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The b.w. and food and water intake of each rat were weighed using an analytical balance with an accuracy of 0.001. No significant b.w. modification was noted during the first week of treatment by TBZ. A week later, the b.w. of the TBZ-treated group was slightly lower than that of the control group and groups co-treated with trace elements. Two-way ANOVA test demonstrated the significant effect of TBZ (P < 0.001).
Meanwhile, the b.w. increased in all co-treated groups, with the most significant change occurring in the fourth week (Table 1). In the TBZ + Cu-treated group, the b.w. increased after four days of treatment and remained almost constant throughout the treatment period. Post hoc comparisons revealed that in the TBZ + (Se + Zn), TBZ + Fe, and TBZ + (Se + Fe + Cu + Zn) groups, a significant increase in b.w. was noted compared with the TBZ-treated rats.
Table 1. Initial and final body weights, absolute and relative testes weights, daily food and water consumption by control and treated rats with TBZ (100 mg/kg) and TBZ + (Cu + Zn + Fe + Se), TBZ + (Se + Zn), TBZ + Cu, or TBZ + Fe
Parameters Groups Control TBZ TBZ + (Se + Fe + Zn + Cu) TBZ + (Se + Zn) TBZ + Cu TBZ + Fe Initial body weights (g) 192 ± 1.08 193.25 ± 2.31 195 ± 0.98 199.75 ± 1.92 194.75 ± 1.95 197.5 ± 4.66 Final body weights (g) 198.75 ± 2.21 172.5 ± 1.19 212.5 ± 1.09 218 ± 2.47 209 ± 4.65 213.05 ± 4.81 Change in body weights (%) +6b −20a +17c +18c +14c +16c Absolute testes weights (g) 2.04 ± 0.51 1.94 ± 0.12 2.24 ± 0.11 2.14 ± 0.10 2.09 ± 0.24 2.15 ± 0.09 Relative testes weight (g/g BW) 1.08 ± 0.13 1.02 ± 0.1 1.08 ± 0.2 1.07 ± 0.15 0.91 ± 0.22 0.96 ± 0.15 Food consumption (g/day/rat) 17.05 ± 3.52 15.72 ± 3.54 16.57 ± 3.75 16.31 ± 3.67 16.57 ± 3.82 16.43 ± 3.70 Drinking water intake (mL/d per/rat) 19.13 ± 2.83 24.45 ± 6.28 18.91 ± 4.24 18.20 ± 6.18 21.56 ± 5.53 22.29 ± 6.51 Note. Values were expressed as means ± SD. The number of determinations was N = 8. Different superscript letters (a, b, c) indicate significant differences at P < 0.05. Similarly, food intake in the first week was comparatively small for all groups. The slight decrease in the second week and distinct decreases in the third and fourth weeks after TBZ treatment were recovered by Se, Zn, Cu, and Fe to a certain extent (Table 1).
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Table 2 shows the concentrations of minerals in the plasma and testis of rats. Two-way ANOVA demonstrated a significant perturbation in mineral composition induced by TBZ. The calcium, iron, zinc, potassium, magnesium, and sodium concentrations significantly increased in the plasma and decreased in the testis of TBZ exposed-rats when compared with the control animals (P < 0.001). Phosphorus concentration was significantly lower in the plasma and higher in the testis.
