MiR-214 Attenuates the Osteogenic Effects of Mechanical Loading on Osteoblasts

Abstract

Exercise is an effective way to prevent osteoporosis, but its mechanism remains unclear. MicroRNAs (miRNAs) play an essential role in bone metabolism. Recently, mechanical loading was reported to induce changes in miRNA expression in osteoblasts. However, the role of miRNAs in bone under exercise and its underlining mechanisms of action still remain unknown. MiR-214 was reported to regulate the process of osteogenesis and is considered a biomarker of osteoporosis. In this study, we aimed to investigate whether exercise could induce changes in miRNA expression in bone and to study the effects of miR-214 on mechanical loading-induced osteogenesis in osteoblasts. The results showed that miR-214 was down-regulated in both tibia from C57BL/6 mice after exercise in vivo and in osteoblasts after mechanical strain in vitro. Mechanical strain could enhance the ALP activity, promote matrix mineralization, up-regulate the expression of osteogenic factors such as ATF4, Osterix, ALP and β-catenin, and down-regulate RANKL and RANK expression. Over-expression of miR-214 not only inhibited the expression of these osteogenic factors but also attenuated mechanical strain-enhanced osteogenesis in osteoblasts. Collectively, our results indicated that miR-214 could attenuate the osteogenic effects of mechanical loading on osteoblasts, suggesting that inhibition of miR-214 may be one of the ways in which exercise prevents osteoporosis.

