姜黄素对Aβ_(25-35)诱导的PC12细胞线粒体内细胞色素C表达的影响  

Effect of Curcumin on Expression of Cytochrome C in Mtochondria of PC12 Cells Induced by Aβ_(25-35)

作　　者：肖韩艳[1] 刘颖[1] 张岩[1] 王冬梅 张云 

机构地区：[1]牡丹江医学院第二附属医院,黑龙江牡丹江157011 [2]牡丹江市第二人民医院,黑龙江牡丹江157013 [3]牡丹江医学院附属红旗医院,黑龙江牡丹江157011

出　　处：《中国实验方剂学杂志》2017年第23期116-121,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：黑龙江省自然科学基金项目(H2015937)

摘　　要：目的:探讨姜黄素对β淀粉样蛋白25-35(β-amyloid protein 25-35,Aβ_(25-35))诱导的大鼠肾上腺嗜铬细胞瘤(PC12)细胞凋亡及线粒体内细胞色素C(cytochrome C,Cyt C)表达的影响,探讨其抗PC12细胞凋亡的线粒体机制。方法:以Aβ_(25-35)作用于PC12细胞,建立阿尔茨海默病(Alzheimer's disease,AD)细胞模型,分别以0(空白组),1.25,2.5,5,10,20,40μmol·L-1不同浓度的姜黄素进行处理,采用噻唑蓝(MTT)比色法检测姜黄素对细胞存活率的影响。实验随机分为空白组,模型组,姜黄素低、高(10,20μmol·L-1)浓度组,采用Annexin V FITC/PI双染流式细胞仪检测凋亡情况;JC-1染色分析检测线粒体膜电位(Δψm)的变化;免疫荧光分析检测Cyt C的分布;蛋白质印迹法(Western blot)检测线粒体内Cyt C蛋白的表达。结果:与空白组比较,模型组PC12细胞存活率显著降低,凋亡率升高(P<0.01);与模型组比较,姜黄素可显著升高PC12细胞存活率,降低凋亡率(P<0.01)。与空白组比较,模型组PC12细胞Δψm降低,Cyt C由线粒体释放至胞浆增多,线粒体内Cyt C的表达下调(P<0.01);与模型组比较,姜黄素可显著升高PC12细胞Δψm,抑制Cyt C由线粒体释放至胞浆并上调线粒体内Cyt C的表达(P<0.05,P<0.01)。结论:姜黄素可抑制线粒体凋亡途径,从而抑制Aβ_(25-35)诱导的PC12细胞凋亡。Objective： To discuss the effect of curcumin on the apoptosis of adrenal pheochromocytoma （PC12） cells induced by β-amyloid protein 25-35 （Aβ25-35） and the expression of cytochrome C （ Cyt C） in mtochondria of rats, in order to analyze the mitoehondrial mechanisms of anti-apoptosis. Method： Aβ25-35 was used to treat PC12 cells, and the cellular model of Alzheimer＇s disease （AD） was established. The PC12 cells were treated with curcumin at concentrations of 0 （blank control group） , 1.25, 2.5, 5, 10, 20, 40 μmol·L^-1, respectively. The survival rate was measured by 3- （4, 5-dimethyl-2-thiazolyl） -2, 5-diphenyl-2-H-tetrazolium bromide （MTT）. The PCI2 cells were randomly divided into the blank control group, the model group, the low-dose （10 μmol·L^-1） curcumin group and the high-dose （20 μmol·L^-1） curcumin group. The effect of curcumin on the apoptosis, the changes in mitochondrial membrane potential （Δψm） , the distribution of Cyt C and the expression of Cyt C in mitochondria were evaluated by Annexin V-FITC/PI flow cytometry, JC-1 staining assay, immunofluorescence analysis and Western blot analysis, respectively. Result： Compared with the blank control group, the survival rate of PC12 ceils was decreased significantly, and the apoptosis rate was increased in model group （P 〈 0.01 ）. Compared with the model group, curcumin significantly increased the cell viability and decreased the apoptosis rate （P 〈 0.01 ）. Compared with the blank control group, Δψm decreased significantly, the release of mitochondrial Cyt C to cytosol increase, and the expression of Cyt C in mitochondria down-regulated in model group （P 〈 0.01）. Compared with the model group, curcumin significantly increased Δψm inhibited the release of mitochondrial Cyt C to cytosol and Up-regulated the expression of Cyt C in the mitochondria （P 〈 0.05, P 〈 0.01 ）. Conclusion： Curcumin can inhibit the apoptosis of PC12 induced by Aβ25-35 by suppressing mitochondrial

关 键 词：阿尔茨海默病 姜黄素 凋亡 线粒体凋亡途径 

分 类 号：R285.5[医药卫生—中药学]