颞叶癫痫大鼠突触后致密物中差异性表达蛋白的筛选和鉴定

Screening and Identification of the Differentially Expressed PSD Proteins of Temporal Lobe Epilepsy Rats

目的筛选和鉴定颞叶癫痫(temporal lobe epilepsy,TLE)大鼠模型突触后致密物(postsynaptic density,PSD)中差异性表达的蛋白质。方法建立氯化锂-毛果芸香碱TLE大鼠模型,收集TLE组和健康对照组大鼠新鲜全脑组织,联合应用蔗糖密度梯度离心和膜顺序提取技术提取PSD蛋白质,进行双向凝胶电泳,通过PDQuest图像分析软件筛选TLE组差异性表达的蛋白质点,并应用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)对其进行鉴定。结果与健康对照组比较,TLE组存在70个差异性表达PSD蛋白质点,其中表达上调的蛋白质主要包括果糖二磷酸醛缩酶、肌酸激酶和热休克蛋白27,表达下调的蛋白质主要包括微管蛋白、肌动蛋白、中间丝蛋白α和脯氨酰顺反异构酶。结论TLE大鼠PSD中存在差异性表达蛋白质,其异常表达可能与TLE的形成、发作或神经元保护有关。

Objective To screen and identify the differentially expressed postsynaptic density （PSD） proteins among temporal lobe epilepsy （TLE） rats and normal ones. Methods Fresh whole brain tissue was immediately taken out. The PSD proteins were extracted and purified by using sucrose gradient centrifugation and membrane sequence extraction and isolated by using 2-DE. After gel were analyzed by PDQuest software. Differentially expressed PSD proteins were partially selected and identified by MALDI-TOF-MS. Results In the group of TLE, the spots number of differentially expressed PSD proteins was 70, and 11 of them were identified. The expressions of heat shock protein-27 （HSP-27）, fructose-bisphosphate aldolase A, creatine kinase （CK） were upregulated, but the expressions of tubulin, actin, intermediate filament proteins-alpha, PHase were downregulated in the group of TLE. Conclusions The differentially expressed PSD＇proteins might be related with the development, seizure attack of TLE or neuroprotection.