A Histopathological, Immunohistochemical and Biochemical Investigation on the in vitro Antioxidant, Myeloprotective, Hematoprotective and Hepatoprotective Effects of Hypericum triquetrifolium Seed Extract Against Cyclophosphamide-Induced Toxicity

Abstract The aim of this study was to investigate in vitro antioxidant properties and in vivo protective effects of the methanol extract of the Hypericum triquetrifolium Turra (HT) seed against acute hepatotoxicity, myelotoxicity and hematotoxicity in rats induced by cyclophosphamide (CP). In order to investigate in vivo protective effects of the HT extract on rat tissues, the rats were divided into nine groups. The toxic effects of CP and the protective effects of HT extract on nucleated cells that are produced by bone marrow, serum alanine transaminase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and oxidative stress index (OSI) levels were investigated biochemically. Additionally, liver tissue samples were examined for histopathological changes and apoptosis by Bcl-2, Bax and caspase-3 immunohistochemistry. The results of this study show that HT seed methanol extract has high total phenolic content (179.52 μg GAE/mg) and antioxidant activity (87.48% in 500 μg/mL concentration). CP administration caused hepatotoxicity, myelotoxicity and hematotoxicity in the rats. Whereas, the groups of rats that were injected with different concentrations of HT (25, 50 and 100 mg/kg) and CP (150 mg/kg) showed significant protective effects on bone marrow nucleated cells and important decreases on serum ALT, ALP, LDH and OSI levels were observed when compared with the CP injected group.

Protective Effect of Carvacrol Against Oxidative Stress and Heart Injury in Cyclophosphamide-Induced Cardiotoxicity in Rat

Possible protective effects of carvacrol (Car) against cyclophosphamide (CP)-induced cardiotoxicity was examined in this study. Experimental groups of the rats were randomly divided into 13 groups,each including seven animals: Group 1 (control) treated with saline; groups 2, 3, and 4 treated with 50, 100, or 150 mg/kg of CP, respectively; group 5 treated with 0.5 mL olive oil; groups 6 and 7 treated with 5.0 and 10 mg/kg of Car, respectively; groups 8, 9, or 10 treated with respective CP plus 5.0 mg/kg of Car; and groups 11, 12, or 13 treated with respective CP plus 10 mg/kg of Car. Serum alanine transaminase (ALT),aspartat transaminase (AST), lactate dehydrogenase (LDH), malondialdehyde (MDA),creatine kinase-MB (CK-MB), total oxidant state (TOS), oxidative stress index (OSI), and levels were high only in the CP groups. There was a dose-dependence on the CP-induced cardiotoxicity. Hemorrhage, inflammatory cell infiltration and the separation of the muscle fibers in the heart tissue supported the biochemical data. With 5.0 and 10 mg/kg Car, there was an important decrease in the CP toxicity and this was related to the oxidative and nitrosative stress in the CP-induced cardiotoxicity. Reduced inflammation and lipid peroxidation in the heart tissue and increase of serum glutathione (GSH) and total antioxidant capacity (TAS) levels were found when carvacrol was applied. Based on these findings, it could be proposed that Car was a strong candidate in preventing the CP-induced cardiotoxicity but further clinical studies should be done in order to verify its application on humans.