三重脑震荡模型大鼠海马神经元铁死亡的变化  

Changes of ferroptosis in hippocampal neurons of rats with multiple cerebral concussion

作　　者：王钱 江晓智 谭贝西 赵乐 王永辉 周然 Wang Qian;Jiang Xiaozhi;Tan Beixi;Zhao Le;Wang Yonghui;Zhou Ran(School of Basic Medicine,Hubei University of Chinese Medicine,Wuhan 430065,China;School of Basic Medicine,Shanxi University of Chinese Medicine,Jinzhong 030600,China)

机构地区：[1]湖北中医药大学基础医学院,武汉430065 [2]山西中医药大学基础医学院,晋中030600

出　　处：《中华行为医学与脑科学杂志》2022年第7期583-590,共8页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：国家自然科学基金(82104972);山西省卫生健康委员会科研项目(2021041);山西中医药大学科技创新能力培育计划"基础研究专项"(2020PY-JC-13)。

摘　　要：目的观察三重脑震荡模型(multiple cerebral concussion,MCC)大鼠海马神经元铁死亡(ferroptosis)的情况。方法90只清洁级雄性Wistar大鼠,体质量(250±10)g,按照随机数字表法分为对照组(12只)和模型组(78只),模型组采用自由落体法撞击大鼠额颞叶部(每天1次,连续3 d)制备大鼠MCC模型。将造模成功的大鼠随机分为12 h组、24 h组、48 h组、72 h组、7 d组、14 d组,每组12只。采用平衡木实验检测大鼠运动协调功能;ELISA法检测大鼠血清谷胱甘肽(glutathione,GSH)、神经元特异性烯醇化酶(neuron-specific enolase,NSE)及炎性因子白介素-1β(interleukin,IL-1β)、白介素-6(interleukin-6,IL-6)含量;比色法检测海马组织铁离子含量;RT-PCR法和Western blot法检测海马组织谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)、铁蛋白重链(ferritin heavy,FTH)和铁蛋白轻链(ferritin light,FTL)mRNA水平和蛋白含量;普鲁士蓝染色观察脑组织内铁沉积情况;透射电子显微镜法(transmission electron microscope,TEM)观察海马神经元及线粒体超微结构的变化。采用SPSS 24.0软件统计软件进行统计分析,多组间比较采用单因素方差分析,进一步两两比较采用LSD检验。结果平衡木实验中各组大鼠平衡木通过时间和运动协调能力评分均差异有统计学意义(F=30.08,60.34,均P<0.05),其中48 h组大鼠[(87.00±4.74)s,(4.75±0.43)分]通过时间和运动协调能力评分明显高于对照组[(35.13±6.99)s,(0.75±0.23)分](均P<0.05)。各组大鼠铁离子总含量、Fe2+含量和Fe3+含量均差异有统计学意义(F=25.20,94.42,40.25,均P<0.05),其中48 h组海马组织Fe2+含量明显高于对照组[(10.17±0.05)ng/μL,(8.65±0.01)ng/μL](P<0.05)。在RT-PCR和Western blot结果中,各组海马组织GPX4、FTH、FTL mRNA水平和蛋白水平均差异有统计学意义(F=37.94,82.09,49.01,71.63,28.94,15.78,均P<0.05),其中24 h组GPX4[(1.09±0.01),(0.23±0.01)]和FTL[(1.60±0.03),(0.64±0.02)]mRNA表达和蛋�Objective To observe the ferroptosis in hippocampal neurons of rats with multiple cerebral concussion(MCC).Methods Ninety clean grade male Wistar rats with a body mass of(250±10)g were randomly divided into control group(12 rats)and model group(78 rats)according to the random number table method.The rat of MCC model was prepared by hitting the frontotemporal lobe of rats with free fall method once a day for 3 consecutive days.Then the MCC model rats were randomly divided into 12 h,24 h,48 h,72 h,7 d and 14 d groups with 12 rats in each group.The balance beam experiment was used to detect the motor coordination function of the rats.The levels of interleukin-β(IL-1β),interleukin-6(IL-6),serum glutathione(GSH),neuron-specific enolase(NSE)in serum of rats were detected by ELISA.The content of iron ion in hippocampus was detected by colorimetry.The mRNA and protein levels of glutathione peroxidase 4(GPX4),ferritin heavy(FTH)and ferritin light(FTL)in the hippocampus were detected by RT-PCR and Western blot.Prussian blue staining was used to observe the iron deposition in brain tissue.The ultrastructural changes of hippocampal neurons and mitochondria were observed by transmission electron microscope.SPSS 24.0 statistical software was used for one-way ANOVA among groups,and LSD test was used for multiple pairwise comparison.Results In the balance beam experiment,the passing time and motor coordination score of rats in each group were significantly different(F=30.08,60.34,both P<0.05).The passing time and motor coordination score of rats in the 48 h group((87.00±4.74)s,(4.75±0.43))were significantly higher than those in the control group((35.13±6.99)s,(0.75±0.23))(both P<0.05).There was significant difference in the total iron ion content,Fe2+content and Fe3+content in hippocampus of rats in each group(F=25.20,94.42,40.25,all P<0.05),and the content of Fe2+in hippocampus of 48 h group was significantly higher than that of the control group((10.17±0.05)ng/μL,(8.65±0.01)ng/μL)(P<0.05).In the results of RT-PCR an

关 键 词：三重脑震荡模型 海马神经元 铁死亡 自由落体法 大鼠 

分 类 号：R-332[医药卫生] R651.15