miR-34a在幼鼠海马神经元细胞增殖和凋亡中的作用  

Effects of miR-34a on Proliferation and Apoptosis of Hippocampal Neuronal Cell from Rats

作　　者：张垚[1] 杨方[2] 王浩[1] 罗建峰[1] 刘雨东[1] 张国成[1] 孙新[1] ZHANG Yao YANG Fang WANG Hao LUO Jian-feng LIU Yu-dong ZHANG Guo-cheng SUN Xin(Department of pacdiatn＇cs, Xijing Hospital of the Fourth Military Medical Univcrsi~ Xi＇an, Shaanxi, 710032, China Depar~cnt ofNeurology, Xijing Hospital of the Fourth Military Medical University, Xi＇an, Shaanxi, 710032, China)

机构地区：[1]第四军医大学西京医院儿科,陕西西安710032 [2]第四军医大学西京医院神经内科,陕西西安710032

出　　处：《现代生物医学进展》2017年第20期3801-3805,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金青年基金项目(81100965)

摘　　要：目的:探讨miR-34a在幼鼠海马神经元细胞增殖凋亡中的作用。方法:分离幼鼠海马神经元细胞,转染miR-34a抑制剂(miR-34a inhibitor)、抑制剂对照(inhibitor control)、miR-34a模拟物(miR-34a mimics)、模拟物对照(mimics control),RT-PCR检测细胞中miR-34a表达水平。MTT检测转染后细胞增殖情况。流式细胞仪检测细胞凋亡情况。Western blot检测细胞中Cleaved-caspase-3、Bcl-2、Bax的表达水平。结果:转染miR-34a inhibitor可以抑制miR-34a的表达,miR-34a mimics可以促进miR-34a的表达。miR-34a mimics对细胞增殖抑制率明显高于mimics control组(P<0.05),miR-34a inhibitor组抑制率明显低于inhibitor control组(P<0.05)。miR-34a inhibitor组神经元细胞凋亡率明显低于inhibitor control组(P<0.05),miR-34a mimics组神经元细胞凋亡率明显高于mimics control组(P<0.01),inhibitor control组和mimics control组神经元细胞凋亡率差异不显著(P>0.05)。miR-34a inhibitor组Cleaved-caspase-3、Bax蛋白表达量低于inhibitor control组,差异显著(P<0.05);miR-34a inhibitor组Bcl-2蛋白表达量高于inhibitor control组,差异显著(P<0.05);miR-34a mimics组Cleaved-caspase-3、Bax蛋白表达量高于mimics control,差异显著(P<0.05);miR-34a mimics组Bcl-2蛋白表达量低于mimics control,差异显著(P<0.05)。结论:miR-34a抑制海马神经元细胞增殖,促进细胞凋亡,其作用机制可能与调控Cleaved-caspase-3、Bcl-2、Bax表达有关。Objective：To investigate the role of miR-34a on proliferation and apoptosis of hippocampal neurons cell.Metlhods：Isolated rats hippocampal neurons were transfected with miR-34a inhibitor,inhibitor control,miR-34a mimics,mimics control,respectively.RT-PCR was used to detect the expression level of miR-34a,and MTT was used to detect cell proliferation.Apoptosis was detected by flow cytometry.Western blot analysis was determined the expression levels of Cleaved-caspase-3,Bcl-2,Bax.Results：Transfection with miR-34a inhibitor could inhibit the expression of miR-34a,miR-34a mimics could promote the expression of miR-34a.The cell proliferation inhibition rate in miR-34a mimics group was significantly higher than that of mimics control group （P ＜0.05）,and the inhibition rate in miR-34a inhibitor group was significantly lower than that of inhibitor control group （P ＜0.05）.The neuronal apoptosis in miR-34a inhibitor group was significantly lower than that of inhibitor control group （P ＜0.05）,and the neuronal apoptosis was significantly higher in miR-34a mimics group than that of mimics control group （P ＜0.01）.The neuronal apoptosis rate was not significantly different between inhibitory control group and mimics control group （P＞ 0.05）.The cleaved-caspase-3,Bax protein expression level lower in miR-34a inhibitor group than that of inhibitor control group,and the difference was significant （P ＜0.05）;and Bcl-2 protein expression was higher in miR-34a inhibitor group than that of inhibitor control group,and the difference was significant （P ＜0.05）.The cleaved-caspase-3,Bax protein expression was higher in miR-34a mimics group than that of mimics control with statistical significance （P ＜0.05）;and Bcl-2 protein expression of miR-34a mimics group was lower than that of mimics control （P ＜0.05）.Conclusions：miR-34a can inhibit proliferation of hippocampal neurons and promote neuronal apoptosis,and its mechanism may be related to Cleaved-caspase-3,Bcl-2,Bax.

关 键 词：海马神经元细胞 细胞增殖 细胞凋亡 MIR-34A 

分 类 号：R-33[医药卫生] R338