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卵巢包裹对玻璃化冷冻小鼠卵巢形态及凋亡相关基因表达的影响

The Effects of Ovarian Encapsulation on Morphology and Expression of Apoptosis-Related Genes in Vitrified Mouse Ovary.

作者信息

Shirazi Tehrani Atefeh, Mazoochi Tahereh, Akhavan Taheri Maryam, Aghadavood Esmat, Salehnia Mojdeh

机构信息

Gametogenesis Research Center, Kashan University of Medical Sciences, Kashan, Iran.

Anatomical Sciences Research Center, Kashan University of Medical Sciences, Kashan, Iran.

出版信息

J Reprod Infertil. 2021 Jan-Mar;22(1):23-31. doi: 10.18502/jri.v22i1.4992.

Abstract

BACKGROUND

The purpose of this study was to determine the effects of alginate hydrogel as a capsule to protect the ovary against possible detrimental effects of vitrification and warming on morphology and expression of apoptosis-related genes in the mouse ovary.

METHODS

In this experimental study, the ovaries from twenty-five female 8-week-old mice were divided into five groups of non-vitrified ovaries, vitrified ovaries, ovaries that were encapsulated with concentrations of 0.5, 0.75 and 1% of alginate hydrogel. The morphological study was performed using hematoxylin and eosin staining. Expression levels of apoptosis-associated genes were quantified in each group by real-time RT-PCR. The one-way ANOVA and post hoc test were used to analyze the data and values of p<0.05 were considered statistically significant.

RESULTS

The results of follicle count showed that the mean of total follicles in all groups was not significantly different. The average number of atretic follicles in vitrified and experimental groups significantly increased in comparison with the nonvitrified group (p=0.001). The results of the evaluation of apoptosis-related genes showed that the ratio of BAX/BCL-2 in experimental groups 1 and 2 was significantly higher than the vitrified group and experimental group 3 (p=0.000). The expression level of caspase 3 gene was not significantly different among all groups.

CONCLUSION

Ovarian encapsulation with used concentrations of alginate hydrogel failed to improve the morphology and molecular aspects of follicles and it was not able to better preserve the intact follicles of vitrified ovaries. However, morphological and molecular findings appear to improve with increasing alginate hydrogel concentration.

摘要

背景

本研究旨在确定海藻酸盐水凝胶作为一种胶囊对卵巢的保护作用,以对抗玻璃化冷冻及复温对小鼠卵巢形态和凋亡相关基因表达可能产生的有害影响。

方法

在本实验研究中,将25只8周龄雌性小鼠的卵巢分为五组:非玻璃化冷冻卵巢组、玻璃化冷冻卵巢组、用浓度为0.5%、0.75%和1%的海藻酸盐水凝胶包裹的卵巢组。使用苏木精和伊红染色进行形态学研究。通过实时逆转录聚合酶链反应对每组中凋亡相关基因的表达水平进行定量分析。采用单因素方差分析和事后检验分析数据,p<0.05的值被认为具有统计学意义。

结果

卵泡计数结果显示,所有组的总卵泡平均数无显著差异。与非玻璃化冷冻组相比,玻璃化冷冻组和实验组的闭锁卵泡平均数显著增加(p=0.001)。凋亡相关基因评估结果显示,实验组1和2中BAX/BCL-2的比值显著高于玻璃化冷冻组和实验组3(p=0.000)。caspase 3基因的表达水平在所有组之间无显著差异。

结论

使用的海藻酸盐水凝胶浓度对卵巢进行包裹未能改善卵泡的形态和分子方面,也无法更好地保存玻璃化冷冻卵巢中的完整卵泡。然而,随着海藻酸盐水凝胶浓度的增加,形态学和分子学结果似乎有所改善。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2a9/7903669/60a21e799cd1/JRI-22-23-g001.jpg

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