TY  - JOUR
T1  - Investigation of the Expression Level of Autophagy Proteins in Infertile Men
AU  - Can, Nazlı
AU  - Ergin, Kemal
PY  - 2025
DA  - September
Y2  - 2025
DO  - 10.33808/clinexphealthsci.1379892
JF  - Clinical and Experimental Health Sciences
PB  - Marmara University
WT  - DergiPark
SN  - 2459-1459
SP  - 463
EP  - 472
VL  - 15
IS  - 3
LA  - en
AB  - Objective: Infertility nowadays affects 10–15% of couples in their reproductive age. Autophagy is a physiologic process responsible for the breakdown of cytoplasmic structures and organelles, and proteins that have lost functionality. In this study, it was aimed to compare the levels of autophagy expression in the spermatozoa of men who have infertility for different reasons.Methods: A total of 120 men were included in the study, and four groups were designed: men who have no infertility disorder and are healthy according to semen parameters (Group 1, control group), patients diagnosed with Grade 2 varicocele (Group 2), with orchitisepididymitis (Group 3), and with unexplained infertility (Group 4). Autophagy proteins (LC3A/B, Beclin-1, p62, ATG5, ATG12, and ATG16) were examined by western blot analysis. LC3A/B and Beclin-1 were also studied by immunohistochemistry. In addition, each patient group specimen was investigated ultrastructurally by Transmission Electron Microscopy.Results: Sperm parameters of the infertility groups decreased compared with the control group. Immunohistochemically, all autophagy proteins were increased in patient groups compared to the control group. Western blot analysis and ultrastructural studies also confirmed these data.Conclusion: These results showed that varicocele, orchitis-epididymitis, and unexplained infertility may cause impairment in the autophagy mechanism. The explanation of this mechanism can lead to the development of new treatment options.
KW  - Autophagy
KW  - Infertility
KW  - Spermatozoon
KW  - Immunohistochemistry
CR  - World Health Organization. Laboratory Manual For The Examination and Processing of Human Semen, 5th edition. [07 September 2025]. https://www.who.int/reproductivehealth/publications/infertility/978.924.1547789/en/
CR  - Agarwal A, Mulgund A, Hamada A, Chyatte MR. A unique view on male infertility around the globe. Reproductive Biology and Endocrinology. 2015;13:37. https://doi.org/10.1186/s12958.015.0032-1
CR  - Hamada A, Esteves SC, Nizza M, Agarwal A. Unexplained male infertility: Diagnosis and management. International
Braz J Urol. 2012;38(5):576-594. https://doi.org/10.1590/S1677.553.8201200.050.0002
CR  - Aparicio IM, Espino J, Bejarano I, Gallardo-Soler A, Campo ML, Salido GM. autophagy-related proteins are functionally active in human spermatozoa and may be ınvolved ın the regulation
of cell survival and motility. Scientific Reports. 2016;6:33647:1-19. https://doi.org/10.1038/srep33647
CR  - Song WH, Yi YJ, Sutovsky M, Meyers S, Sutovsky P. The art and science of sperm mitophagy. Autophagy. 2016;12(12):2510-2511. https://doi.org/10.1080/15548.627.2016.1226735
CR  - Kroemer G, Marino G, Levine B. Autophagy and the ıntegrated stress response. Molecular Cell. 2010;40(2):280-293. https://doi.org/10.1016/j.molcel.2010.09.023
CR  - Zhang YW, Liu J, Ma KH, Tan Y. Effects of autophagy on spermatogenesis and sperm motility. Zhonghua Nan Ke
Xue. 2019;25(1):72-76. https://doi.org/10.13263/j.cnki.nja.2019.01.015
CR  - Can NK. Male reproductive system and autophagy. International Journal of Contemporary Health Sciences
2020;1(1):10-14. https://doi.org/10.37506/ijochs.v1i1.1207
CR  - Komatsu M, Waguri S, Koike M, Sou YS, Ueno T, Hara T, Mizushima N, Iwata J, Ezaki J, Murata S, Hamazaki J, Nishito
Y, Iemura S, Natsume T, Yanagawa T, Uwayama J, Warabi E, Yoshida H, Ishii T, Kobayashi A, Yamamoto M, Yue Z, Uchiyama Y, Kominami E, Tanaka K. Homeostatic levels of p62 control cytoplasmic ınclusion body formation ın autophagy-deficient mice. Cell. 2007;131(6):1149-1163. https://doi.org/10.1016/j.cell.2007.10.035
CR  - Xie Z, Klionsky DJ. Autophagosome formation: core machinery and adaptation. Nature Cell Biology 2007; 9(10):1102-1109. https://doi.org/10.1038/ncb1007-1102
CR  - zmirli M, Ecevit H, Göğebakan B. Autophagy to survive. Archives Medical Review Journal 2014;23(3):411-419. https://
doi.org/10.17827/aktd.22989
CR  - Lessene G, Czabotar PE, Colman PM. BCL-2 Family antagonists for cancer therapy. Nature Reviews Drug Discovery
2008;7(12):989-1000. https://doi.org/10.1038/nrd2658
CR  - Sridharan S, Jain K, Basu A. Regulation of autophagy by kinases. Cancers (Basel). 2011;3(2):2630-2654. https://doi.
