Research Paper Volume 16, Issue 5 pp 4670—4683

Figure 5. Echinatin inhibits sevoflurane-induced ferroptosis via ALOX12. (A) HT22 cells were exposed to either sevoflurane or control conditions. Protein levels of p53, p21, and SLC7A11 were measured by western blot. Compared with the Ctrl group, **P<0.01. (B) HT22 cells were subjected to Echinatin treatment (40 μM) for a duration of 24 hours, followed by exposure to sevoflurane or control conditions. Protein level of ALOX12 was measured by western blot. (C) ALOX12 activity was measured by detecting 12-HETE levels by ELISA. Compared with the Sevo group, *P<0.05, **P<0.01. (D) HT22 cells infected with shRNA-control (si-NC) lentivirus or shRNA-ALOX12 (si-ALOX12) lentivirus, followed by exposure to sevoflurane or control conditions. Protein levels of ALOX12, FPN, TFR1, and DMT1 were measured by western blot. Compared with the Sevo+si-NC group, *P<0.05, **P<0.01. (E) HT22 cells were infected with lentiviruses carrying either the overexpression control (OE-NC) or ALOX12 overexpression (OE-ALOX12) constructs, and subsequently subjected to Echinatin treatment (40 μM) followed by exposure to sevoflurane. Cell apoptosis and ROS levels were measured using flow cytometry. Intracellular Fe²⁺ detected by FerroOrange. Compared with indicated group, **P<0.01, ***P<0.001. The data are presented as the mean ± SD. Ech, Echinatin; Sevo, sevoflurane.