上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

银杏叶提取物保护视网膜缺血-再灌损伤与自噬的关系

袁海虹1，周 薇2，吴国忠1，包辉英1   

  1. 1.上海医药高等专科学校药理学教研室， 上海 201318； 2.上海交通大学 基础医学院药理学教研室， 上海 200025
  • 出版日期:2014-05-28 发布日期:2014-05-30
  • 作者简介:袁海虹（1977—）， 女， 讲师， 硕士; 电子信箱: yuanhaihong@126.com。
  • 基金资助:

    上海市教委科研创新项目（12YZ200）；上海医药高等专科学校基金[(2014zr002),FY(14)703-A5-1-01

Relationship between protective effects of ginkgo biloba extract on retinal ischemia-reperfusion injury and autophagy

YUAN Hai-hong1, ZHOU Wei2, WU Guo-zhong1, BAO Hui-ying1   

  1. 1.Department of Pharmacology, Shanghai Institute of Health Sciences, Shanghai 201318, China; 2.Department of Pharmacology, Basic Medical College, Shanghai Jiao Tong University, Shanghai 200025, China
  • Online:2014-05-28 Published:2014-05-30
  • Supported by:

    Innovation Program of Shanghai Municipal Education Committee, 12YZ200; Shanghai Institute of Health Sciences Fund,2014zr002;FY(14)703-A5-1-01

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摘要：

目的 研究银杏叶提取物（GBE）预处理对大鼠视网膜缺血-再灌注损伤后自噬的影响，探讨GBE保护视网膜缺血-再灌注损伤的作用机制。方法 实验大鼠随机分成5组，正常对照组、缺血-再灌注模型组、GBE低剂量（1 mg/kg）干预组、GBE中剂量（3 mg/kg）干预组和GBE高剂量（10 mg/kg）干预组，采用前房灌注生理盐水升高大鼠眼压至110 mmHg （1 mmHg=0.133 kPa）持续50 min的方法制作实验性视网膜缺血-再灌注损伤模型，观察大鼠视网膜内网状层（IPL）、内核层（INL）厚度以及节细胞层（GCL）细胞数；采用TUNEL法检测视网膜凋亡细胞；采用免疫组织化学和Western blotting法检测自噬相关蛋白LC3和Beclin1的表达变化。结果 与模型组相比，GBE（3 和10 mg/kg）干预组可明显改善大鼠缺血-再灌注损伤引起的IPL、INL和GCL受损，减轻视网膜损伤（P<0.05），降低视网膜细胞凋亡率（P<0.01），增加视网膜LC3和Beclin1阳性细胞数（P<0.01），上调缺血视网膜LC3和Beclin 1蛋白的表达。结论 GBE对大鼠视网膜缺血-再灌注损伤后神经元有保护作用，其机制可能与上调自噬相关基因Beclin1和MAP1-LC3表达有关。

关键词: 缺血-再灌注, 视网膜, 自噬, 银杏叶提取物

Abstract:

Objective To explore the effects of pretreatment by using the ginkgo biloba extract (GBE) on cell autophagy after rat retina being injured by the ischemiareperfusion and the mechanism on which GBE can protect the retina from ischemia-reperfusion injury. Methods Rats were randomly divided into the following five groups: the normal control group, ischemia-reperfusion model group, and groups intervened by low concentration (1 mg/kg), medium concentration (3 mg/kg), and high concentration (10 mg/kg) of GBE. The retinal ischemia-reperfusion injury model was induced by perfusing the anterior chamber with saline to elevate a hydrostatic pressure of 110 mmHg (1 mmHg=0.133 kPa) for 50 min. The thicknesses of the inner plexiform layer (IPL) and inner nuclear layer (INL) of rat retinas and cell numbers of the ganglion cell layer (GCL) were measured. The apoptosis of retinal cells was detected by the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphatebiotin nick end labeling (TUNEL). The expression variations of LC3 and Beclin1 which were related to the autophagy were detected by the immunohistochemical staining and Western blotting. Results Compared to the model group, damages of IPL, INL, GCL, and retina of intervention groups (pretreated by GBE of 3 mg/kg and 10 mg/kg) caused by the ischemia-reperfusion injury were significantly decreased (P<0.05); the apoptosis of retinal cells was decreased (P<0.01); the numbers of positive cells of LC3 and Beclin1 were increased (P<0.01); and the expressions of LC3 and Beclin1 were upregulated. Conclusion The mechanism of the protective effects of GBE on the retinal ischemia-reperfusion injury of rats may be related to the upregulation of expressions of Beclin1 and MAP1-LC3 which are related to the autophagy.

Key words: ischemia-reperfusion, retina, autophagy, ginkgo biloba extract