重组人促红细胞生成素对大鼠癫痫持续状态X-连锁凋亡抑制蛋白的影响及作用机制的实验研究  

Effect and mechanism of recombinant human erythropoietin on X-linked inhibitor of apoptosis protein in status epilepticus rats kindled by Pentylenetetrazol

作　　者：于江华[1] 史志勤 苏旭东[1] 周毅[1] 李彬[1] 王珊[1] 贾丽景[1] 王维平[1] 赵博 朱梦楚 冯晓红 耿丽淳 郭力[1] Yu Jianghua;Shi Zhiqin;Su Xudong;Zhou Yi;Li Bin;Wang Shan;Jia Lijing;Wang Weiping;Zhaobo;Zhumengchu;Feng Xiaohong;Geng Lichun;Guo Li(Department of Neurology,the Second Hospital Hebei Medical University,Shijiazhuang 050000,Chin)

机构地区：[1]河北医科大学第二医院神经内科,石家庄050000 [2]河北医科大学实验诊断教研室 [3]石家庄铁道大学

出　　处：《脑与神经疾病杂志》2018年第6期357-362,共6页Journal of Brain and Nervous Diseases

基　　金：河北省卫计委重点课题(20160658)

摘　　要：目的观察重组人红细胞生成素(r Hu EPO)对戊四氮(PTZ)点燃的癫痫持续状态(SE)的SD大鼠海马神经元的影响,应用磷脂酰肌醇3激酶(PI3K)抑制剂LY294002,观察X连锁凋亡抑制蛋白(XIAP)的变化情况,进一步探讨r Hu EPO作用的可能机制。方法采用PTZ点燃大鼠SE模型,将大鼠随机分为正常对照组(生理盐水,NS)、PTZ组(PTZ+NS)、r Hu EPO组(PTZ+r Hu EPO)、LY294002组(PTZ+LY294002+r Hu EPO)、二甲基亚砜(DMSO)对照组(PTZ+DMSO+r Hu EPO),检测大鼠行为学和脑电图(EEG)的改变及苏木精-伊红染色观察海马病理学的改变;免疫组织化学法观察磷酸化蛋白激酶B(p-PKB/p-Akt)、X-连锁凋亡抑制蛋白(XIAP)的表达;反转录多聚酶链反应(RT-PCR)方法检测各组大鼠海马XIAPm RNA的表达,Western blot方法检测各组大鼠海马Akt、p-Akt、XIAP蛋白的表达。结果 PTZ点燃大鼠SE后下调了p-Akt、XIAP的表达;r Hu EPO可以增加p-Akt、XIAP的表达,发挥神经保护作用;加入PI3K抑制剂LY294002,海马p-Akt、XIAP蛋白、XIAPm RNA的表达较r Hu EPO组减少,减弱了r Hu EPO的保护作用。结论从正反两个方面佐证了PI3K/Akt信号通路是r Hu EPO发挥神经保护作用的通路之一,其作用机制可能是r Hu EPO活化PI3K/Akt通路后,对线粒体凋亡途径的相关调控因子XIAP的表达进行了调控,进而介导线粒体凋亡途经,发挥抗凋亡、促存活的神经保护作用。Objective To observe the effects of recombinant human erythropoietin（r Hu EPO） on the hippocampal neurons of status epilepticus（SE） Sprague-Dawley（SD） rats which kindled by pentylenetetrazol（PTZ） and then explore the possible mechanism of r Hu EPO through the variations of X-linked inhibitor of apoptosis protein（XIAP） post the phosphatidyl inositol 3-kinase（PI3 K） inhibitors LY294002 application. Method The SE rats kindled by the PTZ were randomly divided into normal control group（normal saline, NS）、PTZ group（PTZ＋NS）, r Hu EPO group（PTZ＋r Hu EPO）、LY294002 group（PTZ＋LY294002＋r Hu EPO） and LY294002 solvent dimethyl sulfoxide（DMSO） control group（PTZ＋DMSO＋r Hu EPO）. The behavior changes and electroencephalogram（EEG） recording of rats were detected; The changes of hippocampal pathology were detected through HE staining; The expression of phosphorylated protein kinase B（PKB/p-Akt）、XIAP were detected through immunohistochemical method; The expression of hippocampal XIAPm RNA were detected through reverse transcription polymerase chain reaction（RT-PCR）; The expression of Akt, p-Akt and XIAP protein in hippocampal neurons were detected through Western blot method. Results The expression of p-Akt and XIAP were down-regulated by PTZ. r Hu EPO can increase the expression of p-Akt and XIAP, and then play the neuroprotective role. With the application of PI3 K inhibitor LY294002, the expression p-Akt, XIAP protein and XIAPm RNA decreased than r Hu EPO group in hippocampus, and decreased the protective effect of r Hu EPO. Conclusion The PI3 K/Akt signaling pathway is one of the pathways through that r Hu EPO play neuroprotective effects and be testified from the two aspects of positive and negative. The mechanism may be r Hu EPO activated the PI3 K/Akt pathway, regulated the expression of XIAP and then mediated mitochondrial apoptosis pathway to play the role of antiapoptotic and promote survival effects.

关 键 词：癫痫持续状态 重组人促红细胞生成素 磷脂酰肌醇3激酶/蛋白激酶B X-连锁凋亡抑制蛋白 

分 类 号：R742.1[医药卫生—神经病学与精神病学]