The Effect of SIRT2 on Apoptosis and Mitochondrial Function in Parkinson’s Disease Model Cells Induced by MPP<sup>+</sup>

doi:10.13523/j.cb.2012040

China Biotechnology

2021, Vol. 41

Issue (4): 1-8 DOI: 10.13523/j.cb.2012040

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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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

Received: 21 December 2020 Published: 30 April 2021

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Corresponding Authors: Hai-ning LI E-mail: lhnwww@126.com

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	Yang-yang DUAN
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	Jiang CHENG
	Jin SHI
	Juan YANG
	Hai-ning LI

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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.

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

Fig.1 The HT-22 cells viability treated with MPP⁺ at different concentrations The cell viability in different conditions was detected by CCK-8. Mean ± SD, n = 5. ^** P<0.01, ^*** P<0.001 vs control group

Fig.2 The expression of SIRT2 in HT-22 cells treated with MPP⁺ at different concentrations (a) The expression of SIRT2 protein was detected by Western blot (b) The statistical diagram of SIRT2 protein expression. Mean ± SD, n = 3. ^** P<0.01, ^*** P<0.001 vs control group

Fig.3 The expression of apoptosis-related protein in HT-22 cells treated with MPP⁺ at different concentrations (a)The expression of Caspase-9, Bax and Bcl-2 proteins were detected by Western blot (b) The statistical diagram of Caspase-9 protein expression (c) The statistical diagram of Bax protein expression (d) The statistical diagram of Bcl-2 protein expression. Mean ± SD, n = 3. ^** P<0.01, ^*** P<0.001 vs control group

Fig.4 Transfection with SIRT2-siRNA decreased SIRT2 expression in the HT-22 cells (a) The expression of SIRT2 protein was detected by Western blot (b) The statistical diagram of SIRT2 protein expression. Mean ± SD, n = 3. ^*** P<0.001 vs control group or negative control group

Fig.5 The effect of SIRT2-siRNA on the HT-22 cell’s morphology

Fig.6 The effect of SIRT2-siRNA on apoptosis-related protein (a) The expression of Caspase-9, Bax and Bcl-2 proteins were detected by Western blot (b) The statistical diagram of Caspase-9 protein expression (c) The statistical diagram of Bax protein expression (d) The statistical diagram of Bcl-2 protein expression. Mean ± SD, n = 3. ^***P<0.001 MPP⁺group vs control group or negative control group. ^## P<0.01, ^### P<0.001 MPP⁺+SIRT2-siRNA group vs MPP⁺group

Fig.7 The effect of SIRT2-siRNA on mitochondrial fission related protein (a) The expression of Drp1 and Fis1 proteins were detected by Western blot (b) The statistical diagram of Drp1 protein expression (c) The statistical diagram of Fis1 protein expression. Mean±SD, n = 3. ^** P<0.01, ^*** P<0.001 MPP⁺group vs control group or negative control group. ^# P<0.05, ^### P<0.001 MPP⁺+SIRT2-siRNA group vs MPP⁺group

Fig.8 The effect of SIRT2-siRNA on mitochondrial fusion related protein (a) The expression of Opa1, Mfn1 and Mfn2 proteins were detected by Western blot (b) The statistical diagram of Opa1 protein expression (c) The statistical diagram of Mfn1 protein expression (d) The statistical diagram of Mfn2 protein expression. Mean ± SD, n = 3. ^** P<0.01, ^*** P<0.001 MPP⁺group vs control group or negative control group. ^## P<0.01, ^### P<0.001 MPP⁺+SIRT2-siRNA group vs MPP⁺group


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