32 | Trifolium pratense extract mitigates skeletal muscle atrophy: a nutraceutical approach for the prevention of sarcopenia

Tommaso Raiteri¹, H. Meskine², M. Paiella¹, A. Trabalzini¹, I. Varfaj⁴, L. Salvadori², F. Riuzzi¹, R. Sardella⁴, F. Orlandi³, N. Filigheddu², G. Sorci¹ | ¹Department of Medicine and Surgery, University of Perugia, Italy; ²Department of Translational Medicine, University of Piemonte Orientale, Novara, Italy; ³Department of Civil and Environmental Engineering, University of Perugia, Italy; ⁴Department of Pharmaceutical Sciences, University of Perugia, Italy.

Abstract

Sarcopenia, the age-related loss of muscle mass and function, increases the risk of frailty, fractures, and loss of independence, ultimately reducing quality of life and contributing to higher mortality. With life expectancy rising in Western countries, sarcopenia represents a growing yet unresolved public health challenge, as no pharmacological treatments are currently available. Recent interest in medicinal plants as potential therapeutic agents has highlighted their ability to counteract various pathological conditions, including muscle atrophy. This study explores the protective effects of native Mediterranean on skeletal muscle cells exposed to atrophic stimuli that mimic sarcopenia and disuse (e.g., dexamethasone, starvation). Additionally, we assess whether controlled cultivation protocols can influence the bioactive metabolite content of these plants, potentially enhancing their anti-sarcopenic properties. Among the tested species, hydroalcoholic extract of Trifolium pratense demonstrates promising anti-atrophic effects, effectively preserving C2C12 myotube diameter against TNF- α/IFN-γ-induced muscle wasting. However, its protective efficacy varies depending on cultivation conditions, with some enhancing its bioactivity while others showing no significant effect. Furthermore, Trifolium pratense completely abolishes TNF-α/IFN-γ-induced reactive oxygen species production, underscoring its antioxidant properties. These findings suggest that Trifolium pratense could serve as a promising nutraceutical intervention to counteract muscle atrophy associated with aging. Moreover, optimizing its cultivation conditions may enhance its therapeutic potential, paving the way for targeted agronomic strategies to maximize its bioactivity. By integrating plant-based interventions into sarcopenia management, this study contributes to the development of accessible, nonpharmacological strategies aimed at improving muscle health and quality of life in aging populations.