Table 2. Mineral levels in plasma and testis of controls and rats treated with TBZ (100 mg/kg) and TBZ + (Cu + Zn + Fe + Se), TBZ + (Se + Zn), TBZ + Cu, or TBZ + Fe
Parameters and treatments Controls TBZ TBZ + (Cu + Zn + Fe + Se) TBZ + (Se + Zn) TBZ + Cu TBZ + Fe Plasma levels (mg/L) Calcium 48.42 ± 3.32a 71.43 ± 9.44cd 66.84 ± 1.37b 72.22 ± 9.10d 69.98 ± 2.43bc 68.54 ± 2.15b Phosphorus 45.14 ± 1.30d 12.18 ± 4.80a 56.57 ± 5.15e 32.35 ± 3.65c 56.33 ± 4.80e 15.71 ± 4.56b Iron 0.49 ± 0.07a 0.99 ± 0.05e 0.78 ± 0.03d 0.52 ± 0.02b 0.69 ± 0.06c 1.20 ± 0.01e Zinc 0.020 ± 0.01a 0.44 ± 0.02d 0.049 ± 0.01b 0.92 ± 0.02f 0.36 ± 0.02c 0.24 ± 0.02e Magnesium 0.28 ± 0.02a 1.56 ± 0.01e 0.43 ± 0.03b 0.82 ± 0.11c 1.32 ± 0.01d 0.83 ± 0.02c Potassium 132 ± 0.77b 350.92 ± 5.94f 176.39 ± 0.36d 150 ± 0.82c 227.5 ± 6.37e 83.21 ± 2.89a Sodium 107.09 ± 0.12b 147.21 ± 2.45f 85.01 ± 1.55a 137.08 ± 1.47d 144.20 ± 4.56e 120.93 ± 2.65c Copper 0.09 ± 0.01b 0.11 ± 0.06c 0.16 ± 0.02d 0.22 ± 0.01e 0.18 ± 0.02e 0.04 ± 0.01a Testes levels (mg/L) Calcium 61.32 ± 3.20c 15.71 ± 4.40a 73.58 ± 3.42d 58.80 ± 0.79b 56.33 ± 1.49b 58.58 ± 1.04b Phosphorus 57.48 ± 2.80a 73.38 ± 6.16c 69.98 ± 3.74b 66.84 ± 1.79b 71.34 ± 2.39c 61.32 ± 3.49a Iron 0.60 ± 0.03c 0.030 ± 0.01a 0.73 ± 0.01d 0.32 ± 0.07b 0.56 ± 0.09c 0.89 ± 0.05e Zinc 0.12 ± 0.01b 0.06 ± 0.01a 0.25 ± 0.02c 0.40 ± 0.02a 0.17 ± 0.02b 0.18 ± 0.05b Magnesium 53.35 ± 1.20d 29.48 ± 0.15a 30.05 ± 4.75a 45.78 ± 2.40c 32.76 ± 0.92b 41.35 ± 7.40c Potassium 83.21 ± 0.62d 24.82 ± 3.55a 61.43 ± 2.12c 28.51 ± 2.27a 30.34 ± 0.1ab 34.71 ± 1.23b Sodium 93.83 ± 8.50e 25.21 ± 5.10a 87.67 ± 5.40d 26.07 ± 1.10a 30.82 ± 0.45b 43.65 ± 1.45c Copper 0.04 ± 0.01d 0.01 ± 0.06a 0.03 ± 0.02c 0.02 ± 0.09b 0.03 ± 0.7c 0.02 ± 0.01b Note. Values are expressed as means ± SD for 3 animals in each group.
Different superscript letters (a, b, c, d, e) indicate significant differences at P < 0.05.Post hoc comparisons revealed that trace elements treatment attenuated the changes in mineral levels compared with the TBZ-treated rats. Upon supplementation with Se, Fe, Cu, and Zn, modulations and improvements in mineral contents, especially calcium and phosphorus, were determined compared with the TBZ-treated group (Table 2).
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Two-way ANOVA of sperm mobility, sperm viability, and sperm count in the testes demonstrated the significant effect of TBZ (P < 0.001). Post hoc comparison revealed that sperm motility significantly decreased (P < 0.001) in the TBZ-treated group, along with a significant increase in the abnormal morphology of spermatozoa (Table 3). Trace elements (Se, Cu, Zn, and Fe) associated to TBZ significantly offset the decrease in functional sperm parameters (motility, viability, and count) when compared with the TBZ-treated rats (Table 3).
Table 3. Sperm function in spermatozoa of rats controls and treated with TBZ (100 mg/kg) and TBZ + (Cu + Zn + Fe + Se), TBZ + (Se + Zn), TBZ + Cu, or TBZ + Fe during 30 d
Parameters Groups Control TBZ TBZ + (Se + Fe + Cu + Zn) TBZ + (Se + Zn) TBZ + Cu TBZ + Fe SPZ count X 106 per mL 32.67 ± 7.39a 12.05 ± 1.56a 18.87 ± 5.76bc 19.76 ± 6.76bc 20.54 ± 3.87bc 19.78 ± 6.87ab Motility (%) 76.87 ± 5.75e 43.5 ± 7.87a 59.87 ± 4.87b 62.7 ± 9.4d 55.54 ± 4c 54.67 ± 6.87c Viability (%) 82.5 ± 6.76f 44.7 ± 8.9a 61.4 ± 4,22d 68.9 ± 7.76e 57.87 ± 5.87c 52.78 ± 3.76b Abnormal morphology (%) 7.9 ± 1.56a 22.6 ± 3.89C 13.56 ± 1.45b 12.1 ± 2.6b 12.6 ± 4.6b 11.8 ± 1.4b Note. Values are expressed as means ± SD for 6 animals in each group.
