布美他尼通过PERK/EIF-2a/ATF4信号抑制内质网应激减轻缺血性脑卒中大鼠神经元损伤  

Bumetanib alleviates neuronal injury in ischemic stroke rats by inhibiting endoplasmic reticulum stress via PERK/EIF-2A/ATF4 signaling

作　　者：赵旭[1] 鞠延玲[2] 臧雪莲[1] 童健尔[1] 戚芳 ZHAO Xu;JU Yan-ling;ZANG Xue-lian;TONG Jian-er;QI Fang(Department of Neurology,Jinzhou Central Hospital,Jinzhou 121001;Department of Cardiology,Jinzhou Central Hospital,Jinzhou 121001;Department of Neurology,Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110034,China)

机构地区：[1]锦州市中心医院神经内科,辽宁锦州121001 [2]锦州市中心医院心内科,辽宁锦州121001 [3]辽宁中医药大学附属医院神经内科,辽宁沈阳110034

出　　处：《解剖科学进展》2022年第5期635-638,共4页Progress of Anatomical Sciences

基　　金：辽宁省自然科学基金(20180551070)。

摘　　要：目的探讨布美他尼抑制内质网应激减轻缺血性脑卒中大鼠海马神经元损伤的机制研究。方法30只SD大鼠随机分为假手术组、模型组、布美他尼组,每组10只。通过结扎双侧颈总动脉方法制备缺血性脑卒中大鼠模型。检测各组大鼠神经功能缺损评分;TTC染色检测大鼠脑组织梗死体积;检测脑组织含水量;免疫荧光染色检测神经元标记物NeuN的数量;免疫组织化学染色检测内质网应激相关蛋白葡萄糖调控蛋白78(GRP78)、C/EBP同源蛋白(CHOP)表达;Western blot检测胰腺内质网激酶(PERK)、真核细胞起始因子2α(EIF-2α)、磷酸化PERK(p-PERK)、磷酸化EIF-2α(p-EIF-2α)和转录激活因子4(ATF4)表达。结果与模型组相比,布美他尼组大鼠神经功能缺损评分明显降低,脑梗死体积明显减小,脑组织含水量明显降低,神经元NeuN数量明显增加,GRP78、CHOP蛋白表达明显降低,并且p-PERK、p-EIF-2α和ATF4蛋白表达明显降低。结论布美他尼能够改善缺血性脑卒中大鼠神经元损伤,其作用机制可能与抑制PERK/EIF-2α/ATF4信号通路、抑制内质网应激有关。Objective To investigate the mechanism of bumetanide inhibiting endoplasmic reticulum stress and alleviating hippocampal neurons damage in rats with ischemic stroke.Methods Thirty SD rats were randomly divided into sham group,model group,and bumetanide group,with 10 rats in each group.Rat models of ischemic stroke were prepared by ligating bilateral common carotid arteries.The neurological deficit scores of rats were measured.Infarct volume of rat brain tissue was detected by TTC staining.Brain tissue water content was detected.The number of neuron marker NeuN was observed by immunofluorescence staining.The endoplasmic reticulum stress-related proteins glucose regulated protein 78(GRP78)and C/EBP homologous protein(CHOP)expression was observed by immunofluorescence staining.The expression of pancreatic endoplasmic reticulum kinase(PERK),eukaryotic initiation factor 2α(EIF-2α),phosphorylated PERK(pPERK),phosphorylated EIF-2α(p-EIF-2α)and transcription activator 4(ATF4)was detected by Western blot.Results Compared with the model group,the neurological deficit scores of rats in the bumetanide group were significantly reduced,the volume of cerebral infarction was significantly reduced,the water content of brain tissue was significantly reduced,the number of neurons was significantly increased,and the expression of GRP78 and CHOP was significantly reduced.In addition,the expression of p-PERK,p-EIF-2α and ATF4 protein was significantly reduced.Conclusion Bumetanide can improve neuronal injury in ischemic stroke rats,and its mechanism may be related to inhibiting PERK/EIF-2α/ATF4 signaling pathway and endoplasmic reticulum stress.

关 键 词：布美他尼 缺血性脑卒中 神经元损伤 内质网应激 PERK/EIF-2α/ATF4信号通路 

分 类 号：R743.3[医药卫生—神经病学与精神病学]