El-Dessouki Ahmed M, Kaml Mohamed E, El-Yamany Mohammed F
Pharmacology and Toxicology Department, Faculty of Pharmacy, Ahram Canadian University (ACU), 6th of October City, Giza, 12566, Egypt.
Pharmacology and Toxicology Department, Faculty of Pharmacy, Cairo University, Giza, 11562, Egypt.
Naunyn Schmiedebergs Arch Pharmacol. 2025 Mar 7. doi: 10.1007/s00210-025-03908-3.
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.
本研究调查了埃索美拉唑（ESOM）和卡格列净（CANA）对甲氨蝶呤诱导的肝毒性的肝保护作用，重点关注AMPK调节及其对MAPK/JNK/ERK、JAK1/STAT3和PI3K/Akt信号通路的调控。将50只雄性Wistar大鼠分为五组：对照组、甲氨蝶呤组，以及三个预处理组，分别给予ESOM（30毫克/千克）、CANA（30毫克/千克）或二者联合用药。在第9天单次腹腔注射甲氨蝶呤（20毫克/千克）诱导肝毒性之前8天和之后1天给予ESOM和CANA。使用生化、组织病理学、免疫组织化学、qRT-PCR和蛋白质印迹分析评估肝损伤、氧化应激、炎症和细胞凋亡。数据采用单因素方差分析（ANOVA）和Tukey事后检验进行分析，p < 0.05具有显著性。结果以平均值±标准误差（SE）表示。接受甲氨蝶呤的大鼠表现出明显的肝损伤，表现为ALT、AST、MDA、MPO、iNOS、TNF-α、IL-6和IL-1β水平升高（p < 0.01）以及抗氧化酶（HO-1、Nrf2和GSH）降低。免疫组织化学显示NF-κB p65和caspase-9表达增加（p < 0.01），与组织病理学变化相关。ESOM和CANA预处理降低了肝酶水平，改善了组织学，恢复了抗氧化平衡，并通过p38MAPK/NF-κB p65和JAK1/STAT3抑制炎症信号通路（p < 0.01）。此外，ESOM和CANA保留了PI3K/Akt活性并预防了caspase依赖性细胞凋亡（p < 0.01）。此外，联合治疗显示出协同肝保护作用，所有测量参数均有显著改善。这些发现表明，ESOM和CANA作为减轻甲氨蝶呤诱导的肝毒性的治疗药物具有显著潜力，值得在未来研究中进一步探究。