org/10.3390/cancers3022630
CR  - Chen Y, McMillan-Ward E, Kong J, Israels SJ, Gibson SB. Oxidative stress ınduces autophagic cell death ındependent of apoptosis ın transformed and cancer cells. Cell Death and Differentiation 2008;15(1):171-182. https://doi.org/10.1038/sj.cdd.4402233
CR  - Dikic I, Elazar Z. Mechanism and medical ımplications of mammalian autophagy. Nature Reviews Molecular Cell
Biology 2018;19(6):349-364. https://doi.org/10.1038/s41580.018.0003-4
CR  - Check JH, Adelson HG, Schubert BR, Bollendorf A. Evaluation of sperm morphology using kruger’s strict criteria.
Archives of Andrology 1992;28(1):15-17. https://doi.org/10.3109/014.850.19208987687
CR  - Cilaker Micili S, Ergur BU, Kuscu NK. Immunohistochemical and ultrastructural analysis of the effect of omega-3 on embryonic ımplantation ın an experimental mouse model. Taiwanese Journal of Obstetrics and Gynecology 2016;55(3):351-356. https://doi.org/10.1016/j.tjog.2016.04.034
CR  - Mazières J, Brugger W, Cappuzzo F, Middel P, Frosch A, Bara I, Klingelschmitt G, Klughammer B, Ocaña A, von
Pawel J, Perol M, Ramlau R, Rolski J, Kim JH, Robinet G, Wiewrodt R, Dediu M, Reck M. Evaluation of the egfr protein
expression by ımmunohistochemistry using h-score and the magnification rule: re-analysis of the saturn study. Lung Cancer 2013;82(2):231-237. https://doi.org/10.1016/j.lungcan.2013.08.004
CR  - Springhorn A, Hoppe T. Western blot analysis of the autophagosomal membrane protein lgg-1/lc3 ın caenorhabditiselegans. Methods In Enzymology 2019;619:319-336. https://doi.org/10.1016/bs.mie.2018.12.028
CR  - Eskandari M, Ghalyanchi LA, Zeighami H, Rostami A, Kazemi M, Eyni H, Shokri S. Co-administration of ginseng and ciprofloxacin ameliorates epididymo-orchitis ınduced alterations ın sperm quality and spermatogenic cells apoptosis following ınfection ın rats. Andrologia 2017;49(10):1-11. https://doi.org/10.1111/and.12778
CR  - Lin CY, Chen CY, Yu CH, Yu IS, Lin SR, Wu JT, Wang SH, Tsai YT, Lin SW, Hsu WC, Chang CY, Hsu HM, Wang JY, Hsu PH, Lin YW, Liu KL, Yen PH, Kuo HC, Lin YS, Yang SH, Chen YT. Human x-linked ıntellectual disability factor cul4b ıs required for post-meiotic sperm development and male fertility. Scientific Reports 2016;6:1-17. https://doi.org/10.1038/srep20227
CR  - Aparicio IM, Rojo-Domínguez P, Castillejo-Rufo A, Peña FJ, Tapia JA. The autophagy marker lc3 ıs processed during
the sperm capacitation and the acrosome reaction and translocates to the acrosome where ıt colocalizes with the acrosomal membranes ın horse spermatozoa. International Journal of Molecular Sciences 2023;24(2):937:1-15. https://doi.org/10.3390/ijms24020937
CR  - Wang LH, Zheng L, Jiang H, Jiang T. Research advances in inflammation and oxidative stress in varicocele-induced male infertility: A narrative review. Asian Journal of Andrology 2025;27(2):177-184. https://doi.org/10.4103/aja202514
