Background: Cyclophosphamide (CP) is an oxazaphosphorine- alkylating agent widely used in the treatment of various neoplastic diseases and disorders associated with altered immunity. The optimal clinical usefulness of CP is severely limited by a high incidence of multiple organ toxicity. Many studies suggest that CP through its acrolein radical causes reactive oxygen species (ROS). Thymoquinone (TQ), the main constituent of the volatile oil from Negella sativa seeds. It is reported to possess strong antioxidant properties. Aim of the work: This research evaluates the possible protective effect of Thymoquinone on CP-induced liver toxicity in albino rats model. Material and Methods: Thirty albino rats were divided into three groups each one contains ten rats. Group I (Control group): The rats were received normal saline intraperitoneally. Group II: Each rat received 150 mg/kg cyclophosphamide (CP) as a single dose intraperitoneally. Group III: The rats pretreated with Thymoquinone (TQ) with a dose of 10mg/kg/day intraperitoneal for 7 days before the administration of 150 mg/kg CP a single dose intraperitoneally one week after the last dose. In each group, animals were sacrificed and the liver was identified and dissected out and prepared for histological examination after staining the tissues by Hematoxylin and eosin (H x & E) and Masson's trichrome. Also an Immunohistochemical study was done to detect α-smooth muscle actins (α –SMA). The mean area percentage of (α –SMA) expression was quantified in five images from five non-overlapping fields of each rat. The data were collected from the experiment, recorded and analyzed using IBM SPSS Statistics software. Electromicroscopic examinations: Semi-thin sections approximately 1 μm thickness were cut Sections were stained with toluidine blue. Ultrathin (below 100 nm) sections were collected on copper grids, stained with uranyl acetate and Reynold’s solution (sodium citrate and lead nitrate), then examined with transmission electron microscope (Philips 201C by Netherland) and photographed. Results: The liver sections of rats received cyclophosphamide (CP), showed loss of architecture of liver, dilated central vein and dilated sinusoids , with cellular infiltration around the central vein and in the portal area . The density of hepatocytes reduced with lacking acidophilic cytoplasm and the cells were vacuolated. Inflammatory cells were scattered in the hepatic parenchyma . There were proliferation of bile duct.
Marked increase of the collagen fibers deposition around the central vein and in portal area detected by masson trichrome stain. On examination of liver section treated by TQ prior to CP, there were improvement of the liver tissue. The hepatic cells were normally arranged around the sinusoids, some cells were binucleated and acidophilic. Some sinusoids were dilated and less collagen fibers deposition around central vein than in group II. Immunohistchemical stain (α-SMA stain: Area % of α-SMA immuno- reactivity was highly significant increased in group II as compared to groups I & III (P |
