Ferroptosis Inhibition and ACSL4 Modulation in Ischemia

Liproxstatin-1 (S7699, Selleck, TX, USA), a ferroptosis inhibitor, was administered i.p. at a concentration of 10 mg/kg 1 h before ischemia induction, in accordance with previous study protocols [9 (link)]. Mice were killed at 30 min of reperfusion and serum was collected from the abdominal aorta. In addition, Liproxstatin-1 dissolved to a final concentration of 200 nM was used to treat Caco-2 cells in vitro for 12 h before hypoxia induction. Rosiglitazone (ROSI, S2556, Selleck), a classic peroxisome proliferator-activated receptor-γ agonist that has been used for ACSL4 inhibition, was administered intravenously at a concentration of 0.4 mg/kg 1 h before ischemia induction, as pretreatment of ROSI allows sufficient time for proper phospholipid remodeling in the membranes [21 (link), 22 (link)]. Mice were killed at 45 min of ischemia or at 30 min of reperfusion. Serum was collected from the abdominal aorta. Kits were used to assay the levels of tumor necrosis factor (TNF)-α (ab208348, Abcam, MA, USA), interleukin (IL)-6 (ab100712, Abcam), and ACSL4 activity (ab241005, Abcam).