中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (11): 2330-2344.doi: 10.4103/1673-5374.310691

• 原著:视神经损伤修复保护与再生 • 上一篇    

Magnesium acetyltaurate prevents retinal damage and visual impairment in rats through suppression of NMDA-induced upregulation of NF-κB, p53 and AP-1 (c-Jun/c-Fos)

  

  • 出版日期:2021-11-15 发布日期:2021-04-13
  • 基金资助:

    马来西亚政府高等教育部的资助;马来西亚MARA大学Pengurusan Penyelidikan研究所(RMI)资助项目

Magnesium acetyltaurate prevents retinal damage and visual impairment in rats through suppression of NMDA-induced upregulation of NF-κB, p53 and AP-1 (c-Jun/c-Fos)

Lidawani Lambuk, Igor Iezhitsa, Renu Agarwal, Puneet Agarwal, Anna Peresypkina, Anna Pobeda, Nafeeza Mohd Ismail#br#   

  1. 1Center for Neuroscience Research (NeuRon), Faculty of Medicine, Universiti Teknologi MARA, Sungai Buloh Campus, Selangor, Malaysia; 2School of Medicine, International Medical University, Kuala Lumpur, Malaysia; 3Department of Pharmacology and Clinical Pharmacology, Institute of Medicine, Belgorod State National Research University, Belgorod, Russia
  • Online:2021-11-15 Published:2021-04-13
  • Contact: Igor Iezhitsa, PhD, igoriezhitsa@imu.edu.my.
  • Supported by:
    This study was financially supported by Ministry of Higher education, Government of Malaysia, under the grant No. RAGS/2013/UITM/SKK03/2 [UiTM file no. 600-RMI/RAGS 5/3 (103/2013)] and the Institut Pengurusan Penyelidikan (RMI), Universiti Teknologi MARA, Malaysia, under the grant 600-IRMI/MyRA 5/3/LESTARI (0088/2016).

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摘要：

研究已证实，乙酰牛磺酸镁（MgAT）对N-甲基-D-天冬氨酸（NMDA）诱导的视网膜细胞凋亡具有保护作用，其机制尚不明晰。为此，（1）实验将SD大鼠随机分为3组：对照组，接受玻璃体内注射媒介物；N-甲基-D-天冬氨酸组，接受玻璃体内注射N-甲基-D-天冬氨酸；N-甲基-D-天冬氨酸+乙酰牛磺酸镁组，接受玻璃体内注射N-甲基-D-天冬氨酸前进行乙酰牛磺酸镁预处理；（2）注射后7天，使用荧光金和BRN3A免疫染色的逆行标记检测了视网膜神经节细胞的存活率。使用视网膜电图（ERG）评估视网膜损伤后的功能，并评估核因子κB，p53和AP-1家族成员（c-Jun/c-Fos）在视网膜的基因表达。反转录聚合酶链反应（RT-PCR）和蛋白印迹检测和评估视网膜磷酸化核因子κB，磷酸化p53和AP-1水平。使用视觉对象识别测试（ORT）评估大鼠的视觉功能。逆行标记和BRN3A免疫染色检测；（3）结果均显示，与N-甲基-D-天冬氨酸组相比，N-甲基-D-天冬氨酸+乙酰牛磺酸镁的组中视网膜神经节细胞的数量明显增加；视网膜电图评估结果表明，用乙酰牛磺酸镁进行预处理可以部分保留N-甲基-D-天冬氨酸暴露的视网膜的功能活性；乙酰牛磺酸镁抑制了N-甲基-D-天冬氨酸诱导的视网膜磷酸化核因子κB、磷酸化p53和AP-1水平的增加，并抑制了N-甲基-D-天冬氨酸诱导的核因子κB、p53和AP-1家族成员的转录活性（c-Jun / c-Fos） ；两种视觉对象识别测试结果也显示了乙酰牛磺酸镁改善N-甲基-D-天冬氨酸暴露后识别视觉线索（即具有不同形状的物体）的困难；表明通过乙酰牛磺酸镁预处理，下调核因子κB，p53和AP-1介导的c-Jun/c-Fos的转录活性来抑制N-甲基-D-天冬氨酸诱导的神经元凋亡，从而预防N-甲基-D-天冬氨酸诱导的视网膜损伤，保留大鼠的视觉功能。

https://orcid.org/https://orcid.org/0000-0002-2852-8486 (Igor Iezhitsa)

Abstract: Magnesium acetyltaurate (MgAT) has been shown to have a protective effect against N-methyl-D-aspartate (NMDA)-induced retinal cell apoptosis. The current study investigated the involvement of nuclear factor kappa-B (NF-κB), p53 and AP-1 family members (c-Jun/c-Fos) in neuroprotection by MgAT against NMDA-induced retinal damage. In this study, Sprague-Dawley rats were randomized to undergo intravitreal injection of vehicle, NMDA or MgAT as pre-treatment to NMDA. Seven days after injections, retinal ganglion cells survival was detected using retrograde labelling with fluorogold and BRN3A immunostaining. Functional outcome of retinal damage was assessed using electroretinography, and the mechanisms underlying antiapoptotic effect of MgAT were investigated through assessment of retinal gene expression of NF-κB, p53 and AP-1 family members (c-Jun/c-Fos) using reverse transcription-polymerase chain reaction. Retinal phospho-NF-κB, phospho-p53 and AP-1 levels were evaluated using western blot assay. Rat visual functions were evaluated using visual object recognition tests. Both retrograde labelling and BRN3A immunostaining revealed a significant increase in the number of retinal ganglion cells in rats receiving intravitreal injection of MgAT compared with the rats receiving intravitreal injection of NMDA. Electroretinography indicated that pre-treatment with MgAT partially preserved the functional activity of NMDA-exposed retinas. MgAT abolished NMDA-induced increase of retinal phospho-NF-κB, phospho-p53 and AP-1 expression and suppressed NMDA-induced transcriptional activity of NF-κB, p53 and AP-1 family members (c-Jun/c-Fos). Visual object recognition tests showed that MgAT reduced difficulties in recognizing the visual cues (i.e. objects with different shapes) after NMDA exposure, suggesting that visual functions of rats were relatively preserved by pre-treatment with MgAT. In conclusion, pre-treatment with MgAT prevents NMDA induced retinal injury by inhibiting NMDA-induced neuronal apoptosis via downregulation of transcriptional activity of NF-κB, p53 and AP-1-mediated c-Jun/c-Fos. The experiments were approved by the Animal Ethics Committee of Universiti Teknologi MARA (UiTM), Malaysia, UiTM CARE No 118/2015 on December 4, 2015 and UiTM CARE No 220/7/2017 on December 8, 2017 and Ethics Committee of Belgorod State National Research University, Russia, No 02/20 on January 10, 2020.

Key words: AP-1 (c-Jun/c-Fos), electroretinography, magnesium acetyltaurate, neuroprotection, NF-κB, N-methyl-D-aspartate, object recognition tasks, p53, retinal excitotoxicity, retrograde labelling

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