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Mitohormesis and Regeneration: Natural Compounds Chlorogenic Acid (CGA) and Isovanillic Acid 3-O-sulfate (IVAS) Boost Muscle Cell Recovery in the Equine Athlete Model.
Abstract: Skeletal muscle satellite cells ( SCs), essential for muscle regeneration, are a valuable model for studying exercise-induced stress relevant to human athletes. This study examined the effects of two natural compounds-chlorogenic acid (CGA) and isovanillic acid 3-O-sulfate (IVAS)-increasingly recognized as components of modern, nature-based recovery strategies. Their combination (Hybrid) was also tested on equine model of skeletal muscle satellite cells (ESCs) exposed to heat shock (40 °C, 1 h), mimicking exercise stress. Cells were treated with CGA (0.005%), IVAS (0.0005%), or both for 24 h post-stress. Cell viability (MTS), mitochondrial membrane potential, apoptosis (Annexin V/7-AAD), nitric oxide (NO) production, and gene expression (RT-qPCR) were assessed. CGA significantly improved viability under both normothermia and heat stress (216-227%, p < 0.05), while IVAS was effective only without stress. Only the Hybrid group maintained elongated morphology post-heat shock. CGA increased NO levels (p < 0.05), with no effect from IVAS or Hybrid. Antioxidant gene expression remained unchanged, but proinflammatory cytokines IL-6 and IL-1β were upregulated in the Hybrid group (2.74- and 5.64-fold, p < 0.01), suggesting a controlled, adaptive immune response. Early apoptosis rose in CGA and Hybrid groups (~ 34%, p < 0.05), but total cell death was lowest in the Hybrid group (6.26%). BCL2 was downregulated (p < 0.05), while BAX increased only in the Hybrid group (8.14-fold, p < 0.01). Mitochondrial genes MFN2, TFAM, and PUSL1 were significantly upregulated in the Hybrid group; MIRO1 expression increased in all treated groups. CGA and IVAS synergistically promote mitochondrial stability and ESC survival via mitochondrial activation and inflammation regulation-supporting the growing trend of using natural compounds in muscle recovery strategies.
© 2025. The Author(s).
Publication Date: 2025-09-03 PubMed ID: 40900286PubMed Central: PMC12504121DOI: 10.1007/s12015-025-10959-9Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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Overview
- This study investigates how two natural compounds, chlorogenic acid (CGA) and isovanillic acid 3-O-sulfate (IVAS), individually and combined, affect muscle satellite cells from horses under heat stress, simulating exercise-induced damage.
- The research focuses on cell recovery, mitochondrial function, apoptosis, and inflammatory responses to understand their potential for enhancing muscle regeneration and recovery in athletes.
Background and Purpose
- Skeletal muscle satellite cells (SCs) are crucial for muscle repair and regeneration, making them an important model for exercise-related stress studies.
- Exercise induces cellular stress, including heat shock, which challenges muscle cell viability and function.
- Natural compounds such as CGA and IVAS have gained attention for their possible roles in natural recovery and regeneration strategies.
- The purpose was to test these compounds’ effects on equine skeletal muscle satellite cells (ESCs), a relevant model for human muscle recovery due to physiological similarities.
- The study specifically explored how CGA, IVAS, and their combination (Hybrid) influence cell viability, mitochondrial activity, apoptosis, nitric oxide production, and gene expression after heat-induced stress.
Methods
- ESCs were exposed to heat shock at 40 °C for 1 hour to simulate exercise stress conditions.
- Post-heat shock, cells were treated with CGA (0.005%), IVAS (0.0005%), or their combination for 24 hours.
- Various assays and measurements were conducted:
- Cell viability via MTS assay
- Mitochondrial membrane potential assessment
- Apoptosis detection through Annexin V/7-AAD staining
- Nitric oxide (NO) production measurement
- Gene expression profiling using RT-qPCR for antioxidant, inflammatory, apoptotic, and mitochondrial genes
Key Findings
- Cell Viability:
- CGA significantly enhanced cell viability under both normal and heat stress conditions (increase of ~216-227%).
- IVAS improved viability only in cells not exposed to heat stress.
- The combination (Hybrid) maintained cell viability beyond individual treatments.
- Cell Morphology:
- Only the Hybrid treatment preserved elongated morphology of ESCs after heat shock, indicating better structural preservation.
- Nitric Oxide Production:
- CGA increased NO levels, which are involved in cellular signaling and muscle repair.
- No significant NO change was seen with IVAS or Hybrid treatments.
- Gene Expression:
- Antioxidant gene expression did not significantly change, suggesting these compounds do not primarily act through antioxidant pathways.
- Proinflammatory cytokines IL-6 and IL-1β were significantly upregulated in the Hybrid group (2.74- and 5.64-fold increases), suggesting an adaptive immune response rather than harmful inflammation.
- Apoptotic genes showed differential regulation:
- BCL2 (anti-apoptotic) was downregulated.
- BAX (pro-apoptotic) was strongly upregulated only in the Hybrid group (8.14-fold).
