| 玻璃化冷冻对乳鼠卵巢Dnmt1和Grb10的影响研究 |
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| 引用本文： | 王红燕,周雪,王锐,王燕蓉,裴秀英,马良宏,颜贝,张帆,周岳,田稼.玻璃化冷冻对乳鼠卵巢Dnmt1和Grb10的影响研究[J].生殖与避孕,2017(1):27-32. |
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| 作者姓名： | 王红燕 周雪 王锐 王燕蓉 裴秀英 马良宏 颜贝 张帆 周岳 田稼 |
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| 作者单位： | 1. 750004银川,宁夏医科大学总医院宁夏人类精子库;750004银川,宁夏医科大学生育力保持教育部重点实验室;2. 宁夏医科大学生育力保持教育部重点实验室,银川,750004;3. 宁夏医科大学总医院宁夏人类精子库,银川,750004 |
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| 基金项目： | 国家自然科学基金项目(81460243;81460644)，宁夏回族自治区科技攻关项目(2012ZYS239)，宁夏医科大学生殖与遗传基础与临床创新团队项目(FGCT201505)National Foundation of Natural Science of China(81460243;81460644)，Science and Technology Project of Ningxia(2012ZYS239)，Reproductive and Genetic Basis and Clinical Innovation Project of Ningxia Medical University(FGCT201505) |
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| 摘 要： | 目的探索玻璃化冷冻对乳鼠卵巢Dnmt1和Grb10 mRNA和蛋白表达的影响。方法 26只出生10 d的C57BL/6雌性小鼠,每只乳鼠双侧卵巢均分成新鲜组和玻璃化冷冻组。免疫组织化学法检测Dnmt1蛋白在玻璃化冻融前、后卵巢组织卵泡中的表达变化;通过qRT-PCR检测Dnmt1和Grb10 mRNA在玻璃化冻融前、后卵巢组织中的表达变化;通过Western blotting方法检测Dnmt1和Grb10蛋白在玻璃化冻融前后卵巢组织中的表达变化。结果 Dnmt1在玻璃化冻融前、后的乳鼠卵巢组织各级卵泡的颗粒细胞及卵母细胞胞核表达。与新鲜组相比,玻璃化冷冻组卵巢内Dnmt1蛋白和mRNA表达水平均显著下调(P0.05);Grb10蛋白和mRNA表达水平均显著上调(P0.05)。结论玻璃化冷冻复苏过程导致卵巢内Dnmt1表达降低,使印记基因Grb10过表达,可能使配子糖代谢性表观遗传信息紊乱的的风险增加。
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| 关 键 词： | 乳鼠卵巢 玻璃化冷冻 DNA甲基转移酶1(Dnmt1) 生长因子受体结合蛋白10 (Grb10) |
Study on effection of DNA methyltransferase 1 (Dnmt1) and growth factor receptor-binding protein 10 (Grb10) genes on cryopreservation juvenil mouse ovaries by vitrification |
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| Abstract: | Objective To study the effect of vitrified-warmed juvenil mouse ovaries on the mRNA and protein expression of DNA methyltransferase 1 (Dnmt 1) and growth factor receptor-binding protein 10 (Grb 10).Methods Twenty-six 10-day-old C57BL/6 female mice were used in this experiment.The bilateral ovaries of each juvenile mouse were divided into two groups:the fresh group and the vitrified-warmed group.The expression of Dnmtl in follicles embedding was detected by immunohistochemistry.The expressions of Dnmt1 and Grb10 protein in fresh group and vitrified-warmed group were detected by Western blotting technology.The expressions of Dnmt1 and Grb10 mRNA in fresh groupand vitrified-warmed group were detected by qRT-PCR.Results Dnmtl protein all appeared in the nucleus of granulosa cell and oocytes of flesh and vitrified-warmed juvenile mouse ovaries.The expressions of Dnmtl protein and mRNA in vitrifiedwarmed juvenil mouse ovaries were remarkably decreased than that of fresh group (P＜0.05).The expressions of Grbl0 protein and mRNA in vitrified-warmed juvenil mouse ovaries were remarkably increased than that of fresh group (P＜0.05).Conclusion Decreased expression of Dnmtl in juvenil mouse ovaries induced by vitrified-warmed process may result in the overexpression of Grb 10,which may lead to an increased risk of glucose metabolism epigenetic defects in the oogamete. |
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| Keywords: | Juvenil mouse ovaries Vitrification DNA methyltransferase 1 (Dnmt1) Growth factor receptor-binding protein 10 (Grb10) |
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