淫羊藿苷对H2O2诱导的PC12细胞内质网应激的保护作用
Protection of icariin against H2O2 induced endoplasmic reticulum stress in PC12 cells
张洋洋 1李祖高 1刘义伟 1陈霞 1李菲1
作者信息
- 1. 遵义医科大学药理学教研室暨基础药理教育部重点实验室暨特色民族药教育部国际合作联合实验室,贵州遵义563099
- 折叠
摘要
目的 观察淫羊藿苷(ICA)对H2O2诱导的PC12细胞内质网应激的作用.方法 采用H2O2制备PC12细胞应激损伤模型,观察ICA在6.25、12.5、25 μM不同浓度下的作用.采用MTT和LDH测定法检测细胞活力和细胞死亡率.Western blot检测GRP78、PERK、eIF2α、ATF4和CHOP的蛋白表达,以及PERK和eIF2 α的磷酸化水平.实时荧光定量PCR检测GRP78、PERK、ATF4和CHOP的mRNA水平.结果 结果显示H2O2可诱导PC12细胞损伤和内质网应激,而ICA预处理可增加PC12细胞活力、减轻细胞损伤,并抑制H2O2诱导的GRP78、ATF4和CHOP的mRNA高表达,同时降低了PERK、eIF2α磷酸化水平以及GRP78、ATF4和CHOP的蛋白表达.结论 本研究表明ICA可减轻H2O2诱导的PC12细胞内质网应激损伤,其作用主要与抑制PERK信号通路的激活有关.
Abstract
Objective To investigate the effect of icariin (ICA) on H2O2 induced endoplasmic reticulum (ER) stress in PC12 cells.Methods H2O2 was used as an inducer of ER stress,and PC12 cells were pre-treated with various concentrations of ICA (6.25,12.5 and 25 μM).Cell proliferation and death were detected using MTIand LDH assays.The protein expressions of GRP78,PERK,eIF2α,ATF4 and CHOP,as well as the phosphorylation levels of PERK and eIF2α were tested by Western blot.The mRNA level of GRP78,PERK,ATF4 and CHOP were measured by real-time qPCR analysis.Results The data demonstrated that H2O2 induced PC12 cells loss and ER stress.However,pre-treatment with ICA enhanced PC12 cell viability and repressed cells loss;decreased the levels of PERK,eIF2α phosphorylation and the expressions of GRP78,ATF4,and CHOP protein;as well as reduced GRP78,ATF4 and CHOP mRNA levels in H2O2-injuried PC12 cells.Conclusion The present study revealed that ICA exerts protective effect on H2O2-induced ER stress in PC12 cells,mainly related to suppression of the PERK signaling pathway activation.
关键词
淫羊藿苷/H2O2/内质网应激/蛋白激酶R样内质网激酶/嗜铬细胞瘤Key words
H2O2/endoplasmic reticulum stress/protein kinase R-like endoplasmic reticulum kinase/pheochromocytoma引用本文复制引用
基金项目
国家自然科学基金地区资助项目(81560594)
贵州省国际科技合作计划项目(NO:黔科合外G字[2014]7011)
出版年
2019
NETL
NSTL
维普
万方数据