Fig. 6

The cardiac myocytes protective effect of AA was inhibited by mitophagy and AA-COX-PGI pathway inhibitors. H9C2 cells were pretreated with PINK1 specific siRNA (120 nM), 3-MA (10 mM), Cay10441 (10 μM) or Indomethacin (5 μM) before AA supplement (25 µM) under the condition of HG + OGD. Mitophagy associated proteins LC3II/LC3I (A), p62 (B), PINK1 (C) and Parkin (D); mitochondrial renewal proteins Drp1 (E) and FIS1 (F); cytosolic cytochrome c (cyt c) and mitochondrial cyt c (G, H), as well as cleaved caspase-3 (I) in each group were measured by western blot. GAPDH and VDAC1 were used as internal protein loading controls respectively for cytosolic and mitochondrial protein. Optical densities of the protein bands were quantitatively analyzed with Sigma Scan Pro 5 and normalized with loading control GAPDH or VDAC1. J the ratio of mitochondrial DNA and genomic DNA (mtDNA/gDNA).To detect whether the effect of AA on mitochondria was associated with AA-COX-PGI2 pathway, PGI receptor inhibitor Cay 10,441(K) and COX inhibitor Indomethacin (L) was used. Results are expressed as the mean ± SD. *P < 0.05, **P < 0.01 and ***P < 0.001. Statistical analysis was carried out by a one-way ANOVA analysis