الفهرس 
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Abstract
GSE or FSE extract improved the hazard toxic effect induced by aluminum, lead or cadmium include powerful free radical scavenging property, strong antioxidant enzyme activities by reducing liver tissue MDA and improving antioxidant enzyme activities in liver tissue.
The experiments were carried out in Agricultural Chemistry Department, Faculty of Agriculture, Minia University. The animals were housed in plastic cages. The rats were kept under normal healthy laboratory conditions. A total of one hundred male albino rats, weighing from 145 to 170 g, divided into 20 groups were used as follow: Group1 as control, groups 2-5 administrated with GSE and FSE at two doses (100 and 200 mg/kg b. wt). group 6 was administrated with aluminum chloride 37 mg/kg b.wt. Groups 7-10 were administrated with GSE and FSE at two doses plus Al. group 11 was administrated with Pb as lead acetate, 75 mg/kg b. wt. Groups 12-15 were administrated with GSE and FSE at two doses plus Pb. group 16 was administrated with CdCl2 5 mg/kg b. wt. And groups 17-20 were administrated with GSE and FSE at two doses plus Cd. The experiments continued for 90 days. Animals were weighted every week.
The following parameters were studied:
• Body growth including: average daily weight gain, organs weight and daily food efficiency.
• Some blood parameters including liver function (AST, ALT, total and direct bilirubin, total portion and albumin), kidney function (urea and creatinine), and lipid profile (total cholesterol, TG, HDL-c and LDL-c).
• Hematological investigation as RBC, WBC, HB, HCT, MCV, MCH, MCHC and differential leukocyte count.
• The liver tissues were homogenated and hepatic (MDA, CAT and GSH) were determined.
• Histopathological changes in liver, kidney, lung, spleen, brain, heart and testis.
The obtained results could be summarized as follows:
1. In intoxicated aluminum, lead or cadmium groups after 45 and 90 days, the serum AST and ALT activities were significantly elevated when compared to control group. However, co-administration of GSE at two doses or high dose of FSE plus aluminum or lead restored the level of AST to normal. Whereas, FSE at low dose plus aluminum or lead, decreased the enzyme activities but they were still higher than normal when compared to control group. Simultaneous, administration of GSE at two doses with aluminum reduced ALT serum level elevation by about 27 and 28% respectively compared to aluminum treated group; 63 and 60% respectively compared to normal control after 90 days. Whereas, the level of ALT was restored to normal in group treated with high dose of FSE with aluminum after 90 days while low dose FSE had no effect on ALT activity compared to aluminum treated animal. In addition, administration of GSE (100) or FSE (100, 200) with lead caused a significant reduction in the elevated levels of serum ALT by about 32, 34 and 26% respectively after 90 days when compared to lead alone and 50, 46 and 63% respectively, compared to normal control after 90 days. While administration of high dose of GES recovered the activity of this enzyme to normal control. GSE at high dose plus cadmium reduced ALT activity by about 34% after 90 days when compared to cadmium alone.
2. In intoxicated aluminum, lead or cadmium group after 90 days, the level of serum total protein was significantly elevated by about 29, 38 and 25% respectively when compared to control group. On the other hand, the treatment with either GSE or FSE (100, 200 mg/kg) concurrently with aluminum produced a significant decrease in the level of total protein compared to animal treated with aluminum alone. Moreover, the total protein level was reduced in treated rats with GSE or FSE (100, 200 mg/kg) plus lead when compared to lead alone after 90 days. Whereas, serum total protein in groups treated with GSE or FSE (100, 200 mg/kg) plus cadmium was decreased compared to cadmium alone. In aluminum, lead, cadmium treated groups the serum level of albumin was reduced compared to control group by about 15, 22 and 24% respectively after 45 days while after 90 days they were reduced by about 27, 32 and 32% respectively. Simultaneous administration of GSE or FSE (100, 200 mg/kg) plus aluminum increased the serum level of albumin compared to aluminum treated group about 31, 26, 18 and 30% respectively after 90 days. In addition the presence of GSE (100, 200 mg/kg) or FSE (100, 200 mg/kg) with lead caused a significant increase in serum albumin by about 37, 42, 28 and 41 % respectively after 90 days when compared to lead alone. On the other hand co- administration of GSE or FSE at (100, 200 mg/kg) plus cadmium significantly elevated serum albumin by about 35, 39, 26 and 37% respectively when compared to cadmium alone after 90 days. Although the level of albumin was restored to normal in group treated with GSE at two doses and FSE at high dose plus Al, Pb, Cd. FSE at low dose did not improve the level of albumin.
