Hydrogen Gas Attenuates Hypoxic-Ischemic Brain Injury via Regulation of the MAPK/HO-1/PGC-1a Pathway in Neonatal Rats

Figure 6

H₂ inhibited apoptosis in HIBI rat brain neurons and NGF-differentiated PC12 cells. (a) Apoptotic cells were detected by TUNEL staining (dark brown, indicated by arrows) in the brains of neonatal rats 3 days after HIBI. (b) Images of NGF-differentiated PC12 cells were taken under an inverted microscope immediately after OGD/R. Then, apoptosis analysis was done by Annexin-V/PI double staining. (c) Data of viable (Annexin V-negative/PI-negative), early apoptotic (Annexin V-positive/PI-negative), and late apoptotic (Annexin V-positive/PI-positive) cells from three independent experiments (; ). (d) Representative immunofluorescent signals (green) of Bcl-2, BAX, and caspase-3 with DAPI nuclear counterstain (blue) (scale bar: 20 μm). (e) Western blot analysis of β-actin, Bcl-2, BAX, and caspase-3 proteins in each group. (f, g) Bar graph showing the relative expression of Bcl-2, BAX, and caspase-3 in each group (; ).

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