Iranian Journal of Veterinary Surgery

Iranian Journal of Veterinary Surgery

Metformin Improves BCL-2 Gene Expression, Testosterone, Platelet Indices, and Histopathology of Testis in Varicocele–Induced Rats

Document Type : Original Article

Authors
Fatemeh Ghaderi 1
Hamid Reza Moslemi 1
Mahmood Ahmadi-Hamedani 1
Khatereh Kafshdouzan 2
1 Department of Clinical Sciences, Faculty of Veterinary Medicine, Semnan University, Semnan, Iran.
2 Department of Pathobiology, Faculty of Veterinary Medicine, Semnan University, Semnan, Iran.
10.30500/ivsa.2025.487506.1423
Abstract
The present study attempted to investigate the effects of metformin on B-cell lymphoma 2 (BCL-2) gene expression, testosterone, platelet indices, and the histopathology of testis in varicocele–induced rats. Twenty-four male adult Wistar rats, aged seven weeks, were selected and divided into four groups. Group I: Sham, II: VCL (varicocele–induced rats), III: MET, and IV: VCL-MET (varicocele–induced rats treated with metformin). Koksal surgical method was used to induce varicocele. One week later, the rats in the MET and VCL-MET groups received metformin (100 mg/kg), and the rats in the sham and VCL groups received normal saline orally for four weeks. Then, all rats were anesthetized, body weight was measured, and the left testis was isolated for BCL-2 gene expression. Also, their blood samples were collected to measure platelet indices and evaluate testosterone concentration. The most significant secondary weight loss relative to the initial body weight, lowest platelet count (PLT: 49.6 × 103 µl-1), highest plateletcrit level (PCT: 0.44 %), mean platelet volume (MPV: 6.0 fL), platelet distribution width (PDW: 18.15%), and the lowest BCL-2 gene expression was observed in the VCL group. The lowest MPV/PLT (0.008), and the highest testosterone level (3.21 ng/ml) were observed in the VCl-Met group. Spermatid cells are seen in the VCL-Met group, spermatocyte cells in the VCL group, and spermatozoa cells in the sham and Met groups. Metformin apparently improved testosterone levels and testicular histopathology by increasing BCL-2 gene expression. As an anti-inflammatory, it also mitigated the changes in platelet indices caused by experimental varicocele.
Keywords
Metformin
BCL-2
Platelet indices
Testosterone
Varicocele
Subjects
  1. Pastuszak AW, Wang R. Varicocele and testicular function. Asian Journal of Andrology. 2015; 17(4): 659-667. doi: 10.4103/1008-682X.153539
  2. Peng M, Wang W, Zhu W, Bai Y, Ning N, Huang Q, Pang X, Zhou J, Zhang H, Zhao K. Zishen Yutai Pill improves sperm quality and reduces testicular inflammation in experimental varicocele rats. Heliyon. 2023; 9(6): 1-11. doi: 10.1016/j.heliyon.2023.e17161
  3. Abdelbaki SA, Sabry JH, Al-Adl AM, Sabry HH. The impact of coexisting sperm DNA fragmentation and seminal oxidative stress on the outcome of varicocelectomy in infertile patients: a prospective controlled study. Arab Journal of Urology. 2017; 15(2): 131-139. doi: 10.1016/j.aju.2017.03.002
  4. Talebi‐Yazdabadi Z, Jahanbakhsh N, Dormiani K, Forouzanfar M, Lachinani L, Zohrabi D, Tavalaee M, Nasr-Esfahani MH. Assessment of MUSASHI 1 and MUSASHI 2 expression in spermatozoa and testicular tissue. Andrologia. 2021; 53(10): e14187. doi: 10.1111/and.14187
  5. Roque M, Esteves SC. Effect of varicocele repair on sperm DNA fragmentation: a review. International Urology and Nephrology. 2018; 50(4): 583-603. doi: 10.1007/s11255-018-1839-4
  6. Will MA, Swain J, Fode M, Sonksen J, Christman GM, Ohl D. The great debate: varicocele treatment and impact on fertility. Fertility and Sterility. 2011; 95(3): 841-852. doi: 10.1016/j.fertnstert.2011.01.002
