红景天苷对NMDA介导的小鼠视网膜神经节细胞损伤的保护作用
向圣锦,李童,姜小涵,池在龙
325027 温州医科大学附属眼视光医院
The Neuroprotective Effect of Salidroside on Murine Retinal Ganglion Cell Apoptosis Induced by N-Methyl-D-Aspartate
Shengjin Xiang, Tong Li, Xiaohan Jiang, Zailong Chi
Eye Hospital, Wenzhou Medical University, Wenzhou 325027, China
摘要 目的:探讨红景天苷(Sal)对视网膜神经节细胞(RGCs)凋亡的抑制作用。方法:实验研究。36 只C57BL6J小鼠随机分为空白组、模型对照组、不同浓度Sal实验组,每组6 只,均以右眼为实验眼。空白组仅注射0.9%PBS溶液,模型对照组和实验组玻璃体腔注射N-甲基-D-天(门)冬氨酸(NMDA)1.5 μl建立RGCs损伤模型;建模前48 h,模型对照组注射1.5 μl 0.9% PBS溶液,实验组玻璃体腔注射不同浓度Sal(0.1、0.4、2、8 mmol/L,1.5 μl)。建模5 d后制备视网膜标本,观察视网膜RGCs存活及凋亡情况,Western Blot法检测视网膜中Caspase-3、Caspase-8 蛋白的表达。数据采用单因素方差分析。结果:HE染色显示空白组视网膜RGCs无变化,模型对照组RGCs显著减少,不同浓度Sal实验组RGCs细胞核染色数目较模型对照组逐渐增加,并呈浓度依赖性;经视网膜平铺片测定RGCs
存活情况,发现空白组RGCs正常存活,模型对照组仅少量RGCs存活,不同浓度Sal组RGCs存活率较模型对照组提高,各组小鼠RGCs阳性细胞数比较差异有统计学意义(F=212.0,P < 0.001)。同时,8 mmol/L浓度的Sal实验组Caspase-3、Caspase-8 蛋白表达明显低于模型对照组,差异有统计学意义(F=168.3,P < 0.001)。结论:Sal可以抑制NMDA介导的RGCs凋亡,对RGCs损伤有保护作用。
关键词 :
红景天苷 ,
谷氨酸兴奋性毒性 ,
视网膜神经节细胞 ,
细胞凋亡 ,
视神经保护
Abstract :Objective: To determine the neuroprotective effects of salidroside (Sal) on murine retinal ganglion cell (RGC) apoptosisinduced by N-methyl-D-aspartate (NMDA). Methods: Thirty-six C57BL6J mice were randomized into a blank normal control group, a model control group, and four experimental group (n=6 each). The right eyes of the experimental groups received a 1.5 μl intravitreal injection 0.1, 0.4, 2, or 8 mmol/L Sal (n=6 mice for each dose) 48 hours before NMDA was injected, and phosphate-buffered saline (PBS, 1.5 μl) were injected intravitreally in the model control group 48 hours before NMDA was injected. Intravitreal NMDA injections (1.5 μl at 0.1 mmol/L), were then performed in the model control group and in the experimental groups. PBS (1.5 μl) was injected intravitreally in the blank control group. Retinas derived from the mice were flatmounted and stained with hematoxylin and eosin to identify the RGCs. The number of RGCs was compared between NMDA and PBS-injected eyes for all groups. The levels of caspase-3 and caspase-8 were analyzed by Western Blot. Data were analyzed using ANOVA.Results: Compared with the blank model group, NMDA induced a significant increase of RGC apoptosis in the model control group and in the experimental groups. Compared with the model control group, Sal significantly inhibited RGC apoptosis in a dose-dependent manner (F=212.0, P<0.001), and the protein expression of caspase-3 and caspase-8 was significantly down regulated (F=168.3, P<0.001). Conclusions: Sal inhibited NMDA-induced RGC apoptosis and provided some neuroprotective effect for glutamatemediated RGC excitotoxicity.
Key words :
salidroside
glutamate excitotoxicity
retinal ganglion cells
apoptosis
neuroprotection
收稿日期: 2018-01-09
基金资助: 浙江省自然科学基金资助项目(LY15H270015);温州市科技计划项目(2016Y0379)
通讯作者:
池在龙(ORCID:0000-0002-3617-6425),Email:zailong.chi@mail.eye.ac.cn
引用本文:
向圣锦,李童,姜小涵,池在龙. 红景天苷对NMDA介导的小鼠视网膜神经节细胞损伤的保护作用[J]. 中华眼视光学与视觉科学杂志, 2018, 20(8): 469-474.
Shengjin Xiang, Tong Li, Xiaohan Jiang, Zailong Chi. The Neuroprotective Effect of Salidroside on Murine Retinal Ganglion Cell Apoptosis Induced by N-Methyl-D-Aspartate. Chinese Journal of Optometry Ophthalmology and Visual science, 2018, 20(8): 469-474. DOI: 10.3760/cma.j.issn.1674-845X.2018.08.004.
链接本文:
https://www.cjoovs.com/CN/10.3760/cma.j.issn.1674-845X.2018.08.004. 或 https://www.cjoovs.com/CN/Y2018/V20/I8/469
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