الفهرس 
DedicationOnly 14 pages are availabe for public view
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Abstract
This experiment was carried out to study the haematological, biochemical, histopathological, and immunohistochemical alterations resulting from administration of MTX in adult male albino rats in high and low doses. The histopathological toxic effects of MTX administration were observed in the liver, kidney, testis, and intestine. The main objective of this study is to evaluate the improvement of blood and biochemical parameters associated with FA and PRP treatment. Another goal was to evaluate the possible protective effects of FA and PRP on MTX toxicity in the liver, kidney, testis, and intestine. These objectives were achieved through the estimation of blood pictures, and a lot of biochemical analyses such as AST, ALT, MDA, GPx, TNF-α, and IL-6. As well, as histopathological examination of liver, kidney, testis, and intestine by light microscopy and immunohistochemical evaluation of caspase-3 in liver tissues.
The experiment was carried out on forty-five Sprague-Dawley male albino rats which were divided into nine groups, five animals per each according to the experimental design. The control group was injected intraperitoneally with normal saline three times weekly, the FA group received orally FA 200 µg/kg BW daily for 2 weeks and the PRP group was injected with I.P. PRP 0.5 ml/kg BW three times weekly for 2 weeks. MTX 40 mg/kg group was injected with I.P. MTX 40 mg/kg BW single dose after seven days after the beginning of the experiment. MTX 40 mg/kg & FA was injected with I.P. MTX 40 mg/kg BW single dose seven days after the beginning of the experiment and received orally FA 200 µg/kg BW daily for 2 weeks. MTX 40 mg/kg & PRP were injected with I.P. MTX 40 mg/kg BW single dose seven days after the beginning of the experiment and was injected with I.P. PRP 0.5 ml/kg BW three times weekly for 2 weeks.
Also, the MTX 6 mg/kg group was injected I.P. with MTX 6 mg/kg BW two times; at the beginning of the experiment and after seven days after the beginning of the study. MTX 6 mg/kg & FA was injected with I.P. MTX 6 mg/kg BW two times; at the beginning of the experiment and after seven days after the beginning of the study and received orally FA 200 µg/kg BW daily for 2 weeks. MTX 6 mg/kg & PRP group was injected with I.P. MTX 6 mg/kg BW two times; at the beginning of the experiment and seven days after the beginning of the study and injected with I.P. PRP 0.5 ml/kg BW three times weekly for 2 weeks. At the end of the experiment, rats were sacrificed after anesthesia and samples were whole blood samples collected for CBC, and serum for measurement of AST, ALT, creatinine, and urea. In addition, levels of MDA, GPx, TNF-α, and IL-6 were investigated in the serum. Liver, kidney, testis, and intestine tissue specimens also were removed and prepared for histopathological examination. Immunohistochemical expression of caspase-3 in liver tissue was also evaluated.
The results of this study showed that FA and PRP groups seemed similar to the control group in all haematological and biochemical parameters in addition to the histological structure of hepatic, renal, testicular, and intestinal sections. On the other hand, there was a decrease in the total leucocytic count, differential leucocytic count, erythrocyte count, Hb, PCV, MCV, MCH, MCHC, RDWa, and PLT count in MTX injected groups in comparison with the control group. While there was a partial or complete improvement in all haematological findings in all treated groups. Serum levels of MDA, AST, ALT, creatinine, and urea in addition to TNF-α, and IL-6 increased in both intoxicated groups compared with control groups and there was an improvement in all of these parameters in all treated groups. GPx level also decreased in MTX groups and there was a remarkable increase in its level in all treated groups when compared to the control group.
The MTX injected groups exhibited severe histopathological alterations classified as vascular changes, necrobiotic changes, cellular reactions, and active fibrocytic proliferation although control, FA, and PRP groups showed to a large extent normal histology. The angiopathic changes included congestion, desquamation of the lining endothelial cells of the blood vessels, and focal areas of hemorrhages. The necrobiotic changes were vacuolar degeneration in hepatocytes, variable-sized vacuoles inside cells, apoptotic hepatocytes, and multi-focal necrotic areas. Mononuclear besides polymorphnuclear cells and kupffer cellular hyperplasia were observed in most cases. In addition, there was a massive area of active fibrocytic proliferation which was a very characteristic in these animals. Masson trichrome stain confirmed this finding by staining the C.T. proliferation within the hepatic tissue with a green colouration. Also, MTX induced a strong caspase-3 immunoreaction in hepatic tissues. On the other hand, most of these hepatic alterations were reduced in all treated groups.
Kidneys of control, FA, and PRP groups revealed to a large extent normal histology. Severe microscopic findings were detected in the kidneys of MTX groups in the form of glomerular damage manifested with glomerular congestion, and periglomerular connective tissue proliferation infiltrated with mononuclear cells extending sometimes to the intertubular areas. This was accompanied by thickening of Bowman’s capsule which was confirmed with Masson trichrom stain. Bowman’s space also appeared extremely narrow in some corpuscles and completely absent in others in severe cases. The tubular changes were vacuolation of the tubular epithelium, renal tubular casts, tubular dilatation, and necrosis. Also, congestion of large blood vessels and peritubular capillaries, hemorrhage and mononuclear cell infiltration were observed in the interstitial tissue. FA and PRP showed an ameliorating effect on all these findings in all treated groups especially those treated with PRP.
Testicular tissues of FA and PRP-administered groups were similar to the control rats. MTX-injected groups showed vascular congestion, interstitial edema, and a decrease in spermatogenesis according to JTBS values. However, all treated groups showed an improvement in the histological appearance of testes.
The examination of the intestines of FA and PRP groups revealed normal morphological appearance. There was a marked shortening in the intestinal villi of MTX 40 mg/kg which improved in both MTX 40 mg/kg & FA and MTX 40 mg/kg & PRP groups.