BMAL1通过靶向BMP6并调节卵巢颗粒细胞凋亡来改善多囊卵巢综合征患者的辅助生殖技术结局。

BMAL1 improves assisted reproductive technology outcomes in patients with polycystic ovary syndrome by targeting BMP6 and regulating ovarian granulosa cell apoptosis.

作者信息

Liang Qihui, Wei Chaofeng, Guan Lu, Chen Wen, Ding Shengyong, Wu Haicui

机构信息

First College of Clinical Medicine, Shandong University of Traditional Chinese Medicine, Jinan, 250011, Shandong, China.

Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, 250014, Shandong, China.

出版信息

J Assist Reprod Genet. 2025 Mar;42(3):937-948. doi: 10.1007/s10815-024-03377-2. Epub 2025 Jan 24.

DOI:10.1007/s10815-024-03377-2

PMID:39853686

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11950584/

Abstract

PURPOSE

To investigate BMAL1 and BMP6 expressive differences in ovarian granulosa cells (OGCs) of patients with polycystic ovary syndrome (PCOS), explore regulatory relationship, assess their impacts on OGC proliferation and apoptosis, and analyze their correlations with ART outcomes of patients.

METHODS

A clinical study selected 40 PCOS patients who underwent IVF/ICSI in our hospital from January to October 2022 and 39 controls with male or tubal factor infertility. RT-qPCR and Western blot assessed BMAL1 and BMP6 mRNA/protein levels. The number of oocytes retrieved, 2PN fertilized oocytes, available embryos, and high-quality embryos were compared between groups and analyzed their correlations with BMAL1 and BMP6 expression levels. Cellular experiments were performed by overexpressing or knocking down BMAL1 in KGN cells by plasmid transfection. The dual-luciferase reporter assay was used to identify BMAL1/BMP6 regulatory relationship. CCK-8 and flow cytometry assessed cellular proliferation and apoptosis.

RESULTS

BMAL1 mRNA/protein expression (P < 0.001) in the PCOS group was significantly lower than that in controls, as was the number of high-quality embryos (P = 0.001). Contrastingly, BMP6 (P < 0.001) was significantly higher in the PCOS group. BMAL1 expression levels were negatively correlated with BMP6 (r = - 0.684, P = 0.002) and positively correlated with the number of 2PN fertilized oocytes, available embryos, and high-quality embryos (r = 0.659, P = 0.003; r = 0.623, P = 0.006; and r = 0.738, P < 0.001). Cellular experiments showed that overexpression of BMAL1 significantly decreased relative luciferase activity (P < 0.01). Overexpression of BMAL1 significantly decreased KGN cell apoptosis (P < 0.01) and enhanced proliferation (P < 0.01).

CONCLUSION

BMAL1 regulates OGCs proliferation and apoptosis by targeting BMP6, thereby influencing ART outcomes in patients with PCOS. This study might provide molecular factors that indicate ART outcomes and therapeutic targets for PCOS.

TRIAL REGISTRATION

Registration number: ChiCTR2100052331; registration date: 2021-10-24.

摘要

目的

研究多囊卵巢综合征（PCOS）患者卵巢颗粒细胞（OGCs）中BMAL1和BMP6的表达差异，探索其调控关系，评估它们对OGC增殖和凋亡的影响，并分析它们与患者辅助生殖技术（ART）结局的相关性。

方法

一项临床研究选取了2022年1月至10月在我院接受体外受精/卵胞浆内单精子注射（IVF/ICSI）的40例PCOS患者以及39例因男性因素或输卵管因素不孕的对照者。采用逆转录-定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法评估BMAL1和BMP6的mRNA/蛋白质水平。比较两组间获取的卵母细胞数量、双原核（2PN）受精卵母细胞数量、可用胚胎数量和优质胚胎数量，并分析它们与BMAL1和BMP6表达水平的相关性。通过质粒转染在KGN细胞中过表达或敲低BMAL1进行细胞实验。采用双荧光素酶报告基因检测法鉴定BMAL1/BMP6调控关系。采用细胞计数试剂盒-8（CCK-8）法和流式细胞术评估细胞增殖和凋亡。

结果

PCOS组中BMAL1的mRNA/蛋白质表达（P<0.001）显著低于对照组，优质胚胎数量也显著低于对照组（P=0.001）。相反，PCOS组中BMP6的表达（P<0.001）显著更高。BMAL1表达水平与BMP6呈负相关（r=-0.684，P=0.002），与2PN受精卵母细胞数量、可用胚胎数量和优质胚胎数量呈正相关（r=0.659，P=0.003；r=0.623，P=0.006；r=0.738，P<0.001）。细胞实验表明，BMAL1过表达显著降低相对荧光素酶活性（P<0.01）。BMAL1过表达显著降低KGN细胞凋亡（P<0.01）并增强细胞增殖（P<0.01）。