Different superscript letters (a, b, c, d, e) indicate significant differences at P < 0.05. -
Figure 1 presents the sexual hormone concentrations in the plasma of experimental rats. Statistical analysis of sexual hormones levels in the plasma showed the significant effect of TBZ (P < 0.001). Two-way ANOVA of data showed a significant decrease in testosterone and an increase in FSH and LH levels compared with the control (P < 0.001).
Figure 1. Testosterone, FSH, and LH levels in the plasma of TBZ, TBZ + (Se + Fe + Zn + Cu), TBZ + (Se + Zn), TBZ+ Cu, or TBZ + Fe during 30 d of treatment.
Post hoc comparisons showed that the changes in testosterone levels observed in the plasma of TBZ-treated rats were markedly abrogated by trace elements treatment without reaching the control values (P < 0.05). No significant differences were observed between the trace elements and TBZ-treated group in terms of LH and FSH levels.
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The oxidative stress induced by TBZ was tested initially in the rat models by measuring the MDA and AOPP levels in the testis tissue injected with TBZ (Table 4). Statistical analysis showed an increase in MDA and AOPP levels by TBZ treatment compared with the vehicle group (P < 0.001) (Table 4).
Table 4. Levels of MDA, GSH and activities of SOD, and GPx in the testes of controls and rats treated with TBZ (100 mg/kg) and TBZ + (Cu + Zn + Fe + Se), TBZ + (Se + Zn), TBZ + Cu, or TBZ + Fe in the testes during 30 d
Parameters Groups Control TBZ TBZ + (Se + Fe + Cu + Zn) TBZ + (Se + Zn) TBZ + Cu TBZ + Fe MDA 89.43 ± 7.39a 129.35 ± 6.89e 91.49 ± 8.71ab 96.98 ± 7.88c 99.78 ± 8.27d 92.83 ± 9.19b AOPP 0.53 ± 0.03a 0.84 ± 0.02c 0.58 ± 0.03a 0.70 ± 0.02b 0.82 ± 0.16c 0.67 ± 0.19b GSH 292.65 ± 43.7a 477.2 ± 98.14f 320.57 ± 26.6c 367.6 ± 24.2d 312.49 ± 14.31b 384.32 ± 10.22e GPX 6.94 ± 0.75b 3.96 ± 0.62a 5.50 ± 0.41ab 5.15 ± 0.69ab 5.76 ± 0.78ab 5.23 ± 1.23ab SOD 92.44 ± 5.87d 53.29 ± 6.04a 72.42 ± 6.24c 70.24 ± 8.38c 66.29 ± 6.43b 64.33 ± 7.11b Note. Values are expressed as means ± SD for 6 animals in each group.
Different superscript letters (a,b,c,d,e) indicate significant differences at P < 0.05.
MDA: nmol of MDA/g tissue.
AOPP: μmol/mg protein.
GSH: μg/g tissue.
GPX: nmol of GSH/min/mg of Protein.
SOD: μmol/mg of protein.Post hoc comparisons demonstrated a difference between TBZ- and trace elements-treated rats (P < 0.001). Se, Fe, and Zn addition restored the MDA and AOPP concentrations compared with the TBZ-injected rats (P < 0.001). However, Cu addition caused no effect on the AOPP levels compared with TBZ treatment.
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Two-way ANOVA test showed that injection of 100 mg/kg b.w. TBZ caused a decrease in GPx and SOD activities and an increase in GSH level compared with the untreated group (P < 0.001) (Table 4).
Post hoc comparison demonstrated that trace element administration protected against antioxidant status changes by reversing the increase in SOD and GPx activities and decrease in GSH level compared with the TBZ-treated rats.
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The statistical analysis results presented in Figure 2 show the decrease in DNA contents in the testes of TBZ-treated rats compared with the control animals. Meanwhile, Se, Cu, Zn, and Fe administration showed an increase in DNA contents and reached normal values compared with the TBZ-treated group (Figure 2).
