الفهرس 
ACKNOWLEDGMENTS
Mercuric toxicityOnly 14 pages are availabe for public view
 |  | from | 306 |  |  |
| | | from | 306 | | |
Abstract
Mercury is one of the most common heavy metals, used for more than 3000 years in medicines (as disinfectants, vaccines), industries (fluorescent lamps, batteries, thermostats, thermometers), gold mining and therapeutically as a cathartic, diuretic, anti-inflammatory and in dental amalgams. In recent years, the human exposed widely to mercury in large scale in all over the world causing ecological and health problems.
Sixty Male Wister Albino rats weighting 180 30 g. were used. Rats were divided into six groups(N= 10). G1 : served as control receiving normal diet and tape water. G2 was, considered as positive control ( receiving MC was added as 50 mg/L in drinking water DW ad libium ). G 3.G4, G5 and G6 received the same dose of MC in addition to the therapeutic doses of antioxidants as follows . G3: received MC + Vit.E (Vit. E added in the normal diet 150 mg/100 g), G4: received MC + Se (Selenium was added in DW as 2 mg/ L), G5 received MC + FSPS (Oral fenugreek seed powder; as DW ; 4% W/V in DW and soaked for 24 h and given via drinking ) and G6: sixth group received MC, Se , FSPS and vit. E by the same previous doses. The period of the experiment extended for 30 days , after which the animals were deprived of all treatments food and the drinking water become devoid of MC for one day, Animals were anesthetized with ether, then killed. Blood was collected from anesthetized animals for both hematological and biochemical studies. Liver, kidneys and testes were excised immediately, washed in ice cold saline. Pieces of liver, kidneys and testes were divided into two parts, parts taken for homogenization and the others were fixed in formal alcohols for histological investigation.
The following parameters were measured: (a). blood glucose, serum urea, creatinine, lactate dehydrogenase LDH, Aspartic aminotransferase AST and alanine aminotransferase ALT activities as liver and kidney functions. (b). Oxidative stress biomarkers were measured includes: Lipid peroxidation LPO, total peroxides TPX, nitric acid NO, and carbonyl protein CAR PRO as indices of tissue oxidative damage. (c). Vitamin E Vit. E , Vitamin C Vit. C, ceruloplasmin CP, total thiols T. Thio and GSH content and SOD, CAT, GST activities , copper and Zinc ions as indices of antioxidant defense. (d). measurement of glucose-6-posphate dehydrogenase G-6-PD as indicator of NADPH production. (e). Measurement of tissue glycogen for highlighting carbohydrate metabolism. Also, some serum metabolites were measured such as triglycerides and cholesterol. f. Measurement of total free amino acids TFAA in serum, all examined tissues and erythrocytes for monitoring their incorporation or/and mobilization in tissues for interpreting the sites of expecting synthesis or breakdown protein. (g). Determination of Hepato-somatic index (HSI) , testicular somatic index (TSI) and Renal somatic index (RSI). (i). The normal values of white blood Cells (WBC’s), differential count of (WBC): lymphocytes (LYM), monocyte (MONO), eosenophil (EOS) and neutrophil(NEU)) were determined by using automated technical analyzer (Mindray Bc-2800) in faculty of Veterinary Medicine, Assiut University. Values of hemoglobin content (Hb), hematocrit (HCT), RBCs count, mean corpuscular hemoglobin (MCH) , mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC) and platelets count (PLT) were determined. Histological observation by light microscope for liver, kidney and testes were also done.
Biochemical study where administration of MC in rats led to:
1. Significant decreases in hepatic antioxidants: catalase activity (hCAT) by 68.3%, superoxide dismutase (hSOD) by -57.9%, glutathione-s-transferase (hGST) by -44.4% and reduced glutathione (hGSH) by -53.9% compared with control.
