Journal Article

Modulation of AMPK by esomeprazole and canagliflozin mitigates methotrexate-induced hepatotoxicity: involvement of MAPK/JNK/ERK, JAK1/STAT3, and PI3K/Akt signaling pathways

EL-Yamany, Mohammed F., El-Dessouki, Ahmed M., Kaml, Mohamed E.

2025

Overview

This research investigated the hepatoprotective effects of esomeprazole (ESOM) and canagliflozin (CANA) against methotrexate-induced liver toxicity, focusing on AMPK modulation and its regulation of MAPK/JNK/ERK, JAK1/STAT3, and PI3K/Akt pathways. Fifty male Wistar rats were divided into five groups: control, MTX, and three pretreatment groups receiving ESOM (30 mg/kg), CANA (30 mg/kg), or their combination. ESOM and CANA were administered for 8 days before and 1 day after a single MTX injection (20 mg/kg, intraperitoneally) on day 9 to induce hepatotoxicity. Liver injury, oxidative stress, inflammation, and apoptosis were assessed using biochemical, histopathological, immunohistochemical, qRT-PCR, and western blot analyses. Data were analyzed by one-way analysis of variance (ANOVA) and Tukey’s post hoc test, with significance at p  < 0.05. Results were presented as mean ± standard error (SE). Rats that received MTX showed significant liver damage, marked by elevated ALT, AST, MDA, MPO, iNOS, TNF-α, IL-6, and IL-1β levels ( p  < 0.01) and decreased antioxidant enzymes (HO-1, Nrf2, and GSH). Immunohistochemistry revealed increased NF-kB p65 and caspase-9 expression ( p  < 0.01), correlating with histopathological changes. Pretreatment with ESOM and CANA reduced liver enzyme levels, improved histology, restored antioxidant balance, and inhibited inflammatory pathways via p38MAPK/NF-kB p65 and JAK1/STAT3 ( p  < 0.01). Moreover, ESOM and CANA preserved PI3K/Akt activity and prevented caspase-dependent apoptosis ( p  < 0.01). Additionally, the combination treatment showed synergistic hepatoprotective effects, demonstrated by significant improvements in all measured parameters. These findings suggested that ESOM and CANA had significant potential as therapeutic agents for alleviating MTX-induced hepatotoxicity and warranted further investigation in future research.

• Publisher: Springer Berlin Heidelberg,Springer Nature B.V
• Subject: 1-Phosphatidylinositol 3-kinase / Adenosine triphosphatase / AKT protein / AMP-Activated Protein Kinases - metabolism / Animals / Antioxidants / Apoptosis / Apoptosis - drug effects / Biomedical and Life Sciences / Biomedicine / Canagliflozin - administration & dosage / Canagliflozin - pharmacology / Caspase-9 / Cell death / Chemical and Drug Induced Liver Injury - drug therapy / Chemical and Drug Induced Liver Injury - enzymology / Chemical and Drug Induced Liver Injury - etiology / Chemical and Drug Induced Liver Injury - metabolism / Chemical and Drug Induced Liver Injury - pathology / Chemical and Drug Induced Liver Injury - prevention & control / Cytokines / Design of experiments / Disease / Drug dosages / Hepatotoxicity / Homeostasis / Immunohistochemistry / Inflammation / Janus kinase / Janus Kinase 1 - metabolism / Kinases / Laboratory animals / Liver / Liver - drug effects / Liver - enzymology / Liver - pathology / Male / MAP kinase / MAP Kinase Signaling System - drug effects / Metabolism / Metabolites / Methotrexate / Methotrexate - toxicity / Neurosciences / NF-κB protein / Nitric-oxide synthase / Omeprazole / Oxidative stress / Oxidative Stress - drug effects / Pharmaceutical industry / Pharmacology/Toxicology / Phosphatidylinositol 3-Kinases - metabolism / Proto-Oncogene Proteins c-akt - metabolism / Rats / Rats, Wistar / Signal Transduction - drug effects / Stat3 protein / STAT3 Transcription Factor - metabolism / Toxicity / Tumor necrosis factor-α