Cemre Uçar-Ekin, Department of Physiology, Faculty of Medicine, Dicle University, Diyarbakır, Turkey
Fırat Aşır, Department of Histology and Embryology, Faculty of Medicine, Dicle University, Diyarbakır, Turkey
Fırat Şahin, Department of Histology and Embryology, Faculty of Medicine, Dicle University, Diyarbakır, Turkey
Şehmus Kaplan, Department of Histology and Embryology, Faculty of Medicine, Dicle University, Diyarbakır, Turkey
Objective: This study aimed to investigate the hepatoprotective effects of Rosmarinic acid (RA) against cyclophosphamide (CP)-induced liver injury in rats. Methods: Twenty-one male Wistar Albino rats were divided into three groups: Control, CP, and CP + RA. Hepatotoxicity was induced by administering CP (20 mg/kg/day) intraperitoneally for 14 days. RA (20 mg/kg/day) was administered for 14 days after CP induction. Serum biochemical parameters including malondialdehyde (MDA), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were measured. Liver tissues underwent histological and immunohistochemical analysis for B-cell lymphoma 2 (Bcl-2), apoptotic protease activating factor 1 (Apaf-1), nuclear factor erythroid 2-related factor 2 (Nrf-2), and tumor necrosis factor (TNF)-α. In addition, in silico analysis was performed to explore potential molecular targets of RA and their biological pathways. Results: CP significantly increased liver weight, MDA content, ALT and AST enzyme activities, indicating hepatic oxidative stress and injury. Histologically, CP caused severe hepatocellular damage characterized by hepatocyte degeneration, hemorrhage, and disrupted hepatic architecture. Immunohistochemically, CP exposure upregulated pro-apoptotic (Apaf-1), oxidative stress (Nrf-2), and inflammatory (TNF-α) markers, while downregulating anti-apoptotic (Bcl-2) proteins. RA administration significantly reversed these biochemical and histopathological changes. In silico analysis revealed RA interacts with multiple inflammatory and oxidative stress pathways, reinforcing its hepatoprotective role. Conclusion: RA demonstrates significant hepatoprotective activity against CP-induced liver toxicity by attenuating oxidative stress, inflammation, and apoptosis pathways. RA represents a promising therapeutic agent to manage drug-induced hepatotoxicity.
Keywords: Apoptosis. Cyclophosphamide. Hepatotoxicity. Inflammation. Oxidative stress.