Kapsaisin Katkılı Tris Sulandırıcı Kullanılarak Koç Spermasının Dondurularak Saklanması

Öz

Bu çalışmada, kapsaisinin oksidatif stres ve koç spermasının dondurulabilirliği üzerine etkilerinin ortaya konması tasarlandı. Ejakülatlar Sönmez ırkı koçlardan alınarak beş eşit kısma ayrıldı ve farklı oranlarda (4 mM, 2 mM, 1 mM, 500 µM) kapsaisin içeren ve içermeyen (kontrol) sulandırıcı ile sulandırıldı. Sperma örnekleri çözüm sonu spermatolojik analizler için 37°C sıcaklıklta 30 saniye süre ile çözdürüldü. Çalışma sonunda spermatozoa hareketliliği ve kinetik parametreleri, plazma membran akrozom bütünlüğü (PMAI), mitokondrial membran potansiyeli (MMP), DNA hasarı, oksidan ve antioksidan parametreler analiz edildi. Progresif, total motilite ve kinetik parametreler bakımından kontrol grubuna göre kapsaisin içeren guruplarda azalma görüldü (p<0.05). Ayrıca DNA bütünlüğü, PMAI ve MMP'de olumlu sonuçlar elde edilmedi (p<0.05). Sonuç olarak; Tris sulandırıcısına ilave edilen kapsaisinin, oksidatif stres ve koç spermasının dondurulması üzerine olumlu bir etki göstermediği belirlendi.

Anahtar Kelimeler

• Spermatozaa
• kapsaisin
• koç
• antioksidan

Cryopreservation of Ram Semen Using Capsaicin Supplemented Tris Extender

Öz

In this study, it was designed to reveal the effects of capsaicin on oxidative stress and freezability of ram semen. Ejaculates were taken from Sönmez rams and divided into five specimens and diluted with extender at different rates (4 mM, 2 mM, 1 mM, 500 µM) with and without capsaicin (control; C). Semen samples were thawed with a 37°C water bath for 30 seconds for post-thawed analysis. At the end of the study, sperm motility and kinetic parameters, plasma membrane acrosome integrity (PMAI), mitochondrial membrane potential (MMP), DNA damage, oxidant and antioxidant parameters were analyzed. A decrease was observed in the groups containing capsaicin compared to the C in terms of progressive, total motility and kinetic parameters (p<0.05). Besides, positive results were not obtained DNA integrity, PMAI and MMP (p<0.05). In conclusion; it was determined that capsaicin added to Tris extender did not have a positive effect on oxidative stress and freezing of ram semen.

Anahtar Kelimeler

• Spermatozoa
• capsaicin
• ram
• antioxidant

Kaynakça

1. Aitken, R. J. (2017). Reactive oxygen species as mediators of sperm capacitation and pathological damage. Molecular reproduction and development, 84(10), 1039-1052.
2. Armstrong, J. S., Rajasekaran, M., Chamulitrat, W., Gatti, P., Hellstrom, W. J., & Sikka, S. C. (1999). Characterization of reactive oxygen species induced effects on human spermatozoa movement and energy metabolism. Free Radical Biology and Medicine, 26(7-8), 869-880.
3. Bilodeau, J. F., Blanchette, S., Cormier, N., & Sirard, M. A. (2002). Reactive oxygen species-mediated loss of bovine sperm motility in egg yolk Tris extender: protection by pyruvate, metal chelators and bovine liver or oviductal fluid catalase. Theriogenology, 57(3), 1105-1122.
4. Bucher, K., Malama, E., Siuda, M., Janett, F., & Bollwein, H. (2019). Multicolor flow cytometric analysis of cryopreserved bovine sperm: a tool for the evaluation of bull fertility. Journal of dairy science, 102(12), 11652-11669.
5. Chang, H. C., Chen, S. T., Chien, S. Y., Kuo, S. J., Tsai, H. T., & Chen, D. R. (2011). Capsaicin may induce breast cancer cell death through apoptosis-inducing factor involving mitochondrial dysfunction. Human & experimental toxicology, 30(10), 1657-1665.
6. Chelucci, S., Pasciu, V., Succu, S., Addis, D., Leoni, G. G., Manca, M. E., & Berlinguer, F. (2015). Soybean lecithin–based extender preserves spermatozoa membrane integrity and fertilizing potential during goat semen cryopreservation. Theriogenology, 83(6), 1064-1074.
7. Chen, K. S., Chen, P. N., Hsieh, Y. S., Lin, C. Y., Lee, Y. H., & Chu, S. C. (2015). Capsaicin protects endothelial cells and macrophage against oxidized low-density lipoprotein-induced injury by direct antioxidant action. Chemico-Biological Interactions, 228, 35-45.
8. Chen, Y., Wang, H., Wang, F., Chen, C., Zhang, P., Song, D., & Zeng, X. (2020). Sperm motility modulated by Trpv1 regulates zebrafish fertilization. Theriogenology, 151, 41-51.

