Department of Reproductive Medicine, Binzhou Medical University Hospital, Binzhou, China.
Clinical Medicine Laboratory, Binzhou Medical University Hospital, Binzhou, China.
J Physiol Pharmacol. 2021 Oct;72(5). doi: 10.26402/jpp.2021.5.13. Epub 2022 Mar 10.
The aim of this study was to determine whether endoplasmic reticulum (ER) stress is involved in the apoptosis of granulosa cells in patients with polycystic ovary syndrome (PCOS). A total of 116 patients undergoing in vitro fertilization/intracytoplasmic sperm injection cycles at the Binzhou Medical University Hospital IVF Center between September 2019 and January 2020 were enrolled in the study. Apoptosis of the granulosa cells in each patient was analyzed using flow cytometry, and progesterone and estrogen levels in the cell-culture fluid were measured by enzyme-linked immunosorbent assay. The expressions of X-box-binding protein 1 (XBP1(s)), activating transcription factor 6 (ATF6), C/EBP homologous protein (CHOP), B-cell CLL/lymphoma 2 protein (Bcl-2), and Bcl-2 associated X protein (Bax) at the gene or protein level were analyzed using quantitative real-time polymerase chain reaction and Western blotting, respectively. In patients with PCOS, body mass index and basal serum concentrations of luteinizing hormone, testosterone (T), and anti-Mullerian hormone significantly increased (P < 0.05). The number of oocytes retrieed and the rate of clinical pregnancy after the first frozen embryo transfer also significantly increased (P < 0.05). Although apoptosis rate in the granulosa cells significantly increased in patients with PCOS, progesterone (P) and estrogen (E) levels in the cell-culture fluid significantly decreased (P < 0.05). The apoptosis of granulosa cells was also found to affect blastocyst formation. Furthermore, the messenger ribonucleic acid (mRNA) expressions of XBP1(s), ATF6, CHOP, and Bax significantly increased in patients with PCOS, while that of Bcl-2 significantly decreased (P < 0.05). The protein expressions of CHOP and Bax significantly increased in patients with PCOS, while that of Bcl-2 significantly decreased (P < 0.05). After treatment of the granulosa cells with tauroursodeoxycholic acid, apoptosis rate and mRNA or protein expressions of XBP1(s), CHOP, and Bax significantly decreased, while the expression of Bcl-2 and levels of progesterone and estrogen significantly increased (P < 0.05). We conclude that ER stress could induce the apoptosis of granulosa cells in patients with PCOS. Cell apoptosis may decrease the number of blastocysts formed.
本研究旨在探讨内质网（ER）应激是否参与多囊卵巢综合征（PCOS）患者颗粒细胞的凋亡。2019 年 9 月至 2020 年 1 月，滨州医学院附属医院试管婴儿中心共纳入 116 例行体外受精/胞浆内单精子注射周期的患者。采用流式细胞术分析每位患者颗粒细胞的凋亡情况，酶联免疫吸附试验测定细胞培养液中孕酮和雌二醇水平。采用实时定量聚合酶链反应和 Western 印迹法分别分析 X 盒结合蛋白 1（XBP1(s)）、激活转录因子 6（ATF6）、C/EBP 同源蛋白（CHOP）、B 细胞 CLL/淋巴瘤 2 蛋白（Bcl-2）和 Bcl-2 相关 X 蛋白（Bax）的基因或蛋白表达。结果显示，与对照组相比，PCOS 患者的体重指数和基础血清黄体生成素、睾酮（T）和抗苗勒管激素水平显著升高（P<0.05）。第一次冷冻胚胎移植后的获卵数和临床妊娠率也显著升高（P<0.05）。虽然 PCOS 患者颗粒细胞的凋亡率显著增加，但细胞培养液中的孕酮（P）和雌二醇（E）水平显著降低（P<0.05）。颗粒细胞的凋亡也会影响囊胚的形成。此外，PCOS 患者 XBP1(s)、ATF6、CHOP 和 Bax 的信使核糖核酸（mRNA）表达显著增加，而 Bcl-2 的表达显著降低（P<0.05）。PCOS 患者 CHOP 和 Bax 的蛋白表达显著增加，而 Bcl-2 的蛋白表达显著降低（P<0.05）。用牛磺熊脱氧胆酸处理颗粒细胞后，细胞凋亡率及 XBP1(s)、CHOP 和 Bax 的 mRNA 或蛋白表达显著降低，Bcl-2 的表达及孕酮和雌二醇水平显著升高（P<0.05）。综上所述，内质网应激可诱导 PCOS 患者颗粒细胞凋亡，细胞凋亡可能减少囊胚形成数量。
