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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2021, Vol. 41 Issue (4): 1-8    DOI: 10.13523/j.cb.2012040
研究报告     
SIRT2抑制对MPP+诱导的帕金森病细胞模型凋亡和线粒体动态平衡的影响*
段阳阳,张凤亭,成江,石瑾,杨娟,李海宁()
宁夏医科大学临床医学院 银川 750004
The Effect of SIRT2 on Apoptosis and Mitochondrial Function in Parkinson’s Disease Model Cells Induced by MPP+
DUAN Yang-yang,ZHANG Feng-ting,CHENG Jiang,SHI Jin,YANG Juan,LI Hai-ning()
School of Clinical Medicine, Ningxia Medical University, Yinchuan 750004, China
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摘要：

探究siRNA敲减沉默信息调节因子2(SIRT2)对1-甲基-4-苯基吡啶离子(MPP+)诱导的帕金森病细胞模型细胞损伤的影响和机制。CCK-8法检测不同浓度MPP+处理对体外培养小鼠海马神经元HT-22细胞生存率的影响。将细胞分为对照组、MPP+最佳浓度处理组(1 mmol/L MPP+处理组)、阴性转染组(对照组基础上转染SIRT2阴性序列)、SIRT2 siRNA处理组(损伤组基础上转染SIRT2 siRNA)。观察各组细胞凋亡情况,检测凋亡相关蛋白(Bcl-2、Bax、Caspase-9)、线粒体分裂及融合相关蛋白(Drp1、Fis1、OPA1、Mfn1、Mfn2)。与对照组相比,MPP+处理组细胞抑制率均升高,细胞抑制率随MPP+浓度增加而逐渐增加(P<0.05)。与SIRT2 siRNA转染组相比,损伤组Bax、Caspase-9、Drp1、Fis1蛋白表达和细胞凋亡率升高,Bcl-2、Mfn1、Mfn2蛋白表达降低(P<0.05)。SIRT2在MPP+诱导帕金森病细胞模型中表达升高,抑制SIRT2可减轻MPP+诱导帕金森病细胞模型中细胞凋亡并促进线粒体融合,从而对神经元具有一定的保护作用。

关键词： 帕金森病模型细胞;  沉默信息调节因子2;  凋亡;  线粒体分裂及融合;  siRNA沉默技术    
Abstract:

To investigate the effect and mechanism of cell apoptosis of siRNA knockdown silent information regulator 2 (SIRT2) in Parkinson’s disease model cells induced by 1-methyl-4-phenylpyridinium (MPP+). Immortalized mouse hippocampal neuron HT-22 cells were cultured in vitro and treated with MPP + at different concentrations, and CCK-8 assay was used to detect cell inhibition. The cells were divided into control group, MPP+ optimal concentration group (1 mmol/L MPP+ treatment, injury group), the negative transfection group (based on the control group which was transfected with SIRT2 negative sequence), and the SIRT2-siRNA treatment group (based on the injury group which was transfected with SIRT2-siRNA). The apoptosis of cells in each group was observed, apoptosis-related proteins (Bcl-2, Bax, Caspase-9) and the main proteins mediating fission and fusion of mitochondrial function (Drp1, Fis1, OPA1, Mfn1, Mfn2) were detected by Western blot. Compared with the control group, the cell inhibition rate of MPP+ treatment group increased, and with the concentration increased, the inhibition rate gradually increased (P<0.05). Compared with the SIRT2 siRNA treatment group, the injury group increased the expression of apoptosis and mitochondrial fission factors (Bax, Caspase-9, Drp1, Fis1) and decreased the expression of anti-apoptotic and mitochondrial fusion factors (Bcl-2, Opa1, Mfn1, Mfn2). The expression of SIRT2 significantly increased in a cell model of MPP+-induced Parkinson’s disease, and the inhibition of the SIRT2 was able to decrease apoptosis, promote mitochondrial fusion, inhibit mitochondrial fission and protect neurons.

Key words: Parkinson’s disease model cells    Silent information regulator 2    Apoptosis    Mitochondrial fission and fusion    SiRNA silencing technology
收稿日期: 2020-12-21 出版日期: 2021-04-30
ZTFLH:  Q819  
基金资助: *宁夏重点研发项目资助项目(2019BEB04008)
通讯作者: 李海宁     E-mail: lhnwww@126.com
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段阳阳
张凤亭
成江
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杨娟
李海宁

引用本文:

段阳阳,张凤亭,成江,石瑾,杨娟,李海宁. SIRT2抑制对MPP+诱导的帕金森病细胞模型凋亡和线粒体动态平衡的影响*[J]. 中国生物工程杂志, 2021, 41(4): 1-8.

DUAN Yang-yang,ZHANG Feng-ting,CHENG Jiang,SHI Jin,YANG Juan,LI Hai-ning. The Effect of SIRT2 on Apoptosis and Mitochondrial Function in Parkinson’s Disease Model Cells Induced by MPP+. China Biotechnology, 2021, 41(4): 1-8.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2012040    或    https://manu60.magtech.com.cn/biotech/CN/Y2021/V41/I4/1

图1  不同浓度MPP+处理后HT-22细胞存活率
图2  不同浓度MPP+处理的HT-22细胞SIRT2表达
图3  不同浓度MPP+处理的HT-22细胞凋亡相关蛋白的表达
图4  HT-22细胞转染SIRT2-siRNA后SIRT2蛋白的表达
图5  SIRT2抑制后HT-22细胞形态恢复
图6  SIRT2抑制后凋亡相关蛋白的表达
图7  SIRT2沉默后线粒体分裂蛋白的表达
图8  SIRT2沉默后线粒体融合蛋白的表达
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