氩气后处理降低大鼠脑缺血/再灌注损伤后的铁死亡  

Argon reprocessing to reduce ferroptosis after cerebral ischemia/reperfusion injury in rats

作　　者：张坤 徐科伟 王新军[2] 李天晓 王子亮 李和举 ZHANG Kun;XU Kewei;WANG Xinjun;LI Tianxiao;WANG Ziliang;LI Heju(Department of Cerebrovascular Diseases,Henan Provincial People's Hospital,Zhengzhou University People's Hospital,Zhengzhou,Henan 450003,China;Department of Neurosurgery,the Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou,Henan 450052,China)

机构地区：[1]河南省人民医院脑血管病科郑州大学人民医院,河南郑州450003 [2]郑州大学第五附属医院神经外科,河南郑州450052

出　　处：《中华实用诊断与治疗杂志》2024年第11期1081-1085,共5页Journal of Chinese Practical Diagnosis and Therapy

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20230049);河南省医学科技攻关计划省部共建项目(SBGJ202102026)。

摘　　要：目的观察氩气后处理对大脑中动脉闭塞大鼠重组组织型纤溶酶原激活剂(rtPA)溶栓后的神经保护作用,探讨其与铁死亡的关系。方法SPF级SD雄性大鼠63只,体质量250~350 g,随机分为假手术组、模型组、氩气处理组各21只。模型组采用改良Zea-Longa法制备大脑中动脉闭塞模型,缺血90 min后静脉注射rtPA 0.9 mg/kg溶栓,拔出线栓;氩气处理组模型制备方法同模型组,拔出线栓再灌注开始时吸入体积分数79%氩气和体积分数21%氧气的混合气体3 h;假手术组离断左侧颈外动脉后仅结扎颈总动脉,不置入线栓,不应用rtPA溶栓,不吸入氩气。再灌注3 h后处死3组大鼠取脑组织,行HE染色观察脑组织病理改变,行TTC染色检测脑梗死体积百分比,采用Western blot法检测谷胱甘肽过氧化物酶4(GPX_(4))蛋白相对表达量,采用DTNB比色法检测谷胱甘肽(GSH)含量,采用邻菲啰啉分光光度法检测铁含量。结果(1)HE染色示假手术组神经元、胶质细胞及毛细血管形态正常,结构完整,核仁清晰,无炎细胞渗出;模型组神经元皱缩、核固缩等神经元死亡现象明显;氩气处理组脑皮质缺血区组织和神经元细胞肿胀,神经元死亡现象较模型组减轻。(2)TTC染色示假手术组未见梗死区域;模型组皮层和皮层下见明显梗死区;氩气处理组皮层和皮层下梗死区域较模型组明显缩小。假手术组(0)、模型组[(40.29±2.26)%]、氩气处理组[(32.71±1.98)%]脑梗死体积百分比比较差异有统计学意义(F=3198.570,P<0.001)。氩气处理组脑梗死体积百分比小于模型组(P<0.05)。(3)假手术组、模型组、氩气处理组大鼠脑组织GPX_(4)蛋白相对表达量(30.92±0.85、12.73±0.50、29.08±0.31)、GSH含量[(50.82±0.63)、(25.48±0.75)、(37.45±0.81)mg/g]、铁含量[(44.82±1.01)、(132.37±1.63)、(68.91±0.86)μg/g]比较差异均有统计学意义(F=2252.0、2387.0、12087.0,P均<0.001)。模型组脑组织GPX_(4)蛋白相对表达量、GObjective To observe the neuroprotective effect of argon reprocessing after thrombolytic therapy with recombinant tissue plasminogen activator(rtPA)in rats with middle cerebral artery occlusion(MCAO),and to investigate its relationship with ferroptosis.Methods Sixty-three SPF-grade male SD rats,weighing 250-350 g,were randomly divided into shan-surgery group,model group,and argon treatment group,with 21 rats each.In model group,modified Zea-Longa method was used to prepare MCAO models,0.9 mg/kg rtPA was intravenously injected after 90-min ischemia for thrombolysis,and the suture was removed afterwards.Besides the above procedures in model group,the rats in argon treatment group inhaled a mixture of 79%argon and 21%oxygen for 3 h after removing the suture and starting reperfusion.The rats in sham-surgery group only had the common carotid artery ligated after dissecting the left external carotid artery,without the insertion of a suture and without the application of rtPA or inhalation of argon.After 3-h reperfusion,the rats in all three groups were sacrificed to harvest the brain tissues.HE staining was performed to observe the pathological changes,TTC staining was used to measure the percentage of brain infarct volume,Western blot was used to detect the relative expression of glutathione peroxidase 4(GPX_(4))protein,DTNB colorimetric method was used to measure the content of glutathione(GSH),and the o-phenanthroline spectrophotometric method was used to detect the iron content.Results(1)HE staining showed that sham-surgery group had normal morphology of neurons,glial cells,and capillaries with intact structure and clear nucleoli,no inflammatory cell exudation;model group had noticeable degree of neuronal shrinkage,pyknosis,and other signs of neuronal death;argon treatment group had swelling cerebral cortex of the ischemic area and neuronal cells,with slighter signs of neuronal death in compared with model group.(2)TTC staining showed no infarct area in sham-surgery group,obvious infarct areas in the cortex and sub

关 键 词：脑缺血再灌注损伤 重组组织型纤溶酶原激活剂 静脉溶栓 氩气 后处理 铁死亡 大鼠 

分 类 号：R743.3[医药卫生—神经病学与精神病学]