1 College of Pharmaceutical Sciences, Nanjing Tech University , Nanjing 210009 , People's Republic of China.
2 College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University , Nanjing 211816 , People's Republic of China.
Open Biol. 2019 Apr 26;9(4):180091. doi: 10.1098/rsob.180091.
Asthenozoospermia is a common cause of male infertility, the aetiology of which remains unclear in 50-60% of cases. The current study aimed to characterize metabolic alterations in asthenozoospermic seminal plasma and to explore the signalling pathways involved in sperm motility regulation. At first, high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry was used to detect the targeted metabolic network of arachidonic acid (AA). Metabolomic multivariate data analysis showed significant distinction of AA metabolites between asthenozoospermic and healthy seminal plasma. AA as well as its lipoxygenase (LOX) and cytochrome P450 metabolites were found to be abnormally increased, while cyclooxygenase (COX) metabolites were complicatedly disturbed in asthenozoospermic volunteers compared with those in healthy ones. In vitro experiments and western blot analysis of sperm cells revealed a decrease in sperm motility and upregulation of sperm phosphor-P38 induced by AA. P38 inhibitor could increase AA-reduced sperm motility. Also, all the inhibitors of the three metabolic pathways of AA could block AA-induced P38 mitogen-activated protein kinase (MAPK) activation and further improve sperm motility. We report here for the first time that an abnormal AA metabolic network could reduce sperm motility via P38 MAPK activation through the LOX, cytochrome P450 and COX metabolic pathways, which might be an underlying pathomechanism of asthenozoospermia.
弱精子症是男性不育的常见原因,其病因在 50-60%的病例中仍不清楚。本研究旨在描述弱精子症精液中代谢改变,并探讨参与精子运动调节的信号通路。首先,采用高效液相色谱-电喷雾串联质谱法检测花生四烯酸(AA)的靶向代谢网络。代谢组学多元数据分析显示,弱精子症和健康精液之间 AA 代谢物存在显著差异。与健康对照组相比,弱精子症志愿者的 AA 及其脂氧合酶(LOX)和细胞色素 P450 代谢物异常增加,而环氧化酶(COX)代谢物则复杂紊乱。体外实验和精子细胞 Western blot 分析显示,AA 可降低精子运动能力,并诱导精子磷酸化-P38 上调。P38 抑制剂可增加 AA 降低的精子运动能力。此外,AA 诱导的 P38 丝裂原活化蛋白激酶(MAPK)激活和进一步改善精子运动能力的三种 AA 代谢途径的所有抑制剂均可阻断。我们首次报道,异常的 AA 代谢网络可能通过 LOX、细胞色素 P450 和 COX 代谢途径通过 P38 MAPK 激活降低精子运动能力,这可能是弱精子症的潜在发病机制。