玻璃化冷冻对乳鼠卵巢Dnmt1和Grb10的影响研究  

Study on effection of DNA methyltransferase 1(Dnmt1) and growth factor receptor-binding protein 10(Grb10)genes on cryopreservation juvenil mouse ovaries by vitrification

作　　者：王红燕[1,2] 周雪[2] 王锐[2] 王燕蓉[2] 裴秀英[2] 马良宏[1] 颜贝 张帆[1] 周岳[1] 田稼[1] Wang Hongyan Zhou Xue Wang Rui Wang Yanrong Pei Xiuyin Ma Lianghong Yah Bei Zhang Fan Zhou Yue Tian Jia(Ningxia Human Sperm Bank, General Hospital of Ningxia Medical UniversitY, Yinchuan 750004, Chin Key Laboratory of Fertility Maintenance, Ministry of Education, Ningxia Medical University ＆Key Laboratory of Reproduction and Genetics, Yinchuan 750004, Chin)

机构地区：[1]宁夏医科大学总医院宁夏人类精子库,银川750004 [2]宁夏医科大学生育力保持教育部重点实验室,银川750004

出　　处：《生殖与避孕》2017年第1期27-32,共6页Reproduction and Contraception

基　　金：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)~~

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(P<0.05);Grb10蛋白和mRNA表达水平均显著上调(P<0.05)。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。Objective To study the effect ofvitrified-warmedjuvenil mouse ovaries on the mRNA and protein expression of DNA methyltransferase 1 （Dnmtl） and growth factor receptor-binding protein 10 （Grbl0）. 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 Dnmtl and Grb10 protein in fresh group and vitrified-warmed group were detected by Western blotting technology. The expressions of Dnmtl and GrblO mRNA in fresh group and vitrified-warmed group were detected by qRT-PCR. Results Dnmtl protein all appeared in the nucleus of granulosa cell and oocytes of fresh and vitrified-warmed juvenile mouse ovaries. The expressions of Dnmtl protein and mRNA in vitrified- warmed juvenil mouse ovaries were remarkably decreased than that of fresh group （P〈0.05）. The expressions of Grb 10 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 Grb10, which may lead to an increased risk of glucose metabolism epigenetic defects in the oogamete.

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) 

分 类 号：R714.8[医药卫生—妇产科学]