2. Significant decreases in renal antioxidants: activities of rCAT by -76.4 %, rSOD by -78.3 %, rGST by -50.4 % and the level of rGSH by 60.2 % compared with control. Those parameters were relatively improved in all groups with different percentages after supplementations of Vit. E, (Se), (FSPS) and their combination compared with control values.
3. Significant decreases in testicular antioxidants: activities of tCAT by -59.0 %, tSOD) by -53.8 %, tGST) by -47.8 % and the level of tGSH by -55.4 % compared with control.
4. The administration of MC in rats led to significant decreases in testicular antioxidants: activities of eCAT by -60.5 %, eSOD) by -52.0 %, eGST) by -32.2 % and the level of eGSH by -48.2 % compared with control value.
5. Significant decrease in erythrocytic lysate glucose-6-phosphate dehydrogenase activity (G6PD) in MC-treated rats by -29.9 %.
6. Significant decreases in T. Thio: Hepatic T.Thio by - 44.49 % Renal T.Thio by - 49.7 % Testicular T.Thio by - 67.1 % eT. Thio by - 38.5% compared with control values.
7. Significant decreases in the levels of Vit.C in liver by -41.7 %, in renal Vit.C by -39.4 % and in testicular Vit.C by - 45.9 % compared with corresponding control values. Also, there were significant decreases in the levels of Vit.E in liver Vit.E by -55.9%, in renal Vit. E by-35.3 % and in testicular Vit. E by-30.9 % compared with corresponding control values.
8. Significant decreases in serum CP by -22.48 % compared with control.
9. Significant decreases in the levels of serum free ions of both copper (Cu ) and zinc (Zn) by -33.56 and -45.62%, respectively compared with corresponding control values.
10. Significant increases in LPO: Hepatic LPO by 86.6% (1.86-fold), renal LPO by105.5 % (2.1-fold), testicular LPO by84.9% (1.85-fold) and erythrocytic lysate LPO by250.7 % (3.5-fold) compared with corresponding control values. The adverse effects of MC on LPO levels were almost abolished with Vit. E, Se, FSPS and their combination in testes. However, in the liver and kidney, most alterations of LPO were improved.
11. Significant increases in TPX: Hepatic TPX by 59.5 % (1.6-fold), Renal TPX by 75.1% (1.8-fold), testicular TPX by 75.3 % (1.8-fold) and Erythrocytic lysate TPX by 287.4 % ( 3.9-fold) compared with corresponding control values.
12. Significant increases in CAR PRO: Hepatic CAR PRO by 189.8 % (2.9-fold), Renal CARPRO by 135.2 % (2.35-fold), testicular CARPRO by 123.9 % (2.23-fold) and erythrocytic lysate CAR PRO by 151.1% ( 2.5-fold) compared with corresponding control values.
13. Significant decreases in NO values of all tissues: in hepatic NO by -27.6 %, in renal NO by -42.2% and in testicular NO by -36.1% compared with control values.
14. Significant decreases in hepatic and testicular levels of glycogen by -57.1 and -32.4 %, respectively while, MC significan increase in renal glycogen by 110.4 % were obtained. Blood glucose level was found to be increased by and glycemic level by 92.3 % compared with corresponding control values.
15. Significant increases in the levels of TFAA in all tissues and serum: in hepatic TFAA by 464.8 % (the largest percentage compared with others: 5.6-fold), in renal TFAA by 284.6% (3.8-fold), in testicular TFAA by 68.0 % (1.6-fold), in erythrocytic lysate TFAA by 131.9 % (2.3-fold) and in serum TFAA by 134.8 % (2.3-fold) compared with corresponding control values.
16. Significant decreases in serum triglycerides and cholesterol levels in MC-treated rats by -62.4% and -51.8% respectively compared with control values. Adverse effects of MC on both serum triglycerides and cholesterol levels were relatively abolished with all treatments especially in MC+ Comb and MC+ Vit.E ( - 4.3 % and 6.7% respectively) compared with corresponding control values.
