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Nitrosative stress in human spermatozoa causes cell death characterized by induction of mitochondrial permeability transition-driven necrosis

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Asian Journal of Andrology 2018年第6期20卷 600-607页

作者： Pamela Uribe Maria E Cabrillana Miguel W Forn Favi/m Treulen Rodrigo Boguen vladimir isachenko Evgenia isachenko Ratil S inchez Juana V Villegas Center of Reproductive Biotechnology-Scientific and Technological Bioresource Nucleus（CEBIOR-BIOREN） University of La FronteraTemuco 4811230Chile Center of Excellence in Translational Medicine University of La FronteraTemuco 4810296Chile Department of Internal Medicine Faculty of MedicineUniversity of La FronteraTemuco 4781218Chile %aboratory of Andrology Research of Mendoza（LIAM）Institute of Histology and Embriology of Mendoza（IHEM）Histology and Embryology Area Department of Morphology and PhysiologySchool of MedicineNational University of Cuyo and CCT-MendozaCONICETMendoza 5500Argentin SResearch Institute（CIUDA） Medicine FacultyAcencagua UniversityMendoza 5500Argentina Research Group for Reproductive Medicine Cologne UniversityCologne 50937Germany Department of Preclinical Sciences Faculty of MedicineUniversity of La FrenteraTemuco 4781218Chile.

Peroxynitrite is a highly reactive nitrogen species and a potent inducer of apoptosis and necrosis in somatic cells. Peroxynitrite-induced nitrosative stress has emerged as a major cause of impaired sperm function; however, its ability to trigger cell death has not been described in human spermatozoa. The objective here was to characterize biochemical and morphological features of cell death induced by peroxynitrite-mediated nitrosative stress in human spermatozoa. For this, spermatozoa were incubated with and without （untreated control） 3-morpholinosydnonimine （SIN-l）, in order to generate peroxynitrite. Sperm viability, mitochondrial permeability transition （MPT）, externalization of phosphatidylserine, DNA oxidation and fragmentation, caspase activation, tyrosine nitration, and sperm ultrastructure were analyzed. The results showed that at 24 h of incubation with SIN-l, the sperm viability was significantly reduced compared to untreated control （P〈 0.001）. Furthermore, the MPT was induced （P〈 0.01） and increment in DNA oxidation （P 〈 0.01）, DNA fragmentation （P 〈 0.01）, tyrosine nitration （P 〈 0.0001） and ultrastructural damage were observed when compared to untreated control. Caspase activation was not evidenced, and although phosphatidylserine externalization increased compared to untreated control （P 〈 0.001）, this process was observed in 〈10% of the cells and the gradual loss of viability was not characterized by an important increase in this parameter. In conclusion, peroxynitrite-mediated nitrosative stress induces the regulated variant of cell death known as MPT-driven necrosis in human spermatozoa. This study provides a new insight into the pathophysiology of nitrosative stress in human spermatozoa and opens up a new focus for developing specific therapeutic strategies to better preserve sperm viability or to avoid cell death.

关键词： necrosis nitrosative stress oxidative stress peroxynitrite sperm cell death

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