[1]沈开元a,曲晓力a,代小丽b,等.卵巢功能减退患者卵泡液外泌体差异表达miRNA 筛选及生物学功能分析[J].现代检验医学杂志,2023,38(05):17-22.[doi:10.3969/j.issn.1671-7414.2023.05.004]
　SHEN Kaiyuana,QU Xiaolia,DAI Xiaolib,et al.Screening and Biological Function Analysis of Follicular Fluid-derived Exosomes Differentially Expressed miRNA in Patients with Diminished Ovarian Reserve[J].Journal of Modern Laboratory Medicine,2023,38(05):17-22.[doi:10.3969/j.issn.1671-7414.2023.05.004]
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卵巢功能减退患者卵泡液外泌体差异表达miRNA 筛选及生物学功能分析()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第38卷
期数:
2023年05期
页码:
17-22
栏目:
论著
出版日期:
2023-09-15

文章信息/Info

Title:
Screening and Biological Function Analysis of Follicular Fluid-derived Exosomes Differentially Expressed miRNA in Patients with Diminished Ovarian Reserve
文章编号:
1671-7414（2023）05-017-06
作者:
沈开元1a; 2; 曲晓力1a; 代小丽1b; 黄 萍1a; 邹乾兴1a; 梁媛媛1a; 罗平1a; 2
（1. 柳州市人民医院 a. 生殖医学科；b. 科研科，广西柳州 545006；2. 柳州市生殖与遗传代谢性疾病重点实验室，广西柳州 545006）
Author(s):
SHEN Kaiyuan1a; 2;  QU Xiaoli1a;  DAI Xiaoli1b;  HUANG Ping1a;  ZOU Qianxing1a;  LIANG Yuanyuan1a;  LUO Ping1a; 2
（1a.Reproductive Medicine Center；1b. Research Service Office, Liuzhou People’s Hospital, Guangxi Liuzhou 545006， China；2. Liuzhou Key Laboratory of Reproductive and Genetic Metabolic Diseases, Guangxi Liuzhou 545006 ，China）
关键词:
PCR 阵列芯片法; 卵巢功能减退; 卵泡液外泌体; 微小核糖核酸
分类号:
R711.75；R392.11
DOI:
10.3969/j.issn.1671-7414.2023.05.004
文献标志码:
A
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（microRNA，miRNA），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。
Abstract:
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. Follicular fluid 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 follicular fluid-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.

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备注/Memo

备注/Memo:
基金项目： 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；Z20190175：不同冷冻保护剂对牛体外成熟卵母细胞玻璃化冷冻的影响；Z20200270：HBV-DNA 病毒载量对体外受精-胚胎移植夫妻生育力与妊娠结局的影响。
作者简介： 沈开元（1986-），男，博士，主管技师，研究方向：体外受精机理和人类辅助生殖实验室技术，E-mail: sharkbaozi@163.com。
通讯作者：罗平，男，副主任技师，研究方向：人类辅助生殖实验室技术，E-mail:LUOP521@163.com。
更新日期/Last Update: 2023-09-15