基于铁死亡探讨AMPK抗小鼠脑缺血/再灌注损伤的作用及机制  

Protective Effects and Mechanisms of AMPK Activation Against Cerebral Ischemia/Reperfusion Injury in Mice Based on Ferroptosis

作　　者：胡淼 童旭辉 黄杰 陈蕾 门运政 田成鸿 董淑英 Hu Miao;Tong Xuhui;Huang Jie(Department of Pharmacology,School of Pharmacy,Bengbu Medical College,Bengbu 233030,China)

机构地区：[1]蚌埠医学院药学院药理学教研室,蚌埠233030 [2]蚌埠医学院心脑血管疾病基础与临床重点实验室,蚌埠233030

出　　处：《华中科技大学学报（医学版）》2021年第4期418-423,共6页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.81402930);国家级大学生创新计划项目(No.202010367053);蚌埠医学院心血管损伤与保护基础与临床应用创新团队(No.BYKC20190)。

摘　　要：目的基于铁死亡探讨腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)减轻小鼠脑缺血再灌注(ischemia-reperfusion,I/R)损伤的作用及机制。方法将ICR小鼠随机分为3组:假手术(Sham)组、脑I/R组和脑I/R+AMPK激活剂AICAR(I/R+AICAR)组。采用改良线栓法制备小鼠大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)模型,Longa 5分法进行小鼠的神经行为学评分,TTC染色法检测小鼠的脑梗死体积,丙二醛(malondialdehyde,MDA)试剂盒和Fe^(2+)试剂盒检测脑组织中MDA和Fe^(2+)水平,透射电子显微镜观察脑组织中线粒体形态变化,免疫印迹法检测小鼠脑组织中AMPK、p-AMPK及铁死亡相关蛋白TFR1、SLC7A11、GPX4的表达变化。结果相较于Sham组,I/R组小鼠神经行为学评分、脑梗死体积、MDA及Fe^(2+)水平、TFR1蛋白表达显著增高,脑组织中线粒体损伤明显,p-AMPK/AMPK、SLC7A11和GPX4表达显著降低;相较于I/R组,I/R+AICAR组小鼠神经行为学评分、脑梗死体积、MDA及Fe^(2+)水平、TFR1表达显著性降低,脑组织中线粒体损伤减轻,p-AMPK/AMPK、SLC7A11和GPX4表达显著增高。结论激活AMPK抗小鼠脑I/R损伤的作用可能与其抑制铁死亡有关。Objective To investigate the effects and mechanisms of AMP-activated protein kinase(AMPK)activation on cerebral ischemia-reperfusion(I/R)injury in mice.Methods ICR mice were randomly divided into 3 groups:sham-operated group,I/R group and I/R+AICAR(AMPK activator)group.The mouse model was established by the middle cerebral artery occlusion(MCAO).Longa’s scores were used to assess neurobehavioral scores of mice.The volume of cerebral infarction in mice was detected by TTC staining,and the levels of MDA and Fe^(2+)in cerebral tissue were detected by MDA kits and Fe^(2+)kits.Transmission electron microscope was used to observe the morphological changes of mitochondria in cerebral tissue.Western blotting was performed to detect the expression of AMPK,p-AMPK and ferroptosis related proteins TFR1,SLC7 A11 and GPX4 in cerebral tissue of mice.Results Compared to the sham-operated group,neurobehavioral scores,cerebral infarction volume,levels of MDA and Fe^(2+)and the expression of TFR1 were significantly increased.Mitochondrial damage in brain tissue was obvious,and the expression levels of p-AMPK/AMPK,SLC7 A11 and GPX4 were significantly decreased in I/R group.However,compared to I/R group,the neurobehavioral scores,cerebral infarction volume,levels of MDA and Fe^(2+)and the expression levels of TFR1 were significantly decreased,mitochondrial damage was significantly reduced in I/R+AICAR group,and the expression levels of p-AMPK/AMPK,SLC7 A11 and GPX4 were significantly increased.Conclusion The protective effects of AMPK activation on cerebral I/R injury in mice may be related to its inhibition of ferroptosis.

关 键 词：脑缺血/再灌注损伤 AMPK 铁死亡 

分 类 号：R734.31[医药卫生—肿瘤]