局灶性皮质发育障碍致痫的蛋白质组学研究

目的 寻找皮质发育障碍致痫的疾病相关差异蛋白,以期寻找抗癫痫治疗的新靶点,同时寻找早期十预脑皮质发育障碍新的手段.方法 利用液氮损伤诱导皮质发育障碍,应用比较蛋白质组学方法研究致痫组和对照组皮质蛋白表达图谱差异,并对发现的差异蛋白质进行分析和鉴定.结果皮质发育障碍致痫组筛选到103个差异表达蛋白质斑点,其中64个在致痫组表达上调,39个在致痈组表达下调.有12个蛋白质最终鉴定确认,分别是lissencephaly-lprotein (LIS-1)、synaptotagmin Ⅳ、胶质纤维酸性蛋白(GFAP)、热休克蛋白70(HSP70)、生长相关蛋白-43(GAP-43)、neuronal enolase、tubulin beta chain、谷氨酰胺合成酶、神经元胞浆蛋白、电压依赖性阴离子通道1(VDAC1)、丙酮酸激酶(PK)、neurofilament light polypeptide.结论 12个差异蛋白鉴定有利于进一步研究皮质发育障碍与癫痫关系,该结果也为运用蛋白质组学方法寻找皮质发育障碍致痫治疗新靶点提供了实验数据.

Proteomics of epilepsy induced by focal disorder of cortical development

Objective To explore the proteomics of epilepsy induced by focal disorder of cortical development (DCD) in revealing the molecular mechanisms of epilepsy caused by DCD and looking for the candidate targets and new therapeutic approaches in clinical practice. Methods Animal models of DCD were established and induced by liquid nitrogen in healthy Wistar newborn rats. Animal model of DCD were divided into epilepsy group and control group according to Racine classification. The proteomics maps of the frontal cortex were obtained in the epilepsy group and the control group by two-dimensional electrophoresis and both Coomassie brilliant blue G250 and silver dying. The proteomics profiles of frontal cortex were preliminary analyzed with PD Quest 7.3 analysis package. The differentially expressed protein spots were excised from gel and digested with trypsin under optimal conditions. The masses of tryptic-digested peptides were acquired with a Voyager DA-STR matrix assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometer. The acquired monoisotopic masses of analyzed proteins were matched in silico with theoretical peptide masses of human protein in the swiss-prot database with a mass tolerance of less than 50 ppm. Results One hundred and three proteins of differential expression were observed in the frontal cortex tissues of epilepsy associated with disorder of cortical development in rats, in which 64 were detected to be up-regulated and 39 were down-regulated. Finally, 12 proteins were identified as Lissencephaly-1 protein, synaptotagmin Ⅳ, Glial fibrillary acidic protein, HSP70, growth associated protein 43, neuronal enolase, tubulin beta chain, glutamine synthetase, neuron cytoplasmic protein, voltage-dependent anion channel proteins 1, pyruvate kinase and neurofilament light polypeptide. Conclusion These proteins may play pivotal roles in the pathogenic mechanisms of epilepsy caused by disorder of cortical development and may provide new therapeutic targets for refractory epilepsy in the future.