Figure 2. (A): Agarose gel electrophoresis of DNA fragmentation. Lane 1: control group, lane 2: TBZ-treated group, lane 3: TBZ + (Cu + Zn + Fe + Se)-treated group, lane 4: TBZ + (Se + Zn), lane 5: TBZ + Cu, lane 6: TBZ + Fe. (B): testicle DNA quantity.
Agarose gel electrophoresis demonstrated an undetectable DNA scale in the testes of control rats. The DNA intact band was condensed near the application point with no DNA smearing, suggesting that no DNA fragmentation occurred in the control group, whereas a smear without ladder formation on agarose gel was observed in the TBZ-treated rats. The supplementation of trace elements associated to TBZ resulted in the absence of DNA smearing (Figure 2).
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Figure 3 displays the representative histopathological sections of the testes. H&E-stained testis sections of control rats showed an intact and smooth structure, indicating the normal morphology and organization of seminiferous tubule with all successive stages of spermatogenesis, lumen filled with spermatozoa, and presence of Sertoli cells (Figure 3A). On the contrary, the testes of rats treated with TBZ showed degenerated germinal epithelium and lumen with less sperms. Further, decreases in tubular diameter, numbers of spermatocytes, Leydig and Sertoli cells, and increase of intertubular space and lumen with cellular debris were observed in the testes of TBZ-treated rats. In the same manner, inflammatory infiltration was observed in the interstitium of the group that received TBZ (Figure 3B). These modifications were inhibited by Se, Fe, Zn, and Cu co-administration, resulting in highly regular seminiferous tubules showing spermatogenesis at the level of spermatocytes with normal interstitial tissue (Figures 3C, 3D, 3E, and 3F).
The co-administration of all trace elements, provoked a marked improvement in the testicular structure, especially in the (Zn + Se) + TBZ-treated group (Figures 3C and 3E).
doi: 10.3967/bes2020.101
Efficacy of Essential Trace Elements Supplementation on Mineral Composition, Sperm Characteristics, Antioxidant Status, and Genotoxicity in Testis of Tebuconazole-treated Rats
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Abstract:
Objective This research was performed to evaluate the effect of tebuconazole (TBZ) on reproductive organs of male rats and to assess the protective role of combined essential trace elements in alleviating the detrimental effect of TBZ on male reproductive function. Methods For this purpose, 48 rats were exposed to 100 mg/kg TBZ, TBZ supplemented with zinc (Zn), selenium (Se), copper (Cu), and iron (Fe), TBZ + (Se + Zn); TBZ + Cu; or TBZ + Fe. The experiment was conducted for 30 consecutive days. Results TBZ caused a significant perturbation in mineral levels and reduction in reproductive organs weights, plasma testosterone level, and testicular antioxidant enzyme activities. The TBZ-treated group also showed a significant increase in sperm abnormalities (count, motility, and viability percent), plasma follicle-stimulating hormone and luteinizing hormone concentrations, lipid peroxidation, protein oxidation, and severe DNA degradation in comparison with the controls. Histopathologically, TBZ caused testis impairments. Conversely, treatment with trace elements, in combination or alone, improved the reproductive organ weights, sperm characteristics, TBZ-induced toxicity, and histopathological modifications in testis. Conclusion TBZ exerts significant harmful effects on male reproductive system. The concurrent administration of trace elements reduces testis dysfunction, fertility, and toxicity induced by TBZ. -
Key words:
- Zinc /
- Selenium /
- Iron /
- Copper /
- Testicular toxicity
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Figure 1. Testosterone, FSH, and LH levels in the plasma of TBZ, TBZ + (Se + Fe + Zn + Cu), TBZ + (Se + Zn), TBZ+ Cu, or TBZ + Fe during 30 d of treatment.
Values are expressed as means ± SD. The number of determinations was N = 3. Different superscript letters (a, b, and c) indicate significant differences at P < 0.05.
Figure 2. (A): Agarose gel electrophoresis of DNA fragmentation. Lane 1: control group, lane 2: TBZ-treated group, lane 3: TBZ + (Cu + Zn + Fe + Se)-treated group, lane 4: TBZ + (Se + Zn), lane 5: TBZ + Cu, lane 6: TBZ + Fe. (B): testicle DNA quantity.
Values are expressed as means ± SD. The number of determinations was N = 3. Different superscript letters (a, b, c, and d) indicate significant differences at P < 0.05.
