(P26-067-25) Hesperetin Improves Fatigue and Endurance Performance in Mice by Increasing Muscle Synthesis via the AKT/mTOR Signaling Pathway

Objectives: The present work investigated the molecular mechanism and ergogenic effect of the citrus flavonoid Hesperetin (HES).

Methods: The present work adopted a multifaceted approach involving network pharmacology, which is a systematic approach designed for establishing relationships among drugs, targets, and diseases, thereby improving the comprehension of drug pharmacology and its effects on biological networks, whereas molecular docking, a technique rooted in the properties of receptors and the interactions between drug molecules and receptors, has become pivotal in computer-aided drug discovery. Its ability to accurately predict the binding mechanisms and affinities of molecular interactions positions it as a fundamental method in contemporary drug development research. Furthermore, in our in vitro analysis, C2C12 myotubes were treated with 0, 10, and 20 µM of Hesperetin, respectively. In vivo trials involved the administration of 25 and 50 mg/kg of HES to mice for 12 weeks.

Results: Our network pharmacology analysis revealed 23 common targets between HES and physical fatigue, whereas the top 10 hub targets in the network were AKT1, ESR1, EGFR, PTPN11, BRAF, HRAS, JAK2, CDC42, FGFR1, and PPARG. Molecular docking analysis showed that AKT1 (3o96) was the most potential target for HES with an affinity score of −9.4 kcal/mol. In addition, immunofluorescence staining analyses revealed that HES enhanced both the number and size of myofibers by elevating the expression of type I and type II myofibers through the activation of the AKT/mTOR signaling pathway in C2C12 myotubes (MyHCI (p < 0.05), MyHCIIa (p < 0.01), and MyHCIIb 10 μM (p < 0.05) 20 μM (p < 0.01) and mouse gastrocnemius muscle (MyHCI HES50 group (p < 0.01), and MyHCIIb HES25 (p < 0.05) HES50 (p < 0.01). Besides, HES increased the strength and endurance of mice by increasing grip strength (p < 0.05), running time (p < 0.01), and exhaustion time.

Conclusions: HES treatment improved strength and endurance, thereby alleviating fatigue. Due to its role in skeletal muscle synthesis, HES could be a potential candidate for addressing sarcopenia.

Funding Sources: This study was funded by the National Natural Science Foundation of China and the Natural Science Foundation of Henan.