Abstract: Background　Acute kidney injury (AKI) is a common and critical clinical syndrome with high morbidity and mortality. Currently, clinical management of AKI mainly relies on supportive therapy, and effective targeted pharmacological interventions remain limited. Objective　To investigate the protective effects of paradol in folic acid-induced AKI (FA-AKI) and ischemia/reperfusion-induced AKI (I/R-AKI) mouse models and to explore the underlying mechanisms.Methods　In vivo, the FAAKI model was established by intraperitoneal injection of folic acid (250 mg/kg), while the I/R-AKI model was induced by clamping the unilateral renal pedicle for 30 minutes followed by reperfusion. C57BL/6 mice were randomly divided into four groups: control group, AKI model group, paradol-treated group (AKI + paradol), and paradol-alone group. Renal function was evaluated by measuring serum creatinine (SCr) and blood urea nitrogen (BUN). Histopathological changes were assessed using hematoxylin-eosin (H&E) staining and tubular injury scoring. The mRNA expression levels of kidney injury molecule-1 (KIM-1), interleukin-6 (IL-6), and tumor necrosis factor- α (TNF- α) were determined by RT-qPCR. Renal macrophage infiltration was evaluated by F4/80 immunohistochemical staining. Oxidative stress was assessed by measuring glutathione (GSH) content, total superoxide dismutase (T-SOD) activity, malondialdehyde (MDA) levels, and the mRNA expression of glutathione peroxidase 4 (GPX4) in renal tissues. In vitro, an injury model was established in human kidney-2 (HK-2) cells using RSL3. Potential targets of paradol were predicted using the Swiss Target Prediction database, followed by molecular docking and molecular dynamics simulations to evaluate binding characteristics. An ALOX5-overexpressing HK-2 cell model was constructed to further assess the role of arachidonate 5-lipoxygenase (ALOX5). Cell viability and lactate dehydrogenase (LDH) activity were measured to evaluate cellular injury and the effects of paradol. Results　In vivo, compared with the control group, both FA-AKI and I/R-AKI mice exhibited significantly increased SCr and BUN levels (P＜0.001), markedly upregulated KIM-1 mRNA expression (P＜0.001), and pronounced tubular injury. Paradol treatment significantly reduced SCr and BUN levels (P＜0.05), alleviated histopathological damage, and downregulated KIM-1 mRNA expression (P＜0.05). In addition, AKI mice showed significantly increased renal expression of inflammatory cytokines IL-6 and TNF-α (P＜0.001), enhanced F4/80-positive macrophage infiltration, decreased GSH content (P＜0.001), reduced T-SOD activity (P＜0.001), elevated MDA levels (P＜0.001), and downregulated mRNA expression of GPX4 (P＜0.001). These alterations were markedly ameliorated by paradol treatment (P＜0.01). No significant differences were observed between the paradol-alone group and the control group (P＞0.05). In vitro, molecular docking and molecular dynamics analyses indicated stable binding between paradol and ALOX5. RSL3 significantly decreased HK-2 cell viability (P＜0.001) and increased LDH activity (P＜0.001), whereas paradol effectively reversed these changes (P＜0.001). ALOX5 overexpression exacerbated RSL3-induced cell death (P＜0.05) and attenuated the protective effects of paradol (P＜0.01). Conclusion　Paradol exerts protective effects in both FA-AKI and I/R-AKI mouse models, which may be associated with the alleviation of tubular injury, suppression of inflammatory responses, and improvement of oxidative stress. In vitro findings further suggest that paradol mitigates RSL3-induced HK-2 cell death, and ALOX5 may be involved in mediating its protective effects in AKI.