中国神经再生研究(英文版) ›› 2014, Vol. 9 ›› Issue (5): 481-488.doi: 10.4103/1673-5374.130065

• 原著:退行性病与再生 • 上一篇    下一篇

rAAV/ABAD-DP-6His可有效减轻PC12细胞的氧化应激损伤

  

  • 收稿日期:2014-02-16 出版日期:2014-03-12 发布日期:2014-03-12
  • 基金资助:

    中国国家自然科学基金青年科学基金项目(30800338,30801211)和中国国家自然科学基金面上项目(30872721)

rAAV/ABAD-DP-6His attenuates oxidative stress-induced injury of PC12 cells

Mingyue Jia 1, Mingyu Wang  2, Yi Yang 1, Yixin Chen 3, Dujuan Liu 4, Xu Wang 1, Lei Song 1, Jiang Wu 1, Yu Yang 1   

  1. 1 Department of Neurology, the First Hospital of Jilin University, Changchun, Jilin Province, China
    2 Department of Neurology, People’s Hospital of Jilin Province, Changchun, Jilin Province, China
    3 Radioactive Medicine Specialty, College of Public Health in Jilin University, Changchun, Jilin Province, China
    4 Department of Burn and Plastic Surgery, the General Hospital of CNPC in Jilin, Jilin, Jilin Province, China
  • Received:2014-02-16 Online:2014-03-12 Published:2014-03-12
  • Contact: Jiang Wu, M.D., Ph.D., Department of Neurology, the First Hospital of Jilin University, Changchun 130021, Jilin Province, China, wujiang 540214@126.com. Yu Yang, M.D., Ph.D., Department of Neurology, the First Hospital of Jilin University, Changchun 130021, Jilin Province, China, yy197711@yahoo.com.cn.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China for the Youth, No. 30800338, 30801211; the National Natural Science Foundation of China, No. 30872721.

摘要:

作者前期研究已证实Aβ相关乙醇脱氢酶阻断肽能够拮抗Aβ42的神经毒性作用,但其是否对氧化应激损伤也有改善作用,目前尚缺乏研究报道。实验成功构建能够持续分泌表达Aβ相关乙醇脱氢酶阻断肽的重组腺病毒(rAAV/ABAD-DP-6His),且这种病毒能够增加过氧化氢氧化应激损伤的PC12细胞中超氧化物歧化酶活性,降低丙二醛的含量、细胞内Ca2 +浓度及活性氧水平,并保持线粒体膜电位的稳定性以及保持ATP含量恒定,减少细胞凋亡。表明rAAV/ABAD-DP-6His产生的融合肽Aβ相关乙醇脱氢酶阻断肽能有效减轻过氧化氢诱导PC12细胞的氧化应激损伤,发挥神经保护作用。

关键词: 神经再生, 神经退行性变, 基因治疗, 阿尔茨海默病, Aβ, Aβ相关乙醇脱氢酶, 腺病毒, 过氧化氢, 氧化应激, 线粒体功能障碍, NFSC grant

Abstract:

Our previous studies have revealed that amyloid β (Aβ)-binding alcohol dehydrogenase (ABAD) decoy peptide antagonizes Aβ42-induced neurotoxicity. However, whether it improves oxidative stress injury remains unclear. In this study, a recombinant adenovirus constitutively secreting and expressing Aβ-ABAD decoy peptide (rAAV/ABAD-DP-6His) was successfully constructed. Our results showed that rAAV/ABAD-DP-6His increased superoxide dismutase activity in hydrogen peroxide-induced oxidative stress-mediated injury of PC12 cells. Moreover, rAAV/ABAD-DP-6His decreased malondialdehyde content, intracellular Ca2+ concentration, and the level of reactive oxygen species. rAAV/ABAD-DP-6His maintained the stability of the mitochondrial membrane potential. In addition, the ATP level remained constant, and apoptosis was reduced. Overall, the results indicate that rAAV/ABAD-DP-6His generates the fusion peptide, Aβ-ABAD decoy peptide, which effectively protects PC12 cells from oxidative stress injury induced by hydrogen peroxide, thus exerting neuroprotective effects.

Key words: nerve regeneration, neurodegenerative disease, gene therapy, Alzheimer’s disease, amyloid beta peptide, amyloid beta binding alcohol dehydrogenase, adeno-associated virus, hydrogen peroxide, oxidative stress, mitochondrial dysfunction, NSFC grant, neural regeneration