槲皮素对Aβ25-35介导的线粒体凋亡途径及p38MAPK信号通路的影响
Effects of quercetin on Aβ25-35-mediated mitochondrial apoptosis pathway and p38MAPK signal pathway
戴月英 1姚思凡 1赵雨薇 1沈丽霞1
作者信息
- 1. 河北北方学院药学院/河北省神经药理学重点实验室,河北张家口 075000
- 折叠
摘要
目的:研究槲皮素通过p38MAPK信号通路调节Aβ25-35引起PC12细胞线粒体损伤的作用.方法:20 μmol·L-1的Aβ25-35作用PC12细胞作为AD细胞毒性损伤模型,0.1 μmol·L-1的17β-雌二醇(17β-estradiol,17β-E2)和50μmol·L-1金雀异黄素(Genistein,Gen)作为阳性对照药,分别给予低剂量槲皮素(40 μmol·L-1)、中剂量槲皮素(60 μmol·L-1)和高剂量槲皮素(80μmol·L-1),同时设定SB203580(p38MAPK特异性抑制剂)预处理组(Aβ25-35+10μmol·L-1SB203580).线粒体荧光探针(Mito-Tracker-red)检测线粒体形态;JC-1检测线粒体膜电位;荧光素酶发光法检测ATP含量;免疫荧光染色法检测p-p38MAPK 蛋白的表达;Western blot 法测定 Bcl-2、Bax、p-p38MAPK/p38MAPK 和 Cytochrome C(Cyt C)蛋白的表达.结果:与Aβ模型组相比,槲皮素可显著提高线粒体膜电位和ATP含量,保护线粒体形态;免疫荧光显示,槲皮素显著抑制p38MAPK磷酸化表达.Western blot显示槲皮素上调Bcl-2蛋白表达及下调Bax、p-p38 MAPK/p38MAPK、Cyt C蛋白的表达(P＜0.05);当p38MAPK被SB203580抑制后,Bax、Cyt C蛋白表达降低(P＜0.01),Bcl-2蛋白表达升高(P＜0.01).结论:槲皮素可通过介导p38MAPK信号通路抑制Aβ25-35诱导的线粒体凋亡,进而发挥神经保护效应.
Abstract
OBJECTIVE To investigate the effect of quercetin(Que)on mitochondrial damage induced by Aβ25-35 through p38MAPK signaling pathway in PC12 cells.METHODS PC12 cells were treated with Aβ25.35(20 μmol·L-1)as a model of AD cell toxicity,and 0.1 μmol·L-1 of 17β-estradiol(17β-E2)and 50 μmol·L-1 of genistein(Gen)served as positive controls,low-dose Que(40 μmol·L-1),medium-dose Que(60 μmol·L-1)and high-dose Que(80 μmol·L-1)were used in this experiment.And select the SB203580(p38MAPK specific inhibitor)as pretreatment group(Aβ25-35+10 μmol·L-1 SB203580).Mito-Tracker-red detects Mitochondria morphology;JC-1 detects mitochondrial membrane potential and luciferase luminescence detects ATP;Immunofluorescence staining detects the expression of p-p38MAPK protein.Western blot method determines the expression of Bcl-2,Bax,p-p38MAPK/p38MAPK and Cytochrome C(Cyt C)protein.RESULTS Compared with the model group,quercetin significantly increased mitochondrial membrane potential and ATP content,and protected mitochondrial morphol-ogy.Quercetin significantly inhibited the phosphorylation of p38MAPK.Western blot showed'that quercetin up-regulated the expression of Bcl-2 protein and down-regulated the expression of Bax,p-p38MAPK/p38MAPK and Cyt C protein(P＜0.05).When p38MAPK was inhibited by SB203580,compared with the model group,the expression of Bax and Cyt C protein decreased(P＜0.01),while the expression of Bcl-2 protein increased(P＜0.01).CONCLUSION Quercetin can inhibit A by mediating the p38MAPK signaling pathway β mitochondrial apoptosis induced by 25-35 exerts neuroprotective effects.
关键词
槲皮素/线粒体/PC12细胞/p38MAPK/细胞凋亡/神经保护作用Key words
quercetin/mitochondria/PC12 cells/p38MAPK/apoptosis/neuroprotective effects引用本文复制引用
基金项目
河北省自然科学基金资助项目(H2019405057)
河北省研究生创新资助项目(CXZZSS2022145)
河北省研究生创新资助项目(CXZZSS2022151)
河北省高等学校科学技术研究项目(ZD2020136)
出版年
2024
NETL
NSTL
维普
万方数据