Dexamethasone and Trimetazidine Modulate NLRP3 Activation in Murine C2C12 Myotubes

Murine C2C12 myoblasts were obtained from ATCC and incubated at 37 °C, 5% CO₂ in DMEM with 80U/ml penicillin and 0.08 mg/ml streptomycin and 10% fetal bovine serum (Gibco). For the induction of differentiation into myotubes, sub-confluent myoblasts were switched to DMEM containing 2% horse serum (Biological Industries, Israel), and then cultured for 4 days. The myotubes were treated with 10 μM dexamethasone for 24 h [46 (link)], and 150 μM trimetazidine was added in the last 6 h [19 (link)]. To activate NLRP3, after treated with 10 μM DEX and/or 150 μM TMZ, C2C12 myotubes were cultured in a medium containing 100 ng/mL LPS (Sigma-Aldrich, US) for 2 h, followed by adding 2.5 mM ATP (Solarbio, China) and cultured for another 1 h [24 (link)]. For inhibiting phosphoinositide 3-kinase (PI3K)/AKT pathway, C2C12 myotubes were cultured in a medium containing 2.5 μM picropodophyllin (PPP) (MCE, China) for 24 h [24 (link)].