中文摘要：

目的：利用原子力显微镜等方法,明确α-突触核蛋白（α-synuclein,α-syn）过表达对线粒体的影响。方法：将腺相关病毒（AAV）载体介导表达的α-syn（AAV/α-syn）病毒包装颗粒,给大鼠脑内黑质区定位注射,建立α-syn过表达的大鼠模型,并以AAV介导表达的报告基因LacZ（AAV/LacZ）为动物对照组。提取大鼠黑质脑组织的线粒体,并通过JC-1染色检测线粒体膜电势（ΔΨ）变化和Western blot检测线粒体中α-syn蛋白量的变化;采用原子力显微镜观测线粒体表面结构的变化。结果：实验组大鼠黑质脑组织过表达α-syn基因,第16周时,Western blot显示线粒体中α-syn蛋白量增加约2倍;JC-1染色发现ΔΨ下降;原子力显微镜观测可见线粒体体积增大,线粒体膜表面出现较多孔道样改变。结论：大鼠黑质脑组织过表达α-syn基因,第16周时,可导致α-syn蛋白在线粒体积聚;并可引起线粒体增大、膜形成孔道样改变、ΔΨ下降。

英文摘要：

Objective：To identify the effect of α-synuclein overexpression on mitochondrial membrane structure with atomic force microscopy. Methods：α-syn expression was mediated by AAV （adeno-associated viral vector） and Recombinant AAV/α-syn and AAV/LacZ viral particles were stereotaxically injected in the left side of rat substantia nigra （SN） for rat model of α-synuelein overexpression. Mitochondria were isolated from rats SN of Brain. Mitochondria were analysis with JC-1 staining, atomic force microscopy and Western blot. Results：By 16 weeks post-infection of AAV-α-syn ,the level of α-syn increased about 2 times in mitochondrial fraction with Western blot and mitochondrial membrane potential （△ψ） decreased with JC-1 staining. Furthermore, mitochondria swelling and porous like structure formed on the mitochondrial membrane with atomic force microscopy. Conclusion： The data suggested that α-syn could accumulate in mitocbondria, might form mitochondrial membrane pores and lead to △ψ decreases, α-syn might lead to mitochondrial dysfunction in Parkinson＇ s disease.