Fraczek Monika, Sanocka Dorota, Kamieniczna Marzena, Kurpisz Maciej
Institute of Human Genetics, Polish Academy of Sciences, Strzeszynska 32, 60-479 Poznan, Poland.
J Androl. 2008 Jan-Feb;29(1):85-92. doi: 10.2164/jandrol.107.003319. Epub 2007 Sep 5.
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-1beta (IL-1beta), IL-6, IL-8, IL-12, IL-18, and tumor necrosis factor alpha (TNF-alpha), 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 swim-up 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 +/- 0.21 micromol/L and 0.42 +/- 0.22 micromol/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 +/- 0.13 micromol/L to 0.65 +/- 0.19 micromol/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.
我们在体外条件下研究了白细胞（WBC）、各种促炎细胞因子或二者组合对人精子膜脂质过氧化的影响。分析了六种重组细胞因子，如白细胞介素-1β（IL-1β）、IL-6、IL-8、IL-12、IL-18和肿瘤坏死因子α（TNF-α），单独使用或联合使用。使用密度梯度技术（Histopaque 1.077）从全肝素化血液中分离白细胞。通过上浮技术（上浮部分）和不连续Percoll梯度离心法（90%和47% Percoll部分）从具有正常精子参数的精液样本中分离精子。通过使用高效液相色谱法（HPLC）测定精子裂解物中丙二醛（MDA）的浓度来评估精子膜脂质的过氧化损伤。在与细胞因子孵育的精子部分和各自的对照（仅精子）之间，MDA浓度没有统计学上的显著差异。在上浮技术分离的精子中，与仅与白细胞孵育的精子相比，仅在与IL-6和IL-8加白细胞孵育后，MDA水平显著更高（分别为0.62±0.21μmol/L和0.42±0.22μmol/L；P<.05）。在从47% Percoll中回收的精子中，仅IL-12和IL-18与白细胞一起使用的组合与MDA浓度的显著增加有关（从0.41±0.13μmol/L增加到0.65±0.19μmol/L；P<.05）。获得的结果表明，炎症过程中产生的细胞因子会加剧白细胞引起的氧化应激水平，这可能对精子膜完整性产生严重后果。
