25(OH)D3 通过增加 G2M+S 期细胞来改善子宫内膜异位症患者的颗粒细胞增殖和 IVF 妊娠结局。

25(OH)D3 improves granulosa cell proliferation and IVF pregnancy outcomes in patients with endometriosis by increasing G2M+S phase cells.

机构信息

Lanzhou University, Lanzhou, Gansu, 730000, China.

First Clinical Medical School of Lanzhou University, Lanzhou, Gansu, 730000, China.

出版信息

Reprod Biol Endocrinol. 2023 Dec 5;21(1):115. doi: 10.1186/s12958-023-01165-8.

DOI:10.1186/s12958-023-01165-8

PMID:38053145

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10696887/

Abstract

BACKGROUND

The 25-hydroxyvitamin D3 (25 (OH) D3) is crucial for follicular development. This study aimed to investigate the relationship between the level of 25 (OH) D3 in endometriosis patients, pregnancy outcomes of in vitro fertilization (IVF), and the underlying mechanism.

METHODS

The 25 (OH) D3 levels in serum and follicular Fluid (FF) samples were detected using enzyme-linked immunosorbent assay (ELISA). Clinical features and pregnancy outcomes of endometriosis patients were also compared between the deficient group (< 20 ug/ml) and the adequate group (≥ 20 ug/ml). The effects of 25 (OH) D3 on the proliferation and cell cycle of human ovarian granulosa cells were respectively detected by CCK-8 assay and flow cytometry (FCM). The differentially expressed genes (DEGs) in granulosa cells of endometriosis and tubal infertility patients were screened from GEO database. The effects of 25 (OH) D3 on the expressions of CDKN2D, PPARA, TGFB2 and THBD were determined using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot.

RESULTS

The levels of 25 (OH) D3 in serum and FF samples were decreased in endometriosis patients. The deficient group had fewer embryos that can be transferred, lower quality embryos and lower clinical pregnancy rates. Adequate 25 (OH) D3 levels in FF samples was a protective factor for live birth outcome in endometriosis patients. 25 (OH) D3 enhanced the proliferation capacity of granulosa cells (the concentration of 10 nM was the most significant) and increased the proportion of G2M + S phase cells. The expression of CDKN2D was decreased and TGFB2 and THBD were significantly upregulated.

CONCLUSIONS

25 (OH) D3 deficiency may be associated with poor IVF pregnancy outcomes in endometriosis patients. 25 (OH) D3 promotes ovarian granulosa cell proliferation by promoting the ability of cells to divide, and may accelerate cell cycle progression by up-regulating THBD and down-regulating CDKN2D expression.

摘要

背景

25-羟维生素 D3（25（OH）D3）对卵泡发育至关重要。本研究旨在探讨子宫内膜异位症患者血清和卵泡液（FF）中 25（OH）D3 水平与体外受精（IVF）妊娠结局的关系及其潜在机制。

方法

采用酶联免疫吸附法（ELISA）检测血清和 FF 样本中 25（OH）D3 水平。比较不同 25（OH）D3 水平（<20ug/ml 为缺乏组，≥20ug/ml 为充足组）子宫内膜异位症患者的临床特征和妊娠结局。通过 CCK-8 检测和流式细胞术（FCM）分别检测 25（OH）D3 对人卵巢颗粒细胞增殖和细胞周期的影响。从 GEO 数据库筛选出子宫内膜异位症和输卵管性不孕患者颗粒细胞中的差异表达基因（DEGs）。采用定量逆转录聚合酶链反应（qRT-PCR）和 Western blot 检测 25（OH）D3 对 CDKN2D、PPARA、TGFB2 和 THBD 表达的影响。

结果

子宫内膜异位症患者血清和 FF 样本中 25（OH）D3 水平降低。缺乏组可移植胚胎数减少，优质胚胎数减少，临床妊娠率降低。FF 样本中 25（OH）D3 充足是子宫内膜异位症患者活产结局的保护因素。适当的 25（OH）D3 水平可增强颗粒细胞的增殖能力（10 nM 浓度作用最显著），增加 G2M+S 期细胞的比例。CDKN2D 表达下调，TGFB2 和 THBD 显著上调。