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©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
Quercetin ameliorates oxidative stress-induced senescence in rat nucleus pulposus-derived mesenchymal stem cells via the miR-34a-5p/SIRT1 axis
Wen-Jie Zhao, Xin Liu, Man Hu, Yu Zhang, Peng-Zhi Shi, Jun-Wu Wang, Xu-Hua Lu, Xiao-Fei Cheng, Yu-Ping Tao, Xin-Min Feng, Yong-Xiang Wang, Liang Zhang
Wen-Jie Zhao, Man Hu, Peng-Zhi Shi, Graduate School, Dalian Medical University, Dalian 116044, Liaoning Province, China
Xin Liu, Yu Zhang, Jun-Wu Wang, Yu-Ping Tao, Xin-Min Feng, Yong-Xiang Wang, Liang Zhang, Department of Orthopedics, Clinical Medical College, Yangzhou University, Yangzhou 225001, Jiangsu Province, China
Xu-Hua Lu, Department of Orthopedics, Changzheng Hospital of The Second Military Medical University, Shanghai 200003, China
Xiao-Fei Cheng, Department of Orthopedic Surgery, Shanghai Key Laboratory of Orthopedics Implants, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China
Author contributions: Zhao WJ and Liu X contributed to data curation, Writing-Original draft preparation, contributed equally to this work; Hu M and Zhang Y contributed to Visualization, Validation; Shi PZ and Wang JW performed Investigation; Lu XH, Cheng XF and Tao YP performed conceptualization, methodology; Feng XM, Wang YX and Zhang L performed supervision, writing- reviewing, editing and share corresponding author.
Supported by the National Natural Science Foundation of China, No. 82172462 and No. 81972136; the Traditional Chinese Medicine Science and Technology Development Plan Project of Jiangsu Province, No. YB2020085; and Cross Cooperation Project of Northern Jiangsu People’s Hospital, No. SBJC21014.
Institutional animal care and use committee statement: All animal experiments conformed to the internationally accepted principles for the care and use of laboratory animals, Yangzhou University, No. SYXK (Su) 2017-0044.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article. Figures
12A and
13 were created using Figdraw (
www.figdraw.com).
Data sharing statement: All data during the study are available from the corresponding author by request at
zhangliang6320@sina.com.
ARRIVE guidelines statement: The authors have read the ARRIVE Guidelines, and the manuscript was prepared and revised according to the ARRIVE Guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
https://creativecommons.org/Licenses/by-nc/4.0/ Corresponding author: Liang Zhang, PhD, Professor, Department of Orthopedics, Clinical Medical College, Yangzhou University, No. 98 Nantong West Road, Yangzhou, 225001, Jiangsu Province, China.
zhangliang6320@sina.com
Received: April 18, 2023
Peer-review started: April 18, 2023
First decision: May 17, 2023
Revised: May 25, 2023
Accepted: June 27, 2023
Article in press: June 27, 2023
Published online: August 26, 2023
Processing time: 129 Days and 0.1 Hours
BACKGROUND
Intervertebral disc degeneration (IDD) is a main contributor to low back pain. Oxidative stress, which is highly associated with the progression of IDD, increases senescence of nucleus pulposus-derived mesenchymal stem cells (NPMSCs) and weakens the differentiation ability of NPMSCs in degenerated intervertebral discs (IVDs). Quercetin (Que) has been demonstrated to reduce oxidative stress in diverse degenerative diseases.
AIM
To investigate the role of Que in oxidative stress-induced NPMSC damage and to elucidate the underlying mechanism.
METHODS
In vitro, NPMSCs were isolated from rat tails. Senescence-associated β-galactosidase (SA-β-Gal) staining, cell cycle, reactive oxygen species (ROS), real-time quantitative polymerase chain reaction (RT-qPCR), immunofluorescence, and western blot analyses were used to evaluated the protective effects of Que. Meanwhile the relationship between miR-34a-5p and Sirtuins 1 (SIRT1) was evaluated by dual-luciferase reporter assay. To explore whether Que modulates tert-butyl hydroperoxide (TBHP)-induced senescence of NPMSCs via the miR-34a-5p/SIRT1 pathway, we used adenovirus vectors to overexpress and downregulate the expression of miR-34a-5p and used SIRT1 siRNA to knockdown SIRT1 expression. In vivo, a puncture-induced rat IDD model was constructed, and X rays and histological analysis were used to assess whether Que could alleviate IDD in vivo.
RESULTS
We found that TBHP can cause NPMSCs senescence changes, such as reduced cell proliferation ability, increased SA-β-Gal activity, cell cycle arrest, the accumulation of ROS, and increased expression of senescence-related proteins. While abovementioned senescence indicators were significantly alleviated by Que treatment. Que decreased the expression levels of senescence-related proteins (p16, p21, and p53) and senescence-associated secreted phenotype (SASP), including IL-1β, IL-6, and MMP-13, and it increased the expression of SIRT1. In addition, the protective effects of Que on cell senescence were partially reversed by miR-34a-5p overexpression and SIRT1 knockdown. In vivo, X-ray, and histological analyses indicated that Que alleviated IDD in a puncture-induced rat model.
CONCLUSION
In summary, the present study provides evidence that Que reduces oxidative stress-induced senescence of NPMSCs via the miR-34a/SIRT1 signaling pathway, suggesting that Que may be a potential agent for the treatment of IDD.
Core Tip: In our article, we provide the evidence that quercetin (Que) can prevent oxidative stress induced senescence of nucleus pulposus-derived mesenchymal stem cells via miR-34a/SIRT1 signaling pathway. Moreover, Que could ameliorate the progression of intervertebral disc degeneration (IDD) in rat model. Thus, Que can be considered as a potential agent for the treatment of IDD.
