Transcriptomic analysis of 13.5dpc embryonic ovaries exposed to the acetaminophen (APAP) and ibuprofen (IBU) combination in mice. Transcriptomic analysis of 13.5dpc embryonic ovaries exposed to the acetaminophen (APAP) and ibuprofen (IBU) combination in mice

Nonsteroidal anti-inflammatory drugs such as ibuprofen (IBU) and analgesic drugs, such as acetaminophen (APAP), are the most frequently medications taken by pregnant women, even in combination. They were shown to target fetal gonadal development acting as endocrine disruptors, that might favour genital malformations in new-born boys and reproductive disorders in adults, in an intergenerational manner. However, the consequences on postnatal ovarian development and female reproductive health after in utero exposure are still unknown. Here, we show that in utero exposure to therapeutic doses of the widely used APAP-IBU combination during the sensitive window of sex determination leads to an increased proliferation of female embryonic germ cells and a delayed entry into meiosis in 13.5dpc exposed ovaries. Consequently, the primordial follicle formation is enhanced in postnatal ovaries and is followed by a reduced follicular activation through the Akt/FoxO3 pathway and an increased follicular apoptosis. Subsequently, a reduced ovarian reserve in adult ovaries of exposed animals and in their offspring (F1) is observed. This leads to the subfertility of 6-month-old F1 animals that show an accelerated ovarian aging, a Premature Ovarian Insufficiency-like phenotype, with an abnormal persistence of corpora lutea due to the decreased apoptosis and an increased luteal Akt-mediated cell survival. Our data suggest that the use of APAP+IBU during this critical period of development that occurs during the first trimester of gestation (6 to 10 weeks), could lead in humans to adverse effects that could be passed to the offspring. Overall design: mRNA profiles of 13.5 days post coitum wild type and APAP+IBU exposed ovaries were generated by deep sequencing, in triplicate

非甾体抗炎药（如布洛芬（ibuprofen, IBU））与镇痛药（如对乙酰氨基酚（acetaminophen, APAP））是孕妇最常使用的药物，甚至常联合用药。研究证实，这类药物可作为内分泌干扰物靶向胎儿性腺发育，以跨代际方式增加新生男婴生殖器畸形风险，并可能引发成年个体生殖系统紊乱。然而，宫内暴露此类药物对产后卵巢发育及雌性生殖健康的影响仍不明晰。
本研究发现，在性别决定的敏感窗口期，宫内暴露于临床治疗剂量的广泛使用的APAP-IBU联合用药，可导致妊娠后13.5天（days post coitum, dpc）暴露卵巢内的雌性胚胎生殖细胞增殖增强，且减数分裂启动延迟。产后卵巢的原始卵泡形成增强，随后卵泡激活通过Akt/FoxO3通路受到抑制，同时卵泡凋亡水平升高。进一步观察发现，暴露母鼠及其子代（F1代）的成年卵巢储备均出现降低，进而导致6月龄F1代动物生育力低下，表现出卵巢衰老加速、早发性卵巢功能不全（Premature Ovarian Insufficiency, POI）样表型；同时因细胞凋亡减少、Akt介导的黄体细胞存活增强，出现黄体异常持续的现象。
本研究数据提示，在妊娠首三个月（6至10周）这一关键发育窗口期使用APAP+IBU，可能对人类产生可传递给子代的不良影响。
总体实验设计：采用深度测序技术对妊娠后13.5天的野生型及APAP+IBU暴露组卵巢的mRNA转录组进行测序分析，每组设置3次生物学重复。