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人类卵巢组织玻璃化冷冻/复温对凋亡相关基因的表达影响较小。

Human ovarian tissue vitrification/warming has minor effect on the expression of apoptosis-related genes.

作者信息

Abdollahi Maasoume, Salehnia Mojdeh, Salehpour Saghar, Ghorbanmehr Nassim

机构信息

Dept. of Anatomical Sciences, Medical Sciences Faculty, Tarbiat Modares University, Tehran, Iran.

出版信息

Iran Biomed J. 2013;17(4):179-86. doi: 10.6091/ibj.1243.2013.

Abstract

BACKGROUND

In this study, we evaluated the incidence of apoptosis at the ultrastructural levels and expression of some apoptosis-related genes in vitrified human ovarian tissue just after warming.

METHODS

Human ovarian tissue biopsies from 23 women after caesarean section were transported to the laboratory within 2 hours, and then they were cut into small pieces. Some pieces were vitrified and warmed and the other samples were considered as control. Apoptosis was assessed by a transmission electron microscope and also by molecular analysis of pro-apoptotic (Fas, FasL, Bax, p53, caspase8, and caspase3) and antiapoptotic (Bcl-2 and BIRC5) genem RNA levels using real-time RT-PCR before and after vitrification.

RESULTS

No sign of apoptosis was shown ultrastructurally in vitrified samples. The level of FasL, Bcl-2, Bax, p53, and caspase3 mRNA and Bax:Bcl-2 ratio were similar in non-vitrified and vitrified groups; however, the expression of Fas and caspase8 genes was higher and BIRC5 was lower in vitrified samples compared to non-vitrified group (P<0.05).

CONCLUSION

The fine structure of human vitrified ovarian tissue was well preserved; moreover, vitrification was shown to affect the expression of some apoptosis-related genes. However, additional study is needed to confirm this observation.

摘要

背景

在本研究中,我们评估了复温后玻璃化处理的人卵巢组织在超微结构水平上的凋亡发生率以及一些凋亡相关基因的表达情况。

方法

从23名剖宫产术后妇女获取的人卵巢组织活检标本在2小时内运送至实验室,然后切成小块。部分小块进行玻璃化处理及复温,其他样本作为对照。在玻璃化处理前后,通过透射电子显微镜评估凋亡情况,并使用实时逆转录聚合酶链反应对促凋亡基因(Fas、FasL、Bax、p53、caspase8和caspase3)和抗凋亡基因(Bcl-2和BIRC5)的RNA水平进行分子分析。

结果

玻璃化处理的样本在超微结构上未显示凋亡迹象。非玻璃化组和玻璃化组中FasL、Bcl-2、Bax、p53和caspase3 mRNA水平以及Bax:Bcl-2比值相似;然而,与非玻璃化组相比,玻璃化处理样本中Fas和caspase8基因的表达较高,而BIRC5的表达较低(P<0.05)。

结论

人玻璃化卵巢组织的精细结构保存良好;此外,玻璃化处理显示会影响一些凋亡相关基因的表达。然而,需要进一步研究来证实这一观察结果。

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