cells in the outer nuclear layer arranged loosely, the cells embedded in the inner/outer segment, and the cells in the inner nuclear layer enlarged and arranged loosely. With the increase of MEL dose, the above symptoms were gradually improved. Compared with the normal group, the retinal thickness, orbital venous blood SOD, GSH-Px, CAT activity, and the protein levels of SIRT1 and Ac-FOXO1/FOXO1 in the retina decreased in the model group (P＜0.05). Compared with the model group, the retinal thickness, the activities of SOD, GSH-Px, CAT and the level of SIRT1 protein increased in the three MEL dose groups. The protein levels of ACFoxO1/FoxO1 in retina increased in MEL medium and high dose groups (P＜0.05). Conclusion MEL may protect the retina oxidative damage of AMD model mice by activating SIRT1/FOXO1 pathway.

Key words: melatonin, macular degeneration, age factors, oxidative stress, retinal oxidative damage, SIRT1/FOXO1 pathway