上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2018, Vol. 38 ›› Issue (8): 857-.doi: 10.3969/j.issn.1674-8115.2018.08.002

• 论著·基础研究 • 上一篇    下一篇

卵巢颗粒细胞 11β-羟基类固醇脱氢酶的表达与多囊卵巢综合征卵巢局部皮质醇代谢的相关性研究

李小雪，朱琴玲，王缘，李嘉星，厉心愉，孙赟   

  1. 上海交通大学医学院附属仁济医院生殖医学中心，上海市辅助生殖与优生重点实验室，上海 200135
  • 出版日期:2018-08-28 发布日期:2018-09-11
  • 通讯作者: 孙赟，电子信箱：syun163@163.com。
  • 作者简介:李小雪（1991—），女，硕士生；电子信箱： lxue1124@163.com。
  • 基金资助:
    国家自然科学基金面上项目 (81771648，81571499)；上海市教育委员会高峰高原学科建设计划（ 20161413）；科技部重点专项子课题（ 2017YFC1001403）；国家 973子课题（ 2014CB943300）；上海市卫生系统优秀人才培养计划（ 2017BR015）

Analysis of the correlation between 11β-HSDs in the ovarian granulosa cells and cortisol metabolism in the ovary of PCOS

LI Xiao-xue, ZHU Qin-ling, WANG Yuan, LI Jia-xing, LI Xin-yu, SUN Yun   

  1. Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Centre for Reproductive Medicine, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200135, China
  • Online:2018-08-28 Published:2018-09-11
  • Supported by:
    General Program of National Natural Science Foundation of China, 81771648, 81571499; Shanghai Municipal Education Commission— Gaofeng Clinical Medicine Support, 20161413; National Key R&D Program of China, 2017YFC1001403; Chinese National Key Basic Research Projects, 2014CB943300; Program of Shanghai Academic Research Leader in Shanghai Municipal Commission of Health and Family Planning, 2017BR015

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摘要： 目的 ·探讨卵巢颗粒细胞 11β-羟基类固醇脱氢酶（ 11β-hydroxysteroid dehydrogenases，11β-HSDs）的表达亚型与多囊卵巢综合征（ polycystic ovary syndrome，PCOS）卵巢局部皮质醇（氢化可的松）代谢的相关性。方法 ·按照 2003年鹿特丹诊断标准纳入 PCOS组患者 24例，同时纳入对照组患者 21例。取卵日收集行体外受精 /卵胞质内单精子显微注射（ IVF/ICSI）的 PCOS组患者和对照组患者的卵泡液和黄素化颗粒细胞，采用酶联免疫吸附测定法检测 2组卵泡液中皮质醇、可的松水平，免疫荧光方法验证 11β-HSD1、11β-HSD2在黄素化颗粒细胞上的表达以及实时荧光定量 PCR（real-time quantitative PCR，RT-qPCR）方法检测黄素化颗粒细胞中 11β-HSD1、11β-HSD2的表达差异，并进一步分析卵巢局部皮质醇水平与黄素化颗粒细胞中 11β-HSD1、11β-HSD2表达的相关性。结果 ·与对照组相比， PCOS组患者卵泡液中皮质醇水平显著增高 [（477.3±35.3）nmol/L vs（292.0±36.4）nmol/L， P0.014]，而可的松无显著变化。 11β-HSD1和 11β-HSD2皆在黄素化颗粒细胞上有表达。与对照组相比， PCOS组患者黄素化颗粒细胞 11β-HSD1 mRNA表达 (P0.033) 及还原酶活性（ P0.006）均显著增加，而 11β-HSD2 mRNA表达和氧化酶活性无显著改变。 Pearson相关性分析显示，卵巢局部皮质醇的浓度与黄素化颗粒细胞 11β-HSD1表达成正相关（ r20.347 6，P0.001），而与黄素化颗粒细胞 11β-HSD2表达无相关性。结论 · PCOS患者卵巢局部皮质醇增加可能与卵巢颗粒细胞 11β-HSD1表达及还原酶活性增加有关。

关键词: 皮质醇, 11&, beta, -羟基类固醇脱氢酶, 卵泡液, 黄素化颗粒细胞, 多囊卵巢综合征

Abstract:

Objective · To explore the relationship of 11β-hydroxysteroid dehydrogenases (11β-HSDs) in the ovarian granulosa cells and cortisol regeneration in the ovary of polycystic ovary syndrome (PCOS). Methods · In accordance with the revised Rotterdam inclusion/exclusion criteria (2003), 24 patients in the PCOS group were included, meanwhile 21 patients in the control group were included. Follicular fluid and luteinized granulosa cells were collected patients underwent in vitro fertilization(IVF)or intracytoplasmic sperm injection (ICSI) in theCenterforReproductive Medicine of Renji Hospital. Enzyme linked immunosorbent assay (ELISA) was used to detect the cortisol concentration and the cortisone levels of follicular fluid. Real-time quantitative PCR (RT-qPCR) was used to measure 11β-HSDs abundance in the luteinized granulosa cells. The of 11β-HSD1 and 11β-HSD2 in the luteinized granulosa cells were verifiedimmunofluorescence. The correlation of local cortisol level in the ovary with 11β-HSDs in the luteinized granulosa cells was further analyzed. Results · Compared with the control group, the cortisol level in the follicular fluid was elevated in the PCOS group [(477.3±35.3) nmol/L vs (292.0±36.4)nmol/L, P0.014], while no change was observed in the cortisone levels in two groups. Both 11β-HSD1 and 11β-HSD2 were detected in the luteinized granulosa cells. Compared with the control group, the 11β-HSD1 mRNA abundance (P0.033) and reductase activity (P0.006) were increased significantly in the PCOS group, and there was no significant difference in the 11β-HSD2 and oxidase activity between two groups. Consistently, Pearson correlation analysis showed that the 11β-HSD1 but not 11β-HSD2 mRNA levels in the luteinized granulosa cells was correlated with the cortisol level in the follicular fluid (r20.347 6, P0.001). Conclusion · Elevation of local cortisol in the ovary of PCOS patients may be relatedtotheincreased of 11β-HSD1and reductase activity in the ovariangranulosa cells.

Key words: cortisol, 11&, beta, -hydroxysteroiddehydrogenases(11&, beta, -HSDs), follicular fluid, luteinizedgranulosa cell, polycystic ovary syndrome (PCOS)

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