新型促红细胞生成素衍生肽对毛果云香碱致急性癫小鼠的神经保护作用及机制探究  

Neuroprotection of an Erythropoietin-derived Peptide in an Acute Pilocarpine-induced Epileptic Mice Model and Its Mechanism

作　　者：吴婷婷 赵亚楠 王京 冯宇 丁晶 汪昕 WU Ting-ting;ZHAO Ya-nan;WANG Jing;FENG Yu;DING Jing;WANG Xin(Department of Neurology,Zhongshan Hospital,Fudan University,Shanghai 200032,China)

机构地区：[1]复旦大学附属中山医院神经内科,上海200032

出　　处：《中国临床神经科学》2018年第5期487-494,共8页Chinese Journal of Clinical Neurosciences

基　　金：国家自然科学基金面上项目(编号:31771184;81771308)

摘　　要：目的探究新型促红细胞生成素衍生肽(DEPO)对毛果云香碱致急性癫小鼠的保护作用,借助N-甲基-D-天冬氨酸(NMDA)损伤神经元模型初步探讨DEPO的作用机制。方法 (1)健康成年C57BL/6小鼠随机分为DEPO组(DEPO 500μg·kg^(-1))、EPO组(EPO 50μg·kg^(-1))、溶剂组(ACN+Milliq,0.2 mL/只)、生理盐水组(生理盐水,0.2 mL/只)。腹腔注射毛果云香碱建立急性癫小鼠模型,比较各组干预后癫发作潜伏期、发作严重程度、死亡率的差异。(2)采用NMDA兴奋性毒性损伤神经元模型,分为DEPO组(DEPO 100μg·mL^(-1)干预)、EPO组(EPO 5μg·mL^(-1)干预)、对照组和正常组,检测各组乳酸脱氢酶(LDH)释放率、细胞存活率(MTT法)。(3)拮抗EPOR实验,设置EPO+anti-EPOR组,DEPO+anti-EPOR组,提前4 h加入anti-EPOR(1∶100,biorbyt)中和EPOR,检测LDH释放率和细胞存活率。结果 DEPO组和EPO组癫发作潜伏期分别较溶剂组、生理盐水组显著延长(P<0.05、P<0.001)。各组间死亡率、癫发作严重程度比较,差异无统计学意义。DEPO组和EPO组LDH释放率较对照组显著降低(P<0.01),细胞存活率较对照组显著增高(P<0.001)。DEPO+anti-EPOR组、EPO+anti-EPOR组的LDH释放率分别较DEPO组和EPO组显著增高(P<0.01),细胞存活率分别较DEPO组和EPO组显著降低(P<0.001)。结论新型DEPO可以显著延长毛果云香碱致急性癫小鼠的癫发作潜伏期,并通过EPOR介导其减轻NMDA诱导的神经元兴奋性毒性损伤,发挥神经保护作用。DEPO在癫治疗中存在可能的应用前景。Aim To explore the effects of a novel erythropoietin-derived peptide on the acute pilocarpine-induced epilepsy in mice, and preliminarily study the mechanism. Methods（1）Healthy adult C57 BL/6 mice were randomly divided into a derived peptide of erythropoietin（DEPO） group（500 μg·kg^（-1））, a erythropoietin（EPO） group（50 μg·kg^（-1））, a vehicle group（0.2 mL each） and a saline group（0.2 mL each）. A model of epilepsy in mice was made by intraperitoneal injection of pilocarpine. Measuredparameters included seizure latency, seizure severity and mortality.（2）Establish the NMDA-induced neuronal excitotoxicity model, cultured neurons were divided into DEPO group（100 μg·mL^（-1））, EPO group（5 μg·mL^（-1））, the control group and normal group. The LDH release and cell viability（MTT assay） were measured.（3）To perform EPO receptor（EPOR） blocking, anti-EPOR（1：100, biorbyt） was applied 4 h before the addition of DEPO, EPO in DEPO＋anti-EPOR, EPO＋anti-EPOR group, to inhibit EPO receptor. The LDH release and the cell viability were detected later. Results DEPO, EPO significantly prolonged the seizure latency compared with the vehicle and saline group（P〈0.05, P〈0.001）. There was no significant difference in mortality and seizure severity between each group. Both DEPO and EPO group significantly reduced the LDH release（P〈0.01） and enhanced the cell viability（P〈0.001） compared with vehicle group. The LDH release in DEPO＋anti-EPOR group and EPO＋anti-EPOR group was significantly higher than that in DEPO group and EPO group（P〈0.01）, and the cell viability in DEPO＋anti-EPOR group and EPO＋anti-EPOR group was significantly lower than that in DEPO group and EPO group（P〈0.001）. Conclusion The novel erythropoietin-derived peptide DEPO in our study significantly prolonged the seizure latency in an acute pilocarpine-induced mice model. DEPO attenuated the injury of NMDA-induced neuronal excitotoxicity and promoted the neuron

关 键 词：促红细胞生成素 促红细胞生成素衍生肽 神经保护 癫 抗凋亡 

分 类 号：R741.044[医药卫生—神经病学与精神病学]