Jiang Yan, Zhao Xuehan, Fang Ying, Yang Xiaokui
Department of Maternal Intensive Care Unit, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing Maternal and Child Health Care Hospital, Beijing, China.
Department of Human Reproductive Medicine, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing Maternal and Child Health Care Hospital, Beijing, China.
J Perinat Med. 2025 Apr 14;53(5):604-614. doi: 10.1515/jpm-2024-0509. Print 2025 Jun 26.
Pregnancy-related medical complications such as gestational diabetes mellitus (GDM) are common and associated with several obstetric and neonatal problems. There is growing evidence that microRNAs (miRNAs) are essential players in the pathophysiology of GDM. This study aimed to assess how particular miRNAs and the genes they target are expressed in GDM.
A GDM cell model was created using BeWo cells cultured in hyperglycemic (HG) conditions (25 mM glucose). Low-glucose (LG) conditions (5.5 mM glucose) were used for the BeWo cells in the control group. Differentially expressed genes (DEGs) in BeWo cells were identified by high-throughput sequencing and their levels verified in placental samples from GDM patients and controls using RT-PCR. Furthermore, the target genes of the DEGs were verified using dual-luciferase reporter assays.
High-throughput sequencing revealed 220 DEGs in BeWo cells. Among these, miR-3687 was significantly upregulated, while follistatin-like 3 (FSTL3) was downregulated in BeWo cells under HG conditions. The high-throughput sequencing results were corroborated by RT-PCR, which showed that placental samples from GDM patients had significantly lower levels of FSTL3 expression and substantially higher amounts of miR-3687 expression compared to control samples. FSTL3 was established as a direct target of miR-3687 as shown by dual-luciferase reporter assays.
The increase of miR-3687 might facilitate the onset and advancement of GDM by suppressing FSTL3. This discovery offered a new perspective on the molecular underpinnings of GDM and indicated possible targets for therapeutic intervention.
妊娠相关的医学并发症,如妊娠期糖尿病(GDM)很常见,并与多种产科和新生儿问题相关。越来越多的证据表明,微小RNA(miRNA)在GDM的病理生理学中起着至关重要的作用。本研究旨在评估特定的miRNA及其靶向基因在GDM中的表达情况。
使用在高糖(HG)条件(25 mM葡萄糖)下培养的BeWo细胞建立GDM细胞模型。对照组的BeWo细胞使用低糖(LG)条件(5.5 mM葡萄糖)。通过高通量测序鉴定BeWo细胞中差异表达基因(DEG),并使用逆转录聚合酶链反应(RT-PCR)在GDM患者和对照组的胎盘样本中验证其水平。此外,使用双荧光素酶报告基因测定法验证DEG的靶基因。
高通量测序显示BeWo细胞中有220个DEG。其中,miR-3687在HG条件下的BeWo细胞中显著上调。而卵泡抑素样3(FSTL3)下调。RT-PCR证实了高通量测序结果,表明与对照样本相比,GDM患者的胎盘样本中FSTL3表达水平显著降低,miR-3687表达量显著升高。双荧光素酶报告基因测定法表明FSTL3是miR-3687的直接靶标。
miR-3687的增加可能通过抑制FSTL3促进GDM的发生和发展。这一发现为GDM的分子基础提供了新的视角,并指出了可能的治疗干预靶点。