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SIRT1 Inhibits High Shear Stress-Induced Apoptosis in Rat Cortical Neurons

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Abstract

Introduction

Sirtuin1 (SIRT1), one of NAD+-dependent protein deacetylases, is proved to be neuroprotective in aging diseases, but its effect on neuronal apoptosis has not been clarified. To investigate the role of SIRT1 in inhibiting neuronal apoptosis, SIRT1 was interfered or overexpressed in cortical neurons.

Methods

We exerted overloading laminar shear stress with 10 dyn/cm2 for 4, 8, and 12 h on neurons to cause cortical neuronal apoptosis, and the apoptosis percentage was tested by TUNEL assay. The adenovirus plasmids containing SIRT1 RNA interference or SIRT1 wild type gene were transfected into neurons before shear stress loading. SIRT1 mRNA and protein level were tested by Real-time PCR, immunofluorescence and western blots assay.

Results

SIRT1 was primarily expressed in nucleus of cortical neurons, and its mRNA level was significantly increased after 4 h stimulation. SIRT1 RNAi cortical neurons had higher TUNEL positive cells, while SIRT1 overexpression significantly decreased the percentage of died cells induced by shear stress compared to control group.

Conclusions

SIRT1 plays a neuroprotective role in shear stress induced apoptosis and could be as potential pharmacological targets against neuronal degeneration in future.

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Abbreviations

SIRT1:

Sirtuin1

NAD+ :

Nicotinamide adenine dinucleotide

SIRT1-7:

Sirtuin family isoforms

Sir-2:

Silent information regulator-2

Nkx2.5:

NK2 Homeobox 5

AAA:

Abdominal aortic aneurysms

CR:

Caloric restriction

PASMC:

Pulmonary artery smooth muscle cells

PAH:

Pulmonary arterial hypertension

PDGF-BB:

Platelet-derived growth factor-BB

NRF2:

Nuclear factor (erythroid-derived 2)-like 2

DAI:

Diffuse axonal injury

FSSI:

Fluid shear stress injury

MTT:

3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-[2H]-tetrazolium bromide

TUNEL:

Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling

DAPI:

4,6-Diamidino-2-phenylindole dihydrochloride

BSA:

Albumin from bovine serum

PBS:

Phosphate buffered saline

RT-PCR:

Reverse transcription-polymerase chain reaction

SDS:

Sodium dodecyl sulfate

PAGE:

Polyacrylamide gel electrophoresis

RNAi:

RNA interference

PVDF:

Poly(vinylidene fluoride)

BCA:

Bicinchoninic acid

GFP:

Green fluorescent protein

eNOS:

Endothelial nitric oxide synthase

ECs:

Endothelial cells

Cx40:

Connexin40

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Acknowledgments

This study was supported by funds from National Natural Science Foundation of China (NSFC) Research Grant (31971238, 61871014, 51574246, 31771019, 11472032, 11120101001), and National Basic Research Program of China (973 Program, 2011CB710901), the 111 Project (B13003).

Author Contributions

PL and Y-BF conceived and designed the experiments. WS, M-LL, Z-JZ, C-QH and A-QW performed the experiments. WS, M-LL, J-WX and PL analyzed the data. M-LL contributed reagents/materials/analysis tools. M-LL, PL and Y-BF wrote the paper. All authors read and approved the final manuscript.

Data Availability

The data used to support the findings of this study are included within the article.

Conflict of interest

Wei Song, Mei-Li Liu, Zhi-Jun Zhao, Chong-Quan Huang, Jun-Wei Xu, An-Qing Wang, Ping Li and Yu-Bo Fan declare that they have no conflicts of interest.

Ethical Approval

No human studies were carried out by the authors for this article. All experiments involving the use of animals were in compliance with Provisions and General Recommendation of Chinese Experimental Animals Administration Legislation and were approved by Beijing Municipal Science & Technology Commission (Permit Number: SCXK (Beijing) 2006-0008 and SYXK (Beijing) 2006-0025).

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Author notes

  1. Wei Song and Mei-Li Liu have contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing, 100191, China

    Wei Song, Mei-Li Liu, Zhi-Jun Zhao, Chong-Quan Huang, Jun-Wei Xu, An-Qing Wang, Ping Li & Yu-Bo Fan

  2. Beijing Advanced Innovation Centre for Biomedical Engineering, Beihang University, Beijing, 100191, China

    Wei Song, Mei-Li Liu, Zhi-Jun Zhao, Chong-Quan Huang, Jun-Wei Xu, An-Qing Wang, Ping Li & Yu-Bo Fan

  3. National Research Center for Rehabilitation Technical Aids, Beijing, 100176, China

    Yu-Bo Fan

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  1. Wei Song
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Song, W., Liu, ML., Zhao, ZJ. et al. SIRT1 Inhibits High Shear Stress-Induced Apoptosis in Rat Cortical Neurons. Cel. Mol. Bioeng. 13, 621–631 (2020). https://doi.org/10.1007/s12195-020-00623-2

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