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Human sperm Toll-like receptor 4 (TLR4) mediates acrosome reaction, oxidative stress markers, and sperm parameters in response to bacterial lipopolysaccharide in infertile men

  • Gamete Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

To study the role of Toll-like receptor 4 (TLR4) in human spermatozoa and to assess sperm parameters, oxidative stress markers, and acrosome reaction in response to the stimulation of TLR4 by its ligand, the lipopolysaccharide (LPS), as a major endotoxin of Gram-negative bacteria.

Methods

Our study was carried out in 73 sperm samples from patients undergoing semen analysis for couple infertility investigations. The studied patients were divided into three groups: normozoospermic fertile patients (n = 13), patients with abnormal and leukospermic semen (n = 13), and patients with abnormal and non-leukospermic semen (n = 47). TLR4 expression in human spermatozoa was initially analyzed by western blot. Sperm samples were incubated in the presence of LPS (200 ng/ml) for 18 h. Then, sperm motility and vitality were evaluated by microscopic observation and oxidative stress markers as malondialdehyde (MDA) and carbonyl groups (CG) were spectrophotometrically assessed in neat and selected sperm. A triple-stain technique was also performed to evaluate acrosome reaction in 15 sperm samples from infertile patients.

Results

TLR4 expression was confirmed in human spermatozoa with a molecular weight of 69 kDa. In the normozoospermic group, no significant differences in sperm parameters and oxidative stress markers were shown after incubation with LPS in neat and selected sperms. Regarding samples from the non-leukospermic group, LPS reduced spermatozoa motility and vitality rates in selected sperm (P = 0.003; P = 0.004, respectively). A significant increase of MDA and CG levels was also detected (P = 0.01; P = 0.02, respectively). However, only the MDA levels were significantly increased (P = 0.01) in neat LPS-stimulated sperm. The same results were shown within the leukospermic group. The comparison between the two groups, leukospermic and non-leukospermic, in selected sperms showed a more important LPS effect in the leukospermic group significantly on motility and MDA rates (P = 0.006; P = 0.009, respectively). Furthermore, a significant decrease in reacted spermatozoa rate was detected in response to LPS in selected sperm samples from infertile men (P = 0.03).

Conclusions

These findings indicate that human spermatozoa express TLR4 and respond to LPS stimulation with alterations in viability, motility, and the acrosome reaction implicating reactive oxygen species (ROS) production in sperm samples from infertile patients.

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Acknowledgements

The authors thank all the members of the Higher Institute of Biotechnology, Sfax, and Mrs. Olfa Abida, a member of Autoimmunity and Immunogenetics Research Unit, Habib Bourguiba Hospital, Sfax, who generously contributed to this study. We thank also Mr. Mourad Ben Hmida for his helpful review of the manuscript.

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Authors and Affiliations

  1. Laboratory LR11ES45, Research Group “Biotechnology and Pathology”, Higher Institute of Biotechnology, International School of Engineers, Sfax, Tunisia

    Sana Sahnoun, Malek Mseddi, Hammadi Attia & Saloua Lassoued

  2. Histology Embryology Laboratory and Research Unit, Medicine Faculty, B.P.261, 3038, Sfax, Tunisia

    Afifa Sellami, Nozha Chakroun & Tarek Rebai

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  1. Sana Sahnoun
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  2. Afifa Sellami
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Correspondence to Sana Sahnoun.

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This study was approved by the Institutional Review Board (IRB) of Medicine Faculty of Sfax, Tunisia. Informed written consent was obtained from all patients for being included in the study.

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The authors declare that they have no conflict of interest.

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Sahnoun, S., Sellami, A., Chakroun, N. et al. Human sperm Toll-like receptor 4 (TLR4) mediates acrosome reaction, oxidative stress markers, and sperm parameters in response to bacterial lipopolysaccharide in infertile men. J Assist Reprod Genet 34, 1067–1077 (2017). https://doi.org/10.1007/s10815-017-0957-8

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