Current Issue

Volume 13, Number 4, Jan-Mar 2020 Pages: 301-306

Evaluation of Enkephalin-Degrading Enzymes in Sperm from Heroin-Addicted Men

Samira Rezaei-Mojaz, M.Sc, 1, #, Zohreh Nazmara, Ph.D, 1, #, Mohammad Najafi, Ph.D, 2, 3, Mansoureh Movahedin, Ph.D, 4, Zahra Zandieh, Ph.D, 2, 6, Peymaneh Shirinbayan, Ph.D, 8, Mohsen Roshanpajouh, M.D, 7, Hamid Reza Asgari, Ph.D, 1, Mehdi Abbasi, Ph.D, 8, Morteza Koruji, Ph.D, 1, 2, *,
Department of Anatomical Sciences, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran
Department of Biochemistry, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
Anatomical Sciences Department, School of Medicine, Tarbiat Modares University, Tehran, Iran
Shahid Akbarabadi Clinical Research Development Unit (ShACRDU), Iran University of Medical Sciences, Tehran, Iran
Pediatric Neuro-Rehabilitation Research Center, The University of Social Welfare and Rehabilitation Sciences, Tehran, Iran
School of Behavioral Sciences and mental Health, Tehran Psychiatry Institute, Iran University of Medical Sciences, Tehran, Iran
Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
*Corresponding Addresses: P.O.Box: 14155-5983 Cellular and Molecular Reearch Center Iran University of Medical Sciences Tehran Iran Department of Anatomical Sciences School of Medicine Iran University of Medical Sciences Tehran Iran

The first two authors equally contributed to this work.



The aim of this study was to investigate two enkephalin-degrading enzymes, aminopeptidase N (APN/ CD13) and endopeptidase (NEP/CD10), gene and protein expression levels in sperm samples of fertile and heroin- addicted men, and the correlation between their expressions and semen quality.

Materials and Methods

In this case-controlled study, semen was collected from 24 normozoospermic healthy (as a control group) and 24 heroin-addicted men donors (as case or addiction group). Sperm cells isolated by Cook Medical gradient (40-80%) and followed up by swim-up techniques were used for real-time quantitative polymer- ase chain reaction (qPCR) and flow cytometry techniques to assess APN/CD13 and NEP/CD10 genes and proteins subsequently. Semen parameters were analyzed by computer-assisted sperm analysis.


The findings revealed that there were significant differences in sperm total motility (41.07 ± 3.63 vs. 63.03 ± 3.31 %, P=0.0001), progressive motility (35.21 ± 2.64 vs. 20.93 ± 3.22%, P=0.001) and viability (69.9 ± 4.69 vs. 86.81 ± 1.26 %, P=0.002) in the addicted group vs. control ones. APN and NEP gene expression levels in the addicted group decreased compared with the control ones (1.00 ± 0.67 vs. 0.36 ± 0.13, P= 0.008 and 1.07 ± 0.11 vs. 0.52 ± 0.12 0.002, respectively). Flow cytometry analysis showed that the average percent of APN/CD13 in heroin consumers significantly decreased compared with the healthy ones, while NEP/CD10 rate between two groups was similar. We also observed that duration of drug dependence is correlated with sperm viability (r=-0.627, P=0.016) and motility (r=-0.410, P=0.05), NEP (r=-0.434, P= 0.049), and APN (r=-0.641, P=0.002) gene expression levels.


We conclude that semen quality and enkephalin-degrading enzymes were altered in heroin-addicted men. other confirming the internal validity of our estimates.