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银杏内酯B通过抑制视网膜神经节细胞凋亡促进其轴突生长

王振宇, 莫晓芬, 蒋旭花, 荣先芳, 缪华茂

复旦大学附属眼耳鼻喉科医院；脑科学研究院，上海 200032

摘要

青光眼、视神经炎等疾病的一个共同病理特征是视网膜神经节细胞(retinal ganglion cells, RGCs)的持续、不可逆性凋亡。银杏内酯B对脑组织神经元有很好的保护作用，并对神经元突起的生长和突触形成起重要作用。本文旨在观察银杏叶提取物(EGB761)和银杏内酯B对RGCs轴突生长的影响。采用三维立体培养系统进行视网膜植片培养，观察RCGs轴突数目和长度的变化，并对生长出的轴突进行免疫荧光染色鉴定，培养的视网膜植片用TUNEL法检测RGCs的凋亡，并进行免疫染色观察Caspase-3激活的情况。结果显示，与阴性对照组相比，EGB761或银杏内酯B处理的视网膜植片中轴突数量和长度明显增加。免疫荧光染色证实生长出的突起为RGCs轴突。相对阴性对照组，银杏内酯B处理的RGCs凋亡减少，Caspase-3的激活也被抑制。以上结果提示，银杏内酯B能够促进RGCs轴突的生长，主要机制是抑制RGCs凋亡。

关键词： 银杏叶提取物; 银杏内酯B; 视网膜神经节细胞; 神经保护; 轴突生长; 青光眼

分类号：R775；R338

[Ginkgolide B promotes axonal growth of retina ganglion cells by anti-apoptosis in vitro.] [Article in Chinese]

WANG Zhen-Yu, MO Xiao-Fen, JIANG Xu-Hua, RONG Xian-Fang, MIAO Hua-Mao

Eye and ENT Hospital, Shanghai Medical College； Institutes of Brain Science, Fudan University, Shanghai 200032, China

Abstract

One common feature of glaucoma, optic neuritis and some other optic nerve diseases is sustained and irreversible apoptosis of retinal ganglion cells (RGCs). Ginkgolide B is believed to protect neurons in brain and contribute to neurite outgrowth and synapse formation. The aim of the present study was to explore the effects of Ginkgo biloba extract (EGB761) and ginkgolide B on axonal growth of RCGs. Retina explants were cultured in three-dimensional tissue culture system, and the number and length of neurites were analyzed. Immunohistochemistry staining was performed to confirm that the neurite observed was axon of RGCs. TUNEL and activated caspase-3 staining were also applied to observe RGCs apoptosis. The result shows that neurites of RGCs treated with EGB761 or ginkgolide B were more and longer than those in control. The neurite is proved to be the axon of RGCs by immunostaining. Furthermore, compared with control group, RGCs treated with ginkgolide B showed decreased cellular apoptosis and inhibited caspase-3 activation. These results suggest ginkgolide B can promote RGCs axon growth by protecting RGCs against apoptosis.

Key words: EGB761; ginkgolide B; retina ganglion cells; neuroprotection; axon growth; glaucoma

收稿日期：2012-01-19　　录用日期：2012-03-21

通讯作者：莫晓芬　　E-mail: xfmo@fudan.edu.cn

引用本文：

王振宇, 莫晓芬, 蒋旭花, 荣先芳, 缪华茂. 银杏内酯B通过抑制视网膜神经节细胞凋亡促进其轴突生长[J]. 生理学报 2012; 64 (4): 417-424.

WANG Zhen-Yu, MO Xiao-Fen, JIANG Xu-Hua, RONG Xian-Fang, MIAO Hua-Mao. [Ginkgolide B promotes axonal growth of retina ganglion cells by anti-apoptosis in vitro.] [Article in Chinese]. Acta Physiol Sin 2012; 64 (4): 417-424 (in Chinese with English abstract).