多囊卵巢综合征患者卵泡液中6种miRNAs表达的检测

目的:分析多囊卵巢综合征(PCOS)患者卵泡液外泌体中miRNA-9、miRNA-18b、miRNA-19、miRNA-24、miRNA-199和miRNA-320水平,初步探讨其在PCOS发生、发展中的潜在意义。方法:48例接受体外受精-胚胎移植(IVF-ET)治疗的不孕症患者,分为PCOS组(23例)和对照组(25例),留取其卵泡液。采用超速离心法提取卵泡液外泌体,分别用外泌体粒径分析(NTA)、蛋白质印迹(Western blotting)和透射电镜验证外泌体。通过实时荧光聚合酶链式反应(PCR)法检测卵泡液和外泌体miRNA-9,miRNA-18b,miRNA-19,miRNA-24,...

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Published in国际生殖健康/计划生育杂志 Vol. 37; no. 1; pp. 5 - 10
Main Author 刘凯鲁;胡梦婷;蔡令波;李涵;杨玮杰;刘嘉茵;崔毓桂;千日成
Format Journal Article
LanguageChinese
Published 生殖国家重点实验室, 南京医科大学第一附属医院生殖医学科,210029%同济大学附属第十人民医院生殖医学中心 2018
Subjects
多囊卵巢综合征;卵泡液;微RNAs;外泌体
多囊卵巢综合征
卵泡液
Polycystic ovary syndrome
外泌体
MicroRNAs
Follicular fluid
Exosomes
微RNAs
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ISSN1674-1889
DOI10.3969/j.issn.1674-1889.2018.01.001

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Summary:目的:分析多囊卵巢综合征(PCOS)患者卵泡液外泌体中miRNA-9、miRNA-18b、miRNA-19、miRNA-24、miRNA-199和miRNA-320水平,初步探讨其在PCOS发生、发展中的潜在意义。方法:48例接受体外受精-胚胎移植(IVF-ET)治疗的不孕症患者,分为PCOS组(23例)和对照组(25例),留取其卵泡液。采用超速离心法提取卵泡液外泌体,分别用外泌体粒径分析(NTA)、蛋白质印迹(Western blotting)和透射电镜验证外泌体。通过实时荧光聚合酶链式反应(PCR)法检测卵泡液和外泌体miRNA-9,miRNA-18b,miRNA-19,miRNA-24,miRNA-199,miRNA-320的表达情况。结果:卵泡液中PCOS组miRNA-199的表达水平高于对照组(5.42±0.80 vs.1.02±0.94,P<0.01),miRNA-320的表达水平低于对照组(0.47±0.06 vs.1.23±0.09,P<0.01);2组间miRNA-9,miRNA-18b,miRNA-19和miRNA-24的表达水平差异无统计学意义(P>0.05)。在外泌体中,PCOS组miRNA-9和miRNA-18b的表达水平高于对照组(8.94±0.54 vs.1.01±0.97,2.66±0.19 vs.1.12±0.06,均P<0.01),miRNA-199和miRNA-320的表达水平则低于对照组(0.43±0.11 vs.1.22±0.07,0.53±0.09 vs.1.18±0.15,均P<0.01)。结论:PCOS患者卵泡液和外泌体存在miRNA-9,miRNA-18b和miRNA-199的表达,可能与PCOS发生、发展相关。
Bibliography:Polycystic ovary syndrome;Follicular fluid;MicroRNAs;Exosomes
Objective:To analyze the expressions of miRNA-9,miRNA-18b,miRNA-19,miRNA-24,miRNA-199 and miRNA-320 in the follicular fluid and exosome of patients with polycystic ovary syndrome(PCOS),and to explore the potential roles of these miRNAs in the pathophysiological mechanism of PCOS.Methods:A total of 48 patients undergoing in vitro fertilization and embryo transfer(IVF-ET),including 23 PCOS patients and 25 controls,were enrolled.The follicular fluid was collected.The exosome from the follicular fluid was isolated using ultrafiltration.NTA,Western blotting and TEM were used to identify the exosmoe.The expressions of miRNA-9,miRNA-18b,miRNA-19,miRNA-24,miRNA-199 and miRNA-320 in the follicular fluid and exosome were examined by real-time quantitative PCR.Results:The expression level of miRNA-199 in the follicular fluid of PCOS patients was significantly higher than that in the control group(5.42±0.80 vs.1.02±0.94,P<0.01),while the expression level of
ISSN:1674-1889
DOI:10.3969/j.issn.1674-1889.2018.01.001