Fig. 3
In vivo, MH ameliorates IR-induced neuronal injury by decreasing S100A8 expression. (A) Schematic diagram of the animal experiment. (B) Western blot showing the expression of S100A8, bax, and caspase3 protein, with β-actin serving as the loading control. (C–E) Quantification of the Western blots from (B). Levels of S100A8, bax and cleaved-caspase3/pro-caspase3 normalized to the corresponding loading control across three independent trials (n = 3, per group). (F) Representative images of HE, TUNEL, DHE staining and TEM in the CA1 region of the hippocampus (Scale bars: HE-50 μm, TUNEL-50 μm, DHE-50 μm, TEM-500 nm). (G–I) Quantitative analysis of cell apoptosis ratio, DHE fluorescence intensity, and mitochondrial injury score (n = 5, per group). Sham + AAV9.NC: bilateral lateral ventricle injected with AAV9.NC 28 days before sham-operation; IR + AAV9.NC: bilateral lateral ventricle injected with AAV9.NC 28 days before cerebral IR injury; IR + AAV9.ShS100A8: bilateral lateral ventricle injected with AAV9.ShS100A8 28 days before cerebral IR injury; IR + MH + AAV9.NC: bilateral lateral ventricle injected with AAV9.NC 28 days before cerebral IR injury followed by 4 h of MH (33 ± 0.5 °C). IR + MH + AAV9.S100A8: bilateral lateral ventricle injected with AAV9.S100A8 28 days before cerebral IR injury followed by 4 h of MH (33 ± 0.5 °C). Results are presented as the mean \(\pm\) SD. *p < 0.05, **p < 0.01.
