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Prenatal dexamethasone exposure impairs rat blood-testis barrier function and sperm quality in adult offspring via GR/KDM1B/FSTL3/TGFβ signaling

Acta Pharmacologica Sinica (2024)Cite this article

Abstract

Both epidemiological and animal studies suggest that adverse environment during pregnancy can change the offspring development programming, but it is difficult to achieve prenatal early warning. In this study we investigated the impact of prenatal dexamethasone exposure (PDE) on sperm quality and function of blood-testis barrier (BTB) in adult offspring and the underlying mechanisms. Pregnant rats were injected with dexamethasone (0.1, 0.2 and 0.4 mg·kg-1·d-1, s.c.) from GD9 to GD20. After weaning (PW4), the pups were fed with lab chow. At PW12 and PW28, the male offspring were euthanized to collect blood and testes samples. We showed that PDE significantly decreased sperm quality (including quantity and motility) in male offspring, which was associated with impaired BTB and decreased CX43/E-cadherin expression in the testis. We demonstrated that PDE induced morphological abnormalities of fetal testicle and Sertoli cell development originated from intrauterine. By tracing to fetal testicular Sertoli cells, we found that PDE dose-dependently increased expression of histone lysine demethylases (KDM1B), decreasing histone 3 lysine 9 dimethylation (H3K9me2) levels of follistatin-like-3 (FSTL3) promoter region and increased FSTL3 expression, and inhibited TGFβ signaling and CX43/E-cadherin expression in offspring before and after birth. These results were validated in TM4 Sertoli cells following dexamethasone treatment. Meanwhile, the H3K9me2 levels of FSTL3 promoter in maternal peripheral blood mononuclear cell (PBMC) and placenta were decreased and its expression increased, which was positively correlated with the changes in offspring testis. Based on analysis of human samples, we found that the H3K9me2 levels of FSTL3 promoter in maternal blood PBMC and placenta were positively correlated with fetal blood testosterone levels after prenatal dexamethasone exposure. We conclude that PDE can reduce sperm quality in adult offspring rats, which is related to the damage of testis BTB via epigenetic modification and change of FSTL3 expression in Sertoli cells. The H3K9me2 levels of the FSTL3 promoter and its expression in the maternal blood PBMC can be used as a prenatal warning marker for fetal testicular dysplasia.

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Fig. 1: Changes of testicular morphology and barrier function at PW12 and PW28 caused by PDE.
Fig. 2: Changes in fetal testicular morphology and barrier function caused by PDE.
Fig. 3: Changes of KDM1B, FSTL3 and TGFβ signal pathway in tests induced by PDE before and after birth.
Fig. 4: Dexamethasone inhibits the blood-testis barrier function of TM4 cells.
Fig. 5: GR/KDM1B/FSTL3/TGFβ mediates dexamethasone-induced inhibition of CX43 and N-cadherin expression in TM4 cells.
Fig. 6: Changes of KDM1B, FSTL3 in the placenta, maternal PBMC and Bewo cell after dexamethasone treatment (ADT).
Fig. 7: Placenta and maternal PBMC FSTL3 promoter H3K9me2 and expression level and male neonatal plasma testosterone and the correlations between them respectively.
Fig. 8: Molecular mechanism of impaired blood-testis barrier function induced by prenatal dexamethasone exposure.

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Acknowledgements

This work was supported by grants from the National Key Research and Development Program of China (No. 2020YFA0803900), the National Natural Science Foundation of China (No. 82030111, U23A20407), and Hubei Province’s Outstanding Medical Academic Leader Program.

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  1. These authors contributed equally: Yi Liu, Si-jia Chen

Authors and Affiliations

  1. Department of Pharmacology, Wuhan University School of Basic Medical Sciences, Wuhan, 430071, China

    Yi Liu, Si-jia Chen, Can Ai, Peng-xia Yu, Man Fang & Hui Wang

  2. Hubei Provincial Key Laboratory of Developmentally Originated Disease, Wuhan, 430071, China

    Yi Liu, Si-jia Chen, Can Ai, Peng-xia Yu & Hui Wang

  3. Department of Obstetrics and Gynaecology, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China

    Hui Wang

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YL designed research, performed researches, and wrote the paper. SJC contributed new reagents and analytic tools. CA, PXY, MF Investigated the literatures. HW got the Funding and reviewed, modified the paper.

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Correspondence to Hui Wang.

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Liu, Y., Chen, Sj., Ai, C. et al. Prenatal dexamethasone exposure impairs rat blood-testis barrier function and sperm quality in adult offspring via GR/KDM1B/FSTL3/TGFβ signaling. Acta Pharmacol Sin (2024). https://doi.org/10.1038/s41401-024-01244-5

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