Abstract
Trophoblast cell syncytialization is essential for placental and fetal development. Abnormal trophoblast cell fusion leads to pregnancy pathologies, such as preeclampsia (PE), intrauterine growth restriction (IUGR), and miscarriage. 27-hydroxycholesterol (27-OHC) is the most abundant oxysterol in human peripheral blood synthesized by sterol 27-hydroxylase (CYP27A1) and is considered a critical mediator between hypercholesterolemia and a variety of related disorders. Gestational hypercholesterolemia was associated with spontaneous preterm delivery and low birth weight (LBW) in term infants, yet the mechanism is unclear. In this study, two trophoblast cell models and CD-1 mice were used to evaluate the effects of 27-OHC on trophoblast fusion during placenta development. Two different kinds of trophoblast cells received a dosage of 2.5, 5, or 10 uM 27-OHC. Three groups of pregnant mice were randomly assigned: control, full treatment (E0.5-E17.5), or late treatment (E13.5-E17.5). All mice received daily intraperitoneal injections of saline (control group) and 27-OHC (treatment group; 5.5 mg/kg). In vitro experiments, we found that 27-OHC inhibited trophoblast cell fusion in primary human trophoblasts (PHT) and forskolin (FSK)-induced BeWo cells. 27-OHC up-regulated the expression of the PI3K/AKT/mTOR signaling pathway-related proteins. Moreover, the PI3K inhibitor LY294002 rescued the inhibitory effect of 27-OHC. Inhibition of trophoblast cell fusion by 27-OHC was also observed in CD-1 mice. Furthermore, fetal weight and placental efficiency decreased and fetal blood vessel development was inhibited in pregnant mice treated with 27-OHC. This study was the first to prove that 27-OHC inhibits trophoblast cell fusion by Activating PI3K/AKT/mTOR signaling pathway. This study reveals a novel mechanism by which dyslipidemia during pregnancy results in adverse pregnancy outcomes.
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Data availability
The datasets generated during and analysed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- 27-OHC:
-
27-Hydroxylcholesterol
- CYP27A1:
-
Sterol 27-hydroxylase
- LXR:
-
Liver X receptor
- LBW:
-
Low birth weight
- HDL-c:
-
Higher lipoprotein–cholesterol
- SGA:
-
Small-for-gestational-age
- oxHDL:
-
Oxidized higher lipoprotein
- FSK:
-
Forskolin
- GDM:
-
Gestational diabetes
- IUGR:
-
Intrauterine growth restriction
- STB:
-
Syncytiotrophoblast
- CTB:
-
Cytotrophoblast
- PE:
-
Preeclampsia
- PHT:
-
Primary human trophoblasts
- CCK-8:
-
Counting Kit-8
- DAPI:
-
4′,6-Diamidino-2-phenylindole
- HCG:
-
Human chorionic gonadotropin
- GCM1:
-
Glial cell missing 1
- PLAP:
-
Placental alkaline phosphatase
- GFP:
-
Green fluorescence protein
- p-PI3K:
-
Phospho-PI3 K
- p-AKT:
-
Phospho-AKT
- p-mTOR:
-
Phospho-mTOR
- H&E:
-
Hematoxylin and Eosin
- MHC:
-
Maternal hypercholesterolemia
- PBS:
-
Phosphate-buffered saline
- FBS:
-
Fetal bovine serum
- DMSO:
-
Dimethyl sulfoxide
- MOI:
-
Multiplicity of infection
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This study was supported by the National Natural Science Foundation in China Grants (81872608).
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HY, RX, and XC designed the experiments. HZ, YZ, YZ collected the human samples. XZ, QD performed in experiments in vitro. QY and MZ performed in animal experiments. ZH and YW analyzed the data. XZ wrote the article. HY and XJ revised the manuscript. All authors contributed to the article and approved the submitted version.
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Our studies include human participants, human data, or human tissues. All the patients provided written informed consent before sampling and for the use of their clinical and biological data. This study was approved by the Ethics Committee of Capital Medical University and Fuxing Hospital affiliated to Capital Medical University (2018SY04). All clinical investigations were conducted according to the Declaration of Helsinki principles, and informed consent was obtained from all study participants. Animal experiments were conducted in accordance with the National Institutes of Health guidelines and were approved by the Institutional Animal Care and Use Committee at Capital Medical University (AEEI-2021-163).
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Zhao, X., Cai, X., Zhu, H. et al. 27-Hydroxycholesterol inhibits trophoblast fusion during placenta development by activating PI3K/AKT/mTOR signaling pathway. Arch Toxicol 98, 849–863 (2024). https://doi.org/10.1007/s00204-023-03664-4
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DOI: https://doi.org/10.1007/s00204-023-03664-4