[1]王晓,黄晓平,黎玲,等.氧化应激促进重组腺相关病毒2型体外转导分析[J].华侨大学学报(自然科学版),2022,43(4):489-497.[doi:10.11830/ISSN.1000-5013.202201036]
　WANG Xiao,HUANG Xiaoping,LI Ling,et al.Analysis of Oxidative Stress Promoting In Vitro Transduction of Recombinant Adeno-Associated Virus Type 2[J].Journal of Huaqiao University(Natural Science),2022,43(4):489-497.[doi:10.11830/ISSN.1000-5013.202201036]
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氧化应激促进重组腺相关病毒2型体外转导分析()
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《华侨大学学报（自然科学版）》[ISSN:1000-5013/CN:35-1079/N]

卷:
第43卷
期数:
2022年第4期
页码:
489-497
栏目:
出版日期:
2022-07-18

文章信息/Info

Title:
Analysis of Oxidative Stress Promoting In Vitro Transduction of Recombinant Adeno-Associated Virus Type 2
文章编号:
1000-5013(2022)04-0489-09
作者:
王晓1;  黄晓平2;  黎玲1;  刘嘉1;  刁勇1
1. 华侨大学 医学院, 福建 泉州 362021; 2. 泉州师范学院 化工与材料学院, 福建 泉州 362000
Author(s):
WANG Xiao1;  HUANG Xiaoping2;  LI Ling1;  LIU Jia1;  DIAO Yong1
1. School of Medicine, Huaqiao University, Quanzhou 362021, China; 2. College of Chemical Engineering and Materials Sciences, Quanzhou Normal University, Quanzhou 362000, China
关键词:
重组腺相关病毒2型;  活性氧;  转导;  氧化应激
Keywords:
recombinant adeno-associated virus type 2;  reactive oxygen species;  transduction;  oxidative stress
分类号:
R511.8;R915
DOI:
10.11830/ISSN.1000-5013.202201036
文献标志码:
A
摘要:
为揭示氧化应激在重组腺相关病毒2型(rAAV2)转导中的作用,以过氧化氢(H2O2)和铁过载(Fe-NTA)氧化应激为模型,在293T,LO-2,Hela,A549细胞中,从转基因表达量、阳性细胞数、基因组数和病毒衣壳分布等方面研究氧化应激对rAAV2转导的影响.实验结果表明:H2O2和Fe-NTA能促进rAAV2转导,转基因表达量、阳性细胞数、基因组数与对照组相比的差异具有显著的统计学意义;细胞转导12 h后,氧化应激组核周围衣壳分布数目显著比对照组多,氧化应激能够促使rAAV2长时间聚集在细胞核周围;当氧化应激产生的活性氧(ROS)被N-乙酰-L-半胱氨酸(NAC)消除,则氧化应激促rAAV2转导作用消失,说明氧化应激通过ROS发挥促进rAAV2转导作用.
Abstract:
To reveal the role of oxidative stress in the transduction of recombinant adeno-associated virus type 2(rAAV2), using hydrogen peroxide(H2O2)and iron overload(Fe-NTA)oxidative stress as models, the effect of oxidative stress on rAAV2 transduction was studied from the aspects of transgene expression, positive cell number, genome number and viral capsid distribution in 293T, LO-2, Hela, and A549 cells. The experimental results showed that H2O2 and Fe-NTA could promote rAAV2 transduction, and the differences in transgene expression, positive cell number and genome number were statistically significant compared with those in the control group; after 12 h of cell transduction, the number of capsids distributed around the nucleus in the oxidative stress group was significantly higher than that in the control group. Oxidative stress could promote rAAV2 to accumulate around the nucleus for a long time; when the reactive oxygen species(ROS)produced by oxidative stress were eliminated by N-acetyl-L-cysteine(NAC), the effect of oxidative stress on promoting rAAV2 transduction disappeared, indicating that oxidative stress promotes rAAV2 transduction through ROS.

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备注/Memo

备注/Memo:
收稿日期: 2022-01-21
通信作者: 刁勇(1967-),男,教授,博士,博士生导师,主要从事基因药物的研究.E-mail:diaoyong@hqu.edu.cn.
基金项目: 国家自然科学基金资助项目(81502687, 81371669); 福建省自然科学基金资助项目(2021J01439); 福建省泉州市科技计划项目(2020C061)http://www.hdxb.hqu.edu.cn
更新日期/Last Update: 2022-07-20