摘要
目的:探讨内质网应激后原代海马神经元树突棘密度及突触蛋白表达的变化,以及通过内质网应激分子伴侣4-苯基丁酸(4-phenylbutyric acid,4-PBA)抑制内质网应激对这种神经元损伤的抑制作用。方法:原代培养新生大鼠海马神经元,将表达增强型绿色荧光蛋白的质粒转染到原代培养5～7 d(DIV 5～7)的大鼠海马神经元内持续培养,DIV 20时分为对照组、衣霉素(tunicamycin,Tm)处理组和Tm+4-PBA预处理组(Tm处理前1 h给予4-PBA),采用Western blot法检测内质网应激标志蛋白Bi P和突触蛋白的表达水平,激光共聚焦显微镜下观察神经元,分析树突棘密度,采用MTT法分析细胞活力。结果:Tm处理后使Bi P蛋白水平明显升高,而4-PBA预处理使Bi P蛋白水平显著下降(P <0. 05)。Tm引起的原代海马神经元树突棘密度下降及突触蛋白的表达下降能够被4-PBA抑制。Tm引起的细胞活力下降可被4-PBA抑制。结论:Tm能够通过诱导内质网应激而引起原代海马神经元树突棘密度下降及突触蛋白表达下降,而提前给予4-PBA预处理可明显降低内质网应激反应,抑制树突棘密度下降及突触蛋白表达下降,从而减轻原代海马神经元的损伤。
AIM:To investigate the inhibitory effect of4-phenylbutyric acid(4-PBA),an aromatic short-chain fatty acid with the functions of endoplasmic reticulum(ER)molecule chaperon,on ER stress-induced decreases in both dendritic spine density and synaptic protein expression.METHODS:Primary hippocampal neurons were cultured from neonatal rats.The primary hippocampal neurons were transfected with enhanced green fluorescent protein plasmids at DIV(day in vitro)5~7.At DIV20,the neurons were divided into DMSO group,tunicamycin group and tunicamycin+4-PBA(treated with4-PBA1h before tunicamycin)group.The expression levels of ER stress marker protein BiP and synaptic proteins were detected by Western blot.After immunofluorescence staining,the neurons were morphologically observed under confocal laser scanning microscope,and the density of dendritic spines was analyzed.At last,the cell viability was measured by MTT assay.RESULTS:Tunicamycin induced ER stress in the primary hippocampal neurons,characterized by significantly increased level of BiP in tunicamycin group,which was reduced in tunicamycin+4-PBA group(P<0.05).4-PBA inhibited tunicamycin-induced decreases in both dendritic spine density and synaptic protein expression in the primary hippocampal neurons.4-PBA attenuated tunicamycin-induced decrease in the cell viability.CONCLUSION:4-PBA not only attenuates ER stress of primary hippocampal neurons but also inhibits the decreases in both dendritic spine density and synaptic protein expression induced by tunicamycin.
作者
柳秀平
吕凌云
李夏春
LIU Xiu-ping;L Ling-yun;LI Xia-chun(Department of Clinical Laboratory, Hangzhou Hospital of Traditional Chinese Medicine, Hangzhou 310007, China;Department of Pathophysiology, Medical School of Three Gorges University, Yichang 443002, China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2018年第12期2215-2220,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.31300932)
关键词
内质网应激
阿尔茨海默病
4-苯基丁酸
树突棘
原代海马神经元
Endoplasmic reticulum stress
Alzheimer disease
4-Phenylbutyric acid
Dendritic spines
Primary hippocampal neurons
