| 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. Parkin基因过表达质粒和细胞模型的构建[J].现代生物医学进展英文版,2018,(3):407-412. |
| Parkin基因过表达质粒和细胞模型的构建 |
| Construction of Parkin Overexpression Plasmid and the Cell Model |
| Received:September 25, 2017 Revised:October 18, 2017 |
| DOI：10.13241/j.cnki.pmb.2018.03.002 |
| 中文关键词: 帕金森病 Parkin SH-SY5Y细胞 质粒 转染 |
| 英文关键词: Parkinson's disease Parkin SH-SY5Y cell Plasmid Transfect |
| 基金项目: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） |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| ABSTRACT 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 trans- fected 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). Conclusion: Using lipo- some transduction technology, the Parkin overexpression plasmid was successfully transfected into SH-SY5Y cells by lipo3000, the mR- NA and protein expression of Parkin was significantly increased. A cell model of Parkin gene overexpression was successfully estab- lished. |
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