IL-1RA-PEP融合蛋白的构建及其对脑缺血再灌注后细胞凋亡的抑制作用研究  

Construction of IL-1RA-PEP fusion protein and its inhibitory effect on apoptosis in cerebral ischemia-reperfusion injury

作　　者：张冬冬[1,2,3] 邹民吉[2] 张亚涛[2] 徐涛[2] 付文亮[2] 黄志广[2] 甘向东 刘青[2] 董俊兴[1,3] 徐东刚[1,2] 

机构地区：[1]安徽医科大学研究生学院,合肥230032 [2]军事医学科学院基础医学研究所,北京100850 [3]军事医学科学院放射与辐射医学研究所,北京100850

出　　处：《军事医学》2016年第6期466-470,474,共6页Military Medical Sciences

基　　金：国家科技重大专项资助项目(2012ZX09102301-017)

摘　　要：目的构建白细胞介素-1受体拮抗剂(IL-1RA)与细胞渗透肽(PEP-1)融合蛋白,探讨其对脑缺血再灌注(I/R)后神经细胞凋亡的抑制作用。方法构建IL-1RA与PEP-1融合基因,通过诱导表达和分离纯化,制备IL-1RA-PEP融合蛋白。建立大鼠大脑中动脉阻断模型(MCAo),形成I/R损伤,静脉注射IL-1RA-PEP,24 h后检测其在大鼠脑组织的分布,以及脑组织梗死体积,炎性细胞浸润和神经细胞凋亡情况,并分析相关凋亡蛋白的表达变化。结果 IL-1RA-PEP融合蛋白具有拮抗IL-1的生物学活性,其能有效地渗透进入I/R大鼠脑组织。给药24 h后,缺血半球脑梗死体积和神经细胞凋亡数量减少,胱天蛋白酶(caspase)3和9等凋亡蛋白表达水平下降,同时Bcl-x L抗凋亡蛋白表达水平升高。结论 IL-1RA-PEP融合蛋白能有效地渗透I/R大鼠脑组织,通过对相关凋亡蛋白的调控,抑制了I/R后脑组织神经细胞凋亡,对I/R损伤具有治疗作用。Objective To investigate the inhibitory effect of the IL-1RA-PEP protein on apoptosis in cerebral ischemiareperfusion（ I / R） injury. Methods An IL-1RA-PEP fusion gene was engineered,and then the IL-1RA-PEP fusion protein was prepared by inducing expression and purification. The cerebral I / R injury was induced by transient occlusion of the middle cerebral artery（ MCAo）. At 24 h after intravenous administration with IL-1RA-PEP,the distribution of IL-1RAPEP,the cerebral infarction volume,the infiltration of inflammatory cells and the apoptosis number of neurocytes in the brain tissue were analyzed. Meanwhile,the levels of related apoptosis proteins were assessed. Results IL-1RA-PEP could antagonize the biological activity of IL-1 and infiltrate the ischemic brain effectively. After intravenous injection with IL-1RA-PEP,the cerebral infarction volume was reduced,the neurocyte apoptosis number was decreased,the expression levels of Caspase 3 and Caspase 9 apoptosis protein were reduced,and the expression levels of Bcl-xl anti-apoptotic protein were improved. Conclusion IL-1RA-PEP can efficiently penetrate the ischemic brain tissue. It can inhibit the neurocyte apoptosis and has a therapeutic effect on I / R injury by regulating the expression of related apoptosis proteins.

关 键 词：脑缺血 再灌注损伤 脑组织渗透 IL-1RA-PEP融合蛋白 细胞凋亡 

分 类 号：R743.3[医药卫生—神经病学与精神病学]