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Increased Expression of TGF-β1 Contributes to the Downregulation of Progesterone Receptor Expression in the Eutopic Endometrium of Infertile Women with Minimal/Mild Endometriosis

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Abstract

Endometriosis is a hormone-dependent disease associated with impaired immunoregulation. In our recent study, we have characterized the trascriptomic transformation of eutopic endometrium from patients with minimal/mild endometriosis and controls across the menstrual cycle. However, the regulatory mechanism of altered immune microenvironment in eutopic endometrial stromal cells (ESCs) remains unclear. Here, we want to explore the regulation of immune cell to progesterone resistance and endometrial receptivity in the eutopic ESCs by cytokine (TGF-β1), and to understand the effect of TGF-β1 on the decidualization of the eutopic ESCs. Primary culture of eutopic ESCs was performed to explore the effects of TGF-β1 on the expression of Smad and progesterone receptor (PR) and the in vitro decidualization. Additionally, co-immunoprecipitation (Co-IP) was used to explore the direct interaction between Smad and PR. We found an attenuate expression of PRB protein (p=0.026) after using TGF-β1 in eutopic ESCs, although the difference of PRA before and after treatment was not significant (p=0.678). Similarly, the results of qRT-PCR showed that the mRNA level of PR (p<0.001), PRB (p=0.003) and HOXA10 (p<0.001) decreased significantly after TGF-β1 treatment, but that increased (p<0.023, for all) after SB431542 treatment in the eutopic ESCs. Moreover, TGF-β1 has a negative effect on the in vitro decidualization of eutopic ESCs (p=0.003). And the group with treatment of both TGF-β1 and SB435142 in eutopic ESCs showed significant decidual-like changes with increased prolactin level (p=0.01). We did not observe any physical interaction between the PR and p-Smad3/Smad3 proteins by using Co-IP. By activating TGF-β/Smad signaling in eutopic ESCs, elevated TGF-β1 from CD45+ immune cells could attenuate expression of PR, and further decrease endometrial receptivity.

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Data Availability

The sing cell data of the current study are available in the NCBI’s Gene Expression Omnibus (accession code GSE214411).

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Acknowledgements

We are grateful to all study participants, doctors, and operating room nurses at West China Second University Hospital of Sichuan University.

Funding

This study was supported by the National Natural Science Foundation of China (Grant number: 82071625).

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Author notes

  1. Lukanxuan Wu and Xin Huang have contributed equally to this work and share first authorship.

Authors and Affiliations

  1. Department of Reproductive Medicine, West China Second University Hospital of Sichuan University, Chengdu, 610041, Sichuan, China

    Lukanxuan Wu, Xin Huang, Ruiying Wang, Yujing Li, Huili Zhu, Yunwei Ouyang & Wei Huang

  2. Key Laboratory of Birth Defects and Related Diseases of Women and Children of Ministry of Education, Chengdu, 610041, Sichuan, China

    Lukanxuan Wu, Xin Huang, Ruiying Wang, Yujing Li, Huili Zhu, Yunwei Ouyang & Wei Huang

  3. NHC Key Laboratory of Chronobiology, Sichuan University, Chengdu, 610041, Sichuan, China

    Lukanxuan Wu, Xin Huang, Ruiying Wang, Yujing Li & Wei Huang

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  1. Lukanxuan Wu
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Contributions

LW designed and performed the experiments and analyzed data, drafted the manuscript. XH designed and performed the experiments and analyzed data, and revised the manuscript. RW and JL assisted with experiments and analyzed data. HZ and YO contributed to sample collection and supervised the research. WH designed and supervised the research, revised the manuscript, and approved the final vision.

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Correspondence to Wei Huang.

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The study was approved by the Ethics Committee of West China second University Hospital of Sichuan University (Grant number:2021028).

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Wu, L., Huang, X., Wang, R. et al. Increased Expression of TGF-β1 Contributes to the Downregulation of Progesterone Receptor Expression in the Eutopic Endometrium of Infertile Women with Minimal/Mild Endometriosis. Reprod. Sci. 30, 3578–3589 (2023). https://doi.org/10.1007/s43032-023-01315-8

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