白藜芦醇对小鼠超排卵子质量的影响

Effect of resveratrol on the quality of oocytes after superovulation in mouse models

目的研究白芦藜醇(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(1μmol/L)组和EX-527(SIRT1的抑制剂,10μmol/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。

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(10μmol/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.