中文摘要：

为了检测REST蛋白过表达或干扰对由PrP106-126毒性多肽引起的原代神经元纤维的影响，本试验通过免疫荧光技术和免疫印迹方法分别检测受PrP106-126刺激后的原代神经元中REST表达的变化情况。通过脂质体转染方法将已经构建好的pCMVHA-REST质粒转染进入原代神经元，或利用靶向REST的小干扰siRNA技术干扰原代神经元中REST蛋白的表达。用PrP106-126毒性多肽刺激转染了REST过表达或干扰质粒的原代神经元，利用免疫印迹检测突触后蛋白PSD-95的变化情况；利用免疫荧光观察突触后蛋白和神经纤维的损伤情况。结果显示，受到多肽刺激后，REST从神经元的胞浆向胞核转移的同时表达量也逐渐增加，24h时达到顶峰，之后缓慢减少。生理情况下，过表达REST促进突触后蛋白PSD-95的表达；病理情况下，REST的过表达减轻由毒性多肽引起的突触损伤、神经纤维断裂。相应地，干扰REST后。加剧毒性多肽对神经纤维的损伤。结果表明，REST蛋白的过表达缓解PrP106-126毒性多肽对原代神经细胞造成的病理性损伤，为进一步阐释REST对朊病毒及相关神经退行性疾病引起的病理性损伤的治疗作用和神经保护机制提供了理论依据。

英文摘要：

To investigate effect of overexpression or knockdown of REl-silencing transcription fac- tor （REST） on PrP106-126-induced synaptic damage and neurofibrillary degeneration in primary cortical neurons （PCCN）. To determine whether neurons express REST protein and PrP106-126 alters REST protein expression, REST protein expression was examined by immunofluorescence and Western blot analysis. The potential relationship among overexpression or knockdown of REST,postsynaptie proteins and neurofibrillary in PrP106 126-stimulated PCCN was tested by immunofluorescence and Western blot analysis. After stimulation with the neurotoxic PrP106-126 fragment in primary cortical neurons, REST translocated from the cytoplasm to the nucleus and protected neurons from harmful effects of PrP106-126. REST played an important role in maintaining postsynaptie integrity both in physiological and pathological conditions and protecting primary neurons from PrP106-126-induced neurofibrillary degeneration. REST could protect pri- mary cortical neurons from PrP106-126-induced synaptic damage and neurofibrillary degeneration, which may be important for the basic regulatory mechanism of neuron survival in prion diseases and associated neurodegenerative diseases in vivo.