Areca nut extract exposure disrupts myogenesis and metabolism in C2C12 cells

Areca nut (AN), which is commonly consumed in Southeast Asia, contains bioactive compounds that may influence cellular functions. Accumulating evidence has revealed several health impacts of AN consumption, but the toxicological effects of areca nut extract (ANE) on muscle cells remain largely unexplored. Myogenesis, a critical process for muscle development and regeneration, is closely tied to metabolic activity, which governs the differentiation and function of myocytes. This study aimed to evaluate the effects of ANE on myogenesis in murine C2C12 myoblasts and differentiated myotubes. In ANE-treated C2C12 myoblasts, we observed a significant decrease in the intracellular glutamine level that was accompanied by decreased GSH levels, decreased mTOR signaling, and increased autophagy in myoblasts but not in differentiated myotubes. ANE treatment decreased glutamine and 6-phosphogluconate levels in both myoblasts and myotubes, suggesting the widespread suppression of amino acid and redox-related metabolic pathways. Moreover, ANE significantly altered the metabolomic profile, upregulating the levels of glycolysis and TCA cycle intermediates but reducing ATP levels, indicating impaired energy metabolism in differentiated myotubes. ANE also downregulated the expression of key metabolic genes, including those involved in glycolysis (AKR1B3 and LDHA), glycerol metabolism (GPD1 and GPD2), and nitrogen metabolism (GLUD1, ARG1, GS, and GLS1), indicating that ANE disrupts critical pathways involved in muscle cell metabolism and myogenesis. This study provides new insights into the mechanisms by which AN consumption affects muscle development, emphasizing the need for further research into the dietary and environmental factors influencing myogenesis.