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Overexpression of α-synuclein in SH-SY5Y cells partially protected against oxidative stress induced by rotenone
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- Author:
- No author available
- Journal Title:
- ACTA PHYSIOLOGICA SINICA
- Issue:
- 5
- DOI:
- 10.3321/j.issn:0371-0874.2006.05.004
- Key Word:
- α-突触核蛋白;氧化应激;鱼藤酮;帕金森病
Abstract： 帕金森病(Parkinson's disease,PD)的发病机制涉及到遗传和环境因素.环境因素通过线粒体导致氧化应激和α-突触核蛋白(α-synuclein)聚集,但其确切的作用机制尚不明确.本文利用过表达α-突触核蛋白-增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)的人多巴胺能神经母细胞瘤细胞株SH-SY5Y为模型,研究α-突触核蛋白对鱼藤酮诱导氧化应激的影响,从而进一步了解α-突触核蛋白和细胞存活之间的关系.(1)用荧光显微镜观察融合绿色荧光蛋白的α-突触核蛋白的表达情况;(2)用实时定量PCR检测α-突触核蛋白基因的表达;(3)用免疫细胞化学测定α-突触核蛋白的分布;(4)用不同浓度的鱼藤酮作用细胞后,以MTT法测细胞的活力、DCF法检测细胞的氧化应激状态、黄嘌呤氧化酶法检测超氧化物歧化酶的活力,并用流式细胞仪分析细胞的凋亡.实时定量PCR结果显示,α-突触核蛋白基因表达量在α-突触核蛋白过表达的细胞要高于SH-SY5Y细胞,在荧光显微镜下可见绿色荧光蛋白和α-突触核蛋白的表达.鱼藤酮使细胞活力下降、线粒体complex Ⅰ的活性降低,诱导细胞内氧化应激,而过表达α-突触核蛋白的细胞可以部分抵抗鱼藤酮的毒性作用,表现为细胞抗氧化能力迅速增高(P＜0.05)和鱼藤酮诱导的细胞凋亡数目明显降低.本研究证明α-突触核蛋白对鱼藤酮产生的氧化应激有部分抵抗作用,而使过表达α-突触核蛋白的SH-SY5Y细胞对鱼藤酮的毒性作用表现出一定的耐受性.这种耐受性也可能是细胞对外界损害的一种代偿反应,从而促进细胞的存活.
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