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Transcriptomic and Proteomic Analysis of the Skeletal Muscle Revealed the Effects and Mechanism of Mulberry Leaf Flavonoids on Alleviating Exercise-Induced Muscle Damage in Mongolian Horses
by
Aopan Geng
1,†,
Xuejiao Wang
2,†,
Lianhao Li
1,
Sarah Cowie
3,
Dongyi Bai
1,
Manglai Dugarjaviin
1 and
Xinzhuang Zhang
1,* 1
Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, College of Animal Science and Technology, Inner Mongolia Agricultural University, Hohhot 010018, China
2
Rural Revitalization Research Institute, Inner Mongolia Agricultural University, Hohhot 010018, China
3
School of Psychology, The University of Auckland City Campus, Private Bag 92019, Auckland 1142, New Zealand
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Animals 2026, 16(10), 1548; https://doi.org/10.3390/ani16101548 (registering DOI)
Submission received: 8 April 2026
/
Revised: 8 May 2026
/
Accepted: 13 May 2026
/
Published: 18 May 2026
(This article belongs to the Section Animal Nutrition)
Simple Summary
Prolonged high-intensity training in racehorses induces severe oxidative stress and muscle damage. Dietary supplementation with mulberry leaf extract effectively ameliorates these conditions, as evidenced by reduced blood biomarkers for inflammation and muscle injury. Furthermore, transcriptomic and proteomic analyses of muscle tissue demonstrated significant enrichment in specific cellular signaling pathways, suggesting that the extract potentially modulates protein metabolism and facilitating the restoration of damaged muscle fibers. These findings clarify the molecular mechanisms behind the antioxidant properties of mulberry extract and provide a clearer understanding of its potential as a targeted nutritional intervention for exercise-induced muscle damage.
Abstract
The scientific evidence regarding the use of plant-derived extracts to alleviate exercise-induced muscle damage in horses remains limited. Mulberry leaf flavonoids (MLFs) are the primary bioactive constituents of a traditional medicinal plant and are potent antioxidants. The aim of this study was to investigate the protective effects of MLFs against exercise-induced muscle damage. In this study, twelve Mongolian horses were used in a 3 × 3 Latin square design to investigate the protective effects of MLFs. Our results showed that high-intensity exercise negatively impacted the immune status, metabolic state, myofibrillar structure, and antioxidant capacity of the horses. Conversely, MLFs significantly reduced blood levels of white blood cells (WBC), monocytes (MON), aspartate aminotransferase (AST), creatine kinase (CK), and malondialdehyde (MDA) across various exercise distances and during recovery. Simultaneously, MLFs increased serum glutathione peroxidase (GPx), superoxide dismutase (SOD), and total antioxidant capacity (T-AOC). Mechanistically, transcriptomic analysis revealed that dietary MLFs upregulated genes associated with myofibrillar structural proteins (MYOZ2, MYOM3), the antioxidant defense system (GPX3, SOD3), and skeletal muscle satellite cell proliferation and differentiation (MYOD1, MRF6). Furthermore, quantitative proteomics indicated the enrichment of the PI3K-Akt and TGF-β signaling pathways, as well as ECM–receptor interactions, suggesting their potential involvement in regulating protein metabolism and facilitating myofibrillar restoration. Overall, MLFs effectively alleviated inflammation, metabolic disorder, and exercise-induced muscle damage. Under the tested conditions, a daily dosage of 10 g MLFs provided superior protective effects.
