饱和氢盐水对短暂性脑缺血再灌注损伤小鼠海马PERK/eIF2α/ATF4信号通路的影响

Effects of Hydrogen-rich Saline on Hippocampus PERK/eIF2α/ATF4 Signaling Pathway in Mice After Transient Cerebral Ischemia Reperfusion Injury

目的探讨饱和氢盐水对短暂性脑缺血再灌注损伤小鼠海马PERK/eIF2α/ATF4信号通路的影响。方法选择SPF级健康♂C57BL/6小鼠150只,采用随机数字表法将其分为5组(n=30):对照组、假手术组、脑缺血再灌注组、饱和氢盐水治疗组和生理盐水治疗组。于再灌注12,24 h行神经行为学评分,然后处死小鼠取海马组织,采用TUNEL法测定海马CA1区细胞凋亡指数,分别采用Western blotting和Real-time PCR法测定海马PERK、eIF2α、ATF4、CHOP和caspase-3及其mRNA的表达水平。结果与对照组、假手术组相比,脑缺血再灌注组、饱和氢盐水治疗组和生理盐水治疗组小鼠神经行为学评分升高,海马CA1区神经细胞凋亡指数升高,海马PERK、eIF2α、ATF4、CHOP和caspase-3及其mRNA的表达上调(P均<0.05);与脑缺血再灌注组、生理盐水治疗组相比,饱和氢盐水治疗组小鼠神经行为学评分降低,海马CA1区神经细胞凋亡指数降低,PERK、eIF2α、ATF4、CHOP和caspase-3及其mRNA的表达下调(P均<0.05)。结论饱和氢盐水可改善小鼠短暂性脑缺血再灌注损伤,其机制可能与抑制海马PERK/eIF2α/ATF4信号通路有关。

OBJECTIVE To evaluate the effect of hydrogen-rich saline on hippocampus PERK/eIF2α/ATF4 protein signaling pathways in mice after transient cerebral ischemia reperfusion injury.METHODS One hundred and fifty healthy male C57BL/6 mice were divided into 5 groups(n=30)using a random number table:control group,sham operation group,cerebral ischemia-reperfusion group,hydrogen-rich saline treatment group and normal saline treatment group.Neurobehavioral score was assessed at 12,24 h of reperfusion.Then mice in each group were sacrificed,and the hippocampal tissues were obtained and the apoptosis index of CA1 in hippocampus were determined by TUNEL.The expression of PERK,eIF2α,ATF4,CHOP,caspase-3 protein and mRNA were determined by Western blotting and real-time polymerase chain reaction,respectively.RESULTS Compared with the control group and the sham operation group,the neurobehavioral score and apoptosis index of hippocampal CA1 region were increased,and the expression of PERK,eIF2α,ATF4,CHOP,caspase-3 and their mRNA were up-regulated in the cerebral ischemia-reperfusion group,hydrogen-rich saline treatment group and normal saline treatment group(all P<0.05).Compared with cerebral ischemia-reperfusion group and normal saline treatment group,the neurobehavioral score and apoptosis index in hippocampus CA1 region were decreased and the expression of PERK,eIF2α,ATF4,CHOP,caspase-3 and their mRNA were down-regulated in the hydrogen-rich saline treatment group(all P<0.05).CONCLUSION Hydrogen-rich saline can improve transient cerebral ischemia reperfusion injury in mice,and its mechanism may be associated with inhibited activation of PERK/eIF2α/ATF4 signaling pathway.