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Macrophages Protect Endometriotic Cells Against Oxidative Damage Through a Cross-Talk Mechanism

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Abstract

This aim of this study was to investigate whether macrophages protect endometriotic cells from oxidative injury and to elucidate the underlying mechanisms of any protection. Endometriotic cells cultured with or without differentiated macrophages (dTHP-1 cells) were treated with hydrogen peroxide (H2O2) or methemoglobin, a major component of hemoglobin species in endometriotic cyst fluid. Co-culture experiments, microarray analysis, screening and validation of differentially expressed genes (DEGs), cell proliferation and viability assays, and experiments using a specific inhibitor were conducted to investigate the functional cross-talk between endometriotic cells and macrophages. Microarray analysis revealed that endometriotic cells co-cultured with dTHP-1 differentially express several genes compared with monoculture. Quantitative enzyme-linked immunosorbent assay (ELISA) and Western blotting analysis identified TGF-β1 as a promising candidate gene expressed in endometriotic cells co-cultured with dTHP-1 cells. TGF-β1 stimulated the expression of heme oxygenase-1 (HO-1) in dTHP-1 cells. HO-1 expression was increased in dTHP-1 cells co-cultured with endometriotic cells compared with the dTHP-1 monoculture. Both H2O2 and methemoglobin upregulated the expression of the HO-1 protein in the dTHP-1 monoculture; moreover, co-culture with endometriotic cells further enhanced HO-1 production. The co-culture with dTHP-1 protected endometriotic cells against oxidative injury. Blockade of HO-1 abolished the protective effects of macrophages. In an oxidative stress environment, TGF-β1 produced by endometriotic cells may protect against oxidative injury through the upregulation of macrophage-derived HO-1. The cross-talk between endometriotic cells and macrophages may contribute to the progression and pathogenesis of endometriosis.

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Data Availability

The datasets generated during the current study are available from Hiroshi Kobayashi.

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Acknowledgements

We thank Mrs. Toyomi Kobayashi for creating the figure.

Funding

This work was supported by Japan Society for Promotion of Science Japan (grant nos. 20K09604, 20K09647 and 20K09648).

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Nara Medical University, 840 Shijo-cho, Kashihara, 634-8522, Japan

    Kenji Ogawa, Tingting Liu, Naoki Kawahara & Hiroshi Kobayashi

  2. Department of Obstetrics and Gynecology, Ms.Clinic MayOne, 871-1 Shijo-cho, Kashihara, 634-0813, Japan

    Hiroshi Kobayashi

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  1. Kenji Ogawa
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  2. Tingting Liu
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  3. Naoki Kawahara
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  4. Hiroshi Kobayashi
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Contributions

Hiroshi Kobayashi contributed to the study conception and design. Kenji Ogawa was involved in all the experiments in this study. Naoki Kawahara and Tingting Liu helped with cell culture experiments and western blotting. The data analysis was performed by Kenji Ogawa and Naoki Kawahara. The first draft of the manuscript was written by Hiroshi Kobayashi. The final version of the manuscript has been read and approved by all authors.

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Correspondence to Hiroshi Kobayashi.

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Ethical approval

The research comprises several basic and clinical study designs. This manuscript conforms to the EQUATOR network guidelines. Our study meets checklists of the STROBE Statement (http://www.strobe-statement.org/).

Human and Animal Rights

The study was conducted under the guidelines that had been approved by the medical ethics committee of the Nara Medical University (reference no. 1587). This article does not contain any animal studies that have been performed by any of the authors.

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Written informed consent was obtained from each patient. Human rights statements and informed consent: All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1964 and its later amendments.

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The authors declare no competing interests.

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Ogawa, K., Liu, T., Kawahara, N. et al. Macrophages Protect Endometriotic Cells Against Oxidative Damage Through a Cross-Talk Mechanism. Reprod. Sci. 29, 2165–2178 (2022). https://doi.org/10.1007/s43032-022-00890-6

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