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阿霉素在小鼠离体培养脑片中增强Ⅱ型腺相关病毒的转导
引用本文:阿霉素在小鼠离体培养脑片中增强Ⅱ型腺相关病毒的转导. 阿霉素在小鼠离体培养脑片中增强Ⅱ型腺相关病毒的转导[J]. 首都医科大学学报, 2015, 36(6): 902-907. DOI: 10.3969/j.issn.1006-7795.2015.06.012
作者姓名:阿霉素在小鼠离体培养脑片中增强Ⅱ型腺相关病毒的转导
作者单位:1. 首都医科大学基础医学院生理与病理生理学系, 北京 100069;2. 首都医科大学教育部神经变性病重点实验室, 北京 100069;3. 首都医科大学基础医学院神经生物学系, 北京 100069;4. 北京脑重大疾病研究院, 北京 100069
基金项目:国家重点基础研究发展计划(973计划,2011CB504100),北京市属高等学校创新团队建设与教师职业发展计划项目(IDHT20140514),北京市教育委员会市属高校创新能力提升计划项目(TJSHG201310025006),北京高等学校青年英才计划项目(YETP1667)。
摘    要:目的 在小鼠离体培养脑片中观察Ⅱ型腺相关病毒(adeno-associated virus,AAV-2)载体介导的增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)的感染,并进一步研究阿霉素促进AAV-2转导的效应。方法 离体培养小鼠全脑脑片,在培养基中加入携带EGFP的AAV-2(AV2.EGFP)或阿霉素与AV2.EGFP的混合物,利用荧光显微镜实时观察活细胞中EGFP的表达情况。结果 AV2.EGFP感染脑片6 d后,EGFP稳定表达,同时加入1 μmol/L阿霉素可明显增强EGFP表达,阳性细胞数增加至2.8倍,该效果至少可持续6 d。并且,阿霉素与AV2.EGFP同时给药与在AV2.EGFP感染之后给药效果相似。结论 阿霉素在小鼠离体培养脑片中可明显增强AAV-2的转导,该研究有望促进AAV-2介导的基因治疗在中枢神经系统疾病中的应用。

关 键 词:阿霉素  脑片培养  腺相关病毒  
收稿时间:2015-09-02

Doxorubicin augments adeno-associated-virus type 2 transduction in mice brain slice culture
Gong Xiaoli,Wang Le,Wang Wei,Fu Xia,Zhang Ting,Wang Xiaomin. Doxorubicin augments adeno-associated-virus type 2 transduction in mice brain slice culture[J]. Journal of Capital Medical University, 2015, 36(6): 902-907. DOI: 10.3969/j.issn.1006-7795.2015.06.012
Authors:Gong Xiaoli  Wang Le  Wang Wei  Fu Xia  Zhang Ting  Wang Xiaomin
Affiliation:1. Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China;2. Key Laboratory of Neurodegenerative Disorders of the Ministry of Education, Capital Medical University, Beijing 100069, China;3. Department of Neurobiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China;4. Beijing Institute for Brain Disorders, Beijing 100069, China
Abstract:Objective To investigate whether doxorubicin can be applied to increase the efficacy of AAV-2 transduction in mice brain slice culture system. Methods We co-administrated doxorubicin with AAV-2 carrying enhanced green fluorescent protein(AV2.EGFP) into the mice brain slice culture, and then observed the EGFP expression from live cells in the brain slice by microscope. Results We found that EGFP fluorescence was very low within 6 days post-infection. One micromole doxorubicin augmented AAV-2 transduction dramatically to 2.8 folds compared with control group. This effect can last at least 7 days. Moreover, similar results were obtained when doxorubicin treated at the same time with or after AV2.EGFP. Conclusion Doxorubicin augmented AAV-2 transduction in mice brain slice. Our data provided evidence to facilitate AAV-mediated gene expression in the central nervous system with the treatment of doxorubicin.
Keywords:doxorubicin  brain slice  adeno-associated-virus  
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