Anvari, Morteza
(Department of Biology and Anatomical Sciences, Shahid Sadoughi University of Medical Sciences)
,
Talebi, Ali Reza
(Department of Biology and Anatomical Sciences, Shahid Sadoughi University of Medical Sciences)
,
Mangoli, Esmat
(Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences)
,
Shahedi, Abbas
(Department of Biology and Anatomical Sciences, Shahid Sadoughi University of Medical Sciences)
,
Ghasemi, Mohammad Rasool
(Department of Biology and Anatomical Sciences, Shahid Sadoughi University of Medical Sciences)
,
Pourentezari, Majid
(Department of Biology and Anatomical Sciences, Shahid Sadoughi University of Medical Sciences)
Objective: The present study investigated sperm chromatin quality and testosterone levels in acrylamide-treated mice and the possible protective effects of vitamin E on the fertility potential of spermatozoa. Methods: Thirty-two adult male mice were divided equally into four groups. Group 1 was the ...
Objective: The present study investigated sperm chromatin quality and testosterone levels in acrylamide-treated mice and the possible protective effects of vitamin E on the fertility potential of spermatozoa. Methods: Thirty-two adult male mice were divided equally into four groups. Group 1 was the control, group 2 received acrylamide (10 mg/kg, water solution), group 3 received vitamin E (100 mg/kg, intraperitoneal), and group 4 received both acrylamide and vitamin E. After 35 days, spermatozoa from the right cauda epididymis were analyzed in terms of count, motility, morphology, and viability. Sperm DNA integrity and chromatin condensation were assessed by acridine orange (AO), aniline blue (AB), toluidine blue (TB), and chromomycin A3 (CMA3) staining. Results: In acrylamide-treated mice, significantly lower sperm concentration, viability, motility, and testosterone levels were found in comparison with the control and acrylamide+vitamin E groups (p< 0.05). In the vitamin E group, significantly more favorable sperm parameters and testosterone levels were found than in the other groups (p< 0.05). There were also significantly more spermatozoa with less condensed chromatin in the acrylamide-treated mice than in the other groups. Moreover, significantly more spermatozoa with mature nuclei (assessed by AB, CMA3, AO, and TB staining) were present in the vitamin E group than in the control and acrylamide+vitamin E groups. Conclusion: This study revealed the deleterious effects of acrylamide on sperm parameters and sperm chromatin quality. Vitamin E can not only compensate for the toxic effects of acrylamide, but also improve sperm chromatin quality in mice.
Objective: The present study investigated sperm chromatin quality and testosterone levels in acrylamide-treated mice and the possible protective effects of vitamin E on the fertility potential of spermatozoa. Methods: Thirty-two adult male mice were divided equally into four groups. Group 1 was the control, group 2 received acrylamide (10 mg/kg, water solution), group 3 received vitamin E (100 mg/kg, intraperitoneal), and group 4 received both acrylamide and vitamin E. After 35 days, spermatozoa from the right cauda epididymis were analyzed in terms of count, motility, morphology, and viability. Sperm DNA integrity and chromatin condensation were assessed by acridine orange (AO), aniline blue (AB), toluidine blue (TB), and chromomycin A3 (CMA3) staining. Results: In acrylamide-treated mice, significantly lower sperm concentration, viability, motility, and testosterone levels were found in comparison with the control and acrylamide+vitamin E groups (p< 0.05). In the vitamin E group, significantly more favorable sperm parameters and testosterone levels were found than in the other groups (p< 0.05). There were also significantly more spermatozoa with less condensed chromatin in the acrylamide-treated mice than in the other groups. Moreover, significantly more spermatozoa with mature nuclei (assessed by AB, CMA3, AO, and TB staining) were present in the vitamin E group than in the control and acrylamide+vitamin E groups. Conclusion: This study revealed the deleterious effects of acrylamide on sperm parameters and sperm chromatin quality. Vitamin E can not only compensate for the toxic effects of acrylamide, but also improve sperm chromatin quality in mice.
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문제 정의
To our knowledge, the effect of antioxidants in protecting sperm from acrylamideinduced damage has not yet been established. Accordingly, the purpose of this study was to investigate the beneficial effects of vitamin E on sperm parameters, chromatin condensation, and blood testosterone levels in acrylamide-treated mice.
An intriguing finding of our study was the improvement of sperm viability and motility in the vitamin E group when compared with the control animals. This effect of vitamin E might be due to its antioxidant role against oxidative stress induced by acrylamide.
The use of antioxidants increases sperm quality and DNA integrity [13]. Since compensating for the harmful effects of acrylamide on sperm nuclear structure is of paramount importance, the aim of the present study was to evaluate sperm chromatin condensation and testosterone levels after administration of vitamin E to acrylamide-treated mice.
대상 데이터
Chromatin condensation and DNA integrity assessments were carried out using cytochemical techniques including acridine orange (AO), toluidine blue (TB), aniline blue (AB), and chromomycin A3 (CMA3). All dyes and chemicals were purchased from Sigma Aldrich (St. Louis, MO, USA).
Thirty-two adult male Syrian mice (35 g, 10 weeks old) were assigned to four groups, each comprising eight mice. The mice in group 1 were fed a basal diet and served as the control group, group 2 received the basal diet and acrylamide (Merck, Darmstadt, Germany; 10 mg/kg, water solution) [7]; group 3 received the basal diet and vitamin E (Osvah Pharmaceutical, Tehran, Iran; 100 mg/kg, intraperitoneal) [14]; and group 4 received the basal diet, acrylamide, and vitamin E for 35 days (almost equal to the duration of a cycle of spermatogenesis).
데이터처리
All data are presented as mean ± standard deviation. One-way analysis of variance was applied to evaluate the significance of differences among the groups, and the Tukey post-hoc test was used to evaluate the differences between pairs of groups. A p-value ≤ 0.
이론/모형
Data normalization was performed using the Kolmogorov–Smirnov test.
성능/효과
According to our data from TB– staining, although a difference was observed in the percentage of TB-reacted spermatozoa among the groups, it was not statistically significant.
Table 2 presents the results of the sperm chromatin and DNA integrity evaluations. For AB staining, the percentages of unstained or pale blue-stained (normal spermatozoa) and dark blue-stained (abnormal spermatozoa) were reported. In TB staining, the chromatin quality of sperm was assessed according to metachromatic staining of sperm heads with the following scores: 0, light blue (good chromatin); 1, dark blue (mildly abnormal chromatin); 2, violet; and 3, purple (severely abnormal chromatin).
AB staining detects the presence of histones, and indirectly shows the presence of lower amounts of protamine in the sperm nucleus. In this study, we observed a higher proportion of spermatozoa with residual histones in the acrylamide group than in the control group, but no significant difference was found between the vitamin E and vitamin E+ acrylamide groups and the control mice. Moreover, we identified a higher proportion of CMA3+ (abnormal chromatin packaging) spermatozoa in acrylamidetreated mice than in the other three groups.
According to our data from TB– staining, although a difference was observed in the percentage of TB-reacted spermatozoa among the groups, it was not statistically significant. Moreover, the AO test showed a notably lower percentage of normal sperm in the acrylamide group than in the control, vitamin E, and vitamin E+acrylamide groups. It can thus be concluded that acrylamide causes the denaturation of sperm DNA strands and vitamin E diminishes this effect.
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