3. In intoxicated aluminum, lead or cadmium groups the serum total bilirubin was markedly increased compared to control group by about 2 fold after 45 and 90 days. On the other hand, the treatment with either GSE or FSE (100, 200 mg/kg) concurrently with aluminum produced a significant decrease in serum total bilirubin compared to animal treated with aluminum alone by about 16, 13, 5 and 14% respectively and raised compared to control group by about 32, 39, 50 and 36% respectively after 45 days while, after 90 days the level of total bilirubin was reduced by about 26, 18, 14 and 24% respectively compared to aluminum alone and raised by about 16, 28, 33 and 19% respectively compared to control group. Co- administration of GSE (100 mg/kg) plus lead had no significant effect on total bilirubin level comparing to lead alone after 45 days while after 90 days it was reduced by about 23% when compared to lead alone and 80% when compared to control group . Whereas GSE, FSE at high dose with lead reduced the in total bilirubin by about 10 and 18% respectively when compared to lead alone and raised by about 92 and 73% respectively comparing to control group after 45 days. While, after 90 days the level of total bilirubin were decreased about 29, 5 % comparing to lead alone and increased about 66 and 122 % when compared to control group. Moreover, the serum total bilirubin level was decreased in treated rats with GSE and FSE at two doses plus cadmium when compared to cadmium alone by about 30, 35, 32 and 32% respectively and raised by about 63, 50, 57 and 56% respectively when compared to control group after 90 days. In aluminum, lead, cadmium treated groups, the serum level of direct bilirubin was markedly increased compared to control group table (14) about 1.9, 2.5 and 2.1 fold respectively after 45 days while after 90 days this elevation significantly raised to 2.4, 2.7, 2.4 fold respectively. Simultaneous administration of GSE or FSE at two doses plus aluminum reduced the serum level of direct bilirubin compared to aluminum treated group by about 48, 33, 18 and 41% respectively after 90 days and 26, 63, 100 and 44% respectively compared to normal control. Whereas, in groups treated with GSE or FSE (100, 200 mg/kg) plus lead it was decreased by about 21, 36, 12 and 30 % respectively when compared to lead alone after 90 days and 115,74, 137 and 89% respectively compared to normal control. On the other hand co-administration of GSE or FSE (100, 200 mg/kg) plus cadmium significantly decreased it compared to cadmium alone by about 26, 29, 14 and 32% respectively after 90 days and 81,74,111 and 67% respectively compared to normal control.