17. Significant increases in serum contents of urea, creatinine and plasma free hemoglobin in MC-treated rats by 48.21% (1.48-fold), 83.4% (1.83-fold) and 488.38 % (5.88-fold), respectively compared with control values.
18. Significant elevations in serum transferases: activities of serum AST by 56.2% and serum ALT by 86.9 %, Also increases in serum ALP by 69.2% and the level of serum LDH by 184.5 % compared with corresponding control values.
19. Significant DROP in erythrocytic lysate activity of glucose-6-phosphate dehydrogenase (eG6PD) activity by -29.9% compared with control values.
20. Significant elevations in HIS by25.4% (P < 0.05), significant elevation in RSI by 22.38% (P < 0.001) and significant reduction in TSI level by 37.0% (P < 0.001) compared with control value in response to MC intoxication.
21. Also, MC induced significant increase in both WBC count 181.4 and percentage of lymphocytes (LYM) by 67.7 % accompanied with significant decrease neutrophils (NEU) by -62.2% compared with control values.
22. Hematological parameters: RBCs count, HCT, bl. Hb, MCV, MCH, MCHC and PLT showed alterations in rat-treated with MC.
23. The harmful effects resulted from MC treatment on hepatic antioxidants were relatively improved in all treated groups. The most better responses were in comb group which showed amelioration in the activities of hepatic CAT by 26-25.6%, SOD by -22.8, GST by -5.8 % and in the level of hepatic GSH by -10.7compared with control. The harmful effects of MC on renal antioxidants were relatively improved in activities of CAT , SOD and the level of GSH with better response extents in comb group by -20.4, -25.16 and -31.2%, respectively compared with control values.
24. Testicular antioxidants were also improved in all therapeutic treated groups and with the most better response one of all groups was the in comb group which showed amelioration in the activities of CAT by -6.3%, SOD by -23.7, GST by -4.8 % and in the level of GSH by -29.2 compared with control value.
25. The harmful effects extents of MC-treatment on erythrocytic antioxidants or G6PD were relatively improved in all groups with different percentages after supplementations of vit. E, selenium (Se), fenugreek seed powder (FSPS) and their combination compared with control values. The improvement in activities of eCAT , eSOD and eGST in all treated groups with the better response extent in comb group by -22.1, -19.8 and 4.03 %, respectively.
26. The harmful of MC on the levels of T. Thio were relatively improved compared with control in different treated in all groups. The better improvements of hepatic T.Thio by-0.6 % in MC+ Se group , renal T.Thio by -1.1% in MC+ Combt while, the best recovery of T. Thio in testes and erythrocytic lysate were in MC+ Se group by -2.2 % and 8.7,%, respectively compared with corresponding control values.
27. The harmful effect of MC on the Vit.C levels were relatively ameliorated in all groups after administration with all therapeutic groups (P> 0.05) compared with corresponding control values, respectively. The harmful effect of MC on the Vit.E levels were relatively ameliorated in liver and the better one was in MC+ Comb group by -13.9% compared with control. Also, the adverse effects of MC in kidney and testes were improved with all therapeutic groups and the best recovery was in MC+ Vit.E and Comb groups, by 7.1% and 26.4 % respectively compared with corresponding control values.
28. The adverse changes of MC on serum CP was almost abolished with supplementations of therapeutic nutrients in all groups compared with control values.
29. The adverse effects of MC on serum Zn and Cu free ions were almost abolished in most therapeutic groups compared with control. The best recovery in Cu and Zn free ions levels were MC+ Comb group by 7.45and -11.0 % , respectively compared with corresponding control values.
30. The adverse effects of MC on TPX levels were almost abolished in therapeutic groups in rats-treated with MC.
31. The harmful effect of MC on the eCAR PRO levels was abolished in erythrocytic lysate, while in other organs CAR PRO levels showed relative amelioration after treatments with therapeutic nutrients. The best improvements in hepatic CAR PRO were in group MC+ Se (17.77%), renal CAR PRO were in MC+ Vit.E (-10.6%), testicular CAR PRO were in MC+ Comb (14.6%) and erythrocytic lysate CAR PRO were in MC+ Comb (-9.9%).
