Tehran University of Medical Sciences Iranian Journal of Public Health 2251-6085 54 1 2025 01 05 214 224 Farnoush Naseri Department of Genetics, Faculty of Biological Science, North Tehran Branch, Islamic Azad University, Tehran, Iran Elham Siasy Department of Genetics, Faculty of Biological Science, North Tehran Branch, Islamic Azad University, Tehran, Iran Seyed Abdolhamid Angaji Department of Cell and Molecular Biology, Faculty of Biological Science, Kharazmi University, Tehran, Iran Maryam Peyvandi Department of Genetics, Faculty of Biological Science, North Tehran Branch, Islamic Azad University, Tehran, Iran 2023 12 24 2024 08 07 Background: The cause of oligoasthenoteratozoospermia (OAT) is unclear. In this original study, we examined OAT patients' sperm using RNA-sequencing and studied the impact of curcumin on gene expression during sperm cryopreservation and freeze-thawing. Methods: RNA-seq was performed using the Galaxy Europe server in IKHC (Imam Khomeini Hospital Complex), Tehran, Iran in 2023. Sperm samples were collected from 30 OAT patients and 30 healthy volunteers. Sperm parameters were analyzed, and samples were frozen with 20 μM curcumin at -196 °C for one week. Thawed samples were assessed for sperm parameters, and the expression levels of bax, bak, bcl-2, bclw, casp9, apaf-1, SOD, cat, and GPX4 genes were measured using RT-PCR. Result: RNA-seq analysis showed increased expression of NANOS1, HSPA6, and ALOXE3 genes, while BHLHE41, Hey1, and PPM1D genes were down-regulated in OAT patients' sperm. Curcumin (20 μM) effectively preserves sperm parameters and motility during cryopreservation in healthy subjects and in OAT patients in particular. Cryopreserved sperm from both OAT patients and healthy individuals exhibited reduced expression of pro-apoptotic genes, increased expression of anti-apoptotic genes, and elevated levels of SOD and GPX4 genes. Conclusion: Altered expressions of NANOS1, HSPA6, ALOXE3, BHLHE41, Hey1, and PPM1D genes likely contribute to OAT development. Additionally, curcumin protects sperm parameters during cryopreservation for both healthy individuals and OAT patients. https://ijph.tums.ac.ir/index.php/ijph/article/view/34097 https://ijph.tums.ac.ir/index.php/ijph/article/download/34097/8452