Melatonin affects membrane integrity, intracellular reactive oxygen species, caspase3 activity and AKT phosphorylation in frozen thawed human sperm

Abstract

Cryopreservation is known to induce oxidative stress in spermatozoa. Although melatonin has powerful antioxidant properties, little is known about its effects on human sperm quality during cryopreservation. The present study was undertaken to investigate the effects of melatonin treatment on human sperm parameters essential for fertilization. We first evaluated the effects of various concentrations of melatonin (0-15 mM) on human sperm parameters such as motility, viability and levels of intracellular reactive oxygen species during cryopreservation in order to identify an optimal dose with the greatest effects for further studies. Liquefied semen samples were then divided into three aliquots: cryopreserved without melatonin (control), cryopreserved with 3 mM melatonin and fresh groups. After being thawed, samples were evaluated for motility, viability, membrane integrity, intracellular reactive oxygen species levels, caspase-3 activity and AKT phosphorylation. Treatment of spermatozoa with the various concentrations of melatonin significantly increased their motility and viability and decreased their intracellular reactive oxygen species levels compared with the control group. The optimal melatonin concentration (3 mM) significantly decreased the intracellular reactive oxygen species levels, caspase-3 activity and the percentage of both dead and apoptotic-like sperm cells and increased the vitality, progressive motility and total motility and AKT phosphorylation compared with the control group. Thus, melatonin exerts protective effects against cryodamage during human spermatozoa cryopreservation and may exert its effects via the PI3K/AKT signaling pathway.

Melatonin affects the motility and adhesiveness of in vitro capacitated boar spermatozoa via a mechanism that does not depend on intracellular ROS levels

“…Furthermore, melatonin has been shown to improve motility and other sperm functional parameters in human, ram, equine and boar spermatozoa (reviewed in Cebri an-P erez et al, 2014). In agreement with this hypothesis, in vitro treatment of spermatozoa with melatonin decreases intracellular levels of reactive oxygen (ROS) and nitrogen species (RNS; Rao & Gangadharan, 2008;Du Plessis et al, 2010;Jang et al, 2010;Najafi et al, 2018), membrane lipid peroxidation (Gadella et al, 2008;Du Plessis et al, 2010;Da Silva et al, 2011), apoptosis markers Da Silva et al, 2011;Espino et al, 2011;Najafi et al, 2018) and DNA fragmentation (Sarabia et al, 2009). In agreement with this hypothesis, in vitro treatment of spermatozoa with melatonin decreases intracellular levels of reactive oxygen (ROS) and nitrogen species (RNS; Rao & Gangadharan, 2008;Du Plessis et al, 2010;Jang et al, 2010;Najafi et al, 2018), membrane lipid peroxidation (Gadella et al, 2008;Du Plessis et al, 2010;Da Silva et al, 2011), apoptosis markers Da Silva et al, 2011;Espino et al, 2011;Najafi et al, 2018) and DNA fragmentation (Sarabia et al, 2009).…”

“…While this conclusion could be surprising at first glance, one should note that boar spermatozoa are characterised by a very low ROS production rate, even when subjected to treatments that, such as freeze-thawing, induce the generation of high intracellular ROS in other species (Bilodeau et al, 2000;Guthrie & Welch, 2006;Yeste et al, 2013Yeste et al, , 2015a. As a consequence, in species such as equine, bovine and human, melatonin has a clear antioxidant effect and affects membrane lipid peroxidation and ROS levels (Gadella et al, 2008;Rao & Gangadharan, 2008;Du Plessis et al, 2010;Jang et al, 2010;Da Silva et al, 2011;Najafi et al, 2018). As a consequence, in species such as equine, bovine and human, melatonin has a clear antioxidant effect and affects membrane lipid peroxidation and ROS levels (Gadella et al, 2008;Rao & Gangadharan, 2008;Du Plessis et al, 2010;Jang et al, 2010;Da Silva et al, 2011;Najafi et al, 2018).…”

The efficacy of antioxidants in sperm parameters and production of reactive oxygen species levels during the freeze‐thaw process: A systematic review and meta‐analysis

“…During full‐text screening, no common antioxidants were identified in nineteen articles (Asadmobini, Bakhtiari, Khaleghi, Esmaeili, & Mostafaei, ; Brugnon et al, ; Fontoura et al, ; Isachenko et al, ; Kotdawala et al, ; Lu et al, ; Meamar et al, ; Naeini, Bafrani, & Nikzad, ; Najafi et al, ; Oehninger, Duru, Srisombut, & Morshedi, ; Parameswari et al, ; Pariz & Hallak, ; Saeednia et al, ; Sbracia, Grasso, Sayme, Stronk, & Huszar, ; Sobhani, Eftekhaari, Shahrzad, Natami, & Fallahi, ; Taylor, Roberts, Sanders, & Burton, ; Vickram et al, ; Werner et al, ), twelve articles lacked common sperm parameters (Banihani, Agarwal, Sharma, & Bayachou, ; Branco, Garcez, Pasqualotto, Erdtman, & Salvador, ; Donnelly, McClure, & Lewis, ; Esteves, Spaine, & Cedenho, ; Gadea et al, ; Garcez et al, ; Garcez, Branco, Lara, Pasqualotto, & Salvador, ; Jenkins, Aston, & Carrell, ; Li, Lin, Liu, Xiao, & Liu, ; Minaei et al, ; Nabi et al, ; Shabani Nashtaei et al, ) and one article lacked sufficient data (Karimfar et al, ). Finally, 23 articles (Aliabadi, Jahanshahi, Talaei‐Khozani, & Banaei, ; Askari, Check, Peymer, & Bollendorf, ; Azadi, Tavalaee, Deemeh, Arbabian, & Nasr‐Esfahani, ; Bateni et al, ; Deng et al, ; Duru, Morshedi, Schuffner, & Oehninger, ; Esteves, Sharma, Thomas, & Agarwal, ; Ghorbani et al, ; Kalthur et al, ; Keshtgar, Iravanpour, Gharesi‐Fard, & Kazerooni, ; Martinez‐Soto, Dioshourcade, Gutiérrez‐Adán, Landeras, & Gadea, ; Merino et al, ; Moubasher, Din, Ali, El‐sherif, & Gaber, ; Najafi et al, ; Nekoonam, Nashtaei, Naji, Zangi, & Amidi, ; Rossi, Mazzilli, Delfino, & Dondero, ; Taylor et al, ; Thomson et al, ; Varghese et al, ; Wang, Sikka, Veeraragavan, Bell, & Hellstr...…”

“…In fact, caspase 3 is activated after thawing and alters the outer layer of the sperm plasma membrane. It was shown that caspase 3 activity was decreased in the melatonin group in comparison with the control group (Najafi et al, ). Macromolecules are protected by melatonin against OS.…”

“…Since the 1960s, sperm cryopreservation has been introduced as a routine practice in infertility clinics (Najafi et al, ). This technique is being utilised for several indications, including donor insemination programmes, before treatments which my hurt fertility potential such as chemotherapy, radiotherapy, orchiectomy and vasectomy, testicular sperm retrieval in azoospermic cases and epididymal sperm aspiration (obstructive azoospermia), gender reassignment, spinal cord injury and hypogonadotropic hypogonadism (Banihani & Alawneh, ; Pabon & Meseguer, ; Wierckx et al, ; Yang, Xu, Cui, Wu, & Liao, ).…”