  7. Mallidis C, Czerwiec A, Filippi S, O'Neill J, Maggi M, McClure N. Spermatogenic and sperm quality differences in an experimental model of metabolic syndrome and hypogonadal hypogonadism. Reproduction. 2011; 142(1): 63-71. doi: 10.1530/REP-10-0472
  8. Çayan S, Akbay E, Saylam B, Kadıoğlu A. Effect of varicocele and its treatment on testosterone in hypogonadal men with varicocele: review of the literature. Balkan Medical Journal. 2020; 37(3): 121-124. doi: 10.4274/balkanmedj.galenos.2020.2020.1.85
  9. Luna‐Vargas MP, Chipuk JE. The deadly landscape of pro‐apoptotic BCL‐2 proteins in the outer mitochondrial membrane. The FEBS Journal. 2016; 283(14): 2676-2689. doi: 10.1111/febs.13624
  10. Zhang Y, Wu X, Zhang W, Gao J, Zhang Y, Zhang X. Platelet indices and varicocele: a systematic review and meta‐analysis. Andrologia. 2021; 53(2): e13939. doi: 10.1111/and.13939
  11. Noori Alavije H, Ahmadi‐hamedani M, Moslemi H. Evaluation of platelet indices and mean platelet volume to platelet count ratio in experimentally varicocele‐induced adolescent and adult rats. Andrologia. 2022; 54(3): e14345. doi: 10.1111/and.14345
  12. Erdem E, Güngör Ordueri NE, Usta A, Cenk K, Meriç K. Metformin reduces the extent of varicocele-ınduced damage in testicular tissue. Global Journal of Reproductive Medicine. 2019; 6(4): 70-76. doi: 10.19080/GJORM.2019.06.555691
  13. Karimi H, Asghari A, Jahandideh A, Gh A, Mortazavi P. Effects of metformin on experimental varicocele in rats. Archives of Razi Institute. 2021; 76(2): 371-384. doi: 10.22092/ari.2020.128136.1406
  14. Köksal T, Erdoğru T, Toptaş B, Hakan Gülkesen K, Usta M, Baykal A, Baykara M. Effect of experimental varicocele in rats on testicular oxidative stress status. Andrologia. 2002; 34(4): 242-247. doi: 10.1046/j.1439-0272.2002.00500.x
  15. Abo El Gheit RE, Soliman NA, Nagla SA, El-Sayed RM, Badawi GA, Emam MN, Ghafar MTA, Ibrahim MAA, Elswaidy NRM, Radwan DA, Alshenawy HA, Khaled HE, Kamel S, El-Saka MH, Madi NM, Younis RL. Melatonin epigenetic potential on testicular functions and fertility profile in varicocele rat model is mediated by silent information regulator 1. British Journal of Pharmacology. 2022; 179(13): 3363-3381. doi: 10.1111/bph.15804
  16. Clavijo RI, Carrasquillo R, Ramasamy R. Varicoceles: prevalence and pathogenesis in adult men. Fertility and Sterility. 2017; 108(3): 364-369. doi: 10.1016/j.fertnstert.2017.06.036
  17. Ammar O, Tekeya O, Hannachi I, Sallem A, Haouas Z, Mehdi M. Increased sperm DNA fragmentation in infertile men with varicocele: relationship with apoptosis, seminal oxidative stress, and spermatic parameters. Reproductive Sciences. 2021; 28: 909-919. doi: 10.1007/s43032-020-00311-6
  18. Alasmari WA, Faruk EM, Abourehab MA Elshazly AM, El Sawy NA. The effect of metformin versus vitamin E on the testis of adult diabetic albino rats: histological, biochemical and immunohistochemistry study. Advances in Reproductive Sciences. 2018; 6(4): 113-132. doi: 10.4236/arsci.2018.64010
  19. Karimi H, Asghari A, Jahandideh A, Akbari G, Mortazavi P. Metformin improves semen profile and hormonal levels in experimental varicocele in the rat. Journal of Basic and Clinical Pathophysiology. 2013; 11(1): 1-8. doi: 10.22070/jbcp.2023.18013.1171
  20. Tartarin P, Moison D, Guibert E, Dupont J, Habert R, Rouiller-Fabre V, Frydman N, Pozzi S, Frydman R, Lecureuil C, Froment P. Metformin exposure affects human and mouse fetal testicular cells. Human Reproduction. 2012; 27(11): 3304-3314. doi: 10.1093/humrep/des264
  21. Banihani SA. Effect of metformin on semen quality. Brazilian Journal of Pharmaceutical Sciences. 2016; 52(4): 591-594. doi: 10.1590/S1984-82502016000400002
