重组人红细胞生成素对癫痫持续状态大鼠海马p-Akt和XIAP的影响  

Effects of rHuEPO on p-Akt and XIAP expressions in hippocampus of statural epilepticus rats

作　　者：史志勤[1] 王维平[1] 于江华[2] 郭力[1] 蒋国会[3] 王海祥[1] 范亚林[1] 

机构地区：[1]河北医科大学第二医院神经内科,石家庄050000 [2]北京天坛医院神经介入科,北京100050 [3]重庆市涪陵中心医院神经内科,重庆408000

出　　处：《第三军医大学学报》2009年第15期1453-1457,共5页Journal of Third Military Medical University

基　　金：河北省自然科学基金(C2007000852);河北省科技攻关项目(07276101D-21)~~

摘　　要：目的观察重组人红细胞生成素(recombinant human EPO,rHuEPO)对戊四氮(Pentylenetetrazol,PTZ)点燃的癫痫持续状态(status epilepticus,SE)的SD大鼠海马神经元磷酸化蛋白激酶B(p-PKB/p-Akt)和X-连锁凋亡抑制蛋白(X-linked inhibtor of apoptosis protein,XIAP)表达的影响,应用磷脂酰肌醇3激酶(phosphatidyl inositol3kinase,PI3K)抑制剂LY294002进一步探讨rHuEPO作用的可能机制。方法采用PTZ制作大鼠SE模型,将点燃后的大鼠随机分为PTZ组(PTZ+NS)、rHuEPO组(PTZ+rHuEPO)、LY294002组(PTZ+LY294002+rHuEPO)、LY294002溶剂二甲基亚砜(DMSO)对照组(PTZ+DMSO+rHuEPO),正常对照组腹腔注射生理盐水(n=35)。观察大鼠行为学和脑电图的改变;TUNEL法检测海马神经细胞的凋亡情况;免疫组织化学法观察p-Akt、XIAP的表达;RT-PCR法检测各组大鼠海马XIAPmRNA的表达;Western blot法检测各组大鼠海马Akt、p-Akt蛋白的表达。结果正常对照组仅见少量凋亡细胞,p-Akt和XIAP阳性细胞、p-Akt蛋白、XIAP mRNA均有少量表达;PTZ组与rHuEPO组、LY294002组、LY294002溶剂DMSO对照组比较,p-Akt和XIAP阳性细胞数、p-Akt蛋白表达及XIAP mRNA表达均减少(P<0.05),凋亡细胞数增加(P<0.05);rHuEPO组、LY294002溶剂DMSO对照组与LY294002组比较,p-Akt和XIAP阳性细胞数、p-Akt蛋白表达及XIAP mRNA表达均增加(P<0.05),凋亡细胞数减少(P<0.05)。结论rHuEPO在SE模型中活化了PI3K/Akt,提高p-Akt蛋白的表达,进而对线粒体凋亡途径的相关调控因子XIAP的表达进行了调控,从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 （rHuEPO） on the expressions of p-Akt and XIAP in hippocampus of statural epilepticus SD rats and the influence of LY294002, the inhibitor phosphatidyl inositol 3 kinase. Methods SD rats were divided into normal control group （normal saline, NS）, PTZ group （ PTZ ）, rHuEPO group （ PTZ ＋ rHuEPO）, LY294002 group （ PTZ ＋ LY294002 ＋ rHuEPO） , DMSO control group （PTZ ＋ DMSO ＋ rHuEPO）. Changes of behavior and EEG recordings were observed. The apoptotic neurons of the hippocampal region were detected by TUNEL method. The expressions of p-Akt and XIAP were detected by immunohistochemical method. The expression of XIAP mRNA located in the hippocampus was detected by RT-PCR. The expressions of Akt and p-Akt protein were determined by Western blot. Results A small quantity of apoptotic neurons and p-Akt and XIAP positive neurons but low expressions of protein p-Akt and XIAP mRNA were found in the normal control group. Decreased p-Akt and XIAP positive cells and the expressions of protein p-Akt and XIAP mRNA but increased apoptotic neurons were found in the PTZ group as compared with those in the rHuEPO group, LY294002 group and DMSO control group （ P 〈 0.05 ）. Increased p-Akt and XIAP positive cells and expressions of protein p-Akt and XIAP mRNA but decreased apoptotic neurons were found in rHuEPO group and DMSO control group as compared with those in the LY294002 group （ P 〈 0.05 ）. Conclusion rHuEPO could activate the signal delivery systems of PI3K/ Akt in the SE model and increase the expression of p-Akt protein, and hence mediate the approach for chondriosome apoptosis and neuroprotective effect in anti-apoptosis and enhancement of neuron survival though the regulation of the expression of XIAP, the apoptosis related regulating factors.

关 键 词：癫痫持续状态 重组人红细胞生成素 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：R338.26[医药卫生—人体生理学] R749.1[医药卫生—基础医学]