饥饿对Beclin-1依赖的Ana-1细胞自噬和凋亡的影响  

Effects of starvation on autophagy and apoptosis of Beclin-1-dependent Ana-1 cells

作　　者：杨万霞 苏强[3] 邢爱华 张晓延 YANG- Wanxia;SU Qiang;XING Aihua;ZHANG Xiaoyan(Clinical Laboratory,Second Hospital,Lanzhou University,Lanzhou 730000,China;Department of Medical Laboratory,Fenyang College,Shanxi Medical University,Fenyang 032200,China;Department of Nenrology,First Hospital,Shanxi Medical University,Taiyuan 030001,China;College of Pharmacy,Shanxi Medical University,Taiyuan 030001,China)

机构地区：[1]兰州大学第二医院检验科,甘肃兰州730000 [2]山西医科大学汾阳学院医学检验系,山西汾阳032200 [3]山西医科大学第一医院神经内科,山西太原030001 [4]山西省汾阳医院心内科 [5]山西医科大学药学院,山西太原030001

出　　处：《吉林大学学报（医学版）》2018年第4期704-708,共5页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(81301426);山西省教育厅高等院校大学生创新创业项目资助课题(2016526)

摘　　要：目的:研究饥饿对Ana-1细胞自噬和凋亡的影响,并探讨其作用机制。方法:体外培养Ana-1细胞,将细胞分为对照组(饥饿0h)和饥饿3、6、9、12、24h组(饥饿组),观察饥饿对细胞自噬与凋亡的影响;将Ana-1细胞分为空白对照组、饥饿组、3-甲基腺嘌呤(3-MA)组、3-MA联合饥饿组,孵育24h,采用Western blotting法检测各组细胞中自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ比值及Beclin-1蛋白表达水平和凋亡相关蛋白Caspase-3蛋白水平,采用倒置显微镜观察饥饿6、12和24h组细胞形态表现。结果:与对照组比较,饥饿6、12和24h组Ana-1细胞中LC3-Ⅱ/LC3-Ⅰ比值,饥饿3、6、9、12和24h组Ana-1细胞中Beclin-1蛋白表达水平均明显升高(P<0.05或P<0.01),饥饿3、6、9、12和24h组Ana-1细胞中Caspase-3水平明显降低(P<0.01),呈时间依赖性;倒置显微镜下观察,与对照组比较,不同时间饥饿组细胞形态发生改变,细胞皱缩、出现碎片,随着时间的延长,细胞变化更加明显。与饥饿组比较,3-MA联合饥饿组Ana-1细胞中LC3-Ⅱ/LC3-Ⅰ比值和Beclin-1蛋白表达水平及Caspase-3蛋白水平均明显降低(P<0.05或P<0.01)。结论:饥饿可诱导Beclin-1依赖的Ana-1细胞自噬并引起细胞凋亡。Objective：To study the effects of starvation on the autophagy and apoptosis of Ana-1 cells,and to explore their mechanisms.Methods：The Ana-1 cells were cultured in vitro and starved for 0 h（control group）,3,6,9,12 and 24 h（starvation group）to study the effects of starvation on the autophagy and apoptosis.The Ana-1 cells were divided into blank control group,starvation group,3-methyladenine group（3-MA）and 3-MA combined with starvation group,and they were incubated for 24 hto explore their mechanisms.The ratio of autophagy associated protein LC3-Ⅱ/LC3-Ⅰ,the expression levels of Beclin-1 and apoptosis-related protein Caspase-3 in the cells in various groups were detected by Western blotting method.Cell morphology 6,12,and24 hafter starvation was observed by inverted microscope.Results：Compared with control group,the ratio of LC3-Ⅱ/LC3-Ⅰin the Ana-1 cells in starvation group（6,12,and 24 h）and the expression levels of Beclin-1 in starvation group（3,6,9,12,and 24 h）were significantly increased（P〈0.05 or P〈0.01）,and the Caspase-3 levels in starvation group（3,6,9,12,and 24 h）were decreased significantly（P〈0.05 or P〈0.01）in a timedependent manner.Compared with control group,the cells in starvation group at different time were changed in shape：the cells shrank,the fragments appeared,and the changes of cells became more obvious with the prolongation of time.Compared with starvation group,the ratio of LC3-Ⅱ/LC3-Ⅰ and the expression levels of Beclin-1 and Caspase-3 in the Ana-1 cells in 3-MA combined with starvation group were decreased significantly（P〈0.05 or P〈0.01）.Conclusion：Starvation could induce the autophagy and apoptosis of the Beclin-1-dependent Ana-1 cells.

关 键 词：Ana-1细胞 饥饿 BECLIN-1 细胞自噬 细胞凋亡 

分 类 号：Q255[生物学—细胞生物学]