Fig. 7
Inhibition of the CAMKK2/AMPK pathway reduces the protective effect of MH on apoptosis and oxidative stress in SH-SY5Y cells after OGDR injury. (A) Schematic diagram of the cell experiment; (B) Western blot analysis shows the expression of S100A8, CAMKK2, p-AMPK, AMPK, bax and caspase3 protein expression, with β-actin serving as the loading control; (C–G) Quantification of the Western blots from (B). Normalized S100A8, CAMKK2, p-AMPK/AMPK, bax and cleaved-caspase3/pro-caspase3 to corresponding loading control are summarized across three independent trials (n = 3, per group). (H) Representative images of TUNEL, DHE and TMRE staining (Scale bar: 50 μm) of SH-SY5Y cells; (I–K) Quantitative analysis of apoptosis ratio, DHE and TMRE fluorescence intensity (n = 5, per group). Control: Control group; OGDR: oxygen-glucose deprivation 2 h and reoxygenation 24 h; OGDR + MH: OGDR injury followed by 4 h of MH (33 ± 0.5 °C); OGDR + MH + STO-609: SH-SY5Y were treated with STO-609 for inhibiting the activation of CAMKK2 30 min before OGDR injury followed by 4 h of MH (33 ± 0.5 °C); OGDR + MH + Dor: SH-SY5Y were treated with Dor for inhibiting the activation of AMPK 30 min before OGDR 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.
