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<p>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&#xa0;°C, 1&#xa0;h), mimicking exercise stress. Cells were treated with CGA (0.005%), IVAS (0.0005%), or both for 24&#xa0;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%, <i>p</i> &lt; 0.05), while IVAS was effective only without stress. Only the Hybrid group maintained elongated morphology post-heat shock. CGA increased NO levels (<i>p</i> &lt; 0.05), with no effect from IVAS or Hybrid. Antioxidant gene expression remained unchanged, but proinflammatory cytokines <i>IL-6</i> and <i>IL-1β</i> were upregulated in the Hybrid group (2.74- and 5.64-fold, <i>p</i> &lt; 0.01), suggesting a controlled, adaptive immune response. Early apoptosis rose in CGA and Hybrid groups (~ 34%, <i>p</i> &lt; 0.05), but total cell death was lowest in the Hybrid group (6.26%). <i>BCL2</i> was downregulated (<i>p</i> &lt; 0.05), while <i>BAX</i> increased only in the Hybrid group (8.14-fold, <i>p</i> &lt; 0.01). Mitochondrial genes <i>MFN2</i>,<i> TFAM</i>, and <i>PUSL1</i> were significantly upregulated in the Hybrid group; <i>MIRO1</i> 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.</p>

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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

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年9月3日

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  • Olga Witkowska-Piłaszewicz,
  • Małgorzata Nowicka-Kazmierczak,
  • Patrycja Pietrzak,
  • Krzysztof Marycz

摘要

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.