中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2018, Vol. 13 ›› Issue (9): 1665-1672.doi: 10.4103/1673-5374.237140

• 原著:视神经损伤修复保护与再生 • 上一篇    

新型查尔酮类似物L2H17对氧化应激诱导视网膜神经节细胞的保护作用

  

  • 收稿日期:2018-05-29 出版日期:2018-09-15 发布日期:2018-09-15
  • 基金资助:

    中国国家自然科学基金项目（81473295, 81373312, 81371028）

The novel chalcone analog L2H17 protects retinal ganglion cells from oxidative stress-induced apoptosis

Lei Wang1, 2, Huai-Cheng Chen1, 2, Xi Yang1, 2, Jian-Jian Tao1, 2, Guang Liang1, Jian-Zhang Wu1, Wen-Can Wu2, Yi Wang1, Zong-Ming Song2, Xin Zhang1   

  1. 1 Chemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang Province, China
    2 The Eye Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, China
  • Received:2018-05-29 Online:2018-09-15 Published:2018-09-15
  • Contact: Zong-Ming Song, Ph.D. or Xin Zhang, Ph.D., szmeyes@126.com or wzmczhangxin@163.com.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81473295 (to ZMS), 81373312 (to XZ) and 81371028 (to WCW).

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摘要：

查耳酮是一种广泛存在于水果、蔬菜、香料和茶叶中的植物代谢产物，具有抗肿瘤、抗炎、免疫调节、抗菌、抗氧化等多种药理和生物活性。作者所在团队最近的研究显示其类似物具有抗氧化活性，而氧化应激是视网膜缺血再灌注损伤发生的病理学标志，可导致视网膜损伤和视力丧失，实验设计从大鼠视网膜神经节细胞系RGC-5细胞观察查耳酮类似物的作用。首先将RGC-5细胞经L2H7，L2H17，L3H6，L6H9和L43H2预处理，再以叔丁基氢氧化物进行氧化损伤，以MTT法检测细胞存活率，见5种查耳酮类似物中仅L2H17能明显增加氧化损伤RGC-5细胞的存活率；因而后续实验仅对L2H17干预的结果进行分析。以流式细胞仪测量细胞凋亡，以Caspase-3活性试剂盒测量Caspase-3活性，以WST-8试剂盒检测细胞内超氧化物歧化酶活性，以流式细胞术检测细胞内活性氧水平，以Western blot分析凋亡相关蛋白Bcl2和Bad及内质网应激/UPR通路蛋白激活转录因子4、磷酸化真核起始因子2和CHOP表达水平，以免疫荧光染色检测控制氧化应激的Nrf2免疫阳性反应。数据结果为L2H17可减少氧化损伤细胞的凋亡，抑制Caspase-3活性，增加Bcl2表达，减少Bad表达，增加超氧化物歧化酶活性，抑制活性氧产生，增加Nrf2免疫阳性反应，减少激活转录因子4、磷酸化真核起始因子2和CHOP表达。L2H17通过调节Nrf2对氧化应激诱导视网膜神经节细胞发挥了保护作用，告诉我们其具有成为治疗视网膜缺血再灌注药物的潜力。

orcid:0000-0001-7495-7049(Lei Wang)
        0000-0003-4342-1203(Xin Zhang)

 

关键词: 视网膜缺血再灌注损伤, 氧化应激, 活性氧, 细胞凋亡, Nrf2, 内质网应激, 查耳酮类似物, 视网膜神经节细胞, 神经再生

Abstract:

Chalcone is a plant metabolite widely found in fruits, vegetables, spices and tea, and has anti-tumor, anti-inflammation, immunomodulation, antibacterial and anti-oxidation activities, as well as many other pharmacological and biological effects. Our team has shown that its analogs have antioxidant activity, and oxidative stress is a pathological hallmark of retinal ischemia/reperfusion injury that can lead to retinal damage and visual loss. This investigation aims to identify a chalcone that protects retinal ganglion cells in vitro from the effects of oxidative stress and examine its mechanism. Rat retinal ganglion cell-5 cells were pretreated with chalcones and then exposed to tert-butyl hydroperoxide that causes oxidative damage. Controls received dimethyl sulfoxide only or tert-butyl hydroperoxide in dimethyl sulfoxide. Only (E)-3,4-dihydroxy-2′-methylether ketone (L2H17), of the five chalcone analogs, markedly increased the survival rate of oxidatively injured RGC-5 cells. Thus, subsequent experiments only analyzed the results of the L2H17 intervention. Cell viability and apoptosis were measured. Intracellular superoxide dismutase and reactive oxygen species levels were used to assess induced oxidative stress. The mechanism of action by L2H17 was explored by measuring the ER stress/UPR pathway and the expression and localization of Nrf2. All results demonstrated that L2H17 could reduce the apoptosis of oxidatively injured cells, inhibit caspase-3 activity, increase Bcl-2 expression, decrease Bad expression, increase the activity of superoxide dismutase, inhibit the production of reactive oxygen species, increase Nrf2 immunoreactivity, and reduce the activating transcription factor 4, phospho-eukaryotic initiation factor 2 and CHOP expression. L2H17 protects retinal ganglion cells induced by oxidative stress by regulating Nrf2, which indicates that it has the potential to become a drug for retinal ischemia/reperfusion.

Key words: nerve regeneration, retinal ischemia/reperfusion injury, oxidative stress, reactive oxygen species, apoptosis, nuclear erythroid-related factor-2, endoplasmic reticulum stress, chalcone analogs, retinal ganglion cells, neural regeneration