Potts R J, Notarianni L J, Jefferies T M
School of Pharmacy and Pharmacology, University of Bath, UK.
Mutat Res. 2000 Feb 14;447(2):249-56. doi: 10.1016/s0027-5107(99)00215-8.
Exposure of spermatozoa to reactive oxygen species (ROS) has been associated with cellular injury, that includes DNA damage and lipid peroxidation. In addition, sperm preparation techniques such as centrifugation, commonly used prior to in vitro fertilization and scientific studies, are associated with the generation of ROS and an increase in the level of DNA damage. The preservation, therefore, of sperm in vitro that might decrease the potential for oxidative DNA damage to arise and allow for an improvement in semen quality used for artificial insemination, is of importance. Seminal plasma is a rich source of antioxidants, which, potentially, safeguards sperm from oxidative attack during storage and once ejaculated. We have investigated the protection of human spermatozoa from ROS afforded by seminal plasma. Sperm were exposed to exogenous ROS by incubating the cells with hydrogen peroxide in the presence of ferrous sulfate and ADP. Aliquots of seminal plasma were added to the incubation mixture in differing amounts, and the generation of DNA strand breaks and thiobarbituric acid reactive species (TBARS), indicative of lipid peroxidation, determined. Incubation of sperm with exogenous ROS resulted in a significant generation of DNA strand breaks and lipid peroxidation compared to basal levels of damage (P<0.05). Addition of seminal plasma to the incubation media produced a significant decrease in DNA strand breaks and TBARS (P<0. 05), when the amount of plasma added exceeded 60% v/v. The results indicate that spermatozoal oxidative damage induced by exogenous ROS, specifically DNA damage and lipid peroxidation, is reduced by the presence of seminal plasma.
精子暴露于活性氧(ROS)与细胞损伤有关,包括DNA损伤和脂质过氧化。此外,体外受精和科学研究之前常用的精子制备技术,如离心,与ROS的产生以及DNA损伤水平的增加有关。因此,在体外保存精子以降低氧化DNA损伤的可能性,并改善用于人工授精的精液质量,具有重要意义。精浆是抗氧化剂的丰富来源,可能在储存期间和射精后保护精子免受氧化攻击。我们研究了精浆对人类精子免受ROS损伤的保护作用。通过在硫酸亚铁和ADP存在的情况下用过氧化氢孵育细胞,使精子暴露于外源性ROS。将不同量的精浆等分试样加入孵育混合物中,并测定DNA链断裂和硫代巴比妥酸反应性物质(TBARS)的生成,TBARS是脂质过氧化的指标。与基础损伤水平相比,精子与外源性ROS孵育导致DNA链断裂和脂质过氧化显著增加(P<0.05)。当加入的精浆量超过60%(v/v)时,向孵育培养基中添加精浆可使DNA链断裂和TBARS显著减少(P<0.05)。结果表明,精浆的存在可减少外源性ROS诱导的精子氧化损伤,特别是DNA损伤和脂质过氧化。