XIAO Han-yan, LIU Ying, ZHANG Yan, et al. Effect of Curcumin on Expression of Cytochrome C in Mtochondria of PC12 Cells Induced by A[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(23): 116-121.
XIAO Han-yan, LIU Ying, ZHANG Yan, et al. Effect of Curcumin on Expression of Cytochrome C in Mtochondria of PC12 Cells Induced by A[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(23): 116-121. DOI: 10.13422/j.cnki.syfjx.2017230116.
目的:探讨姜黄素对β淀粉样蛋白25-35(β-amyloid protein 25-35,Aβ25-35)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Aβ25-35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),1.25,2.5,5,10,20,40 μmol·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(10,20 μmol·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Aβ25-35诱导的PC12细胞凋亡。
Abstract
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 mitochondrial 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 PC12 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 cells 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 apoptosis pathway.