Background: Sarcopenia is the age-induced, progressive loss of skeletal muscle mass and function. This phenomenon is observed in patients with chronic kidney disease (CKD). However, the intracellular mechanism underlying the progressive sarcopenia in CKD has not been completely elucidated. Although hyperphosphatemia contributes to cellular senescence, it is unclear whether this condition induces skeletal muscle atrophy. The aim of this study was to determine the effect of hyperphosphatemia on skeletal muscle. Methods: Differentiated rat myoblast cells (L6) were exposed to normal (0.9 mM; CON), medium (2.5 mM), and high (3.8mM; HPi) phosphate conditions for 10 days. We measured the protein levels of myosin heavy chain (MHC) and myostatin and determined the ratio of phosphorylated p70S6K and cleaved caspase-3 by western blot. The expression levels of the myogenic transcriptional regulators MyoD and myogenin were measured by qPCR. Results: The levels of MHC were gradually downregulated depending on the phosphate concentration. Myostatin in HPi was about 20 times higher than in CON (P<0.001). In myotubes cultured in HPi, protein synthesis was significantly lower, and degradation was significantly higher than in CON (P<0.01). The mRNA expression of MyoD in HPi was significantly lower than in CON. Conclusions: This study showed that hyperphosphatemia strongly induced muscle atrophy with the accumulation of myostatin. This mechanism might include the downregulation of protein synthesis, upregulation of proteolysis, and attenuated muscle regeneration.