Cardioprotective Effects of AMPK Activation

H9C2 cells (Chinese Academy of Sciences Cell Bank, Shanghai, China) were cultured in high glucose DMEM (GIBCO, USA) supplemented with 1% cyan streptomycin double-antibody and 10% (v/v) FB Sat 37°C under a 5% CO₂ atmosphere. For the present study, H9C2 cells were incubated overnight to reach 70-80% confluence at 37°C before experimentation. H9C2 cells were treated with H₂O₂ at a concentration of 100 μM for 4 hours to construct an apoptosis model [26 (link)]. AMPK was activated after exercise [27 (link)], so AICAR, an AMPK agonist, was used to simulate the exercise effect of H9C2 cells. Furthermore, H9C2 cells were treated with recombinant human FGF21 (rhFGF21, 75 ng/ml, 15 h; Selleck, USA), FGFR1 inhibitor (PD166866, 100 ng/ml, 15 h, Selleck Chemicals) [8 (link)], PI3K inhibitor (LY294002, 10 μM, 1 h, Selleck Chemicals), AICAR (1 mM, 1 h, Selleck Chemicals) [28 (link)], and lentiviral vectors carrying alcat1 gene (ALCAT1 OE, MOI = 1; Brain VTA, China).