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Effect of Combined Resistance-Aerobic Exercise Training on the Wnt/Beta-Catenin Signaling Pathway and Bone Remodeling in a Rat Model of Aging
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Vajihe Varzandeh1    , Yaser Kazemzadeh *2 , Sanaz Mirzayan Shanjani3 , Hossein Shirvani4 |
1- Ph.D Candidate in Exercise Physiology, Department of Exercise Physiology, Islamshahr Branch, Islamic Azad University, Islamshahr, Iran.
2- Assistant Professor, Department of Exercise Physiology, Islamshahr Branch, Islamic Azad University, Islamshahr, Iran. , yaser.kazemzadeh@yahoo.com
3- Assistant Professor, Department of Exercise Physiology, Islamshahr Branch, Islamic Azad University, Islamshahr, Iran.
4- Associate Professor, Sports Physiology Research Center, Lifestyle Research Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran. |
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Keywords: Exercise [MeSH], beta Catenin [MeSH], T Cell Transcription Factor 1 [MeSH], Aging [MeSH]
Article ID: Vol27-31 |
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Type of Study: Original Articles |
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Exercise Physiology
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Abstract: (122 Views) |
Extended Abstract
Introduction
Bone tissue quality is influenced by age-related alterations in bone minerals, collagen composition, cross-linking profiles, hydration levels, and even non-collagenous proteins. Musculoskeletal health represents a critical determinant of the aging process, with bones serving as a fundamental component of the musculoskeletal system. While age-associated structural changes in bone are inevitable, the impact is further exacerbated by diseases that directly or indirectly affect osseous tissue, as well as age-related lifestyle modifications. Compromised bone integrity and strength predispose the elderly population to functional impairment, disability, dependence, frequent falls, chronic pain, substantial healthcare expenditures, and elevated mortality rates.
Beta-catenin (β-catenin) is a multifunctional and evolutionarily conserved molecule that plays a pivotal role in numerous developmental and homeostatic processes within metazoans. Specifically, β-catenin serves as both an integral structural component of cadherin-based adherens junctions and the primary nuclear effector of the wingless-type mouse mammary tumor virus (MMTV) integration site family (Wnt) signaling pathway. Research indicates that β-catenin is indispensable for normal skeletal development. In the human embryo, β-catenin is critical for the early stages of osteoblast differentiation. Postnatally, β-catenin within mature osteoblasts and osteocytes indirectly suppresses osteoclast differentiation. Furthermore, it regulates bone homeostasis by enhancing osteoblast activity and inhibiting osteoblast turnover, while simultaneously impeding the formation of osteoclasts and bone marrow adiposity.
T-cell factor 1 (TCF1) serves as a primary target of the Wnt signaling pathway and functions as a classical transcription factor for β-catenin. Upon activation by various upstream signals, TCF activates the transcription of downstream target genes and regulates biological processes, such as the differentiation, proliferation, and apoptosis of osteoblasts, which play a pivotal role in bone development and remodeling. Animal models have demonstrated that mutations in TCF1 result in loss of function, which can consequently cause numerous pathologies, most notably skeletal abnormalities and bone mineral deficiencies.
Physical training represents a beneficial non-pharmacological intervention for enhancing muscle mass and strength, as well as stimulating increases in bone density. The osteogenic effects of exercise on bone tissues are directly correlated with the magnitude of the mechanical strain applied. Scientific evidence demonstrates that physical training is highly effective in preventing bone mass attrition, fractures, and osteoporosis. Furthermore, bone metabolism is significantly influenced by physical training. During physical activities, bone tissue undergoes deformation, triggering mechanosensors located within the cells, such as ion channels and integrins, to alter their conformational structure. This process activates multiple signaling pathways, most notably the Wnt/β-catenin pathway, which subsequently induces the process of osteogenesis. The phenomenon of aging, in conjunction with intrinsic and extrinsic factors, including genetics, hormones, sedentary lifestyle, and poor nutrition, accelerates age-related bone mass attrition, which is of critical clinical significance. Consequently, the present study was conducted to investigate the effects of combined resistance-aerobic exercise training on the Wnt/β-catenin signaling pathway and bone remodeling in a rat model of aging.
Methods
This experimental study was conducted on 16 aged male Wistar rats (22 months old, weighing approximately 300–350 g) and 8 young male rats (3 months old, weighing approximately 190–230 g) as a young control group.
Following a two-week period of animal acclimatization and habituation to the laboratory environment, the rats were randomly assigned to three groups as follows:
Combined resistance-aerobic exercise training group: This group consisted of 8 aged male rats that performed combined resistance-aerobic exercise training for eight weeks, with five sessions per week. These rats were dissected 48 hours after the final training session.
Aged group: This group involved 8 aged male rats that received no intervention. They were dissected concurrently with the experimental group.
Young control group: This group comprised 8 young male Wistar rats that had no physical training and were merely included in the study to facilitate a comparative analysis of the aging process against the aged groups. This group was also dissected concurrently with the other research cohorts.
The combined resistance-aerobic exercise training protocol was adapted from the modified protocol developed by Dotzert et al. This protocol comprised a combination of resistance and aerobic training performed five days per week over an eight-week period. In each session, the resistance training was conducted first; following a one-hour rest period, the rats proceeded to the aerobic training. The resistance training involved climbing a one-meter ladder with weights attached to the tails. To prevent inflammation and muscle soreness, only concentric contractions were utilized. The rats were weighed weekly, and the resistance level (training intensity) was adjusted based on a specific percentage of their body weight.
