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

doi:10.5312/wjo.v15.i4.363

Exercise promotes osteogenic differentiation by activating the long non-coding RNA H19/microRNA-149 axis

World J Orthop. 2024 Apr 18;15(4):363-378. doi: 10.5312/wjo.v15.i4.363.

Authors

Xu-Chang Zhou¹, Dong-Xue Wang¹, Chun-Yu Zhang¹, Ya-Jing Yang², Ruo-Bing Zhao¹, Sheng-Yao Liu³, Guo-Xin Ni⁴

Affiliations

¹ School of Sport Medicine and Rehabilitation, Beijing Sport University, Beijing 100084, China.
² Department of Acupuncture and Moxibustion, Hubei University of Chinese Medicine, Wuhan 430065, Hubei Province, China.
³ Department of Spinal Surgery, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 510260, Guangdong Province, China.
⁴ Department of Rehabilitation Medicine, The First Affiliated Hospital of Xiamen University, Xiamen 361003, Fujian Province, China. nigx@xmu.edu.cn.

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