大黄素抗Aβ25-35诱导原代大鼠皮层细胞氧化应激损伤的机制  

Emodin anti-oxidative stress damage of rat cortical neurons induced by Aβ25-35

作　　者：刘涛[1] 胡海涛[2] 

机构地区：[1]西安体育学院运动人体科学系,陕西西安710068 [2]西安交通大学医学院解剖与组织胚胎学系

出　　处：《中国老年学杂志》2013年第18期4475-4478,共4页Chinese Journal of Gerontology

基　　金：陕西省教育厅资助项目(No.11JK0654)

摘　　要：目的探讨大黄素对抗Aβ25-35诱导的原代大鼠皮层细胞的氧化应激损伤机制i方法将原代培养的大鼠皮质神经元分为对照组、模型组及大黄素预处理组。采用比色法测试超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH—Px）和脂质过氧化产物丙二醛（MDA）的活性。2’，7’-二氯二氢荧光素二乙酯（H2DCF-DA）染色后，荧光显微镜和流式细胞仪（FCM）检测细胞内活性氧（ROS）的含量。罗丹明123（Rh123）染色后，FCM检测线粒体膜电位。Western印迹检测细胞色素c（CytC）蛋白表达水平。结果30μmol／L的Aβ25-35孵育皮层细胞24h后显著降低SOD、GSH-Px的活性和线粒体膜电位，同时增加MDA和ROS的产生。大黄素预处理皮层细胞后可明显抑制AB25q引起的上述细胞毒性效应，并减少CytC的表达。结论大黄素通过恢复皮层细胞抗氧化酶的活性，稳定线粒体膜电位，同时抑制MDA、ROS的聚集和CytC的释放来对抗Aβ25-35引起的氧化应激损伤。Objective To investigate the mechanisms of emodin against Aβ25-35-inducing oxidative stress-triggered damage in primary culture of rat cortical neurons. Methods Primary cultured rat cortical neurons were randomly divided into 5 groups ：control, model, 3 doses of emodin groups treated with different concentrations. The activities of superoxide dismutase （SOD） and glutathione peroxidase （GSH- Px） and the content of malondialdehyde （MDA） were assayed by eolorimetric assay. To study the change of reactive oxygen species （ROS） content in cortical neurons, the DCF fluorescence intensity of H2DCF-DA-stained cells was observed by fluorescence microscope and detected by FCM. The change of mitochondrial membrane potential （ASm） , the rhodamine123 （Rh123） fluorescence intensity of cortical neurons with Rh123 stain was detected by FCM. Release of cytochrome C from mitochondrial membrane was determined by Western blot analysis. Results Exposure of cortical neurons to 30μmol/L Aβ25-35 decreased activities of SOD and GSH-Px as well as increased levels of MDA production. In parallel, Aβ25-35 significant increased the intracellular ROS elevation and decreased the level of △ψm. However, pretreat- ment of the cells with emodin prior to Aβ25-35 exposure suppressed these Aβ25-35-induced cellular events noticeably. In addition, emodin reduced the level of cytochrome C. Conclusions The emodin prevents Aβ25-35-induced cell damage in primarily cultured cortical neurons by rescuing loss of antioxidant enzyme activities and decreasing the level of △ψm and inhibiting an increase in MDA, ROS and cytochrome C.

关 键 词：大黄素 Β-淀粉样蛋白 阿尔茨海默病 皮层细胞 神经毒性 氧化应激 

分 类 号：R285.5[医药卫生—中药学]