Jiang Shengyuan, Wang Yixiao, Chen Haiyan, Teng Yuanyuan, Zhu Qiaoying, Xie Kaipeng
Department of Obstetrics and Gynecology, Women's Hospital of Nanjing Medical University, Nanjing Women and Children's Healthcare Hospital, Nanjing 210004, China.
Department of Public Health, Women's Hospital of Nanjing Medical University, Nanjing Women and Chidren's Healthcare Hospital, Nanjing 210004, China.
Toxics. 2025 May 29;13(6):454. doi: 10.3390/toxics13060454.
Bifenthrin (BF) is a widely used pyrethroid pesticide recognized as an endocrine-disrupting chemical (EDC). Previous studies have confirmed that chronic exposure to BF is associated with various health risks. However, its potential association with recurrent implantation failure (RIF) and recurrent pregnancy loss (RPL) remains unclear. In this study, the potential targets of BF were identified using several databases, including the Comparative Toxicogenomics Database (CTD), TargetNet, GeneCards, SwissTargetPrediction, and STITCH. Differentially expressed genes (DEGs) associated with RIF were obtained from bulk RNA-seq datasets in the GEO database. Candidate targets were identified by intersecting the predicted BF-related targets with the RIF-associated DEGs, followed by functional enrichment analysis using the DAVID and g:Profiler platforms. Subsequently, hub genes were identified based on the STRING database and Cytoscape. A diagnostic model was then constructed based on these hub genes in the RIF cohort and validated in an independent recurrent pregnancy loss (RPL) cohort. Additionally, we performed single-cell type distribution analysis and immune infiltration profiling based on single-cell RNA-seq and bulk RNA-seq data, respectively. Molecular docking analysis using AutoDock Vina was conducted to evaluate the binding affinity between BF and the four hub proteins, as well as several hormone-related receptors. Functional enrichment results indicated that the candidate genes were mainly involved in apoptotic and oxidative stress-related pathways. Ultimately, four hub genes-BCL2, HMOX1, CYCS, and PTGS2-were identified. The diagnostic model based on these genes exhibited good predictive performance in the RIF cohort and was successfully validated in the RPL cohort. Single-cell transcriptomic analysis revealed a significant increase in the proportion of myeloid cells in RPL patients, while immune infiltration analysis showed a consistent downregulation of M2 macrophages in both RIF and RPL. Moreover, molecular docking analysis revealed that BF exhibited high binding affinity to all four hub proteins and demonstrated strong binding potential with multiple hormone receptors, particularly pregnane X receptor (PXR), estrogen receptor α (ESRα), and thyroid hormone receptors (TR). In conclusion, the association of BF with four hub genes and multiple hormone receptors suggests a potential link to immune and endocrine dysregulation observed in RIF and RPL. However, in vivo and in vitro experimental evidence is currently lacking, and further studies are needed to elucidate the mechanisms by which BF may contribute to RIF and RPL.
联苯菊酯（BF）是一种广泛使用的拟除虫菊酯类农药，被认为是一种内分泌干扰化学物质（EDC）。先前的研究证实，长期接触BF与多种健康风险有关。然而，其与反复种植失败（RIF）和反复流产（RPL）的潜在关联仍不清楚。在本研究中，利用多个数据库，包括比较毒理基因组学数据库（CTD）、TargetNet、GeneCards、SwissTargetPrediction和STITCH，确定了BF的潜在靶点。从GEO数据库中的批量RNA测序数据集中获得与RIF相关的差异表达基因（DEG）。通过将预测的BF相关靶点与RIF相关的DEG进行交叉，确定候选靶点，随后使用DAVID和g:Profiler平台进行功能富集分析。随后，基于STRING数据库和Cytoscape确定了枢纽基因。然后在RIF队列中基于这些枢纽基因构建诊断模型，并在独立的反复流产（RPL）队列中进行验证。此外，我们分别基于单细胞RNA测序和批量RNA测序数据进行了单细胞类型分布分析和免疫浸润分析。使用AutoDock Vina进行分子对接分析，以评估BF与四种枢纽蛋白以及几种激素相关受体之间的结合亲和力。功能富集结果表明，候选基因主要参与凋亡和氧化应激相关途径。最终，确定了四个枢纽基因——BCL2、HMOX1、CYCS和PTGS2。基于这些基因的诊断模型在RIF队列中表现出良好预测性能，并在RPL队列中成功得到验证。单细胞转录组分析显示，RPL患者中髓样细胞比例显著增加，而免疫浸润分析显示，RIF和RPL中M2巨噬细胞均一致下调。此外，分子对接分析显示，BF对所有四种枢纽蛋白均表现出高结合亲和力，并与多种激素受体，特别是孕烷X受体（PXR）、雌激素受体α（ESRα）和甲状腺激素受体（TR）表现出强结合潜力。总之，BF与四个枢纽基因和多种激素受体的关联表明其与RIF和RPL中观察到的免疫和内分泌失调存在潜在联系。然而，目前缺乏体内和体外实验证据，需要进一步研究以阐明BF可能导致RIF和RPL的机制。
