Figure 4.

Sirt1 mediates hydrogen sulfide-induced cytoprotection effect in neonatal mouse cardiomyocytes. Neonatal mouse cardiomyocytes were pretreated in free-serum medium with or without 100 μmol/L NaHS and 10 μmol/L Ex 527 for 12 h as indicated group. Consequently, cells were washed twice with PBS and were recovered for 24h by changing into FBS-MEM, followed by exposure to free-serum medium with 600 μmol/L H₂O₂ for another 4 h. (a) Sod2 protein expression level is determined using Western blot analysis. (b) LDH release was detected. (c) ROS production was tested by flow cytometry. (d) Mitochondrial DNA number was detected by quantitative polymerase chain reaction. The data shown are mean ± SE of three independent experiments (*P < 0.05 vs. Con, ^†P < 0.05 vs. NaSH). FBS: Fetal bovine serum; MEM: Modified Eagle's medium; LDH: Lactate dehydrogenase, ROS: Reactive oxygen species; SE: Standard error; Con: Control; NaHS: Sodium hydrosulfide; Sod2: Superoxide dismutase; Sirt1: Sirtuin 1.