Table 1. Initial and final body weights, absolute and relative testes weights, daily food and water consumption by control and treated rats with TBZ (100 mg/kg) and TBZ + (Cu + Zn + Fe + Se), TBZ + (Se + Zn), TBZ + Cu, or TBZ + Fe
Parameters Groups Control TBZ TBZ + (Se + Fe + Zn + Cu) TBZ + (Se + Zn) TBZ + Cu TBZ + Fe Initial body weights (g) 192 ± 1.08 193.25 ± 2.31 195 ± 0.98 199.75 ± 1.92 194.75 ± 1.95 197.5 ± 4.66 Final body weights (g) 198.75 ± 2.21 172.5 ± 1.19 212.5 ± 1.09 218 ± 2.47 209 ± 4.65 213.05 ± 4.81 Change in body weights (%) +6b −20a +17c +18c +14c +16c Absolute testes weights (g) 2.04 ± 0.51 1.94 ± 0.12 2.24 ± 0.11 2.14 ± 0.10 2.09 ± 0.24 2.15 ± 0.09 Relative testes weight (g/g BW) 1.08 ± 0.13 1.02 ± 0.1 1.08 ± 0.2 1.07 ± 0.15 0.91 ± 0.22 0.96 ± 0.15 Food consumption (g/day/rat) 17.05 ± 3.52 15.72 ± 3.54 16.57 ± 3.75 16.31 ± 3.67 16.57 ± 3.82 16.43 ± 3.70 Drinking water intake (mL/d per/rat) 19.13 ± 2.83 24.45 ± 6.28 18.91 ± 4.24 18.20 ± 6.18 21.56 ± 5.53 22.29 ± 6.51 Note. Values were expressed as means ± SD. The number of determinations was N = 8. Different superscript letters (a, b, c) indicate significant differences at P < 0.05. Table 2. Mineral levels in plasma and testis of controls and rats treated with TBZ (100 mg/kg) and TBZ + (Cu + Zn + Fe + Se), TBZ + (Se + Zn), TBZ + Cu, or TBZ + Fe
Parameters and treatments Controls TBZ TBZ + (Cu + Zn + Fe + Se) TBZ + (Se + Zn) TBZ + Cu TBZ + Fe Plasma levels (mg/L) Calcium 48.42 ± 3.32a 71.43 ± 9.44cd 66.84 ± 1.37b 72.22 ± 9.10d 69.98 ± 2.43bc 68.54 ± 2.15b Phosphorus 45.14 ± 1.30d 12.18 ± 4.80a 56.57 ± 5.15e 32.35 ± 3.65c 56.33 ± 4.80e 15.71 ± 4.56b Iron 0.49 ± 0.07a 0.99 ± 0.05e 0.78 ± 0.03d 0.52 ± 0.02b 0.69 ± 0.06c 1.20 ± 0.01e Zinc 0.020 ± 0.01a 0.44 ± 0.02d 0.049 ± 0.01b 0.92 ± 0.02f 0.36 ± 0.02c 0.24 ± 0.02e Magnesium 0.28 ± 0.02a 1.56 ± 0.01e 0.43 ± 0.03b 0.82 ± 0.11c 1.32 ± 0.01d 0.83 ± 0.02c Potassium 132 ± 0.77b 350.92 ± 5.94f 176.39 ± 0.36d 150 ± 0.82c 227.5 ± 6.37e 83.21 ± 2.89a Sodium 107.09 ± 0.12b 147.21 ± 2.45f 85.01 ± 1.55a 137.08 ± 1.47d 144.20 ± 4.56e 120.93 ± 2.65c Copper 0.09 ± 0.01b 0.11 ± 0.06c 0.16 ± 0.02d 0.22 ± 0.01e 0.18 ± 0.02e 0.04 ± 0.01a Testes levels (mg/L) Calcium 61.32 ± 3.20c 15.71 ± 4.40a 73.58 ± 3.42d 58.80 ± 0.79b 56.33 ± 1.49b 58.58 ± 1.04b Phosphorus 57.48 ± 2.80a 73.38 ± 6.16c 69.98 ± 3.74b 66.84 ± 1.79b 71.34 ± 2.39c 61.32 ± 3.49a Iron 0.60 ± 0.03c 0.030 ± 0.01a 0.73 ± 0.01d 0.32 ± 0.07b 0.56 ± 0.09c 0.89 ± 0.05e Zinc 0.12 ± 0.01b 0.06 ± 0.01a 0.25 ± 0.02c 0.40 ± 0.02a 0.17 ± 0.02b 0.18 ± 0.05b Magnesium 53.35 ± 1.20d 29.48 ± 0.15a 30.05 ± 4.75a 45.78 ± 2.40c 32.76 ± 0.92b 41.35 ± 7.40c Potassium 83.21 ± 0.62d 24.82 ± 3.55a 61.43 ± 2.12c 28.51 ± 2.27a 30.34 ± 0.1ab 34.71 ± 1.23b Sodium 93.83 ± 8.50e 25.21 ± 5.10a 87.67 ± 5.40d 26.07 ± 1.10a 30.82 ± 0.45b 43.65 ± 1.45c Copper 0.04 ± 0.01d 0.01 ± 0.06a 0.03 ± 0.02c 0.02 ± 0.09b 0.03 ± 0.7c 0.02 ± 0.01b Note. Values are expressed as means ± SD for 3 animals in each group.
