Basic Study
Copyright ©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Orthop. Apr 18, 2024; 15(4): 363-378
Published online Apr 18, 2024. doi: 10.5312/wjo.v15.i4.363
Exercise promotes osteogenic differentiation by activating the long non-coding RNA H19/microRNA-149 axis
Xu-Chang Zhou, Dong-Xue Wang, Chun-Yu Zhang, Ya-Jing Yang, Ruo-Bing Zhao, Sheng-Yao Liu, Guo-Xin Ni
Xu-Chang Zhou, Dong-Xue Wang, Chun-Yu Zhang, Ruo-Bing Zhao, School of Sport Medicine and Rehabilitation, Beijing Sport University, Beijing 100084, China
Ya-Jing Yang, Department of Acupuncture and Moxibustion, Hubei University of Chinese Medicine, Wuhan 430065, Hubei Province, China
Sheng-Yao Liu, Department of Spinal Surgery, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 510260, Guangdong Province, China
Guo-Xin Ni, Department of Rehabilitation Medicine, The First Affiliated Hospital of Xiamen University, Xiamen 361003, Fujian Province, China
Co-first authors: Xu-Chang Zhou and Dong-Xue Wang.
Author contributions: Ni GX and Zhou XC designed and coordinated the study; Wang DX and Yang YJ performed the experiments, acquired and analyzed data; Zhao RB interpreted the data; Zhou XC and Liu SY acquired the Fund; Zhou XC and Wang DX wrote the manuscript; all authors approved the final version of the article.
Supported by the Key Laboratory of Sports and Physical Health of the Ministry of Education (Beijing Sport University), the Fundamental Research Funds for the Central Universities, No. 20221021; the Guangzhou Science and Technology Plan Project, No. 202102020536; and the Plan on Enhancing Scientific Research in GMU.
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the Beijing Sport University (Approval Number: 2023026A).
Conflict-of-interest statement: The authors declare no conflict of interest.
Data sharing statement: No additional data are available.
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: Guo-Xin Ni, MD, PhD, Chief Doctor, Chief Physician, Professor, Department of Rehabilitation Medicine, The First Affiliated Hospital of Xiamen University, No. 55 Zhenhai Road, Siming District, Xiamen 361003, Fujian Province, China. nigx@xmu.edu.cn
Received: November 22, 2023
Peer-review started: November 22, 2023
First decision: January 24, 2024
Revised: February 4, 2024
Accepted: March 19, 2024
Article in press: March 19, 2024
Published online: April 18, 2024
Processing time: 144 Days and 0.3 Hours
Abstract
BACKGROUND

Regular physical activity during childhood and adolescence is beneficial to bone development, as evidenced by the ability to increase bone density and peak bone mass by promoting bone formation.

AIM

To investigate the effects of exercise on bone formation in growing mice and to investigate the underlying mechanisms.

METHODS

20 growing mice were randomly divided into two groups: Con group (control group, n = 10) and Ex group (treadmill exercise group, n = 10). Hematoxylin-eosin staining, immunohistochemistry, and micro-CT scanning were used to assess the bone formation-related indexes of the mouse femur. Bioinformatics analysis was used to find potential miRNAs targets of long non-coding RNA H19 (lncRNA H19). RT-qPCR and Western Blot were used to confirm potential miRNA target genes of lncRNA H19 and the role of lncRNA H19 in promoting osteogenic differentiation.

RESULTS

Compared with the Con group, the expression of bone morphogenetic protein 2 was also significantly increased. The micro-CT results showed that 8 wk moderate-intensity treadmill exercise significantly increased bone mineral density, bone volume fraction, and the number of trabeculae, and decreased trabecular segregation in the femur of mice. Inhibition of lncRNA H19 significantly upregulated the expression of miR-149 and suppressed the expression of markers of osteogenic differentiation. In addition, knockdown of lncRNA H19 significantly downregulated the expression of autophagy markers, which is consistent with the results of autophagy-related protein changes detected in mouse femurs by immunofluorescence.

CONCLUSION

Appropriate treadmill exercise can effectively stimulate bone formation and promote the increase of bone density and bone volume in growing mice, thus enhancing the peak bone mass of mice. The lncRNA H19/miR-149 axis plays an important regulatory role in osteogenic differentiation.

Keywords: Exercise; Osteogenic differentiation; Bone formation; Bone mesenchymal stem cells; Autophagy

Core Tip: Adolescence is a critical period for laying the foundation for optimal peak bone mass in adulthood. Studies have shown that pre-puberty and early puberty are the periods when bones are most responsive to mechanical loading. It is essential to explore the effect of exercise on promoting bone formation in adolescence and its underlying mechanisms. In this paper, we explored the promotional effects of treadmill exercise on bone formation in growing mice and further explored the underlying mechanisms. Our results validate that moderate intensity running exercise is effective in stimulating bone formation and promoting increases in bone mineral density and bone mass, thereby enhancing peak bone mass in growing mice. Notably, the long non-coding RNA H19 (lncRNA H19)/microRNA-149 (miR-149) axis plays an important regulatory role in the osteogenic differentiation of bone mesenchymal stem cells. Overall, this paper emphasizes that exercise may promote bone formation in mice through the lncRNA H19/miR-149 axis, which may be closely related to the activation of autophagy.