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Parkin基因过表达质粒和细胞模型的构建 被引量:1

Construction of Parkin Overexpression Plasmid and the Cell Model
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摘要 目的:构建Parkin基因过表达质粒并转染SH-SY5Y细胞,为进一步研究帕金森病的发病机制及中药的作用环节奠定基础。方法:首先构建Parkin基因过表达质粒,采用脂质体转导技术,将Parkin过表达质粒应用Lipo3000转染SH-SY5Y细胞。荧光显微镜观察细胞绿色荧光蛋白的表达;RT-PCR检测其Parkin mRNA的表达;Western Blot技术检测其Parkin蛋白的表达。结果:转染组可观测到较多的绿色荧光蛋白表达;Parkin过表达细胞Parkin mRNA和蛋白的表达均显著提高(P<0.01)。结论:通过脂质体转导技术,应用Lipo3000可将Parkin过表达质粒成功转染入SH-SY5Y细胞,转染的基因和蛋白表达均较高,提示此法可成功构建Parkin基因过表达的细胞模型。 Objective: Construct Parkin overexpression plasmid and transfect into human neuroblastoma SH-SY5Y cells, with the aim of laying a foundation for studying the pathogenesis of Parkinson's disease and the role of traditional Chinese medicine. Methods:Firstly, construct Parkin overexpression plasmid. Using liposome transduction technology, the Parkin overexpression plasmid was transfected into SH-SY5Y cells by lipo3000. The green fluorescent protein was observed by fluorescence microscopy; RT-PCR technique was used to detect the expression of Parkin at mRNA level; Western-blot technique was used to detect the expression of Parkin at protein level.Results: More expression of enhanced green fluorescent protein(EGFP) was observed in the transfected group; the mRNA and protein expression of Parkin was significantly increased after transfection with Parkin overexpression plasmid(P〈0.01). Conclusions: Using liposome transduction technology, the Parkin overexpression plasmid was successfully transfected into SH-SY5Y cells by lipo3000, the mRNA and protein expression of Parkin was significantly increased. A cell model of Parkin gene overexpression was successfully established.
作者 马浩洁 冯琬迪 盖聪 强天遥 张淑静 胡京红 高誉珊 毛颖秋 郭振宇 孙红梅 MA Hao-jie1, FENG Wan-di1, GAI Cong1, QIANG Tian-yao1, ZHANG Shu-jing2, HU Jing-hong2, GAO Yu-shan2, MAO Ying-qiu3, GUO Zhen-yu1, SUN Hong-mei1(1 Depatment of Anatomy, Beijing University of Chinese Mechcine, Beijing, 100029, China; 2 Department ofscientitic research center, Beijing University of Chinese Medicine, Beijing, 100029, China; 3 Chinese traditional medicine research institute, Beijing University of Chinese Medicine, Beijing, 100029, Chin)
出处 《现代生物医学进展》 CAS 2018年第3期407-412,406,共7页 Progress in Modern Biomedicine
基金 国家自然科学基金项目(81573773) 北京中医药大学自主课题(2015-JYB-JSMS017)
关键词 帕金森病 PARKIN SH-SY5Y细胞 质粒 转染 Parkinson's disease Parkin SH-SY5Y cell Plasmid Transfect
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