- Mitochondrial-related genes MFN2, TFAM, and PUSL1 were significantly increased in the Hybrid group, indicating enhanced mitochondrial function and biogenesis.
- MIRO1, involved in mitochondrial transport, was upregulated in all treated groups.
- Apoptosis and Cell Death:
- Early apoptosis increased in CGA and Hybrid groups (~34%), implying activation of controlled cell turnover processes.
- The Hybrid group showed the lowest total cell death (6.26%), reflecting a protective effect possibly through mitochondrial and inflammatory regulation.
Interpretation and Implications
- The study suggests that CGA and IVAS, especially when combined, synergistically enhance the recovery and survival of muscle satellite cells after stress.
- The combination supports mitochondrial stability and function, crucial for energy production and cell survival during recovery.
- The controlled upregulation of proinflammatory cytokines may represent a beneficial, adaptive immune response that facilitates regeneration rather than damaging inflammation.
- Activation of apoptotic pathways alongside increased viability points to a fine-tuned balance between removing damaged cells and promoting muscle repair.
- These findings support the use of natural bioactive compounds in muscle recovery therapies and interventions, potentially applicable to human athletes based on the equine model.
Conclusion
- Natural compounds CGA and IVAS promote muscle cell recovery after exercise-like stress by enhancing mitochondrial function and regulating inflammatory and apoptotic responses.
- Their combined use (Hybrid) provides superior benefits, maintaining cell viability and morphology, and activating adaptive mechanisms involved in muscle regeneration.
- This research underlines the potential of nature-based compounds as effective elements in muscle recovery strategies for athletes.
Cite This Article
APA
Witkowska-Piłaszewicz O, Nowicka-Kazmierczak M, Pietrzak P, Marycz K.
(2025).
Mitohormesis and Regeneration: Natural Compounds Chlorogenic Acid (CGA) and Isovanillic Acid 3-O-sulfate (IVAS) Boost Muscle Cell Recovery in the Equine Athlete Model.
Stem Cell Rev Rep, 21(8), 2654-2666.
https://doi.org/10.1007/s12015-025-10959-9 Publication
Researcher Affiliations
- Department of Large Animals Diseases and Clinic, Institute of Veterinary Medicine, Warsaw University of Life Sciences, Nowoursynowska 166, Warsaw, 02- 787, Poland. olga_witkowska_pilaszewicz@sggw.edu.pl.
- International Institute of Translational Medicine (MIMT), ul. Jesionowa 11, Malin Wisznia Mała, 55-114, Poland.
- International Institute of Translational Medicine (MIMT), ul. Jesionowa 11, Malin Wisznia Mała, 55-114, Poland.
- International Institute of Translational Medicine (MIMT), ul. Jesionowa 11, Malin Wisznia Mała, 55-114, Poland. krzysztofmarycz@gmail.com.
- Veterinary Institute for Regenerative Cures, School of Veterinary Medicine, University of California-Davis, Davis, CA, USA. krzysztofmarycz@gmail.com.
MeSH Terms
- Animals
- Horses
- Chlorogenic Acid / pharmacology
- Apoptosis / drug effects
- Membrane Potential, Mitochondrial / drug effects
- Regeneration / drug effects
- Cell Survival / drug effects
- Mitochondria / drug effects
- Mitochondria / metabolism
- Muscle, Skeletal / drug effects
- Nitric Oxide / metabolism
Grant Funding
- INNOGLOBO/2/myo(miR)egen/10/2022 / Narodowe Centrum Badań i Rozwoju
Conflict of Interest Statement
Declarations. Ethics Approval: Muscles sampling procedures were conducted post mortem. Following the European Directive 2010/63/EU and Polish regulations on animal experimentation, ethical approval was not required and are therefore exempt from the directive. Consent to Participate: n/a. Consent for Publication: n/a. Competing Interests: The authors declare no competing interests. Conflict of interest: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
References
This article includes 32 references
- Suchomel TJ, Nimphius S, Stone MH. The importance of muscular strength in athletic performance.. (10), 1419–1449.
- Reis IL, Lopes B, Sousa P, Sousa AC, Caseiro AR, Mendonça CM, Santos JM, Atayde LM, Alvites RD, Maurício AC. Equine musculoskeletal pathologies: Clinical approaches and therapeutical perspectives-A review.. (5), Article 190.
- Kawai M, Aida H, Hiraga A, Miyata H. Muscle satellite cells are activated after exercise to exhaustion in thoroughbred horses.. , (4), 512–517.
- Ceusters J, Lejeune JP, Sandersen C. From skeletal muscle to stem cells: An innovative and minimally-invasive process for multiple species.. , 696.
- DeBoer ML, Martinson KM, Pampusch MS, Hansen AM, Wells SM, Ward C, Hathaway M. Cultured equine satellite cells as a model system to assess leucine stimulated protein synthesis in horse muscle.. , (1), 143–153.
- Gonzalez ML, Busse NI, Waits CM, Johnson SE. Satellite cells and their regulation in livestock.. , (5), skaa081.
