TIGAR基因过表达对低氧复氧损伤大鼠海马神经元的影响

EFFECT OF OVEREXPRESSION OF TP53-INDUCED GLYCOLYSIS AND APOPTOSIS REGULATOR ON HIPPOCAMPAL NEURONS IN A RAT MODEL OF HYPOXIA/REOXYGENATION INJURY

目的探讨TP53介导的糖酵解和凋亡诱导因子(TIGAR)过表达对低氧复氧损伤大鼠海马神经元的影响。方法采用随机数字表法将提取的原代海马神经元细胞随机分为正常组(A组)、低氧复氧损伤组(B组)、过表达对照组(C组)、TIGAR过表达组(D组)、沉默对照组(E组)、TIGAR沉默组(F组)6组。建立低氧复氧损伤模型,然后各组采用相应的方法处理,使用激光共聚焦显微镜检测不同组神经元细胞内活性氧(ROS)的浓度,流式细胞仪检测各组细胞凋亡率,Western blot方法检测各组神经元中TIGAR表达量及凋亡诱导因子(AIF)、线粒体分裂相关蛋白Drp1、Fis1和Rho A蛋白1(ROCK1)的表达。结果与B组比较,D组神经元细胞内TIGAR的表达量明显上调(P<0.05),ROS浓度、细胞凋亡率及AIF、Drp1、Fis1、ROCK1蛋白的表达水平明显降低(P<0.05);与B组比较,F组TIGAR的表达量下调,ROS浓度、细胞凋亡率以及AIF、Drp1、Fis1、ROCK1蛋白的表达明显升高(P<0.05);与B组比较,C组和E组上述各指标差异无显著性(P>0.05)。结论海马神经元低氧复氧模型中,TIGAR通过调控线粒体分裂对神经元起保护作用。

Objective To investigate the effect of TP53-induced glycolysis and apoptosis regulator（ TIGAR） overexpression on hippocampal neurons in a rat model of hypoxia/reoxygenation injury. Methods Primary cultured hippocampal neurons were randomly divided into normal group（ group A）,hypoxia/reoxygenation injury group（ group B）,overexpression control group（ group C）,TIGAR overexpression group（ group D）,silencing control group（ group E）,and TIGAR silencing group（ group F）,using a random number table. A model of hypoxia/reoxygenation injury was established,and each group was given the correspon-ding treatment. A laser scanning confocal microscope was used to measure the concentration of reactive oxygen species（ ROS） in neurons,flow cytometry was used to determine the apoptosis rate,Western blot was used to measure the expression of TIGAR,apoptosis-inducing factor（ AIF）,and proteins associated with mitochondrial fission（ Drp1,Fis1,and ROCK1） in neurons. Results Compared with group B,group D had a significant increase in the expression of TIGAR and significant reductions in ROS concentration,apoptosis rate,and the expression of AIF,Drp1,Fis1,and ROCK1（ all P〈0.05）. Compared with group B,group F had a significant reduction in the expression of TIGAR and significant increases in ROS concentration,apoptosis rate,and the expression of AIF,Drp1,Fis1,and ROCK1（ all P〈0.05）. There were no significant differences in these index between groups C/E and group B（ P〈0.05）. Conclusion TIGAR exerts a protective effect on neurons by regulating mitochondrial fission in a rat model of hypoxia/reoxygenation injury.