4. The levels of serum urea and creatinine were increased in groups treated with aluminum, cadmium and lead after 45 and 90 days of the experiment. Whereas, the level of urea was restored to normal in group treated with FSE (100, 200 mg/kg) concurrently with aluminum, lead or cadmium. On other hand, simultaneous treatment with GSE (100 and 200 mg/kg) plus cadmium normalized urea level. While, co-administration of GSE at two doses plus lead acetate restored urea level to nearly normal control. Simultaneous administration of GSE (100 and 200 mg/kg) with aluminum reduced the elevated serum creatinine level by about 19 and 27% respectively after 45 days while after 90 days this reduction was increased to reach 36 and 42% respectively when compared to aluminum alone and 27, 15, 24 and 12 % respectively comparing to normal control after 45 and 90 days. Also, co administration of GSE (100, 200 mg/kg) with lead reduced the raised serum creatinine level by about 22 and 34% respectively after 45 and 26 and 33% after 90 days when compared to lead alone and 33, 13, 26 and 13% respectively compared to normal control after 45 and 90 days. Whereas in groups treated with GSE (100, 200 mg/kg) with cadmium, it was reduced by about 23 and 31% respectively after 45 days while after 90 days it was decreased by about 37 and 41% when compared to cadmium alone and 41, 27, 26 and 18% respectively compared to normal control after 45 and 90 days. The groups treated with FSE (100, 200 mg/kg) plus cadmium significantly reduced serum creatinine level by about 15 and 21% respectively after 45 days, while after 90 days it was reduced by about 35 and 42% respectively when comparing to aluminum alone and 32, 24, 26 and 13% respectively compared to normal control. Also, group treated with FSE (100, 200 mg/kg) plus lead decreased it by about 23 and 25% respectively after 45 days, while after 90 days it was reduced by about 23 and 27% respectively when compared to lead alone and 32, 28, 32 and 24% respectively comparing to normal control. In the groups which received cadmium plus FSE (100, 200 mg/kg) the elevated level of serum creatinine was by about 18 and 26% respectively after 45 days while after 90 days it was decreased by about 34 and 37% respectively when compared to cadmium alone and 51,35,33 and 26% respectively comparing to normal control.
5. In aluminum, lead or cadmium treated group after 45 and 90 days the serum total cholesterol level was significantly elevated by about 1.9 fold when compared to control group. On the other hand, the treatment with either GSE or FSE (100, 200 mg/kg) concurrently with aluminum produced a decrease in the serum total cholesterol compared to animal treated with aluminum alone by about 40, 42, 37 and 26% respectively after 90 days and 10, 9, 17 and 38% respectively compared to normal control. Moreover, the serum total cholesterol was reduced in treated rats with GSE or FSE (100, 200 mg/kg) plus lead when compared to lead alone after 90 days by about 27, 43, 29 and 36% respectively and 45, 14, 42 and 30% respectively compared to normal control. Whereas, in groups treated with GSE or FSE (100, 200mg/kg) plus cadmium it was decreased by about 27, 29, 38 and 46% respectively when compared to cadmium alone after 90 days and 43, 39,22 and 6% respectively compared to normal control.
6 Intoxicated aluminum, lead or cadmium group after 45 days significantly elevated the serum triglyceride level by about 1.5 fold while after 90 days this elevation significantly raised again to 1.8 fold when compared to control group. On contrast, treatment with either GSE or FSE (100, 200 mg/kg) concurrently with aluminum recovered the level of serum triglyceride to normal control.
7 Aluminum, lead or cadmium after 45 days of treatment significantly decreased the serum HDL level by about 46, 46 and 48% respectively while after 90 days they reduced it by about 48, 51 and 53% respectively when compared to control group. On the other hand, treatment with either GSE or FSE (100, 200 mg/kg) concurrently with aluminum produced significant increase in serum HDL compared to animal treated with aluminum alone by about 66, 62, 45 and 67 % respectively after 90 days. Moreover, the serum HDL was markedly raised in treated rats with GSE or FSE (100, 200 mg/kg) plus lead when compared to lead alone after 90 days by about 75, 76, 61 and 83 % respectively. Whereas, groups treated with GSE or FSE (100, 200 mg/kg) plus cadmium were increased by about 74, 87, 66 and 99 % respectively when compared to cadmium alone after 90 days.
8 In intoxicated aluminum, lead or cadmium group after 45days, the serum LDL level was significantly elevated by about 2.5 fold while after 90 days this elevation significantly raised again to 3 fold when compared to control group. On the other hand, the treatment with either GSE or FSE (100, 200 mg/kg) concurrently with aluminum produced a decrease in the serum LDL compared to animal treated with aluminum alone about 55, 54, 55 and 58% respectively after 90 days. Moreover, the serum LDL was reduced in treated rats with GSE or FSE (100, 200 mg/kg) plus lead when compared to lead alone after 90 days by about 54, 58, 50 and 47% respectively. Whereas, in groups treated with GSE or FSE (100, 200mg/kg) plus cadmium it was decreased by about 54, 57, 59 and 64% respectively when compared to cadmium alone after 90 days.