- Witkowska-Piłaszewicz O, Malin K, Dąbrowska I, Grzędzicka J, Ostaszewski P, Carter C. Immunology of physical exercise: Is an appropriate animal model for human athletes??. (10), Article 5210.
- Martín-Rodríguez A, Belinchón-deMiguel P, Rubio-Zarapuz A. Advances in understanding the interplay between dietary practices, body composition, and sports performance in athletes.. (4), 571.
- Nguyen V, Taine EG, Meng D, Cui T, Tan W. Chlorogenic acid: A systematic review on the biological functions, mechanistic actions, and therapeutic potentials.. , , 924.
- Wang Y, Sun J, Xue L. Chlorogenic acid improves high-fat diet-induced skeletal muscle metabolic disorders by regulating mitochondrial function and lactate metabolism.. ..
- Ommati MM, Farshad O, Mousavi K. Chlorogenic acid supplementation improves skeletal muscle mitochondrial function in a rat model of resistance training.. , , 1221–1230.
- Dąbrowska I, Grzędzicka J, Niedzielska A, Witkowska-Piłaszewicz O. Impact of chlorogenic acid on peripheral blood mononuclear cell proliferation, oxidative stress, and inflammatory responses in racehorses during exercise.. (11), Article 1924.
- Przewłócka K, Folwarski M, Kaźmierczak-Siedlecka K. Gut-muscle axis exists and may affect skeletal muscle adaptation to training.. , 1451.
- Houghton MJ, Kerimi A, Mouly V, Tumova S, Williamson G. Gut microbiome catabolites as novel modulators of muscle cell glucose metabolism.. (2), 1887–1898.
- Mthembu SXH, Dludla PV, Ziqubu K. The potential role of polyphenols in modulating mitochondrial bioenergetics within the skeletal muscle: A systematic review of preclinical models.. , , 2791.
- Merry TL, Ristow M. Mitohormesis in exercise training. 123–130.
- Fennel ZJ, Ducharme JB, Berkemeier QN. Effect of heat stress on heat shock protein expression and hypertrophy-related signaling in the skeletal muscle of trained individuals. (6), R735–R749.
- Eberhardt EL, Ludlam AV, Tan Z, Cianfrocco MA. Miro: A molecular switch at the center of mitochondrial regulation. (6), 1269–1284.
- Endoni BT, Koval OM, Allamargot C. MIRO1 is required for dynamic increases in mitochondria-ER contact sites and mitochondrial ATP during the cell cycle. (7), 482.
- Votion DM, Gnaiger E, Lemieux H. Physical fitness and mitochondrial respiratory capacity in horse skeletal muscle. (4), e34890.
- Ahmad T, Mukherjee S, Pattnaik B. Miro1 regulates intercellular mitochondrial transport & enhances mesenchymal stem cell rescue efficacy. (9), 994–1010.
- Musci RV, Hamilton KL, Linden MA. Exercise-induced mitohormesis for the maintenance of skeletal muscle and healthspan extension. (7), Article 170.
- Powers SK, Bomkamp M, Ozdemir M, Hyatt H. Mechanisms of exercise-induced preconditioning in skeletal muscles. 101462.
- Kwak HB, Song W, Lawler JM. Exercise training attenuates age-induced elevation in Bax/Bcl-2 ratio, apoptosis, and remodeling in the rat heart. (6), 791–793.
- Kaczmarek A, Kaczmarek M, Ciałowicz M. The role of satellite cells in skeletal muscle regeneration-The effect of exercise and age. (10), Article 1056.
- Kistner TM, Pedersen BK, Lieberman DE. Interleukin 6 as an energy allocator in muscle tissue. 170–179.
- Langston PK, Mathis D. Immunological regulation of skeletal muscle adaptation to exercise. (6), 1175–1183.
- Zhang J, Gao B, Ye B, Sun Z, Qian Z, Yu L, Bi Y, Ma L, Ding Y, Du Y, Wang W, Mao Z. Mitochondrial-Targeted delivery of Polyphenol-Mediated antioxidases complexes against pyroptosis and inflammatory diseases. (11), e2208571.
- Rodriguez-Mateos A, Le Sayec M, Cheok A. Dietary (poly)phenols and cardiometabolic health: from antioxidants to modulators of the gut microbiota. Published online. 1–11.
- Li Y, Xu Y, Le Sayec M, Yan X, Spector TD, Steves CJ, Bell JT, Small KS, Menni C, Gibson R, Rodriguez-Mateos A. Development of a (poly)phenol metabolic signature for assessing (poly)phenol-rich dietary patterns. (23), 13439–13450.
- Leal Reis I, Lopes B, Sousa P, Sousa AC, Branquinho M, Caseiro AR, Pedrosa SS, Rêma A, Oliveira C, Porto B, Atayde L, Amorim I, Alvites R, Santos JM, Maurício AC. Allogenic synovia-derived mesenchymal stem cells for treatment of equine tendinopathies and desmopathies-Proof of concept. (8), Article 1312.
- Wall BT, Morton JP, van Loon LJC. Strategies to maintain skeletal muscle mass in the injured athlete: Nutritional considerations and exercise mimetics. (1), 53–62.
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