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Microcystin–leucine–arginine causes blood–testis barrier disruption and degradation of occludin mediated by matrix metalloproteinase-8

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Abstract

Microcystin–leucine–arginine (MC-LR) can cause male reproductive disorders. However, the underlying mechanisms are not yet fully understood. In this study, we aimed to investigate the effects of MC-LR on the integrity of blood–testis barrier (BTB) and the related molecular mechanisms. Both transepithelial electrical resistance measurement in vitro and electron microscope observation ex vivo revealed that MC-LR caused disruption of the tight junction between Sertoli cells, which was paralleled by the degradation of occludin. We observed increased expression of matrix metalloproteinase-8 (MMP-8) upon exposure to MC-LR, and confirmed that abrogation of MMP-8 activity by specific inhibitors as well as transfection with MMP-8 shRNA could abolish the degradation of occludin. Our data demonstrated that MC-LR up-regulated nuclear levels of c-Fos and c-Jun through activating ERK and JNK, and increased NF-κB levels by activating the phosphatidylinositol 3-kinase (PI3K)/AKT cascades. Enhanced binding of c-Fos and NF-κB to the promoter of MMP-8 promoted the transcription of MMP-8 gene. Furthermore, miR-184-3p was significantly downregulated in SC following exposure to MC-LR through targeting MMP-8 expression. Together, these results confirmed that MC-LR-induced MMP-8 expression was regulated at both transcriptional and post-transcriptional levels, which was involved in MC-LR-induced degradation of occludin and BTB destruction. This work may provide new perspectives in developing new diagnosis and treatment strategies for MC-induced male infertility.

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Abbreviations

AP-1:

Activator protein-1

BTB:

Blood–testis barrier

ChIP:

Chromatin immunoprecipitation

Co-IP:

Coimmunoprecipitation

ECM:

Extracellular matrix

ER:

Endoplasmic reticulum

ES:

Ectoplasmic specialization

FBS:

Fetal bovine serum

MC-LR:

Microcystin–leucine–arginine

MMPs:

Matrix metalloproteinases

PI3K:

Phosphatidylinositol 3-kinase (PI3K)

SC:

Sertoli cells

TER:

Transepithelial electrical resistance

TJ:

Tight junction

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Acknowledgements

This work was supported by National Natural Science Foundation of China (31370524, 21377052, and 31670519) and Fundamental Research Funds for the Central Universities (021414380332).

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  1. Yabing Chen and Jing Wang contributed equally to this manuscript.

Authors and Affiliations

  1. Immunology and Reproduction Biology Laboratory & State Key Laboratory of Analytical Chemistry for Life Science, Medical School, Nanjing University, Nanjing, 210093, China

    Yabing Chen, Jing Wang, Chun Pan, Dongmei Li & Xiaodong Han

  2. Jiangsu Key Laboratory of Molecular Medicine, Nanjing University, Nanjing, 210093, China

    Yabing Chen, Jing Wang, Chun Pan, Dongmei Li & Xiaodong Han

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  1. Yabing Chen
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  2. Jing Wang
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Correspondence to Dongmei Li or Xiaodong Han.

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Chen, Y., Wang, J., Pan, C. et al. Microcystin–leucine–arginine causes blood–testis barrier disruption and degradation of occludin mediated by matrix metalloproteinase-8. Cell. Mol. Life Sci. 75, 1117–1132 (2018). https://doi.org/10.1007/s00018-017-2687-6

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