饱和氢盐水对短暂性脑缺血再灌注损伤小鼠海马PERK/eIF2a/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

关键词:  饱和氢盐水  脑缺血  再灌注损伤  海马  磷酸化的细胞外信号调节激酶  真核翻译起始因子2a  活化转录因子4

Key words:  hydrogen-rich saline  brain ischemia  reperfusion injury  hippocampus  PERK  eIF2a  ATF4

摘要:

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/eIF2a/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、eIF2a、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、eIF2a、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、eIF2a、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/eIF2a/ATF4信号通路有关。

Abstract:

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.