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，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。

Expressions of Six MiRNAs in Follicular Fluid of Patients with Polycystic Ovary Syndrome

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 miRNA-320 was significantly lower (0.47±0.06 vs. 1.23±0.09, P＜0.01). The expressions of miRNA-9, miRNA-18b, miRNA-19 and miRNA-24 were showed no significant differences between the two groups (P＞0.05). In exosomes, the expressions of miRNA-9 and miRNA-18b in the PCOS group were significantly higher than those in the control group (8.94±0.54 vs. 1.01±0.97，2.66±0.19 vs. 1.12±0.06, both P＜0.01), while the expressions of miRNA-199 and miRNA-320 were significantly lower (0.43±0.11 vs. 1.22±0.07, 0.53±0.09 vs. 1.18±0.15, both P＜0.01). Conclusions: It is helpful to analyze the expressions of miRNA-9, miRNA-18b and miRNA-199 in the follicular fliud and exosome of PCOS patients for us to explore the roles of miRNAs in the pathophysiological mechanism of PCOS.