重组腺病毒介导的缺氧诱导因子-1α对脑缺氧后神经元中缺氧诱导因子-1α的调控  

Modulation of hypoxia inducible factor-1 alpha in neurons after cerebral hypoxia by recombinant adenovirus-mediated hypoxia inducible factor-1 alpha

作　　者：邹哲华[1] 陶陶[2] 晋明璐 徐坚 Zou Zhehua;Tao Tao;Jin Minglu;Xu Jian(Department of General Practice,The First Hospital of Qinhuangdao,Qinhuangdao,066000,China;Department of Rehabilitation Medicine,Guizhou Provincial People's Hospital,Guiyang,550002,China;Neurology,Chinese Medical Hospital of Liang Jiang New Area Chongqing,Chongqing,440020,China;Department of Neurology,The Affiliated Hospital of Guizhou Medical University,Guiyang,550001,China)

机构地区：[1]秦皇岛市第一医院全科医疗科,秦皇岛066000 [2]贵州省人民医院康复医学科,贵阳550004 [3]重庆两江新区中医院神经内科,重庆440020 [4]贵州医科大学附属医院神经内科,贵阳550004

出　　处：《中华老年医学杂志》2023年第8期970-973,共4页Chinese Journal of Geriatrics

基　　金：贵州省科学技术资金项目[黔科合J字(2007)2096];贵州省优秀科技教育人才省长专项资金[黔省专合字(2006)60号];贵州省卫生计生委科学技术基金[黔卫计发(2014)41号]。

摘　　要：目的探讨重组腺病毒介导的缺氧诱导因子-1α(HIF-1α)转染对缺氧后的大鼠大脑皮层神经元中HIF-1α的调控作用及可能机制。方法(1)制备大鼠大脑皮层神经元原代培养模型及缺氧模型。重组腺病毒缺氧诱导因子-1α(AdHIF-1α)转染和重组腺病毒空载体(Ad)转染正常及缺氧细胞;将其分为正常对照组(Normal)、缺氧组(Hypoxia)、AdHIF-1α基因转染组、Ad组。(2)荧光显微镜下观察AdHIF-1α/Ad转染情况;采用蛋白印迹(Westernblot)法分别在12h、24h、48h及72 h观察各实验组HIF-1α蛋白的表达。结果用Ad/AdHIF-lα转染缺氧的神经元后,在荧光显微镜下观察转染48h时荧光表达最明显。Westernblot检测经AdHIF-1α基因转染组各时间点HIF-1α的表达明显增加,12h有阳性表达,24h表达有所增加,48h达高峰,持续到72h开始下降,与缺氧组相比差异均有统计学意义(P<0.01,P<0.05),Ad组与缺氧组相比无统计学意义。结论重组腺病毒介导的HIF-1α基因转染能明显提高缺氧神经元中HIF-1α的表达量,有可能对缺氧后的神经元发挥保护作用。Objective To investigate the regulatory effects and possible mechanisms of recombinant adenovirus-mediated hypoxia inducible factor-1 alpha(HIF-1α)transfection on HIF-1αin post-hypoxic rat cortical neurons.Methods(1)Preparation of rats'cerebral cortical neurons in primary culture and hypoxia model.Recombinant adenovirus-mediated HIF-1α(AdHIF-1α)transfection and recombinant Ad transduction in normal and hypoxic cells.Then they will be divided into normal control group,hypoxia group,the recombinant adenovirus hypoxia-inducible factor-lα(AdHIF-lα)gene transfection group and recombinant adenovirus empty vector(Ad)transfection group.(2)To observe the transfection of AdHIF-lα/Ad under fluorescence microscopy.The expression of hypoxia-inducible factor-1 alpha(HIF-1α)were observed by Western blot analysis in each group of neurons at 12 h、24 h.48hand72h.Results Hypoxic neurons transfected with Ad/AdHIF-lαshowed the obvious expression 48 h under fluorescence microscopy.At each time of the AdHIF-lαgene transfection group,the expression of HIF-lαis significantly increased,the positive expression emerges at 12 h,the expression increased at 24 h,and it reaches the peak at 48 h,until 72 h declines,and it has statistical significance compared with the hypoxia group(P<0.01,P<0.05),but the Ad group has no statistically significant differences compared with the hypoxia group.Conclusions Thetransfection of recombinant adenovirus-mediated HIF-lαgene can significantly increase the expression levels of HIF-lαin hypoxic neurons,and it may play a protective role in the neurons after hypoxia.

关 键 词：腺病毒科感染 缺氧诱导因子1 Α亚基 年缺氧 脑 

分 类 号：R743[医药卫生—神经病学与精神病学]