阿尼古力·沙比尔?甄雅惠?付玉霞?盛娟?阿依努尔·斯迪克?李瑞岐

【摘要】目的 研究白蘆藜醇（RSV）对小鼠超排卵效果及胚胎发育的影响。方法 以2～3月龄的CD-1小鼠为研究对象，在超排卵同时腹腔注射RSV（RSV组）或二甲基亚砜（DMSO组），观察RSV对小鼠超排卵子质量的影响。选取了小鼠胚胎成纤维细胞（MEF）、卵巢和睾丸组织的模板DNA，通过实时荧光定量PCR（qRT-PCR）分别检测沉默信息调节因子（SIRT1） mRNA在这三者中的表达情况。另外获取小鼠P1卵丘细胞，设置DMSO组、RSV（1 μmol/L）组和EX-527（SIRT1的抑制剂，10 μmol/L）组，使用荧光免疫染色检测各组SIRT的相对荧光强度、qRT-PCR检测各组的mtDNA相对表达量。结果 RSV组的卵裂率和囊胚率均高于DMSO组（P均< 0.05）。体外实验结果显示，SIRT1 mRNA的相对表达量在卵巢是MEF的15～25倍（P < 0.001）。小鼠P1卵丘细胞在经RSV处理后SIRT1荧光相对强度升高（P < 0.001）。在用RSV和EX-527分别处理小鼠卵丘细胞48 h后，RSV组mtDNA相对表达量高于DMSO组（P < 0.001），而EX-527组mtDNA相对表达量低于DMSO组（P < 0.01）。结论 小鼠超排卵时补充RSV能够显著改善卵子的发育潜能。

【关键词】白藜芦醇;线粒体;线粒体DNA;卵子质量;辅助生殖技术

Effect of resveratrol on the quality of oocytes after superovulation in mouse models Aniguli·Shabier， Zhen Yahui， Fu Yuxia， Sheng Juan， Ayinuer·Sidike， Li Ruiqi. Reproductive Medicine Center， the First Peoples Hospital of Kashgar， Kashgar 844000， China

Corresponding author， Li Ruiqi， E-mail： wxszzx@ 163. com

【Abstract】Objective To evaluate the effect of resveratrol （RSV） on the superovulation and embryonic development in mouse models. Methods In this study， CD-1 mice （2-3 months old） were selected. RSV （RSV group） or dimethyl sulfoxide （DMSO group） was injected intraperitoneally during superovulation to observe the effect of RSV on the quality of superovulated oocytes in mice. Template DNA of mouse embryonic fibroblasts （MEF）， ovary and testis tissues was extracted. The expression level of SIRT1 mRNA in these three tissues was detected by quantitative PCR. In addition， mouse P1 cumulus cells were obtained and divided into the DMSO， RSV （1μmol/L） and EX-527 （10 μmol/L inhibitor of SIRT1） groups. The relative fluorescence intensity of SIRT in each group was detected by immunofluorescence staining. The relative expression of mtDNA in each group was detected by real-time fluorescence quantitative PCR. Results The cleavage rate and blastocyst rate in the RSV group were significantly higher than those in the control group （both P < 0.05）. The results of in vitro experiments showed that the relative expression of SIRT1 mRNA in the ovary was 15-25 times of that in the MEF （P < 0.001）. The relative fluorescence intensity of SIRT1 in the P1 mouse cumulus cells was significantly increased after RSV treatment （P < 0.001）. After the mouse cumulus cells were treated with RSV and SIRT1 inhibitor （EX-527） for 48 h， the relative expression level of mtDNA in the RSV group was significantly higher than that in the DMSO group （P < 0.001）， whereas the relative expression level of mtDNA in the EX-527 group was significantly lower compared with that in the DMSO group （P < 0.01）. Conclusion RSV supplementation during superovulation can significantly improve the developmental potential of oocytes in mouse models.

【Key words】Resveratrol;Midtochondria;mtDNA;Oocyte quality;Assisted reproductive technique

改善卵子质量，提高胚胎发育潜能一直是辅助生殖技术（ART） 所追求的目标。近年来，越来越多的研究显示，线粒体功能下降是造成卵子质量下降的重要原因。白藜芦醇 （RSV）又称为芪三酚，是一类增强线粒体功能的药物。有研究显示，在卵母细胞体外成熟 （IVM） 时补充 RSV能够提高胚胎的囊胚率[1]。超排卵过程是卵泡快速生长的时期，也是卵母细胞内的线粒体和线粒体DNA（mtDNA）快速增殖的时期[2]。在此时补充 RSV会对超排卵产生什么样作用，笔者尚未见有相关报道。为此，本研究在小鼠超排卵同时补充RSV，观察其对超排效果的影响，并初步探讨其可能的作用机制，现报告如下。

材料与方法

一、实验动物

本实验所用小鼠为CD-1品系，超排卵实验为12只2 ～ 3月龄的雌性小鼠，体质量约30 g。获取颗粒细胞所用小鼠为1月龄小鼠，体质量约20 g。……