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Fondaparinux attenuates methotrexate-induced hepatotoxicity by regulating coagulation, endothelial dysfunction, and
Asmaa Saleh1, Nahed A Raslan2,3, Heba Mohammed Refat M Selim4,5
1Department of Pharmaceutical Sciences, College of Pharmacy, Princess Nourah bint Abdulrahman University, P.O. Box 84428, Riyadh, 11671, Saudi Arabia.
Abstract:
This study evaluates the protective potential of Fondaparinux (Fond), a selective antithrombin-mediated Factor Xa inhibitor, in methotrexate-induced hepatotoxicity. The work explores its ability to correct coagulation imbalance, improve endothelial function, and attenuate oxidative and inflammatory cascades (TLR4/NLRP3, NF-κB p65/IL-1β/MCP-1). Animals were allocated into 4 groups. A control group was given distilled water via the intraperitoneal route (i.p.); an MTX group was given a single intraperitoneal injection of MTX (20 mg/kg) on the seventh experimental day; and two groups received prior prophylactic administration of Fondaparinux (at doses of 5 or 10 mg/kg, intraperitoneally) throughout seven consecutive days before as well as for an additional four-day period following MTX administration. MTX significantly elevated hepatic injury markers (AST, ALT, ALP), induced oxidative stress with depleted antioxidants (SOD, GSH), and activated TLR4/NLRP3 signaling, resulting in upregulation of inflammatory mediators (TNF-α, NF-κB p65, IL-18, IL-1β, MCP-1, caspase-1, iNOS, ICAM-1, MPO) and suppression of IL-10 (p < 0.05). Endothelial dysfunction was evidenced by reduced eNOS. MTX also triggered marked coagulation disturbances, including enhanced Factor Xa-dependent thrombin generation, increased tissue factor, fibrin deposition, and elevated PAI-1. Mitochondrial apoptotic signaling was promoted, as indicated by elevated expression of cytochrome c along with induced caspase-3 and caspase-9 activation . Histologically, MTX caused extensive hepatic damage characterized by periportal fibrosis, inflammatory infiltration, bile duct proliferation, hepatocellular necrosis, vacuolation, and vascular congestion. Fondaparinux pretreatment dose-dependently restored hemostatic balance, improved endothelial function, suppressed oxidative and inflammatory responses, attenuated apoptosis, and markedly ameliorated histopathological alterations. Fondaparinux limits methotrexate-associated liver damage through inhibition of Factor Xa-dependent coagulation pathways while providing antioxidant, anti-inflammatory, anti-apoptotic, and hepatoprotective actions.
Insights
Fondaparinux protects the liver from methotrexate damage by inhibiting Factor Xa, reducing inflammation, and restoring coagulation. This study shows Fondaparinux offers significant hepatoprotective effects against methotrexate-induced liver injury.
Area of Science:
- Hepatology and Pharmacology
- Coagulation and Inflammation Research
- Drug-Induced Liver Injury Models
Background:
- Methotrexate (MTX) is a widely used chemotherapy agent with known hepatotoxicity.
- MTX-induced liver injury involves complex mechanisms including oxidative stress, inflammation, endothelial dysfunction, coagulation imbalance, and apoptosis.
- Selective Factor Xa inhibition is a potential therapeutic strategy for mitigating drug-induced liver damage.
Purpose of the Study:
- To evaluate the protective effects of Fondaparinux (Fond), a selective antithrombin-mediated Factor Xa inhibitor, against methotrexate-induced hepatotoxicity.
- To investigate Fondaparinux's ability to correct coagulation imbalance, improve endothelial function, and attenuate oxidative and inflammatory cascades.
- To assess Fondaparinux's impact on apoptotic signaling and histopathological alterations in MTX-treated animals.
Main Methods:
- Animals were administered MTX (20 mg/kg, i.p.) to induce hepatotoxicity.
- Prophylactic administration of Fondaparinux (5 or 10 mg/kg, i.p.) was given for 11 consecutive days (7 days prior to and 4 days after MTX).
- Hepatic injury markers, oxidative stress parameters, inflammatory mediators, endothelial function markers, coagulation profiles, apoptotic markers, and histopathological changes were assessed.
Main Results:
- MTX significantly increased liver injury markers, oxidative stress, and inflammatory responses (TLR4/NLRP3, NF-κB p65/IL-1β/MCP-1), while decreasing antioxidants and IL-10.
- MTX induced endothelial dysfunction (reduced eNOS), coagulation disturbances (enhanced Factor Xa activity, fibrin deposition), and promoted mitochondrial apoptosis.
- Fondaparinux pretreatment dose-dependently ameliorated liver damage, restored hemostatic balance, improved endothelial function, suppressed oxidative/inflammatory cascades, attenuated apoptosis, and improved histopathology.
Conclusions:
- Fondaparinux demonstrates significant dose-dependent hepatoprotective effects against MTX-induced liver injury.
- Fondaparinux limits MTX-associated liver damage by inhibiting Factor Xa-dependent coagulation pathways.
- The protective mechanism involves antioxidant, anti-inflammatory, and anti-apoptotic actions, alongside improved endothelial function and hemostasis.
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