Different superscript letters (a, b, c, d, e) indicate significant differences at P < 0.05.Table 3. Sperm function in spermatozoa of rats controls and treated with TBZ (100 mg/kg) and TBZ + (Cu + Zn + Fe + Se), TBZ + (Se + Zn), TBZ + Cu, or TBZ + Fe during 30 d
Parameters Groups Control TBZ TBZ + (Se + Fe + Cu + Zn) TBZ + (Se + Zn) TBZ + Cu TBZ + Fe SPZ count X 106 per mL 32.67 ± 7.39a 12.05 ± 1.56a 18.87 ± 5.76bc 19.76 ± 6.76bc 20.54 ± 3.87bc 19.78 ± 6.87ab Motility (%) 76.87 ± 5.75e 43.5 ± 7.87a 59.87 ± 4.87b 62.7 ± 9.4d 55.54 ± 4c 54.67 ± 6.87c Viability (%) 82.5 ± 6.76f 44.7 ± 8.9a 61.4 ± 4,22d 68.9 ± 7.76e 57.87 ± 5.87c 52.78 ± 3.76b Abnormal morphology (%) 7.9 ± 1.56a 22.6 ± 3.89C 13.56 ± 1.45b 12.1 ± 2.6b 12.6 ± 4.6b 11.8 ± 1.4b Note. Values are expressed as means ± SD for 6 animals in each group.
Different superscript letters (a, b, c, d, e) indicate significant differences at P < 0.05.Table 4. Levels of MDA, GSH and activities of SOD, and GPx in the testes of controls and rats treated with TBZ (100 mg/kg) and TBZ + (Cu + Zn + Fe + Se), TBZ + (Se + Zn), TBZ + Cu, or TBZ + Fe in the testes during 30 d
Parameters Groups Control TBZ TBZ + (Se + Fe + Cu + Zn) TBZ + (Se + Zn) TBZ + Cu TBZ + Fe MDA 89.43 ± 7.39a 129.35 ± 6.89e 91.49 ± 8.71ab 96.98 ± 7.88c 99.78 ± 8.27d 92.83 ± 9.19b AOPP 0.53 ± 0.03a 0.84 ± 0.02c 0.58 ± 0.03a 0.70 ± 0.02b 0.82 ± 0.16c 0.67 ± 0.19b GSH 292.65 ± 43.7a 477.2 ± 98.14f 320.57 ± 26.6c 367.6 ± 24.2d 312.49 ± 14.31b 384.32 ± 10.22e GPX 6.94 ± 0.75b 3.96 ± 0.62a 5.50 ± 0.41ab 5.15 ± 0.69ab 5.76 ± 0.78ab 5.23 ± 1.23ab SOD 92.44 ± 5.87d 53.29 ± 6.04a 72.42 ± 6.24c 70.24 ± 8.38c 66.29 ± 6.43b 64.33 ± 7.11b Note. Values are expressed as means ± SD for 6 animals in each group.
Different superscript letters (a,b,c,d,e) indicate significant differences at P < 0.05.
MDA: nmol of MDA/g tissue.
AOPP: μmol/mg protein.
GSH: μg/g tissue.
GPX: nmol of GSH/min/mg of Protein.
SOD: μmol/mg of protein. -
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