32. The adverse effects of MC on hepatic NO, renal NO and testicular NO levels were almost improved in most therapeutic groups and the best improvements of hepatic NO was in MC+ Vit.E group by 4.24 % , renal NO were in MC+ Se group by -5.51% and testicular NO were in MC+ Comb group by -6.7 % compared with corresponding control values.
33. The harmful effect of MC on the carbohydrate levels were relatively ameliorated in all therapeutic groups compared with corresponding control values, respectively.
34. The harmful effect of MC on the TFAA levels were relatively ameliorated in all therapeutic groups compared with corresponding control values. The better improvements of TFAA were in the liver, kidney and serum were in MC+ Comb group by 62.7, 63.7 and 24.5 %, respectively compared with control values
35. The adverse effects of MC on both serum contents of urea and creatinine were almost completely abolished and the best extent of improvements were in groups of MC+ FSPS and MC+ Comb by 5.99 and -2.9 % respectively compared with control values.
36. The harmful extents of MC in activities of transferases were improved with better extents of sAST by 1.6 % in MC+ FSPS group and sALT by19.3 % in MC+ Comb group , also better ameliorations in the activities of sALK by 2.9% in MC+ Vit.E group and LDH by 35.9 % in MC+ FSPS group, compared with control values.
37. The adverse effects of MC on eG6PD activity was almost completely abolished in all therapeutic groups and the best extent of improvement was in group of MC+ Comb by 6.65 % compared with corresponding control value.
38. The rises in HSI and RSI in rat-treated with MC, was improved with better extents after treatment with MC+ Comb by 10.24 % (P > 0.05) and 1.02 % (P > 0.05), respectively compared with corresponding control values. While the adverse effect of MC on RSI was improved with better extent after treatment with MC+ FSPS by % -0.48 (P > 0.05) compared with control value.
39. The adverse effect of MC on WBC and their differentiation, showed significant relative improvements compared to control in all therapeutic groups and the best results in COMB group were observed. Also, The hematological alterations resulted from MC toxicity were improved with different extents after administration of vit. E, Se, FSPS and their combination. The most better improvement of hematological parameters was in COMB group.
40. Vit. E, Se, FSPS and their combination, decreased the DNA damage that induced by mercuric chloride.
Histological studies:
41. Marked histological changes was induced in both liver and kidney including hydropic degeneration and necrosis in liver, while kidney structures showed atrophy of the glomeruli and dilated renal spaces, apical border degeneration in tubules with luminal casts and exfoliation of cells in the tubular lumen in mercury treated groups. In testes, MC led to distortion of tubules, vaculation of germ cells, detachment in and desquamation into seminiferous lumina, and Also, necrosis ,disorganization of most spermatogenic stages, lost of mature sperms and intertubular space widening due to vascular dilation and associated edema .
42. The histological observation of liver and kidney of rats showed significant improvements after treatments with Vit. E, S E, FSP either solely or with their combination in MC-treated rats with the best response in COMB group compared with control. The histological alterations in testes resulted from MC treatment, were relatively improved particularly spermatogenesis and formation of mature sperms after supplementation with Vit. E, Se , FSPS and their combination with the best result in COMB group.
In conclusion
Both biochemical and histological studies showed that combination of FSPS, Se and Vit.E was found to protect liver and kidney against MC toxicity in the rats most probably due to integrated antioxidant powers exist in these supplementations. The result showed a close relationship of oxidative damage between erythrocytes and testicular tissue in MC-treated rats. In turn, the testicular biochemical, histological and hematological alterations resulted from MC toxicity were improved after administration of Vit. E, Se, FSPS and their combination ; the later showed better extent of improvement.