基于2-Cl-MGV-1/BDNF-TrkB通路探讨脑梗死后认知功能改善的研究

A study on improvement in cognitive function after cerebral infarction based on 2⁃Cl⁃MGV⁃1/BDNF⁃TrkB pathway

目的基于2-(2-氯苯基)喹唑啉-4-基二甲基氨基甲酸酯(2-Cl-MGV-1)/脑源性神经营养因子(BDNF)-原肌球蛋白受体激酶B(TrkB)通路探讨脑梗死后认知功能改善的研究。方法成年雄性Sprague-Dawley大鼠随机分为3组,每组20只,分别为对照组、大脑中动脉栓塞(MCAO)组和2-Cl-MGV-1组。除对照组外,其他组建立MCAO模型,2-Cl-MGV-1组在模型建立后采用2-Cl-MGV-1治疗,连续给药7 d。评估各组大鼠脑梗死面积、空间学习记忆障碍、线粒体损伤和BDNF/TrkB信号通路。体外将PC12神经元细胞系分为以下3组:对照(Con)组、氧气和葡萄糖剥夺/再灌注模型(OGD/R)组和2-Cl-MGV-1组。除Con组,其他组建立OGD/R模型,2-Cl-MGV-1组加入25μmol/L 2-Cl-MGV-1处理细胞24 h。通过CCK-8评估细胞活力。结果与MCAO组相比,2-Cl-MGV-1组梗死面积显著减少(P<0.05),和尼氏体的数量显著增加(P<0.05)。与对照组相比,MCAO组大鼠的逃避潜伏期显著增加(P<0.001),而2-Cl-MGV-1组的逃避潜伏期显著低于MCAO组(P<0.01)。在第7天,MCAO组大鼠穿过平台的次数显著低于对照组(P<0.001),而2-Cl-MGV-1组大鼠穿过平台的次数较MCAO组显著增加(P<0.05)。与对照组相比,MCAO组皮质中的线粒体膜电位和ATP产量显著降低(P<0.01),而2-Cl-MGV-1组线粒体膜电位和ATP产量较MCAO组显著增加(P<0.05)。与对照组相比,MCAO组大鼠皮层神经元中的BDNF、TrkB蛋白水平显著增加(P<0.05),并且2-Cl-MGV-1组BDNF、TrkB蛋白水平显著高于MCAO组(P<0.05)。与Con组相比,OGD/R组细胞活力和ATP产量显著降低(P<0.05),而2-Cl-MGV-1组细胞活力和ATP产量较OGD/R组显著增加(P<0.05)。与Con组相比,OGD/R组PC12细胞中BDNF、TrkB蛋白水平显著增加(P<0.05),并且2-Cl-MGV-1组BDNF、TrkB蛋白水平显著高于MCAO组(P<0.05)。结论2-Cl-MGV-1对MCAO诱导的大鼠脑缺血/再灌注损伤具有线粒体保护作用,并改善大鼠的认知功能。2-Cl-MGV-1可能通过调节BDNF/TrkB信号通路发挥神经保护作用。

Objective To explore the improvement in cognitive function after cerebral infarction based on the 2⁃(2⁃chlorophenyl)quinazolin⁃4⁃yl dimethylcarbamate(2⁃Cl⁃MGV⁃1)/brain⁃derived neurotrophic factor(BDNF)⁃tropomyosin receptor kinase B(TrkB)pathway.Methods Adult Sprague⁃Dawley rats were randomly divided into a control group,middle cerebral artery embolism(MCAO)group,and 2⁃Cl⁃MGV⁃1 group,20 in each group.Except the control group,other groups established MCAO models.2⁃Cl⁃MGV⁃1 group was treated with 2⁃Cl⁃MGV⁃1 for 7 days after the model was established.The cerebral infarction area,spatial learning and memory impairment,mitochondrial damage and BDNF/TrkB signal pathway were assessed in each group.In vitro PC12 neuron cell lines were divided into the following three groups:control group(Con),oxygen and glucose deprivation/reperfusion model(OGD/R)group,and 2⁃Cl⁃MGV⁃1 group.Except Con group,other groups established OGD/R models,and 2⁃Cl⁃MGV⁃1 group was treated with 25μmol/L 2⁃Cl⁃MGV⁃1 for 24 hours.Cell viability was evaluated by CCK⁃8.Results As compared with MCAO group,the infarct area in 2⁃Cl⁃MGV⁃1 group decreased significantly(P<0.05),and the number of Nissl bodies increased significantly(P<0.05).As compared with the control group,the escape latency was significantly increased in MCAO group(P<0.001),while it was significantly shorter in 2⁃Cl⁃MGV⁃1 group than in MCAO group(P<0.01).On day 7,the number of crossing the platform in MCAO group was significantly lower than that in the control group(P<0.001),while the number of crossing the platform in 2⁃Cl⁃MGV⁃1 group was significantly higher than that in MCAO group(P<0.05).As compared with the control group,cortical mitochondrial membrane potential and ATP production decreased significantly in MCAO group(P<0.01),while increased markedly in 2⁃Cl⁃MGV⁃1 group(P<0.05).As compared with the control group,lev⁃els of BDNF and TrkB protein in cerebral cortex neurons in MCAO group were significantly increased(P<0.05),and levels of BDNF and TrkB protein in 2⁃Cl⁃MGV⁃1 group were significantly higher than those in MCAO group(P<0.05).As compared with Con group,cell viability and ATP production decreased significantly in OGD/R group(P<0.05),while increased significantly in 2⁃Cl⁃MGV⁃1 group(P<0.05).As compared with Con group,levels of BDNF and TrkB protein in OGD/R group increased significantly(P<0.05),and levels of BDNF and TrkB protein in 2⁃Cl⁃MGV⁃1 group were significantly higher than those in MCAO group(P<0.05).Conclusions 2⁃Cl⁃MGV⁃1 can protect mitochondria from MCAO⁃induced cerebral ischemia/reperfusion injury and improve cogni⁃tive function in rats.It may play a neuroprotective role by regulating BDNF/TrkB signaling pathway.