中国神经再生研究(英文版) ›› 2018, Vol. 13 ›› Issue (6): 1081-1086.doi: 10.4103/1673-5374.233452

• 原著:视神经损伤修复保护与再生 • 上一篇    下一篇

黄芪甲苷对抗视网膜神经节细胞氧化应激损伤

  

  • 收稿日期:2018-02-04 出版日期:2018-06-15 发布日期:2018-06-15
  • 基金资助:

    黑龙江省教育厅基金(12541398)

Astragaloside IV protects RGC-5 cells against oxidative stress

Ming Hao1, Yu Liu2, Ping Chen3, Hong Jiang2, Hong-Yu Kuang1   

  1. 1 Department of Endocrinology, The First Clinical Hospital of Harbin Medical University, Harbin, Heilongjiang Province, China
    2 Department of Endocrinology, Heilongjiang Provincial Hospital, Harbin, Heilongjiang Province, China
    3 Department of Endocrinology, The First Hospital of Harbin, Harbin, Heilongjiang Province, China
  • Received:2018-02-04 Online:2018-06-15 Published:2018-06-15
  • Contact: Hong-Yu Kuang, M.D.,kuanghongyu1114@163.com.
  • Supported by:

    This study was supported by a grant from the Education Department of Heilongjiang Province of China, No. 12541398

摘要:

黄芪甲苷是中药黄芪的主要活性成分,具有较强的抗氧化应激和延缓周围神经病变进展的作用,为此实验拟观察黄芪甲苷是否对视网膜神经节细胞氧化应激损伤也有保护作用。实验设计传代培养鼠RGC-5细胞系,采用过氧化氢诱导细胞氧化应激损伤,并用黄芪甲苷干预。以CCK-8检测及DAPI染色检测显示,黄芪甲苷可提高RGC-5细胞存活率,降低细胞凋亡数量;以流式细胞仪检测发现,黄芪甲苷可降低过氧化氢引起的RGC-5细胞内ROS水平升高;以JC-1染色激光共聚焦显微镜观察显示,黄芪甲苷可以抑制过氧化氢引起的RGC-5细胞内线粒体膜电位的升高;以Western blot 检测发现,黄芪甲苷可以下调过氧化氢诱导的RGC-5细胞内的细胞色素C的释放、抑制细胞内促凋亡因子Bax及caspase-3蛋白的表达,并且可以增加抑凋亡因子Bcl-2蛋白的表达。上述数据可证实,黄芪甲苷可以保护视网膜神经节细胞对抗因过氧化氢诱导的氧化应激损伤。

orcid:0000-0003-1682-7013(Hong-Yu Kuang)

关键词: 黄芪, 过氧化氢, H2O2, 视网膜神经病变, 神经保护, 视网膜神经节细胞, 凋亡, 活性氧, 线粒体膜电位, 线粒体通路, 神经再生

Abstract:

Astragaloside IV is the main active compound of Astragalus membranaceus. Astragaloside IV has strong anti-oxidative activities and protective effects against progression of peripheral neuropathy. In this study, we determined whether astragaloside IV protects retinal ganglion cells (RGC) from oxidative stress injury using the rat RGC-5 cell line. Hydrogen peroxide (H2O2) was used to induce oxidative stress injury, with the protective effect of astragaloside IV examined. Cell Counting Kit-8 and 4′,6-diamidino-2-phenylindole staining showed that astragaloside IV increased cell survival rate and decreased apoptotic cell number. Flow cytometry showed that astragaloside IV decreased H2O2-induced reactive oxygen species levels. While laser confocal microscopy showed that astragaloside IV inhibited the H2O2-induced decrease of mitochondrial membrane potential. Western blot assay showed that astragaloside IV reduced cytochrome c release induced by H2O2, inhibited Bax and caspase-3 expression, and increased Bcl-2 expression. Altogether, these results indicate that astragaloside IV has potential protective effects against H2O2-induced oxidative stress in retinal ganglion cells.

Key words: nerve regeneration, Astragalus membranaceus, hydrogen peroxide, H2O2, retinopathy, neuroprotective effects, retinal ganglion cells, apoptosis, reactive oxygen species, mitochondrial membrane potential, mitochondrial pathway, neural regeneration