Nodakenin attenuates cerebral ischemia–reperfusion injury by modulating the PI3K/AKT/NF-κB signaling pathway

The research aimed to explore the protective effect of Nodakenin against cerebral ischemia–reperfusion (I/R) injury and its underlying mechanism, with a particular focus on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/nuclear factor kappa B (NF-κB) signaling axis, neuroinflammation, oxidative stress and ferroptosis. The function of Nodakenin was investigated in a rat middle cerebral artery occlusion and reperfusion (MCAO/R) model and a PC12 cell oxygen–glucose deprivation and reoxygenation (OGD/R) model. Infarct size was evaluated by 2,3,5-triphenyltetrazolium chloride (TTC) staining, brain edema was quantified using the wet–dry weight method, and histological changes were examined by hematoxylin and eosin (HE) staining. Biochemical assays and Western blotting was detected by malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase 4 (GPX4), ferritin heavy chain 1 (FTH1), tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β), and interleukin 6 (IL-6), along with PI3K/AKT–NF-κB pathway activity. Nodakenin treatment reduced infarct volume, histological damage and brain edema in cerebral I/R models. Nodakenin treatment suppressed proinflammatory cytokines, decreased MDA, restored SOD activity and attenuated ferroptosis-related alterations including Fe2+ accumulation and downregulation of GPX4 and FTH1. Consistent effects were observed in PC12 cells subjected to OGD/R. Mechanistically, Nodakenin activated the PI3K/AKT signaling pathway but suppressed NF-κB activation. These protective effects were abolished by PI3K inhibitor LY294002. Nodakenin exerted neuroprotective effects against cerebral I/R injury by regulating the PI3K/AKT/NF-κB pathway to reduce neuroinflammation, oxidative stress and ferroptosis. These findings identify Nodakenin as a promising candidate for limiting secondary injury after ischemic stroke. Nodakenin attenuated cerebral ischemia–reperfusion injury in vivo and in vitro. Nodakenin reduced neuroinflammation, oxidative stress and ferroptosis. Nodakenin modulated the PI3K/AKT–NF-κB signaling pathway. The protective role of Nodakenin in cerebral ischemia/reperfusion injury. Nodakenin regulated the PI3K/AKT/NF-κB signaling pathway by decreasing the phosphorylation levels of PI3K, AKT and NF-κB, thereby suppressing NF-κB nuclear translocation. In OGD/R-induced PC12 cells, Nodakenin promoted proliferation while reducing inflammation and ferroptosis. In MCAO rats, Nodakenin alleviated brain damage, neuroinflammation and ferroptosis.