Aicar

We obtained 24 mongrel dogs (age 8–12 months, 11 females and 13 males, weight 11.05±0.76 kg) from the Experimental Animal Surgery Department of Zunyi Medical University (Zunyi, China). All experimental procedures in this study were approved by the Animal Ethics Committee of Zunyi Medical University (Permission No.2015[2-015]) and were in compliance with the China’s State Ethics Code for Animal Care for Scientific Experimentation. The dogs were randomly assigned to 4 groups: the control group (n=6) did not undergo aortic cross-clamping; the model group (n=6) underwent 60 min of aortic cross-clamping with 150 ml cardioplegic solution; the treatment group (n=6) underwent 60 min of aortic cross-clamping with 0.11 mg/kg 5-aminoimidazole-4-carboxamide riboside [10 (link)] (AICAR, an AMPK agonist, Beyotime, China) in 150 ml cardioplegic solution; and the inhibition group (n=6) underwent 60 min of aortic cross-clamping with 0.11 mg/kg Compound C [10 (link)] (a specific inhibitor of AMPK, Beyotime, China) in 150 ml cardioplegic solution. All research was conducted in compliance with the rules of animal experimental ethics.

Cells were cultured at 37°C in RPMI-1640, or DMEM supplemented with 10% FBS. Experiments were performed on 70–80% confluent cells. The cells were treated with acadesine (AICAR; Beyotime Institute of Biotechnology) or ND. ND was established by incubating cells in Hank's balanced salt solution (HBSS)-4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) (0.185 g/l CaCl₂•2H₂O, 0.2 g/l MgSO₄•7H₂O, 0.4 g/l KCl, 0.06 g/l KH₂PO₄, 0.35 g/l NaHCO₃, 8 g/l NaCl, 0.09 g/l Na₂HPO₄•7H₂O, 20 mM HEPES, pH 7.4) (11 (link)) containing neither glucose nor amino acids.