Liu Can, Liu Hongya, Li Hongyu, Yang Deguang, Li Ye, Wang Rui, Zhu Jiashu, Ma Shuqin, Guan Suzhen
School of Public Health, Ningxia Medical University, Yinchuan, Ningxia, China.
Key Laboratory of Environmental Factors and Chronic Disease Control, Yinchuan, Ningxia, China.
PLoS One. 2025 Jul 18;20(7):e0313705. doi: 10.1371/journal.pone.0313705. eCollection 2025.
This study aimed to investigate the association between prenatal stress (PS) and corticosterone levels, and its influence on DNA methylation of genes related to the placental glucocorticoid (GC) barrier, including 11β-HSD2, ABCB1 (P-gp), NR3C1, and FKBP5. The PS model was established through chronic unpredictable mild stress (CUMS). DNA methylation of GC-related genes was analyzed by reduced representation bisulfite sequencing (RRBS), and the results were confirmed using MethylTarget™ sequencing. The mRNA and protein expression levels of these genes were detected through qRT-PCR and Western blotting, respectively. Plasma corticosterone levels were elevated in pregnant female rats exposed to PS conditions and their offspring. Compared to the offspring of the prenatal control (OPC) group, the offspring of the prenatal stress (OPS) group exhibited down-regulation in both mRNA and protein expression of DNA methyltransferases (DNMT 3A and DNMT 3B), while up-regulation was observed in the expression of DNMT1. RRBS analyses identified ABCB1 and FKBP5 as hypermethylated genes, including a total of 43 differentially methylated sites (DMS) and 2 differentially methylated regions (DMR). MethylTarget™ sequencing further confirmed 15 differentially methylated CpG sites in these genes. This study provides preliminary evidence that PS disrupts the placental GC barrier through abnormal gene expression caused by hypermethylation of GC-related genes, resulting in elevated corticosterone levels in offspring and affecting their growth and development.
本研究旨在探讨产前应激（PS）与皮质酮水平之间的关联，及其对与胎盘糖皮质激素（GC）屏障相关基因（包括11β-HSD2、ABCB1（P-糖蛋白）、NR3C1和FKBP5）DNA甲基化的影响。通过慢性不可预测轻度应激（CUMS）建立PS模型。采用简化代表性亚硫酸氢盐测序（RRBS）分析GC相关基因的DNA甲基化，并使用MethylTarget™测序进行结果验证。分别通过qRT-PCR和蛋白质免疫印迹法检测这些基因的mRNA和蛋白质表达水平。暴露于PS条件下的怀孕雌性大鼠及其后代的血浆皮质酮水平升高。与产前对照组（OPC）的后代相比，产前应激组（OPS）的后代DNA甲基转移酶（DNMT 3A和DNMT 3B）的mRNA和蛋白质表达均下调，而DNMT1的表达上调。RRBS分析确定ABCB1和FKBP5为高甲基化基因，共有43个差异甲基化位点（DMS）和2个差异甲基化区域（DMR）。MethylTarget™测序进一步证实了这些基因中的15个差异甲基化CpG位点。本研究提供了初步证据，表明PS通过GC相关基因的高甲基化导致的异常基因表达破坏胎盘GC屏障，导致后代皮质酮水平升高，并影响其生长发育。
