Figure 3
Protective effects of HCH following I/R in experiment 1. Mice were sacrificed after 4 h of reperfusion and the hearts were harvested for biochemical examination (n = 224). The expression of the following proteins was detected using western blotting: p-Akt, Akt, p-JNK, JNK, p-ERK, ERK, p-p38 MAPK, p38 MAPK, p-GSK-3β (Ser 9), GSK-3β, p-IκB, IκB, p-BAD, BAD, FOXO1, FOXO3, Fas-L, Fas, Bax, Bcl-2, cytoplasmic Cyto-c, mitochondrial Cyto-c, e-NOS, caspase-3, and NF-κBp65 (A,C). HCH significantly reduced the protein expression of p-JNK, p-IκB, FOXO1, FOXO3, Fas, Fas-L, Bax, cytoplasmic Cyto-c, caspase-3, and NF-κB, but markedly increased the protein expression of p-Akt, p-ERK, p-p38 MAPK, p-GSK-3β, p-BAD, Bcl-2, e-NOS, and mitochondrial cyt-c (B), Supplementary Fig. 4). HCH significantly reduced caspase-3 mRNA expression (G), MDA content (H), and caspase-3 activity (I), and markedly increased SOD activity (H). HCH significantly decreased the contents of TNF-α (L), IL-1β (J), and MPO (K) in the heart. PI3K inhibitor treatment markedly abolished the protective effects of HCH. The time course of Akt phosphorylation during ischemia and after reperfusion (D). Akt phosphorylation occurred earlier in HCH-treated animals (45 min after reperfusion) than in untreated I/R mice (90 min) (E). *P < 0.05 vs. sham; # P < 0.05 vs. I/R; § P < 0.05 vs. I/R + HCH.
