Research Article
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Year 2025, Volume: 15 Issue: 3, 463 - 472, 30.09.2025
https://doi.org/10.33808/clinexphealthsci.1379892

Abstract

Project Number

TPF-17038

References

  • 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/
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • Sridharan S, Jain K, Basu A. Regulation of autophagy by kinases. Cancers (Basel). 2011;3(2):2630-2654. https://doi. org/10.3390/cancers3022630
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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

Investigation of the Expression Level of Autophagy Proteins in Infertile Men

Year 2025, Volume: 15 Issue: 3, 463 - 472, 30.09.2025
https://doi.org/10.33808/clinexphealthsci.1379892

Abstract

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.

Supporting Institution

Adnan Menderes University Scientific Research Projects Department

Project Number

TPF-17038

Thanks

We would like to thank Adnan Menderes University Scientific Research Projects Unit.

References

  • 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/
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • Sridharan S, Jain K, Basu A. Regulation of autophagy by kinases. Cancers (Basel). 2011;3(2):2630-2654. https://doi. org/10.3390/cancers3022630
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
There are 33 citations in total.

Details

Primary Language English
Subjects Urology
Journal Section Articles
Authors

Nazlı Can 0000-0003-2023-8246

Kemal Ergin 0000-0002-0222-2710

Project Number TPF-17038
Early Pub Date September 30, 2025
Publication Date September 30, 2025
Submission Date October 23, 2023
Acceptance Date July 10, 2025
Published in Issue Year 2025 Volume: 15 Issue: 3

Cite

APA Can, N., & Ergin, K. (2025). Investigation of the Expression Level of Autophagy Proteins in Infertile Men. Clinical and Experimental Health Sciences, 15(3), 463-472. https://doi.org/10.33808/clinexphealthsci.1379892
AMA Can N, Ergin K. Investigation of the Expression Level of Autophagy Proteins in Infertile Men. Clinical and Experimental Health Sciences. September 2025;15(3):463-472. doi:10.33808/clinexphealthsci.1379892
Chicago Can, Nazlı, and Kemal Ergin. “Investigation of the Expression Level of Autophagy Proteins in Infertile Men”. Clinical and Experimental Health Sciences 15, no. 3 (September 2025): 463-72. https://doi.org/10.33808/clinexphealthsci.1379892.
EndNote Can N, Ergin K (September 1, 2025) Investigation of the Expression Level of Autophagy Proteins in Infertile Men. Clinical and Experimental Health Sciences 15 3 463–472.
IEEE N. Can and K. Ergin, “Investigation of the Expression Level of Autophagy Proteins in Infertile Men”, Clinical and Experimental Health Sciences, vol. 15, no. 3, pp. 463–472, 2025, doi: 10.33808/clinexphealthsci.1379892.
ISNAD Can, Nazlı - Ergin, Kemal. “Investigation of the Expression Level of Autophagy Proteins in Infertile Men”. Clinical and Experimental Health Sciences 15/3 (September2025), 463-472. https://doi.org/10.33808/clinexphealthsci.1379892.
JAMA Can N, Ergin K. Investigation of the Expression Level of Autophagy Proteins in Infertile Men. Clinical and Experimental Health Sciences. 2025;15:463–472.
MLA Can, Nazlı and Kemal Ergin. “Investigation of the Expression Level of Autophagy Proteins in Infertile Men”. Clinical and Experimental Health Sciences, vol. 15, no. 3, 2025, pp. 463-72, doi:10.33808/clinexphealthsci.1379892.
Vancouver Can N, Ergin K. Investigation of the Expression Level of Autophagy Proteins in Infertile Men. Clinical and Experimental Health Sciences. 2025;15(3):463-72.

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