人参皂甙Rb1对马桑内酯癫痫的作用及海马蛋白组表达的影响  

EFFECTS OF GINSENOSIDE Rb1 ON RAT EPILEPSY INDUCED BY CORIARIA LACTONE AND PROTEOME CHANGES IN HIPPOCAMPUS

作　　者：赵春玲[1] 张春来[1] 徐倩[1] 杨烨[1] 张春燕[1] 

机构地区：[1]泸州医学院生理教研室

出　　处：《泸州医学院学报》2011年第5期539-545,共7页Journal of Luzhou Medical College

基　　金：四川省教育厅资助项目(2006C010)

摘　　要：目的:观察人参皂甙Rb1(GRb1)对马桑内酯(CL)所致大鼠癫痫及海马神经元损伤的作用,运用双向电泳(2-DE)技术获得可能与CL癫痫发作及GRb1神经保护作用有关的蛋白质成分,探讨海马组织蛋白质在癫痫发生发展过程中的作用和地位及GRb1对其的影响。方法:雄性成年SD大鼠随机均分为对照组、CL组和GRb1+CL组(n=10)。GRb1+CL组在实验前3d,大鼠每日灌胃GRb1(1.5mg/ml,30mg/kg),另2组灌胃等量生理盐水;实验日CL组和GRb1+CL组腹腔注射CL(1mg/ml,4mg/kg),对照组腹腔注射等量生理盐水。观察各组大鼠的行为学变化3h。各组半数动物麻醉后灌注固定,HE染色观察海马组织学改变。半数动物,断头处死剥离海马组织并提取蛋白,双向电泳(2-DE)分离,ImageMaster 2D Platinum v5.0软件差异表达蛋白质组分析,基质辅助激光解吸附离子化飞行时间质谱(MALDI-TOF-MS)鉴定蛋白质。结果:①癫痫发作的行为表现:对照组大鼠未见痫性发作;CL组痫性发作程度较高(Ⅲ～Ⅴ级);与CL组相比,GRb1+CL组大鼠的痫性发作程度明显减低,潜伏期变长,持续时间变短(P<0.01)。②海马组织学(HE染色)改变:对照组大鼠海马组织结构正常;CL组大鼠可见海马神经元变性坏死,尤以CA3区和齿状回最明显;GRb1+CL组大鼠海马神经元结构无明显改变。③获得大鼠海马组织蛋白质组双向电泳图谱;差异表达蛋白质组分析显示GRb1+CL组与对照组差异蛋白斑点84个,GRb1+CL组与CL组差异蛋白斑点120个。④鉴定出6个蛋白质分别是:脑肌酸激酶(brain creatine kinase)、吞蛋白A1(endophilin-A1)、UPF0628蛋白C10orf96同源物(UPF0628 proteinC10orf96 homolog)、细胞色素P-450(cytochrome P-450)、磷导素样蛋白(phosducin-like protein)及桥整合蛋白3(bridging inte-grator 3)。其中前3个蛋白质在GRb1+CL组表达低于CL组,后3个蛋白质在GRb1+CL组表达低于对照组。结论:GRb1可减轻CL所致大鼠癫痫的发作程度及海马神经元损伤Objective：To observe the effects of ginsenoside Rb1（GRb1） on rat epilepsy seizures and the chan-ges of protein expression in hippocampus induced by Coriaria Lactone（CL）,and to explore the molecular mechani-sm in development of CL induced epilepsy.Methods： Thirty adult male SD rats were randomly divided into 3 groups ： control group,CL epileptic group and GRb1 ＋CL group.The GRb1 ＋CL group was administered orally with GRb1（1.5mg/ml,30mg/kg） once a day for 3 days.The others were administered orally with same volume of NS.On the fourth day,the rats in CL and GRb1＋CL group were administered by intraperitoneal injection with CL（1mg/ml,4mg/kg）,while the others were administered by intraperitoneal injection with same volume of NS.The behavior changes were observed for 3 hours.One half rats of each group were anesthetized with 2% pentobarbital sodium,then fixed by infusion with 4% paraformaldehyde.Rat brain was taken and put into 4% paraformaldehyde for one night.Then the microstructure of hippocampal neurons of rats was observed with hematoxylin-eosin staining.The other half were decapitated and their hippocampus were stripped.Two-dimensional electrophoresis was applied to separate the proteins of hippocampus tissue from each group.Analysis of 2-DE maps was used to determine differential expression of proteins by ImageMaster 2D Platinum v5.0,and some protein spots expressed differently were picked up for further identification by matrix-assisted laser sorption ionization time of flight mass spectrometry（MALDI-TOF-MS）.Results： ①Behavior changes of epileptogenic rats： control rats had no seizures;CL epileptic rats showed Ⅲ～Ⅴattack;in GRb1 ＋ CL group,the degree of epileptic seizures and latency and duration were significantly lower and less than those of CL epileptic rats（P〈0.01）.②Morphologic changes in hippocampus： the structure of neurons in hippocampus in control group was normal.Degeneration or necrosis of neurons was observed in CL epileptic group rats,particularl

关 键 词：马桑内酯 癫痫 人参皂甙RB1 双向电泳 蛋白质组 

分 类 号：R338[医药卫生—人体生理学] Q593[医药卫生—基础医学]