摘要
目的 筛选卵巢功能减退(diminished ovarian reserve,DOR)患者卵泡液外泌体差异表达微小核糖核酸(micro RNA,mi RNA),探索其与DOR相关的潜在分子机制。方法 收集2021年12月~2022年3月于柳州市人民医院生殖医学科辅助生殖助孕18例患者的卵泡液为研究样本,根据卵巢功能评估结果分为卵巢功能正常(Normal)组和卵巢功能减退(DOR)组。miRNA PCR阵列芯片法检测两组卵泡液外泌体miRNA的表达,实时荧光定量聚合酶链式反应(qRT-PCR)验证芯片结果,最后通过生物信息学分析差异表达miRNA的靶基因及其富集的相关生物学功能和分子通路。结果 DOR组筛选出5条差异表达miRNA:let-7d-3p,let-7e-5p,miR-25-3p和miR-30c-5p表达下调,miR-483-3p表达上调;qRT-PCR验证结果显示与Normal组比较,DOR组let-7d-3p,let-7e-5p,miR-25-3p和miR-30c-5p表达降低,miR-483-3p的表达升高,差异具有统计学意义(t=3.147,4.405,3.95,3.147,-3.198,均P<0.05);差异表达miRNA的靶基因主要富集于p53信号通路和FoxO信号通路,靶基因CDKN1A, RRM2, CCND1, SESN3, MDM4, BCL2L11,SMAD4, IGF1及IGF1R参与p53信号通路和FoxO信号通路,靶基因METTL3,METTL6及METTL8参与RNA m6A甲基化。结论 DOR患者卵泡液外泌体中的差异表达miRNA参与p53和FoxO信号通路及RNA m6A甲基化的调控,是DOR发生机制和诊疗的潜在标靶。
Objective To conduct a screening analysis of differential micro RNA(miRNA)present in the follicular fluid exosomes in patients diagnosed with diminished ovarian reserve(DOR),and investigate the potential molecular mechanisms associated with the regulation of DOR.Methods From December 2021 to March 2022,a total of 18 patients undergoing assisted reproduction at the Reproductive Medicine Center of Liuzhou People’s Hospital were recruited for this study.Follicularfluid samples were collected from these patients,and they were subsequently categorized into two groups based on the evaluation results of their ovarian function:Normal group and DOR group.The miRNA PCR array was employed to identify the differential expression of miRNAs in the follicular fluid-derived exosome,which was subsequently confirmed through qRT-PCR.Additionally,bioinformatics analysis was conducted to ascertain the target genes of the differentially expressed miRNAs,as well as their enriched biological functions and molecular pathways.Results Five differentially expressed miRNA were identified:let-7d-3p,let-7e-5p,miR-25-3p and miR-30c-5p were downregulated,while miR-483-3p was upregulated.Compared to the Normal group,the DOR group showed decreased expression of let-7d-3p,let-7e-5p,miR-25-3p and miR-30c-5p and increased expression of miR-483-3p,these differences were statistically significant(t=3.147,4.405,3.95,3.147,-3.198,all P<0.05).The differentially expressed miRNA were mainly enriched in the p53 signaling pathway and FoxO signaling pathway.Target genes CDKN1A,RRM2,CCND1,SESN3,MDM4,BCL2L11,SMAD4,IGF1 and IGF1R were involved in the p53 signaling pathway and FoxO signaling pathway,while target genes METTL3,METTL6 and METTL8 were involved in RNA m6A methylation.Conclusion The differentially expressed miRNA in follicularfluid-derived exosome of patients with DOR participate in the regulation of p53,FoxO signaling pathway and RNA m6A methylation,and are potential targets for the pathogenesis and diagnosis of DOR.
作者
沈开元
曲晓力
代小丽
黄萍
邹乾兴
梁媛媛
罗平
SHEN Kaiyuan;QU Xiaoli;DAI Xiaoli;HUANG Ping;ZOU Qianxing;LIANG Yuanyuan;LUO Ping(Reproductive Medicine Center,Liuzhou People’s Hospital,Guangxi Liuzhou 545006,China;Research Service Office,Liuzhou People’s Hospital,Guangxi Liuzhou 545006,China;Liuzhou Key Laboratory of Reproductive and Genetic Metabolic Diseases,Guangxi Liuzhou 545006,China)
出处
《现代检验医学杂志》
CAS
2023年第5期17-22,共6页
Journal of Modern Laboratory Medicine
基金
广西自然科学基金项目(2018GXNSFBA281048):人卵泡液外泌体对小鼠卵母细胞体外成熟及胚胎发育的影响
柳州市科技计划项目(2022SB004):卵巢功能减退卵泡液外泌体miRNA靶点筛选及其对辅助生殖治疗结局相关性的研究
广西壮族自治区卫生健康委员会自筹经费科研课题(Z20180308):人卵泡液来源外泌体对小鼠卵母细胞体外成熟影响的研究
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