Proinflammatory Cytokines as an Intermediate Factor Enhancing Lipid Sperm Membrane Peroxidation in In Vitro Conditions

Abstract

ABSTRACT:We have examined the effect of white blood cells (WBCs), various proinflammatory cytokines, or a combination of the two on the peroxidation of human sperm membrane lipids in in vitro conditions. Six recombinant cytokines, such as interleukin-1b (IL-1b), IL-6, IL-8, IL-12, IL-18, and tumor necrosis factor alpha (TNF-a), used singly or in combinations, were analyzed. WBCs were isolated from the whole heparinized blood using a density gradient technique (Histopaque 1.077). Spermatozoa were isolated from semen samples with normal sperm parameters by both the swim-up technique (swim-up fraction) and by a discontinuous Percoll gradient centrifugation (90% and 47% Percoll fractions). Peroxidative damage to sperm membrane lipids was assessed by determining the concentration of malondialdehyde (MDA) in lysates of spermatozoa using high-performance liquid chromatography (HPLC). There were no statistically significant differences in MDA concentrations between sperm fractions incubated with cytokines and respective controls (spermatozoa alone). In spermatozoa isolated by the swimup technique, the MDA level was significantly higher only after incubation with IL-6 and IL-8 plus WBCs when compared to sperm incubated with leukocytes alone (0.62 6 0.21 mmol/L and 0.42 6 0.22 mmol/L, respectively; P , .05). In spermatozoa recovered from the 47% Percoll, only a combination of IL-12 and IL-18 used together with WBCs was linked with a significant increase in MDA concentration (from 0.41 6 0.13 mmol/L to 0.65 6 0.19 mmol/L; P , .05). The results obtained suggest that cytokines produced during the inflammatory process intensify the level of oxidative stress caused by leukocytes, which may have serious consequences for sperm membrane integrity.

Mechanisms of the harmful effects of bacterial semen infection on ejaculated human spermatozoa: potential inflammatory markers in semen

“…This has been confirmed by numerous reports that have documented correlations between seminal leukocyte count and proinflammatory cytokine concentrations [140,[143][144][145][146][147][148]. Also in our own in vitro observations, the peroxidative damage of sperm membrane lipids was markedly enhanced in the simultaneous presence of cytokines and leukocytes [149]. Most probably, oxidative stress during semen inflammation and infection is modulated by the levels of certain cytokines; www.fhc.viamedica.pl even more, during leukocytospermia, the ROS and cytokines produced by the leukocytes cooperate with each other, provoking a destructive effect on sperm membranes.…”

Recent scenario of obesity and male fertility

“…Role of cytokines is documented in fertility and they are secreted by different parts of the male genital tract and exert effects on steroidogenesis, spermatogenesis, and sperm functions (Shukla et al, 2013a,b). Proinflammatory cytokines such IL-1, IL-6, and TNFa are capable of reducing the ova-penetrating ability of spermatozoa (Gruschwitz et al, 1996).These cytokines being mediator of inflammation are also increased in obese males (Fraczek et al, 2008) as supported by earlier studies that have documented positive correlation of cytokines like IL-6 with BMI and body fat mass (Berthier et al, 2003). Low testosterone in young as well as old men may be associated with low grade systemic inflammation which might be a part of the mechanism underlying adverse health outcomes of male hypogonadism as connection is not exclusively dependent on the amount of adipose tissue, but may also be an independent effect of androgens (Bobjer et al, 2013), (Fig.…”

Effect of Interleukin-1β on Lipid Peroxidation in Emotiogenic Structures of the Brain in Rats during Acute Stress

“…One of the mechanisms for intensifi cation of lipid metabolism is probably associated with the cytokine-induced inhibition of some antioxidant enzymes in the brain [9].…”