Figure 4
Apelin treatment reduces hypoxia-induced ROS production and apoptosis in cardiomyocytes through the FoxO3 pathway.
Primary cardiomyocytes isolated from HFD-fed mice were treated with apelin for 15 minutes and submitted to hypoxia for 2 h. (A–F) Mitochondrial O2− (A,B) and H2O2 (C,D) production were followed using the fluorescent probes MitoSOX Red and MitoPY1 respectively. Apoptosis was measured by TUNEL labeling (E,F). Arrowheads highlight TUNEL-positive cells. Bar is 20μm in all panels. (G,H) H9C2 cardiomyoblasts were transfected with siRNA targeting FoxO3 or with scramble control siRNA, treated or not (C) with apelin (A, 10−7M for 15 minutes) and subjected to hypoxia (H) or normoxia (N) for 2 hours. Mitochondrial ROS production was measured by MitoSOX Red fluorescence (in red). (I,J) H9C2 cells were treated as in (G) and apoptotic cells were labeled with TUNEL staining. Bar is 20μm. *p < 0.05 vs N; ***p<0.001 vs N; #p < 0.001 vs H; §p < 0.05 between indicated conditions.
