Cottonseed Oil Attenuates Traumatic Brain Injury by Inhibiting AKT/GSK-3β-mediated Neuroinflammatory Responses

Traumatic brain injury (TBI) is a major cause of long-term neurological disability worldwide, and effective therapeutic strategies remain limited. Cottonseed oil (CSO), a plant-derived oil rich in polyunsaturated fatty acids, has been reported to exert protective effects in ischemia-related injuries; however, its role and underlying mechanisms in TBI remain unclear. Here, we established a controlled cortical impact (CCI) model in mice and administered CSO by oral gavage prior to injury. Neurological function was assessed using the modified neurological severity score, rotarod, and balance beam tests, and neuronal injury was evaluated by histological and biochemical analyses. CSO administration significantly improved neurological and motor function, attenuated neuronal damage and apoptosis, enhanced cerebral blood perfusion, preserved blood-brain barrier (BBB) integrity, and reduced brain edema. CSO modulated the expression of AQP4, VEGFA, and BDNF, and markedly suppressed glial activation and pro-inflammatory cytokine production. In vitro, CSO alleviated H₂O₂-induced cellular injury in HT22 cells by reducing reactive oxygen species accumulation and restoring antioxidant defenses. Mechanistically, the neuroprotective effects of CSO were associated with activation of the AKT/GSK-3β signaling pathway, characterized by enhanced phosphorylation of AKT (Ser473) and inhibitory phosphorylation of GSK-3β (Ser9). Pharmacological inhibition of AKT with MK2206 abolished CSO-induced AKT/GSK-3β pathway activation, reduced cell viability, and restored the elevation of IL-1β and IL-6 levels in vitro, confirming that the anti-neuroinflammatory effects of CSO are mediated at least in part through the AKT/GSK-3β signaling axis. Collectively, these findings demonstrate that CSO exerts significant neuroprotective effects in experimental TBI by preserving neurovascular integrity, attenuating neuroinflammation, and activating the AKT/GSK-3β pathway, highlighting CSO as a promising preventive strategy for TBI.