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 (5): 923-929.doi: 10.4103/1673-5374.232489

• 原著:视神经损伤修复保护与再生 • 上一篇    下一篇

抑制视网膜神经节细胞凋亡：PACAP调节了维持线粒体的功能？

  

  • 收稿日期:2017-10-27 出版日期:2018-05-15 发布日期:2018-05-15
  • 基金资助:

    广东省医学科学基金；广东省自然科学基金，编号：2016A030313208；广东省科学技术规划项目，编号：2014A02021239

Inhibition of retinal ganglion cell apoptosis: regulation of mitochondrial function by PACAP

Huan-Huan Cheng1, Hui Ye1, Rui-Ping Peng1, Juan Deng1, Yong Ding2   

  1. 1 Department of Ophthalmology, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China
    2 Department of Ophthalmology, The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong Province, China
  • Received:2017-10-27 Online:2018-05-15 Published:2018-05-15
  • Contact: Juan Deng, Ph.D. or Yong Ding,Ph.D., viviadeng@163.com or dingyongjnu@163.com
  • Supported by:

    This study was supported by grants from the Medical Scientific Research Foundation of Guangdong Province of China, No.A2016271; the Natural Science Foundation of Guangdong Province of China, No. 2016A030313208; and the Science and Technology Planning Project of Guangdong Province of China, No. 2014A020212393

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摘要：

垂体腺苷酸环化酶激活多肽(PACAP)是具有保护视网膜神经元作用的内源性多肽，但具体保护机制尚不明确。在视网膜神经节细胞中富含有丰富的线粒体，鉴于此，实验假设PACAP的视网膜神经保护作用与调节线粒体功能有关。将视网膜神经节细胞(RGC-5)去血清培养48 h诱导建立细胞凋亡模型，同时予以100 nM的PACAP干预培养。细胞活性检测结果见，PACAP能显著增加RGC-5细胞活性并抑制去血清引起的过度产生活性氧。流式细胞结果显示，PACAP抑制去血清诱导的RGC-5细胞凋亡率和降低线粒体去极化细胞的比例。Western blot检测显示，PACAP可上调抗凋亡基因Bcl-2蛋白表达，并且抑制PACAP特异性受体PAC1蛋白的代偿性升高。上述数据客观显示了，PACAP抑制去血清诱导的离体视网膜神经节细胞的凋亡，其机制与维持线粒体功能有关。

orcid:0000-0002-1536-1840(Juan Deng)
        0000-0003-0538-9951(Yong Ding)

关键词: 垂体腺苷酸环化酶激活多肽, 垂体腺苷酸环化酶激活多肽特异性受体, 血清剥夺, 凋亡, 视网膜神经节细胞, 视网膜神经节细胞-5, 青光眼, 线粒体, 神经再生

Abstract:

Pituitary adenylate cyclase-activating polypeptide (PACAP) is an endogenous peptide with neuroprotective effects on retinal neurons, but the precise mechanism underlying these effects remains unknown. Considering the abundance of mitochondria in retinal ganglion cells (RGCs), we postulate that the protective effect of PACAP is associated with the regulation of mitochondrial function. RGC-5 cells were subjected to serum deprivation for 48 hours to induce apoptosis in the presence or absence of 100 nM PACAP. As revealed with the Cell Counting Kit-8 assay, PACAP at different concentrations significantly increased the viability of RGC-5 cells. PACAP also inhibited the excessive generation of reactive oxygen species in RGC-5 cells subjected to serum deprivation. We also showed by flow cytometry that PACAP inhibited serum deprivation-induced apoptosis in RGC-5 cells. The proportions of apoptotic cells and cells with mitochondria depolarization were significantly decreased with PACAP treatment. Western blot assays demonstrated that PACAP increased the levels of Bcl-2 and inhibited the compensatory increase of PAC1. Together, these data indicate protective effects of PACAP against serum deprivation-induced apoptosis in RGCs, and that the mechanism of this action is associated with maintaining mitochondrial function.

Key words: nerve regeneration, pituitary adenylate cyclase-activating polypeptide, pituitary adenylate cyclase-activating polypeptide receptor type 1, serum deprivation, apoptosis, retinal ganglion cell, retinal ganglion cell-5, glaucoma, mitochondria, neural regeneration