Department of Obstetrics and Gynecology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan 250021, Shandong, China; Department of Obstetrics and Gynecology, Liao Cheng People's Hospital, 67 Dongchang West Road, Liaocheng 252000, Shandong, China; Shandong Key Laboratory of Reproductive Medicine, Jinan 250021, Shandong, China.
Department of Obstetrics, Shandong Provincial Hospital Affiliated to Shandong First Medical University, 324 Jingwu Road, Jinan 250021, Shandong, China.
Reprod Toxicol. 2022 Dec;114:22-31. doi: 10.1016/j.reprotox.2022.10.001. Epub 2022 Oct 4.
Di-2-ethylhexyl phthalate (DEHP) has been proven to boost miR-155-5p level in trophoblasts, but how DEHP-induced miR-155-5p regulates trophoblastic functions is unclear. For in vivo experiments, DEHP was administered to pregnant Sprague-Dawley (SD) rats at various dosages. After birth, the development of rat fetuses was evaluated. The morphology of the placentae was evaluated using HE staining. miR-155-5p level in placentae was measured utilizing RT-qPCR. Placental cAMP/PKA pathway activation and lipid metabolism levels were assessed using WB, RT-qPCR, and ELISA. For in vitro experiments, DEHP, miR-155-5p inhibitor, or cAMP/PKA inhibitor were applied to treat HTR-8/Svneo cells. Cell viability and functions were investigated utilizing WB, RT-qPCR, CCK-8, transwell assay, plate colony formation assay, and flow cytometry. Besides, the cAMP/PKA pathway activation and lipid metabolism levels in HTR-8/Svneo cells were detected via ELISA, WB, and RT-qPCR. DEHP resulted in fetal malformations and abnormal placental histopathology. DEHP also promoted placental miR-155-5p expression, the cAMP/PKA inactivation, and lipid metabolism in placentae. miR-155-5p knockdown abrogated DEHP-induced proliferative, migrative, and invasive inhibition in HTR-8/Svneo cells. Moreover, miR-155-5p downregulation abolished DEHP-induced inactivation of the cAMP/PKA pathway and enhanced lipid metabolism. Additionally, DEHP-induced miR-155-5p facilitated lipid metabolism by inhibiting the cAMP/PKA signaling pathway in HTR-8/Svneo cells. The current study reveals that miR-155-5p plays an indispensable role in DEHP-induced trophoblastic toxicity by promoting lipid metabolism via inhibiting the cAMP/PKA signaling pathway, indicating miR-155-5p might be a promising therapeutic target for DEHP exposure during pregnancy.
邻苯二甲酸二(2-乙基己基)酯(DEHP)已被证明可提高滋养细胞中的 miR-155-5p 水平,但 DEHP 诱导的 miR-155-5p 如何调节滋养细胞功能尚不清楚。在体内实验中,将 DEHP 以不同剂量给予怀孕的 Sprague-Dawley(SD)大鼠。出生后,评估大鼠胎儿的发育情况。用 HE 染色评估胎盘形态。利用 RT-qPCR 测量胎盘 miR-155-5p 水平。利用 WB、RT-qPCR 和 ELISA 评估胎盘 cAMP/PKA 通路激活和脂质代谢水平。在体外实验中,将 DEHP、miR-155-5p 抑制剂或 cAMP/PKA 抑制剂应用于 HTR-8/Svneo 细胞。利用 WB、RT-qPCR、CCK-8、Transwell assay、平板集落形成 assay 和流式细胞术研究细胞活力和功能。此外,通过 ELISA、WB 和 RT-qPCR 检测 HTR-8/Svneo 细胞中 cAMP/PKA 通路激活和脂质代谢水平。DEHP 导致胎儿畸形和胎盘组织病理学异常。DEHP 还促进胎盘 miR-155-5p 表达、cAMP/PKA 失活和胎盘脂质代谢。miR-155-5p 敲低消除了 DEHP 诱导的 HTR-8/Svneo 细胞增殖、迁移和侵袭抑制。此外,miR-155-5p 下调消除了 DEHP 诱导的 cAMP/PKA 通路失活和增强的脂质代谢。此外,DEHP 诱导的 miR-155-5p 通过抑制 cAMP/PKA 信号通路促进 HTR-8/Svneo 细胞中的脂质代谢。本研究揭示,miR-155-5p 通过抑制 cAMP/PKA 信号通路促进脂质代谢,在 DEHP 诱导的滋养细胞毒性中发挥不可或缺的作用,表明 miR-155-5p 可能是怀孕期间 DEHP 暴露的有前途的治疗靶点。