Ajwa dates extract mitigates methotrexate-induced cardiotoxicity in rats via antioxidant and anti-inflammatory modulation.

Methotrexate (MTX) is a widely used immunosuppressive and chemotherapeutic agent known to induce cardiotoxicity through mechanisms involving oxidative stress, inflammation, and apoptosis. The present study investigated the cardioprotective effects of Ajwa date methanolic extract (AJDAE) against MTX-induced cardiac injury in rats, with emphasis on biochemical alterations, histopathological changes, immunohistochemical markers, and exploratory molecular docking. Forty male rats were randomly divided into four groups: control, AJDAE alone, MTX alone, and AJDAE pretreated with MTX. Phytochemical profiling of AJDAE using liquid chromatography-mass spectrometry (LC-MS) in both positive- and negative-ion modes revealed a complex composition rich in phenolic acids, flavonoids, vitamins, and minerals, as shown by total ion chromatograms (TICs) and base peak chromatograms (BPCs). MTX administration significantly increased cardiac injury biomarkers, pro-inflammatory cytokines, oxidative stress markers, and the apoptotic marker caspase-3, together with marked histopathological alterations, including myocardial necrosis. In addition, antioxidant defense parameters, including reduced glutathione (GSH), superoxide dismutase (SOD), and paraoxonase 1 (PON1), were significantly decreased. Pretreatment with AJDAE markedly ameliorated these changes by improving antioxidant status, reducing inflammatory and apoptotic markers, and preserving myocardial architecture. AJDAE also restored nuclear factor erythroid 2-related factor 2 (Nrf2) immunoreactivity, suggesting improved cellular antioxidant defense and tissue protection. Molecular docking provided complementary exploratory evidence of favorable predicted interactions between selected AJDAE bioactive compounds and proteins related to apoptosis and oxidative stress, including caspase-3 and the Keap1 Kelch domain as an Nrf2-regulatory target. In conclusion, these findings suggest that AJDAE exerts cardioprotective effects against MTX-induced cardiac injury through attenuation of oxidative stress, inflammation, and apoptosis. The restoration of Nrf2 immunoreactivity, together with improved antioxidant markers, suggests possible involvement of Nrf2-related antioxidant defense; however, direct activation of the canonical Nrf2/ARE pathway was not confirmed and requires further validation using downstream molecular targets.