Chen Hong, Chow Pak Ham, Cheng So Kwan, Cheung Annie L M, Cheng Lydia Y L, O Wai-Sum
Department of Anatomy, Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, P. R. China.
J Androl. 2003 Sep-Oct;24(5):704-11. doi: 10.1002/j.1939-4640.2003.tb02730.x.
Recently, we reported that male accessory sex gland (ASG) secretions protect sperm genomic integrity by demonstrating that DNA damage was more extensive in sperm not exposed to the secretions. The present study was conducted to find out if ASGs secrete the main antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GPx or GSH-Px), and catalase (CAT) and if the most abundant one, SOD, can protect those sperm that were not exposed to ASG secretions against NADPH-induced oxidative stress. Four experimental groups of male golden hamsters were used: intact animals with proven fertility, animals with all major ASGs removed (TX), animals that were bilaterally vasectomized, and sham-operated controls. SOD, CAT, and GPx activities were measured in secretions from all 5 ASGs and sperm-free uterine flushing from virgin females and those mated with the experimental males. The alkaline comet assay was used to analyze DNA integrity of the TX group sperm after incubation in a medium containing 50 U/mL of SOD along with 0 to 20 mmol/L NADPH. The main antioxidant enzyme in ASGs was SOD from coagulating glands (P <.05) and GPx together with CAT from ampullary glands (P <.05). Uterine flushing of ejaculates that contained ASG secretions had more SOD and CAT activities than those with epididymal secretions alone (P <.05 and P <.001, respectively), whereas activity of GPx was the same (P >.05). Addition of SOD in vitro dose dependently decreased the incidence of single-strand DNA damage in sperm not exposed to ASG secretions incubated in the presence of 0 to 20 mmol/L NADPH (P <.001). These results indicated that, in terms of abundance, SOD was the main antioxidant enzyme secreted by male ASGs, whereas CAT was the second one. The GPx activity came from both epididymis and ASGs. We conclude that ASG secretions play a significant role in protecting sperm against oxidative stress.
最近,我们报告称雄性附属性腺(ASG)分泌物可保护精子基因组完整性,证据是未接触该分泌物的精子中DNA损伤更为广泛。本研究旨在查明ASG是否分泌主要抗氧化酶超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GPx或GSH - Px)和过氧化氢酶(CAT),以及最丰富的一种酶SOD是否能保护未接触ASG分泌物的精子免受NADPH诱导的氧化应激影响。使用了四组雄性金黄地鼠进行实验:经证实有生育能力的完整动物、切除所有主要ASG的动物(TX)、双侧输精管结扎的动物以及假手术对照组。测定了所有5种ASG分泌物以及未交配雌性和与实验雄性交配的雌性的无精子子宫冲洗液中的SOD、CAT和GPx活性。采用碱性彗星试验分析TX组精子在含有50 U/mL SOD以及0至20 mmol/L NADPH的培养基中孵育后的DNA完整性。ASG中的主要抗氧化酶是来自凝固腺的SOD(P <.05)以及来自壶腹腺的GPx和CAT(P <.05)。含有ASG分泌物的射精子宫冲洗液中的SOD和CAT活性高于仅含附睾分泌物的冲洗液(分别为P <.05和P <.001),而GPx活性相同(P >.05)。在体外添加SOD可剂量依赖性降低在0至20 mmol/L NADPH存在下孵育的未接触ASG分泌物的精子中单链DNA损伤的发生率(P <.001)。这些结果表明,就丰度而言,SOD是雄性ASG分泌的主要抗氧化酶,其次是CAT。GPx活性来自附睾和ASG两者。我们得出结论,ASG分泌物在保护精子免受氧化应激方面发挥着重要作用。