  22. Hollander-Cohen L, Golan M, Levavi-Sivan B. Differential regulation of gonadotropins as revealed by transcriptomes of distinct LH and FSH cells of fish pituitary. International Journal of Molecular Sciences. 2021; 22(12): 6478. doi: 10.3390/ijms22126478
  23. Pourheydar B, Azarm F, Farjah G, Karimipour M, Pourheydar M. Effect of silymarin and metformin on the sperm parameters and histopathological changes of testes in diabetic rats: an experimental study. International Journal of Reproductive Biomedicine. 2021; 19(12): 1091-1104. doi: 10.18502/ijrm.v19i12.10060
  24. .Yilmaz M, Karaaslan M, Ceylan C, Tonyali S. The evaluation of platelet volume indices in patients with varicocele. Grand Journal of Urolology. 2021; 1(1): 6-8.
  25. Bozkurt Y, Soylemez H, Sancaktutar AA, Islamoglu Y, Kar A, Penbegul N, Atar M, Penbegul N, Atar M, Nuri Bodakci M, Hatipoglu NK. Relationship between mean platelet volume and varicocele: a preliminary study. Urology. 2012; 79(5): 1048-1051. doi: 10.1016/j.urology.2012.01.019
  26. Rokni, GR, Shiran M, Abounoori M, Houshmand G, Babakhanian M, Godazandeh G, Bayat S, Pazyar N, Abedi M, Khorshidi F, Yari F, Ghafouri Z, Patil A, Goldust M, Mirmohammadi Langari L. Effects of metformin on autoimmune immunoglobins and interferon-c in patients with early diagnosed pemphigus vulgaris: a prospective clinical trial. Clinical and Experimental Dermatology. 2021; 47(1): 110-113. doi: 10.1111/ced.14832
  27. Taher I, El‑Masry E, Abouelkheir M, Taha AE. Anti‑inflammatory effect of metformin against an experimental model of LPS‑induced cytokine storm. Experimental and Therapeutic Medicine. 2023; 26(3): 1-7. doi: 10.3892/etm.2023.12114
  28. Babaei A, Asadpour R, Mansouri K, Sabrivand A, Kazemi-Darabadi S. Lycopene improves testicular damage and sperm quality in experimentally induced varicocele: Relationship with apoptosis, hypoxia, and hyperthermia. Food Science and Nutrition. 2022; 10(5): 1469-1480. doi: 10.1002/fsn3.2762
  29. Zhao W, Liu J, Wang D, Wang Y, Zhang F, Jin G, Yuan C, Wang X, Qin Q. Effect of silencing HIF-1α gene on testicle spermatogenesis function in varicocele rats. Cell and Tissue Research. 2019; 378: 543-554. doi: 10.1007/s00441-019-03064-0
  30. Cho CL, Esteves SC, Agarwal A. Novel insights into the pathophysiology of varicocele and its association with reactive oxygen species and sperm DNA fragmentation. Asian Journal of Andrology. 2016; 18(2): 186-193. doi: 10.4103/1008-682X.170441
  31. Wang K, Gao Y, Wang C, Liang M, Liao Y, Hu K. Role of oxidative stress in varicocele. Frontiers in Genetics. 2022; 13: 850114. doi: 10.3389/fgene.2022.850114
  32. Shokoohi M, Khaki A, Roudi Rasht Abadi, MohammadZadeh Boukani L, Hassanpour Khodaie S, Kalarestaghi H, Khaki AA, Moghimian M, Niazkar HA, Shoorei H. Minocycline can reduce testicular apoptosis related to varicocele in male rats. Andrologia. 2022; 54(4): e14375. doi: 10.1111/and.14375
  33. Kuwana T, Newmeyer DD. Bcl-2-family proteins and the role of mitochondria in apoptosis. Current Opinion in Cell Biology. 2003; 15(6): 691-699. doi: 10.1016/j.ceb.2003.10.004
  34. Sadraie M, Tavalaee M, Nasr-Esfahani MH, Forouzanfar M. Evaluation of sperm and morphometric parameters of testicular tissue following curcumin administration in rats after varicocele induction. Qom University of Medical Sciences Journal. 2021; 15(4): 284-295.
  35. Yan WJ, Mu Y, Yu N, Yi TL, Zhang Y, Pang XL, Cheng D, Yang J. Protective effects of metformin on reproductive function in obese male rats induced by high-fat diet. Journal of Assisted Reproduction and Genetics. 2015; 32: 1097-1104. doi: 10.1007/s10815-015-0506-2
Iranian Journal of Veterinary Surgery
Volume 20, Issue 2 - Serial Number 43
October 2025
Pages 122-129

  • Receive Date 06 November 2024
  • Revise Date 12 December 2024
  • Accept Date 07 January 2025
  • First Publish Date 07 January 2025