中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2013, Vol. 8 ›› Issue (1): 39-48.doi: 10.3969/j.issn.1673-5374.2013.01.005

• 原著:退行性病与再生 • 上一篇    下一篇

新型淀粉样β肽相关适配子抑制细胞内淀粉样β肽

  

  • 收稿日期:2012-09-08 修回日期:2012-11-07 出版日期:2013-01-05 发布日期:2013-01-05

The novel amyloid-beta peptide aptamer inhibits intracellular amyloid-beta peptide toxicity

Xu Wang1, Yi Yang1, Mingyue Jia1, Chi Ma2, Mingyu Wang1, Lihe Che3, Yu Yang1, Jiang Wu1   

  1. 1 Department of Neurology, First Hospital of Jilin University, Changchun 130021, Jilin Province, China
    2 Department of Neurosurgery, First Hospital of Jilin University, Changchun 130021, Jilin Province, China
    3 Department of Infection, First Hospital of Jilin University, Changchun 130021, Jilin Province, China
  • Received:2012-09-08 Revised:2012-11-07 Online:2013-01-05 Published:2013-01-05
  • Contact: Jiang Wu, M.D., Ph.D., Professor, Department of Neurology, First Hospital of Jilin University, Changchun 130021, Jilin Province, China, sjnkwujiang@sina.com.cn. Yu Yang, M.D., Ph.D., Associate professor, Department of Neurology, First Hospital of Jilin University, Changchun 130021, Jilin Province, China, yy197711@yahoo.com.cn.
  • About author:Xu Wang☆, Studying for doctorate.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 30872721; the National Natural Science Foundation for the Youth, No. 30801211, 30800338; and the Scientific Research Foundation for New Teachers of High Institutes, No. 200801831073, 200801831072.

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摘要：

淀粉样β肽相关乙醇脱氢酶阻断肽（ABAD-DP）可在线粒体内竞争性拮抗淀粉样β肽与ABAD的结合，抑制淀粉样β肽的细胞毒性作用。实验以肽适配子为基础，利用分子克隆技术，将ABAD-DP插入到人硫氧化还原蛋白（TRX）的双硫键中间，构建了能够表达阻断肽适配子TRX1-ABAD-DP-TRX2的融合基因，利用腺相关病毒实现其稳定表达。免疫荧光染色可见转染的融合基因TRX1-ABAD-DP-TRX2及淀粉样β肽的表达均位于NIH-3T3细胞内，提示TRX1-ABAD-DP-TRX2肽适配子具有与细胞内淀粉样β肽结合的能力。细胞形态学及MTT结果显示TRX1-ABAD-DP-TRX2可减轻淀粉样β肽对SH-SY5Y细胞的损伤，提高细胞活力。实验结果证明了将ABAD-DP构建成以人TRX为支架的肽适配子的可能性，且TRX1-ABAD-DP-TRX2对细胞淀粉样β肽的毒性有抑制作用。

关键词: 神经再生, 神经退行性疾病, 阿尔茨海默病, 适配子, 淀粉样β肽, 淀粉样β肽相关乙醇脱氢酶, 阻断肽, 硫氧化还原蛋白, 线粒体功能障碍, 分子克隆, 基因治疗, 基金资助文章, 图片文章

Abstract:

Amyloid β peptide binding alcohol dehydrogenase (ABAD) decoy peptide (DP) can competitively antagonize binding of amyloid β peptide to ABAD and inhibit the cytotoxic effects of amyloid β peptide. Based on peptide aptamers, the present study inserted ABAD-DP into the disulfide bond of human thioredoxin (TRX) using molecular cloning technique to construct a fusion gene that can express the TRX1-ABAD-DP-TRX2 aptamer. Moreover, adeno-associated virus was used to allow its stable expression. Immunofluorescent staining revealed the co-expression of the transduced fusion gene TRX1-ABAD-DP-TRX2 and amyloid β peptide in NIH-3T3 cells, indicating that the TRX1-ABAD-DP-TRX2 aptamer can bind amyloid β peptide within cells. In addition, cell morphology and MTT results suggested that TRX1-ABAD-DP-TRX2 attenuated amyloid β peptide-induced SH-SY5Y cell injury and improved cell viability. These findings confirmed the possibility of constructing TRX-based peptide aptamer using ABAD-DP. Moreover, TRX1-ABAD-DP-TRX2 inhibited the cytotoxic effect of amyloid β peptide.

Key words: neural regeneration, neurodegenerative disease, gene therapy, Alzheimer’s disease, aptamer, amyloid &beta, peptide, amyloid &beta, peptide binding alcohol dehydrogenase, decoy peptide, thioredoxin, mitochondrial dysfunction, molecular cloning, grants-supported paper, photographs-containing paper, neuroregeneration