Overexpression of SIRT1 prevents hypoxia‑induced apoptosis in osteoblast cells

Zhou,Lu; Wang,Sung Il; Moon,Young Jae; Kim,Kyoung Min; Lee,Kwang Bok; Park,Byung‑Hyun; Jang,Kyu Yun; Kim,Jung Ryul

doi:10.3892/mmr.2017.6917

Overexpression of SIRT1 prevents hypoxia‑induced apoptosis in osteoblast cells

Authors:
- Lu Zhou
- Sung Il Wang
- Young Jae Moon
- Kyoung Min Kim
- Kwang Bok Lee
- Byung‑Hyun Park
- Kyu Yun Jang
- Jung Ryul Kim
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Affiliations: Department of Sports Medicine, Taishan Medical University, Taian, Shandong 271000, P.R. China, Department of Orthopaedic Surgery, Chonbuk National University Medical School, Research Institute of Clinical Medicine of Chonbuk National University, Biomedical Research Institute of Chonbuk National University Hospital and Research Institute for Endocrine Sciences, Jeonju 54896, Republic of Korea, Department of Biochemistry, Chonbuk National University Medical School, Research Institute of Clinical Medicine of Chonbuk National University, Biomedical Research Institute of Chonbuk National University Hospital and Research Institute for Endocrine Sciences, Jeonju 54896, Republic of Korea, Department of Pathology, Chonbuk National University Medical School, Research Institute of Clinical Medicine of Chonbuk National University, Biomedical Research Institute of Chonbuk National University Hospital and Research Institute for Endocrine Sciences, Jeonju 54896, Republic of Korea
Published online on: July 4, 2017 https://doi.org/10.3892/mmr.2017.6917
Pages: 2969-2975

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Abstract

Hypoxic‑ischemic injury of the bone results in osteonecrosis. Nicotinamide adenosine dinucleotide (NAD)‑dependent deacetylase sirtuin‑1 (SIRT1), a type of NAD‑dependent deacetylase, is involved in multiple biological functions, particularly in anti‑apoptosis. However, the effects of SIRT1 in osteoblasts remain unclear and whether SIRT1 protects osteoblasts in hypoxic conditions remains to be elucidated. In the present study, the role of SIRT1 in the osteoblast cells under hypoxia and the underlying mechanism of the anti‑apoptotic activity of SIRT1 were investigated. MC3T3‑E1 osteoblast cells were used for the present study and oxygen‑absorbing packs were used to induce cell hypoxia and apoptosis. MC3T3‑E1 cells were overexpressed SIRT1 by transfection with a SIRT1 adenovirus. The small interfering RNA of SIRT1 to was used to transfect cells to decrease the protein level. An MTT assay was used to estimate cell viability. Apoptosis was examined with the APOPercentage apoptosis assay kit and the activity of caspases was measured by a caspase 3 and 7 activity kit. Co‑immunoprecipitation was used to investigate protein binding ability. The mRNA and protein expression levels were quantified with reverse transcription‑quantitative polymerase chain reaction and immunoblotting. It was demonstrated that the expression of SIRT1 mRNA and protein were elevated, and peaked at 12 h under hypoxic conditions. The data demonstrated that SIRT1 overexpression in cells significantly increased cell viability and markedly decreased the percentage of apoptosis compared with the control and knockdown groups. Furthermore, overexpression of SIRT1 significantly activated anti‑apoptotic effects by deacetylating lysine residue binding to protein kinase B and decreasing the activity of caspases 3, 9 and subsequent pathways. The results from the present study suggested that SIRT1 may serve a protective function in hypoxia‑induced apoptosis in MC3T3‑E1 cells, and that SIRT1 intervention may potentially aid in the treatment of ischemic bone disease.

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