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LPS impairs steroidogenesis and ROS metabolism and induces PPAR transcriptional activity to disturb estrogen/androgen receptor expression in testicular cells

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Abstract

Inflammation can deregulate the testicular functions of steroidogenesis and spermatogenesis, consequently contributing to male infertility. Animals and cells treated with lipopolysaccharide (LPS) exhibit infection- and inflammation-induced testicular dysfunction. However, the precise mechanisms affecting steroidogenesis and spermatogenesis in response to LPS-treatment remain poorly understood. We isolated distinct testicular cells including spermatocytes, round spermatids and late spermatids to analyze distribution of peroxisome proliferator-activated receptor (PPAR) family, plays central roles in the regulation of metabolism. Our results suggested Pparα/Pparγ mRNA was highly expressed in late spermatids, while Pparβ mRNA was highly expressed in round spermatids. To analyze the effect of LPS on testicular cells, we established an LPS infection model using primary Sertoli cells and testicular cell lines (TM4, GC2 and MLTC1). We observed that PPARγ and SIRT1 were concentrated in the nuclear region and that the mRNA expression levels of antioxidative enzymes (Cat and Homx1) and PPARγ were upregulated in primary Sertoli cells after LPS-treatment. Moreover, luciferase reporter gene assays of the testicular cell lines revealed that the activity of the PPAR response element (PPRE) was significantly increased. Importantly, the transcriptional activity of the androgen response element was significantly reduced, whereas activity of estrogen response element was strongly induced in LPS-treated TM4 cells, consistent with the RT-PCR results. Meanwhile, the qRT-PCR results revealed that the LPS-induced upregulation of Ar mRNA in MLTC1 cells and Erβ mRNA in TM4 cells were significantly recovered after treatment with the specific PPARγ-antagonist GW9662. In addition, we also found that LPS induced alterations in enzymes involved in steroidogenesis in testicular cell lines. Taken together, our results revealed that LPS may induce PPAR transcriptional activity to disturb estrogen/androgen receptor expression and impair steroidogenesis and ROS metabolism in testicular cells.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Abbreviations

ANOVA:

Analysis of variance

AR:

Androgen receptor

ARE:

Antioxidant response element

ELISA:

Enzyme-linked immunosorbent assay

ER:

Estrogen receptor

ERE:

Estrogen response element

FBS:

Fetal bovine serum

FSH:

Follicle-stimulating hormone

HO-1:

Heme oxygenase-1

Lhr:

Luteinizing hormone receptor

LPS:

Lipopolysaccharide

NRs:

Nuclear receptors

OS:

Oxidative stress

PCRs:

Polymerase chain reactions

PFA:

Paraformaldehyde

PPAR:

Peroxisome proliferator-activated receptor

PPRE:

PPAR response element

PUFAs:

Polyunsaturated fatty acids

qRT-PCR:

Quantitative real-time PCR

ROS:

Reactive oxygen species

RT:

Room temperature

SCs:

Sertoli cells

SIRT1:

Silent information regulator type 1

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Acknowledgements

The excellent technical assistance of Yayun Fang and Danni Shan is gratefully acknowledged.

Funding

This work was supported by grants from the Health Commission of Hubei Province Scientific Research Project (Grant Nos. WJ2019H013 and WJ2019H023) and the Fundamental Research Funds for the Central Universities (Grant No. 2042019kf0150). The funders had no role in the study design, data collection, data analysis, decision to publish, or preparation of the manuscript.

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Author notes

  1. Gang Wang and Songtao Cheng have contributed equally to this work.

Authors and Affiliations

  1. Department of Biological Repositories, Zhongnan Hospital of Wuhan University, Wuhan, China

    Gang Wang, Shanshan Zhang, Yuan Zhu, Yu Xiao & Lingao Ju

  2. Human Genetics Resource Preservation Center of Wuhan University, Wuhan, China

    Gang Wang, Shanshan Zhang, Yuan Zhu, Yu Xiao & Lingao Ju

  3. Human Genetics Resource Preservation Center of Hubei Province, Wuhan, China

    Gang Wang, Shanshan Zhang, Yuan Zhu, Yu Xiao & Lingao Ju

  4. Department of Urology, Zhongnan Hospital of Wuhan University, Wuhan, China

    Songtao Cheng & Yu Xiao

  5. Cancer Precision Diagnosis and Treatment and Translational Medicine Hubei Engineering Research Center, Wuhan, China

    Songtao Cheng, Yu Xiao & Lingao Ju

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  1. Gang Wang
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  4. Yuan Zhu
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Contributions

GW, YX and LJ conceived of and designed the experiments; GW, SC and SZ performed the experiments; GW, YZ and YX analyzed the results; GW and SC contributed reagents/materials/analysis tools; GW, YX and LJ wrote the manuscript. All authors reviewed the manuscript.

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Correspondence to Lingao Ju.

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Wang, G., Cheng, S., Zhang, S. et al. LPS impairs steroidogenesis and ROS metabolism and induces PPAR transcriptional activity to disturb estrogen/androgen receptor expression in testicular cells. Mol Biol Rep 47, 1045–1056 (2020). https://doi.org/10.1007/s11033-019-05196-6

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  • DOI: https://doi.org/10.1007/s11033-019-05196-6

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