Effect of simvastatin lactone (SVL) and simvastatin hydroxy acid (SVA) on lactate dehydrogenase (LDH) release from undifferentiated and differentiated C2C12 cells maintained under different pH levels. C2C12 cells were cultured at a density of 4,000 cells/well and allowed to attach for 24 hours or to differentiate for 4 days, then exposed to increasing concentrations of SVL or SVA under acidic, neutral, and alkaline medium pH. Undifferentiated cells were treated for 72 hours, whereas differentiated cells were maintained for 24 hours. Data are presented as mean ± standard deviation, (n = 3) and analyzed by 2-way analysis of variance followed by Bonferroni's post hoc test. (A) LDH release from undifferentiated C2C12 cells treated with SVL. a = pH 6.8 vs pH 7.4 (P < 0.001); b = pH 6.8 vs pH 7.4, 7.8 (P < 0.001); c = pH 6.8 vs pH 7.4, 7.8; pH 7.4 vs pH 7.8 (P < 0.001). (B) LDH release from differentiated C2C12 cells treated with SVL. a = pH 6.8 vs pH 7.4, 7.8; pH 7.4 vs pH 7.8 (P < 0.001). (C) LDH release from undifferentiated C2C12 cells treated with SVA. a = pH 6.8 vs pH 7.8 (P < 0.05); b = pH 6.8 vs pH 7.4, 7.8; pH 7.4 vs pH 7.8 (P < 0.001); c = pH 6.8 vs pH 7.4 (P < 0.01); pH 7.8 vs pH 6.8, 7.4 (P < 0.001). (D) LDH release from differentiated C2C12 cells treated with SVA. a = pH 6.8 vs pH 7.4 (P < 0.01); pH 6.8 vs pH 7.8 (P < 0.05); b = pH 6.8 vs pH 7.4, 7.8 (P < 0.0001).