Key words

exercise - mechanical strain - miR-214 - osteoblasts - osteoporosis

• References

• 1 Johnell O, Kanis JA. An estimate of the worldwide prevalence and disability associated with osteoporotic fractures. Osteoporos Int 2006; 17: 1726-1733
• 2 Hernlund E, Svedbom A, Ivergard M, Compston J, Cooper C, Stenmark J, McCloskey EV, Jonsson B, Kanis JA. Osteoporosis in the European Union: medical management, epidemiology and economic burden. A report prepared in collaboration with the International Osteoporosis Foundation (IOF) and the European Federation of Pharmaceutical Industry Associations (EFPIA). Arch Osteoporos 2013; 8: 136
• 3 Shepstone L, Lenaghan E, Cooper C, Clarke S, Fong-Soe-Khioe R, Fordham R, Gittoes N, Harvey I, Harvey N, Heawood A, Holland R, Howe A, Kanis J, Marshall T, O'Neill T, Peters T, Redmond N, Torgerson D, Turner D, McCloskey E, Team SS. Screening in the community to reduce fractures in older women (SCOOP): A randomised controlled trial. Lancet 2018; 391: 741-747
• 4 Yuan Y, Chen X, Zhang L, Wu J, Guo J, Zou D, Chen B, Sun Z, Shen C, Zou J. The roles of exercise in bone remodeling and in prevention and treatment of osteoporosis. Prog Biophys Mol Biol 2016; 122: 122-130
• 5 Chen X, Li L, Guo J, Zhang L, Yuan Y, Chen B, Sun Z, Xu J, Zou J. Treadmill running exercise prevents senile osteoporosis and upregulates the Wnt signaling pathway in SAMP6 mice. Oncotarget 2016; 7: 71072-71086
• 6 Harding AT, Weeks BK, Watson SL, Beck BR. The LIFTMOR-M (Lifting Intervention For Training Muscle and Osteoporosis Rehabilitation for Men) trial: Protocol for a semirandomised controlled trial of supervised targeted exercise to reduce risk of osteoporotic fracture in older men with low bone mass. BMJ Open 2017; 7: e014951
• 7 Watson SL, Weeks BK, Weis LJ, Harding AT, Horan SA, Beck BR. High-intensity resistance and impact training improves bone mineral density and physical function in postmenopausal women with osteopenia and osteoporosis: The LIFTMOR randomized controlled trial. J Bone Miner Res 2018; 33: 1-10
• 8 Damaraju S, Matyas JR, Rancourt DE, Duncan NA. The effect of mechanical stimulation on mineralization in differentiating osteoblasts in collagen-I scaffolds. Tissue Eng Part A 2014; 20: 3142-3153
• 9 Li R, Liang L, Dou Y, Huang Z, Mo H, Wang Y, Yu B. Mechanical strain regulates osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells. Biomed Res Int 2015; 2015; 873251:
• 10 Qin TW, Sun YL, Thoreson AR, Steinmann SP, Amadio PC, An KN, Zhao C. Effect of mechanical stimulation on bone marrow stromal cell-seeded tendon slice constructs: A potential engineered tendon patch for rotator cuff repair. Biomaterials 2015; 51: 43-50
• 11 Vishal M, Vimalraj S, Ajeetha R, Gokulnath M, Keerthana R, He Z, Partridge NC, Selvamurugan N. MicroRNA-590-5p stabilizes Runx2 by targeting Smad7 during osteoblast differentiation. J Cell Physiol 2017; 232: 371-380
• 12 Xiao WZ, Gu XC, Hu B, Liu XW, Zi Y, Li M. Role of microRNA-129-5p in osteoblast differentiation from bone marrow mesenchymal stem cells. Cell Mol Biol 2016; 62: 95-99
• 13 Li JP, Zhuang HT, Xin MY, Zhou YL. MiR-214 inhibits human mesenchymal stem cells differentiating into osteoblasts through targeting beta-catenin. Eur Rev Med Pharmacol Sci 2017; 21: 4777-4783
• 14 Shi K, Lu J, Zhao Y, Wang L, Li J, Qi B, Li H, Ma C. MicroRNA-214 suppresses osteogenic differentiation of C2C12 myoblast cells by targeting Osterix. Bone 2013; 55: 487-494
• 15 Wang X, Guo B, Li Q, Peng J, Yang Z, Wang A, Li D, Hou Z, Lv K, Kan G, Cao H, Wu H, Song J, Pan X, Sun Q, Ling S, Li Y, Zhu M, Zhang P, Peng S, Xie X, Tang T, Hong A, Bian Z, Bai Y, Lu A, Li Y, He F, Zhang G, Li Y. miR-214 targets ATF4 to inhibit bone formation. Nat Med 2013; 19: 93-100
• 16 Yang L, Ge D, Cao X, Ge Y, Chen H, Wang W, Zhang H. MiR-214 Attenuates osteogenic differentiation of mesenchymal stem cells via targeting FGFR1. Cell Physiol Biochem 2016; 38: 809-820
• 17 Zhao C, Sun W, Zhang P, Ling S, Li Y, Zhao D, Peng J, Wang A, Li Q, Song J, Wang C, Xu X, Xu Z, Zhong G, Han B, Chang YZ, Li Y. miR-214 promotes osteoclastogenesis by targeting Pten/PI3k/Akt pathway. RNA Biol 2015; 12: 343-353
• 18 Chang M, Lin H, Fu H, Wang B, Han G, Fan M. MicroRNA-195-5p regulates osteogenic differentiation of periodontal ligament cells under mechanical loading. J Cell Physiol 2017; 232: 3762-3774
