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. 2020 Sep-Oct;14(5):1557988320970053.
doi: 10.1177/1557988320970053.

Reactive Oxygen Species Secreted by Leukocytes in Semen Induce Self-Expression of Interleukin-6 and Affect Sperm Quality

Xiaoping Li  1   2 , Mengxia Ni  3 , Shiyu Xing  3 , Yi Yu  3 , Yan Zhou  3 , Shenmin Yang  3 , Hong Li  3 , Rui Zhu  3 , Mutian Han  4   3
Affiliations

Reactive Oxygen Species Secreted by Leukocytes in Semen Induce Self-Expression of Interleukin-6 and Affect Sperm Quality

Xiaoping Li et al. Am J Mens Health. 2020 Sep-Oct.

Abstract

Reproductive tract inflammation is considered an important cause of male infertility. Increased leukocytes in semen can produce many reactive oxygen species (ROS), which affect sperm function. The aim of this study is to identify the main source of ROS in seminal plasma and to assess the effect of ROS on leukocytes. Semen samples (n = 20) with leukocyte concentration >1 × 106 were collected from a male infertility clinic. This study mainly compares the sperm function parameters of the normal group and the semen white blood cell group >1 × 106. The results identified that ROS in semen was closely related to sperm function parameters, and CD45+ leucocytes were the main source of ROS. Compared with the control group, the concentration of IL-2, IL-4, IL-6, IFN-γ, and TNF-α was higher in the experimental group. Leukocytes in semen may regulate the secretion of ROS through the mammalian target of rapamycin (mTOR) pathway. A considerable amount of ROS can upregulate the expression of IL-6 in leukocytes via the nuclear factor kappa-B (NF-kB) pathway.

Keywords: IL-6; Leukocytes; NF-kB; mTOR; reactive oxygen species.

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Conflict of interest statement

Declaration of Conflicting Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Figures

Figure 1.
Figure 1.
Sperm motility parameters decreased in the group with a high level of leucocytes. Note. The age, semen volume, sperm concentration, total sperm count, sperm motility rate, fast forward motility sperm rate, forward motility sperm rate, forward motility sperm total number, nonforward motility sperm rate, and inactive sperm rate of the two groups (Leuk. n = 20, Con. n = 20) were counted. *p < .05. **p < .01. *** p < .001.
Figure 2.
Figure 2.
Leukocytes in semen are the main source of ROS. Note. Semen samples of the two groups (Leuk. n = 12, Con. n = 12) were washed with PBS, incubated with anti-human CD45 antibody for 20 min, and then tested for ROS. (A) ROS levels in all cells in semen, statistical analysis of average fluorescence intensity. (B) The dark part is the ROS level in CD45+ leucocytes and the light part is the ROS level in all cells. (C) Comparison of ROS levels between leukocytes in semen and leukocytes in peripheral blood. ROS = reactive oxygen species.
Figure 3.
Figure 3.
Rapamycin, an inhibitor of mTOR, inhibits the secretion of ROS by leucocytes in semen. Note. 500 ng/mL rapamycin was added to semen specimens and the content of ROS was detected by flow cytometry after incubation for 4 hr. (A) Levels of ROS in total cells in semen treated with rapamycin (Rapa. n = 12, Con. n = 12). (B) Levels of ROS in CD45+ leucocytes after rapamycin treatment. mTOR = mammalian target of rapamycin; ROS = reactive oxygen species.
Figure 4.
Figure 4.
Increased levels of IL-2, IL-4, IL-6, IFN-γ, and TNF-α in seminal plasma with a high concentration of leucocytes. Note. The seminal plasma of the two groups (Leuk. n =12, Con. n = 19) was collected, and the concentrations of IL-2, IL-4, IL-6, IFN-γ, and TNF-α were measured by the immuno-magnetic bead method. *p < .05. **p < .01. ***p < .001.
Figure 5.
Figure 5.
The increase of ROS promotes the upregulation of IL-6 expression in semen leucocytes, which is related to the NF-kB pathway. Note. Leukocyte concentration >1 × 106/mL specimens were divided into five groups, each group contains 1 × 107 cells (n = 12): control group; Dimethyl sulfoxide (DMSO) solvent control group; H2O2 group (adding 1% H2O2); BAY11-7082 group (1 mM, MCE, USA); and H2O2 + BAY11-7082 group. After incubation for 2 hr, the expression of IL-6 in CD45+ leucocytes was detected by flow cytometry. H2O2 = hydrogen peroxide; NF-kB = nuclear factor kappa-B; ROS = reactive oxygen species
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References

    1. Agarwal A., Mulgund A., Alshahrani S., Assidi M., Abuzenadah A. M., Sharma R., Sabanegh E. (2014). Reactive oxygen species and sperm DNA damage in infertile men presenting with low level leukocytospermia. Reproductive Biology and Endocrinology, 12(126), 1–8. - PMC - PubMed
    1. Agarwal A., Virk G., Ong C., Plessis S. S. (2014). Effect of oxidative stress on male reproduction. The World Journal of Men’s Health, 32(1), 1–17. - PMC - PubMed
    1. Aitken R. J., Buckingham D. W., Brindle J., Gomez E., Baker H. W., Irvine D. S. (1995). Analysis of sperm movement in relation to the oxidative stress created by leukocytes in washed sperm preparations and seminal plasma. Human Reproduction, 10(8), 2061–2071. - PubMed
    1. Aitken R. J., Smith T. B., Jobling M. S., Baker M. A., De Iuliis G. N. (2014). Oxidative stress and male reproductive health. Asian Journal of Andrology, 16(1), 31–38. - PMC - PubMed
    1. Anderson D. J., Politch J. A., Martinez A., Van Voorhis B. J., Padian N. S., O’Brien T. R. (1991). White blood cells and HIV-1 in semen from vasectomised seropositive men. Lancet, 338(8766), 573–574. - PubMed

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