To implement the aerobic training protocol, the maximum running speed of the rats was first determined using a specialized rodent treadmill. The training intensity was subsequently established based on a percentage of the maximum speed attained.
For bone tissue sampling, 48 hours after the final training session, the rats were anesthetized via intraperitoneal injection of ketamine (30–50 mg/kg/bw) and xylazine (3–5 mg/kg/bw) and then sacrificed. Femoral bone tissue samples were isolated under sterile conditions. After meticulously removing the surrounding soft tissues while ensuring the periosteum remained intact and undamaged, the bone tissues were transferred to ribonuclease (RNase)-free and deoxyribonuclease (DNase)-free microtubes. Finally, the samples were frozen by immersion in liquid nitrogen.
For ribonucleic acid (RNA) extraction, the femoral bone tissue was first homogenized using a homogenizer, followed by RNA isolation according to the manufacturer's instructions. The quality and quantity of the extracted RNA were assessed using a NanoDrop spectrophotometer. Subsequently, complementary deoxyribonucleic acid (cDNA) synthesis was performed following the manufacturer's protocol. The expression levels of the target genes were evaluated using real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR).
Results
Following eight weeks of combined resistance-aerobic exercise training, a significant increase was observed in the gene expression of β-catenin and TCF1 compared to the aged control group. Statistically significant differences in the expression levels of both β-catenin and TCF1 were identified among the study groups (P<0.001).
Regarding TCF1 gene expression, significant differences were found between the aged group and the training group (P<0.028), as well as between the aged group and the young group (P<0.001). However, no significant difference was observed between the training group and the young group in the TCF1 expression. Similarly, for β-catenin gene expression, significant differences were observed between the aged group and the training group (P<0.024), and between the aged group and the young group (P<0.001). Moreover, a significant difference in β-catenin expression was also observed between the training group and the young group (P<0.024).
Conclusion
According to the findings of this study, a significant increase was observed in the expression of β-catenin and TCF1 genes following eight weeks of training compared to the aged group. Given the pivotal role of these genes in bone development, the results of the present research demonstrate the positive effects of combined resistance-aerobic exercise training on bone development in an aged animal model.
Bone aging is a progressive process typically characterized by an imbalance between osteoclast bone resorption and osteoblast bone formation. This process is influenced by genetic factors and lifestyle-induced epigenetic modifications. The loss of bone mass associated with skeletal aging is driven by oxidative stress, which negatively impacts signaling pathways involved in bone cell survival and the osteogenesis process. Moreover, the production of mitochondrial superoxide anions in aged osteocytes enhances bone resorption, while reactive oxygen species (ROS) suppress β-catenin signaling. Furthermore, the activation of Forkhead Box O (FOXO) transcription signaling, which plays a pivotal role in aging and longevity, induces apoptosis in osteoblasts and osteocytes. Conversely, physical training positively modulates bone metabolism through various mechanisms, such as the activation of inflammatory cascades involving immune cells and inflammatory mediators, the stimulation of metabolic responses due to skeletal muscle-derived interleukin-6 (IL-6) increase, and the induction of the Wnt signaling pathway. Moreover, mechanical loading induced by physical training suppresses the adipogenic differentiation of mesenchymal stem cells by facilitating the β-catenin-mediated release of FOXO transcription. Additionally, physical training has been shown to attenuate osteocyte apoptosis.
Physical training and physical activities serve as a non-pharmacological preventive strategy against osteoporosis in the elderly. The interplay among mechanical loading, hormones or cytokines, and physical training-induced signaling pathways enhances bone formation and attenuates bone resorption, thereby maintaining skeletal integrity. Furthermore, dysregulation of bone angiogenesis is associated with various bone pathologies, including osteoporosis; physical training facilitates the improvement of bone angiogenesis by modulating key angiogenic mediators.
Ethical Statement
This study was approved by the Research Ethics Committee at Islamic Azad University, Parand Branch (IR.IAU.PIAU.REC.1403.034).
Authors' Contributions
Vajihe Varzandeh: Project execution, Data collection, Data analysis, Interpretation of the results, Drafting of the initial manuscript, Approval of the final manuscript.
Yaser Kazemzadeh (Ph.D): Project administration and design, Interpretation of the results, Approval of the final manuscript.
Sanaz Mirzayan Shanjani (Ph.D): Data analysis, Approval of the final manuscript.
Hossein Shirvani (Ph.D): Project administration and design, Project execution, Data collection, Data analysis, Interpretation of the results, Approval of the final manuscript.
Conflicts of Interest
No conflicts of interest.
Acknowledgement
This study has been derived from the doctoral dissertation by Vajihe Varzandeh in Exercise Physiology at the Faculty of Physical Education at Islamic Azad University, Islamshahr Branch. The authors would like to thank everyone who contributed to the implementation of this study.
Key Message: Performing combined resistance-aerobic exercise training for eight weeks resulted in a significant increase in the expression of β-catenin and TCF1 genes within aged male Wistar rats. As a non-pharmacological, low-cost, and safe intervention, this protocol exerted positive effects on bone development in the aged animal model.
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Varzandeh V, Kazemzadeh Y, Mirzayan Shanjani S, Shirvani H. Effect of Combined Resistance-Aerobic Exercise Training on the Wnt/Beta-Catenin Signaling Pathway and Bone Remodeling in a Rat Model of Aging. J Gorgan Univ Med Sci 2025; 27 (4) :1-10 URL: http://goums.ac.ir/journal/article-1-4579-en.html
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