• 19 Li J, Hu C, Han L, Liu L, Jing W, Tang W, Tian W, Long J. MiR-154-5p regulates osteogenic differentiation of adipose-derived mesenchymal stem cells under tensile stress through the Wnt/PCP pathway by targeting Wnt11. Bone 2015; 78: 130-141
• 20 Liu L, Liu M, Li R, Liu H, Du L, Chen H, Zhang Y, Zhang S, Liu D. MicroRNA-503-5p inhibits stretch-induced osteogenic differentiation and bone formation. Cell Biol Int 2017; 41: 112-123
• 21 Wang H, Sun Z, Wang Y, Hu Z, Zhou H, Zhang L, Hong B, Zhang S, Cao X. miR-33-5p, a novel mechano-sensitive microRNA promotes osteoblast differentiation by targeting Hmga2. Sci Rep 2016; 6: 23170
• 22 Yuan Y, Zhang L, Tong X, Zhang M, Zhao Y, Guo J, Lei L, Chen X, Tickner J, Xu J, Zou J. Mechanical stress regulates bone metabolism through microRNAs. J Cell Physiol 2017; 232: 1239-1245
• 23 Drummond GB, Paterson DJ, McGrath JC. ARRIVE: new guidelines for reporting animal research. Exp Physiol 2010; 95: 841
• 24 Harriss DJ, Macsween A, Atkinson G. Standards for ethics in sport and exercise science research: 2018 update. Int J Sports Med 2017; 38: 1126-1131
• 25 Jilka RL. Parathyroid hormone-stimulated development of osteoclasts in cultures of cells from neonatal murine calvaria. Bone 1986; 7: 29-40
• 26 Zhang L, Chen X, Wu J, Yuan Y, Guo J, Biswas S, Li B, Zou J. The effects of different intensities of exercise and active vitamin D on mouse bone mass and bone strength. J Bone Miner Metab 2017; 35: 265-277
• 27 Li L, Chen X, Lv S, Dong M, Zhang L, Tu J, Yang J, Zhang L, Song Y, Xu L, Zou J. Influence of exercise on bone remodeling-related hormones and cytokines in ovariectomized rats: a model of postmenopausal osteoporosis. PLoS One 2014; 9: e112845
• 28 McCabe LR, Irwin R, Tekalur A, Evans C, Schepper JD, Parameswaran N, Ciancio M. Exercise prevents high fat diet-induced bone loss, marrow adiposity and dysbiosis in male mice. Bone 2019; 118: 20-31
• 29 Chen X, Guo J, Yuan Y, Sun Z, Chen B, Tong X, Zhang L, Shen C, Zou J. Cyclic compression stimulates osteoblast differentiation via activation of the Wnt/beta-catenin signaling pathway. Mol Med Rep 2017; 15: 2890-2896
• 30 Gu Q, Tian H, Zhang K, Chen D, Chen D, Wang X, Zhao J. Wnt5a/FZD4 Mediates the mechanical stretch-induced osteogenic differentiation of bone mesenchymal stem cells. Cell Physiol Biochem 2018; 48: 215-226
• 31 Grunhagen J, Bhushan R, Degenkolbe E, Jager M, Knaus P, Mundlos S, Robinson PN, Ott CE. MiR-497 approximately 195 cluster microRNAs regulate osteoblast differentiation by targeting BMP signaling. J Bone Miner Res 2015; 30: 796-808
• 32 You L, Pan L, Chen L, Gu W, Chen J. MiR-27a is essential for the shift from osteogenic differentiation to adipogenic differentiation of mesenchymal stem cells in postmenopausal osteoporosis. Cell Physiol Biochem 2016; 39: 253-265
• 33 Zuo B, Zhu J, Li J, Wang C, Zhao X, Cai G, Li Z, Peng J, Wang P, Shen C, Huang Y, Xu J, Zhang X, Chen X. microRNA-103a functions as a mechanosensitive microRNA to inhibit bone formation through targeting Runx2. J Bone Miner Res 2015; 30: 330-345
• 34 Desvignes T, Contreras A, Postlethwait JH. Evolution of the miR199-214 cluster and vertebrate skeletal development. RNA Biol 2014; 11: 281-294
• 35 Watanabe T, Sato T, Amano T, Kawamura Y, Kawamura N, Kawaguchi H, Yamashita N, Kurihara H, Nakaoka T. Dnm3os, a non-coding RNA, is required for normal growth and skeletal development in mice. Dev Dyn 2008; 237: 3738-3748
• 36 Sun Y, Kuek V, Liu Y, Tickner J, Yuan Y, Chen L, Zeng Z, Shao M, He W, Xu J. MiR-214 is an important regulator of the musculoskeletal metabolism and disease. J Cell Physiol 2018; 234: 231-245
• 37 Xi Y, Chen Y. PTEN plays dual roles as a tumor suppressor in osteosarcoma cells. J Cell Biochem 2017; 118: 2684-2692
• 38 Shen WC, Lai YC, Li LH, Liao K, Lai HC, Kao SY, Wang J, Chuong CM, Hung SC. Methylation and PTEN activation in dental pulp mesenchymal stem cells promotes osteogenesis and reduces oncogenesis. Nat Commun 2019; 10: 2226
• 39 Roberto VP, Gavaia P, Nunes MJ, Rodrigues E, Cancela ML, Tiago DM. Evidences for a new role of miR-214 in chondrogenesis. Sci Rep 2018; 8: 3704
• 40 Cao F, Zhan J, Chen X, Zhang K, Lai R, Feng Z. miR-214 promotes periodontal ligament stem cell osteoblastic differentiation by modulating Wnt/betacatenin signaling. Mol Med Rep 2017; 16: 9301-9308