Cellular Anti-Apoptotic Effects of Dapagliflozin in Methotrexate-Induced Liver Toxicity: Bax/Bcl-2/Cyt-C/Cas-9/Cas-3 Signaling Pathway

Methotrexate (MTX), an effective immunosuppressive and antiproliferative agent, is clinically restricted by its hepatotoxic potential through oxidative stress, inflammation, and apoptosis. Dapagliflozin (DAPA), a sodium–glucose cotransporter 2 inhibitor, exhibits antioxidant and anti-inflammatory actions. This study investigated the hepatoprotective effects of DAPA against MTX-induced acute liver injury. Thirty-two female Wistar albino rats were divided into four groups (n = 8): Control, MTX (20 mg/kg), MTX + DAPA (MTX + DAPA 10 mg/kg/day for 10 days), and DAPA. Liver samples were examined histologically, immunohistochemically (Nuclear factor NF-kappa-B p65 subunit (NF-κB p65), Tumor necrosis factor alpha (TNF-α), Interleukin 1 beta (IL-1β), Caspase (Cas)-3, Vascular endothelial growth factor (VEGF)), molecularly (Reverse transcription–polymerase chain for Bcl-2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), Cytochrome C (Cyt-C), Apoptotic peptidase activating factor 1 (Apaf-1), Cas-9, Cas-3, Cas-12), and biochemically (total oxidant status (TOS), total antioxidant status (TAS) and oxidative stress index (OSI)). MTX induced severe hepatic injury with congestion, sinusoidal dilatation, and inflammatory infiltration, accompanied by upregulation of NF-κB, TNF-α, IL-1β, Bax, Cyt-C, Apaf-1, Cas-9, Cas-3, and Cas-12 and reduced Bcl-2. DAPA co-treatment significantly restored hepatic structure, suppressed inflammatory and apoptotic markers, and normalized VEGF expression, indicating reduced pathological angiogenesis. Although DAPA did not fully reverse MTX-induced weight loss, it effectively mitigated hepatocellular damage. DAPA protects against MTX-induced liver injury by inhibiting NF-κB/TNF-α/IL-1β-mediated inflammation, modulating Bax/Bcl-2–Cyt-C–Cas-dependent apoptosis, and balancing VEGF-driven angiogenesis. DAPA may thus serve as a promising hepatoprotective adjunct in MTX therapy.