Abstract
Introduction Freezing of ovarian tissue is used for preservation of fertility. The freezing-thawing process is accompanied by oxidative stress and induction of apoptosis. Apoptosis is a complex process that has been studied in animal models. The present study was aimed at investigating the effect of selenium on suppression of apoptosis during vitrification-thawing process of mice ovary via studying expression of apoptosis-related genes, and also, we aimed to design statistical models for the roles of single genes and gene-gene interactions in suppression of apoptosis. Methods A total of 10 right ovary samples from 10 mice were randomly divided into two groups of selenium treatment (at dose 5 μg/ml sodium selenite, through adding to the media) and control group. Vitrification-thawing process was done according to the existed protocols. Real-time PCR was used for gene expression study. The apoptosis gene profile included P53, Bax, Fas, and Bcl-2. General linear model was applied to study single gene associations and gene-gene interactions. Results From the studied genes, P53 showed a significant downregulation in the selenium group in comparison to the control group (∆∆CT = 1.96; P = 0.013; relative expression (RE) = 0.28). Bcl-2 showed a significant upregulation in the selenium group in comparison to the control group (∆∆CT = −2.49; P < 0.001; RE = 3.49). No significant result was found for other genes. According to the multiple models, Bcl-2 showed a protective single gene association (beta = −0.33; P = 0.032), and Fas∗Bcl-2 interaction was significantly positive (beta = 0.19; P = 0.036). Conclusion Addition of selenium to cryomedia of vitrification-thawing process could reduce the apoptosis induced by freezing-thawing stress in mice ovary via downregulation of P53 and upregulation of Bcl-2 at transcription level. Multivariable statistical models should be performed in future researches to study biological systems.
Highlights
Freezing of ovarian tissue is used for preservation of fertility
Since there was a gap in evidence on the role of supplementation of nutritional elements to cryomedia of ovarian freezing-thawing process, this study was aimed at evaluating the role of selenium supplementation on reduction of apoptosis after vitrification of mice ovarian tissue through study of apoptosis-related genes including P53, Bax, Bcl-2, and Fas
Relative expression (RE) were calculated based on ΔCT(sample-control) amounts adjusted with the efficiencies while statistical analyses were done based on ΔCT(target-reference) and ΔΔCT amounts (Tables 2 and 3)
Summary
Freezing of ovarian tissue is used for preservation of fertility. The freezing-thawing process is accompanied by oxidative stress and induction of apoptosis. The present study was aimed at investigating the effect of selenium on suppression of apoptosis during vitrification-thawing process of mice ovary via studying expression of apoptosis-related genes, and we aimed to design statistical models for the roles of single genes and gene-gene interactions in suppression of apoptosis. P53 showed a significant downregulation in the selenium group in comparison to the control group (ΔΔCT = 1:96; P = 0:013; relative expression ðREÞ = 0:28). Addition of selenium to cryomedia of vitrification-thawing process could reduce the apoptosis induced by freezing-thawing stress in mice ovary via downregulation of P53 and upregulation of Bcl-2 at transcription level. Ovaries may be affected by chemical and physical cytotoxic agents [2] To solve this problem, freezing of ovarian samples was suggested [3]. Using additive antioxidants in cellular or tissue preservation media has been suggested [4]
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