CR  - Zhu SH, Rao T, Yang X, Nin JZ, Yu WM, Ruan Y, Yuan R, Li C, Jiang K, Huang W, Cheng F. Autophagy may play an important role in varicocele. Molecular Medicine Reports 2017;16(5):5471-5479. https://doi.org/10.3892/mmr.2017.7274
CR  - Agarwal A, Sharma RK, Desai NR, Prabakaran S, Tavares A, Sabanegh E. Role of oxidative stress in the pathogenesis of varicocele and infertility. Urology 2009;73(3):461-469. https://doi.org/10.1016/j.urology.2008.07.041
CR  - Wu H, Wang ZF, Jiang Q, Shi L, Gong M, Liu W, Gao B, Song C, Li Q, Chen Y, Han D. Mouse testicular cell type-specific
antiviral response against mumps virus replication. Frontiers In Immunology 2017;8:117:1-12. https://doi.org/10.3389/
fimmu.2017.00117
CR  - Xiong YW, Li DX, Ling ZJ, Tan LL, Zhang YF, Zhang J, Li H, Chang W, Zhu HL, Zhang J, Gao L, Xu DX, Yang L, Wang H. Loss of atg5 in sertoli cells enhances the susceptibility of cadmium-impaired testicular spermatogenesis in mice. Food and Chemical Toxicology 2023;179:113967:1-12. https://doi.org/10.1016/j.fct.2023.113967
CR  - Aslani F, Sebastian T, Keidel M, Fröhlich S, Elsasser HP, Schuppe HC, Fijak M, Mayerhofer A, Klug J, Meinhardt A. Resistance to apoptosis and autophagy leads to enhanced survival ın sertoli cells. Molecular Human Reproduction 2017;23(5):370-380. https://doi.org/10.1093/molehr/gax017
CR  - Scherz-Shouval R, Shvets E, Fass E, Shorer H, Gil L, Elazar Z. Reactive oxygen species are essential for autophagy
and specifically regulate the activity of atg4. EMBO Journal 2007;26(7):1749-1760. https://doi.org/10.1038/
sj.emboj.7601623
CR  - Uribe P, Merino J, Matus CE, Schulz M, Zambrano F, Villegas JV, Conejeros I, Taubert A, Hermosilla C, Sánchez R. Autophagy ıs activated ın human spermatozoa subjected to oxidative stress and its inhibition impairs sperm quality and promotes cell death. Human Reproduction 2022;37(4):680-695. https://doi.org/10.1093/humrep/deac024
CR  - Shojaedini M, Hemadi M, Saki G, Fakhredini F, Khodayar MJ, Khorsandi L. Thymoquinone effects on autophagy,
apoptosis, and oxidative stress in cisplatin-induced testicular damage in mice. Journal of Assisted Reproduction and Genetics 2024;41(7):1881-1891. https://doi.org/10.1007/s10815.024.03295-1
CR  - Li X, Qiao F, Guo J, Jiang T, Lou H, Li H, Shi L, Yang Q, Xu W, Zhang Y, Chen M, Wang S, Liu X, Sun J, Liu G, Zhao Y, Wang
H. In situ architecture of the intercellular organelle reservoir between epididymal epithelial cells by volume electron microscopy. Nature Communications 2023;14:4195:1-12. https://doi.org/10.1038/s41467.023.39634-4
CR  - Sidjanin DJ, Park AK, Ronchetti A, Martins J, Jackson WT. TBC1D20 mediates autophagy as a key regulator of
autophagosome maturation. Autophagy 2016;12(10):1759-1775. https://doi.org/10.1080/15548.627.2016.1203505
UR  - https://doi.org/10.33808/clinexphealthsci.1379892
L1  - https://dergipark.org.tr/en/download/article-file/3491081
ER  -