Research Article
Hydrogen Gas Attenuates Hypoxic-Ischemic Brain Injury via Regulation of the MAPK/HO-1/PGC-1a Pathway in Neonatal Rats
Figure 9
Knockdown of HO-1 expression in the HO-differentiated PC12 cells was followed by OGD and reperfusion with or without H2 gas. β-NGF-differentiated PC12 cells were pretreated with HO-1 siRNA for 24 h followed by OGD (6 h) and then reperfusion with or without H2 gas. (a) Percentage of apoptotic cells (provided for each condition) was determined by combined Annexin V/PI staining and FACS analysis. (b) Data of early apoptotic (Annexin V-positive/PI-negative), late apoptotic (Annexin V-positive/PI- positive), and dead (Annexin V-negative/PI-negative) cells are summarized for three independent experiments. (c) Western blot analysis of β-actin, HO-1, PGC-1α, and SIRT1 proteins in each group. (d–f) The protein expression of HO-1, PGC-1α, and SIRT1 is shown as the ratio of the protein to β-actin for each sample. (g–i) The mRNA expression of each gene was normalized with 18S to determine the relative expression ratio (log2 fold) for each group (; ; ).
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