9 Chronic administration of aluminum, lead or cadmium in to rats resulted in markedly oxidative stress as indicated by increase the lipid peroxidation by about 2.3, 2.3 and 2.4 fold respectively, while they decreased reduced glutathione levels by about 82, 78 and 90% respectively and catalase by 82, 76 and 87% respectively when compared to control group.
Concomitant GSE or FSE (100, 200 mg/kg) administration to aluminum chloride treated rats caused a significant decrease in the level of MDA compared to animal treated with aluminum alone about 22, 29, 42 and 38 % respectively, and higher than normal control by about 80, 62, 34 and 42% respectively. Simultaneous administration of GSE or FSE (100, 200 mg/kg) plus lead acetate reduced the elevated hepatic MDA by about 28, 32, 12, and 16% respectively and by 68, 59, 105 and 95% comparing to control. Whereas, GSE or FSE (100, 200 mg/kg) treatment did not offer any protection on lipid peroxidation in cadmium treated rats.
The results of this study revealed that oral administration of GSE or FSE (100, 200 mg/kg) for three months significantly attenuated the reduction in GSH and CAT activities in rat treated with aluminum, lead or cadmium. Concurrent administration of GSE or FSE (100, 200 mg/kg) with aluminum chloride significantly raised hepatic GSH activity by about 182, 35, 107 and 209 % respectively compared to aluminum alone and by 51, 77, 64 and 46% respectively compared to control group. Similarly, simultaneous administration of GSE or FSE (100, 200 mg/kg) with lead acetate significantly increased hepatic GSH activity by about 207, 212, 85 and 72 % respectively compared to lead alone and by 33, 32, 59 and 62% respectively compared to control group. Whereas, concurrent administration of GSE or FSE (100, 200 mg/kg) with cadmium significantly raised hepatic GSH activity by about 187, 247, 252 and 229 % respectively compared to cadmium alone While lower than control 71, 66, 64 and 65% respectively compared to control group. In the same line, the treatment with either GSE (100, 200mg/kg) or FSE (100 mg/kg) concurrently with aluminum produced significant increase in CATactivity by about 85, 107 and 161% respectively compared to aluminum alone and by about 69, 65 and 45% respectively compared to control group. Whereas, FSE (200 mg/kg) restored CAT activity to normal value. Similarly, simultaneous administration of FSE (100, 200 mg/kg) with lead acetate significantly increased hepatic CAT activity by about 200 and 147% respectively compared to lead alone and by 27, and 40% respectively compared to control group. Whereas, GSE (100, 200mg/kg) plus lead restored CAT activity to normal. Concurrent administration of GSE or FSE (100, 200 mg/kg) with cadmium significantly raised hepatic CAT activity by about 220, 290 110 and 130 % respectively compared to cadmium alone and by 59, 50 73 and 71% respectively compared to control group.
The changes in total hemoglobin (Hb), RBCs, WBCs PCV and differential leucocytes count indicated narrow ranges in group treated with grape seeds and flaxseeds extracts comparing to the normal values in animals control. Statistical studies of the data revealed no significant differences between them. Significantly decreasing effects were observed on the of total hemoglobin concentration, RBCs and PCV between aluminum, cadmium or lead compared with normal control. But the levels of WBCs were higher by administration of aluminum and lead and decreased in rat administrated cadmium compared with normal control. While, total hemoglobin concentration, RBCs and PCV in rats treated with grape seeds, flaxseeds with heavy metals showed a significant decrease when compared with control group but higher than that found in rats treated with heavy metals alone.
Neutrophil ratio was decreased in rat administrated aluminum, lead and cadmium compared with control group, while there were no significant differences in other groups compared with control, but lymphocyte ratio was increased in rat treated with aluminum, lead and cadmium compared with control group.