Investigation of molecular cryopreservation, fertility potential and microRNA-mediated apoptosis in Oligoasthenoteratozo

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Investigation of molecular cryopreservation, fertility potential and microRNA-mediated apoptosis in Oligoasthenoteratozoospermia men Maryam Ezzati . Dariush Shanehbandi . Behzad Bahramzadeh . Kobra Hamdi . Maryam Pashaiasl

Received: 16 January 2020 / Accepted: 4 October 2020 Ó Springer Nature B.V. 2020

Abstract Investigation of the cryo-injury mechanism can provide novel insight into cryopreservation. The objective of this study is to assess the effect of cryopreservation on fertility potential, motility, oxidative stress (OS), DNA fragmentation, microRNAs (miRNAs), and apoptotic target genes in the infertile men compared to the fertile men. All 40 samples were divided into two leading groups of fresh and cryopreserved sperms. Each main group was subdivided into three groups including, Normozoospermia, and Mild, and Severe Oligoasthenoteratozoospermia (OAT). In all collected samples the following were assessed: microRNA-34c (miR-34c) and miR-184, P53 and Caspase9 using Quantitative real-time polymerase chain reaction (RT-PCR), malondialdehyde (MDA), Superoxide dismutase (SOD) using imaging

multi-mode reader, and DNA fragmentation using Sperm DNA Fragmentation Assay Test (SDFA). Within the studied groups, immotile spermatozoa were increased due to cryopreservation. We observed an increasing levels of SOD, MDA, and DNA fragmentation. Also, cryopreservation was associated with decreasing the expression of P53, mir-43c, and miR-184 while capase9 was showed enhancing expression after freeze-thawing of sperm cells. During cryopreservation, sperm fertility and motility were influenced via apoptosis cascade-mediated mitochondrial dysfunctions such as caspase9. Also, we found that miR-34c, miR184, and P53 could impact fertility potential. In Addition, there was a meaningful correlations between microRNAs and motility post freeze-

M. Ezzati  D. Shanehbandi  M. Pashaiasl Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

K. Hamdi  M. Pashaiasl (&) Women’s Reproductive Health Research Center, Tabriz University of Medical Sciences, P.O. Box 51376563833, Tabriz, Iran e-mail: [email protected]

M. Ezzati  M. Pashaiasl Department of Anatomical Sciences, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran B. Bahramzadeh Bachelor of Laboratory Sciences, Al-Zahra Hospital, Tabriz University of Medical Sciences, Tabriz, Iran

K. Hamdi  M. Pashaiasl Department of Reproductive Biology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran

B. Bahramzadeh Member of Nursing Research Committee of Al-Zahra Hospital, Tabriz University of Medical Sciences, Tabriz, Iran

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Cell Tissue Bank

thawing process in Severe Oligoasthenoteratozoospermia men. Keywords Cryopreservation  Oligoasthenoteratozoospermia  MicroRNA  DNA fragmentation

Introduction Research studies have revealed that the cryopreservation of human semen is useful for different purposes such as chemotherapy and infertility cases (