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

  • Gang Wang
  • Songtao Cheng
  • Shanshan Zhang
  • Yuan Zhu
  • Yu Xiao
  • Lingao JuEmail author
Original Article

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.

Keywords

Lipopolysaccharide (LPS) Estrogen/androgen receptor Peroxisome proliferator-activated receptor (PPAR) Sirtuin Testis 

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

Notes

Acknowledgements

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

Author 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.

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.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

11033_2019_5196_MOESM1_ESM.docx (489 kb)
Supplementary material 1 (DOCX 489 kb)

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Biological RepositoriesZhongnan Hospital of Wuhan UniversityWuhanChina
  2. 2.Human Genetics Resource Preservation Center of Wuhan UniversityWuhanChina
  3. 3.Human Genetics Resource Preservation Center of Hubei ProvinceWuhanChina
  4. 4.Department of UrologyZhongnan Hospital of Wuhan UniversityWuhanChina
  5. 5.Cancer Precision Diagnosis and Treatment and Translational Medicine Hubei Engineering Research CenterWuhanChina

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