Prenatal exposure to diethylhexyl phthalate (DEHP) has been linked with a decline in testosterone levels in adult male rats, but the underlying mechanism remains unclear. We investigated the potential epigenetic regulation, particularly focusing on N6-methyladenosine (m6A) modification, as a possible mechanism. Dams were gavaged with DEHP (0, 10, 100, and 750 mg/kg/day) from gestational day 14 to day 21. The male offspring were examined at the age of 56 days. Prenatal DEHP administration at 750 mg/kg/day caused a decline in testosterone concentrations, an elevation in follicle-stimulating hormone, a downregulated expression of CYP11A1 HSD3B2, without affecting Leydig cell numbers. Interestingly, Methyltransferase Like 4 (METTL4), an m6A methyltransferase, was downregulated, while there were no changes in METTL3 and METTL14. Moreover, CYP11A1 showed m6A reduction in response to prenatal DEHP exposure. Additionally, METTL4 expression increased postnatally, peaking in adulthood. Knockdown of METTL4 resulted in the downregulation of CYP11A1 and HSD3B2 and an increase in SCARB1 expression. Furthermore, the increase in autophagy protection in adult Leydig cells induced by prenatal DEHP exposure was not affected by 3-methyladenosine (3MA) treatment, indicating a potential protective role of autophagy in response to DEHP exposure. In conclusion, prenatal DEHP exposure reduces testosterone by downregulating CYP11A1 and HSD3B2 via m6A epigenetic regulation and induction of autophagy protection in adult Leydig cells as a response to DEHP exposure.

中文翻译：

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产前邻苯二甲酸二乙酯暴露通过表观遗传下调雄性大鼠 METTL4 表达扰乱成年 Leydig 细胞功能


产前接触邻苯二甲酸二乙基己酯 (DEHP) 与成年雄性大鼠睾酮水平下降有关，但其潜在机制仍不清楚。我们研究了潜在的表观遗传调控，特别关注 N6-甲基腺苷 (m6A) 修饰，作为一种可能的机制。从妊娠第14天到第21天，对母鼠灌胃DEHP（0、10、100和750mg/kg/天）。雄性后代在56日龄时进行检查。产前给予 750 毫克/公斤/天的 DEHP 会导致睾酮浓度下降、卵泡刺激素升高、CYP11A1 HSD3B2 表达下调，但不影响 Leydig 细胞数量。有趣的是，甲基转移酶样 4 (METTL4)（一种 m6A 甲基转移酶）下调，而 METTL3 和 METTL14 没有变化。此外，CYP11A1 表现出 m6A 因产前 DEHP 暴露而减少。此外，METTL4 表达在出生后增加，并在成年期达到顶峰。 METTL4 的敲除导致 CYP11A1 和 HSD3B2 的下调以及 SCARB1 表达的增加。此外，产前 DEHP 暴露诱导的成年 Leydig 细胞自噬保护的增加不受 3-甲基腺苷 (3MA) 治疗的影响，表明自噬对 DEHP 暴露具有潜在的保护作用。总之，产前 DEHP 暴露通过 m6A 表观遗传调节下调 CYP11A1 和 HSD3B2，并诱导成年 Leydig 细胞的自噬保护，从而降低睾酮水平，作为对 DEHP 暴露的反应。