9. Choucair, F., Saliba, E., Abou Jaoude, I., & Hazzouri, M. (2018). Antioxidants modulation of sperm genome and epigenome damage: Fact or fad? Converging evidence from animal and human studies. Middle East Fertility Society Journal, 23(2), 85-90.
10. Claudia, C., Horatiu, S., Iudith, I., & Constanta, B.V.S. (2014). Research regarding the role of TRPV1 and capsaicin (CPS) implication for capacitation and acrosome reaction. Romanian Biotechnological Letters, 19(3), 9437.
11. De Toni, L., Garolla, A., Menegazzo, M., Magagna, S., Di Nisio, A., Šabović, I., & Foresta, C. (2016). Heat sensing receptor TRPV1 is a mediator of thermotaxis in human spermatozoa. PloS one, 11(12), e0167622.
12. De, A. K., Ghosh, J. J., & Mandal, T. K. (1993). Ultraviolet radiation‐induced lipid peroxidation in liposomal membrane: Modification by capsaicin. Phytotherapy Research, 7(1), 87-89.
13. Draper, H. H., & Hadley, M. (1990). Malondialdehyde determination as index of lipid Peroxidation. In Methods in enzymology. Academic press, 186, 421-431.
14. Esen, Ç., Alkan, B. A., Kırnap, M., Akgül, Ö., Işıkoğlu, S., & Erel, Ö. (2012). The effects of chronic periodontitis and rheumatoid arthritis on serum and gingival crevicular fluid total antioxidant/oxidant status and oxidative stress index. Journal of periodontology, 83(6), 773-779.
15. Francavilla, F., Battista, N., Barbonetti, A., Vassallo, M. R. C., Rapino, C., Antonangelo, C., & Maccarrone, M. (2009). Characterization of the endocannabinoid system in human spermatozoa and involvement of transient receptor potential vanilloid 1 receptor in their fertilizing ability. Endocrinology, 150(10), 4692-4700.
16. Gervasi, M. G., Osycka-Salut, C., Caballero, J., Vazquez-Levin, M., Pereyra, E., Billi, S., & Perez-Martinez, S. (2011). Anandamide capacitates bull spermatozoa through CB1 and TRPV1 activation. PLoS One, 6(2), e16993.
17. Gundogan, M., Yeni, D., Avdatek, F., & Fidan, A. F. (2010). Influence of sperm concentration on the motility, morphology, membrane and DNA integrity along with oxidative stress parameters of ram sperm during liquid storage. Animal Reproduction Science, 122(3-4), 200-207.
18. Hissin, P. J., & Hilf, R. (1976). A fluorometric method for determination of oxidized and reduced glutathione in tissues. Analytical biochemistry, 74(1), 214-226.
19. Hosseini, M., Tavalaee, M., Rahmani, M., Eskandari, A., Shaygannia, E., Kiani‐Esfahani, A., & Nasr‐Esfahani, M. H. (2020). Capsaicin improves sperm quality in rats with experimental varicocele. Andrologia, 52(11), e13762.
20. İnanç, M. E., Güngör, Ş., Öztürk, C., Korkmaz, F., Baştan, İ., & Çil, B. (2019). Cholesterol-loaded cyclodextrin plus trehalose improves quality of frozen-thawed ram sperm. Veterinarni Medicina, 64,(03), 118–124.
21. İnanç, M. E., Güngör, S, Avdatek, F., Yeni, D., Gülhan, M. F., Olgaç, K. T., Denk, B., & Taşdemir, U. (2022). Thymoquinone improves motility, plasma membrane integrity and DNA integrity of frozen–thawed ram semen. Andrologia, 54(10), e14547.
22. Joo, J. I., Kim, D. H., Choi, J. W., & Yun, J. W. (2010). Proteomic analysis for antiobesity potential of capsaicin on white adipose tissue in rats fed with a high fat diet. Journal of proteome research, 9(6), 2977-2987.
23. Kim, J. G., & Parthasarathy, S. (1998, December). Oxidation and the spermatozoa. In Seminars in reproductive endocrinology. Thieme Medical Publishers, Inc., 16,(04),235-339.
24. Korkmaz, F., & Çil, B. (2020). Akış Sitometrisinin (Flow Cytometry) Sperma Kalite Analizlerinde Kullanımı. Atatürk Üniversitesi Veteriner Bilimleri Dergisi, 15(1), 76-83.
25. Krzyzosiak, J., Evenson, D., Pitt, C., Jost, L., Molan, P., & Vishwanath, R. (2000). Changes in susceptibility of bovine sperm to in situ DNA denaturation during prolonged incubation at ambient temperature under conditions of exposure to reactive oxygen species and nuclease inhibitor. Reproduction, Fertility and Development, 12(6), 251-261.
26. Kulaksız, R., & Daşkın A. (2009). Farklı antioksidanlarla dondurulan Saanen teke spermasının in vitro ve in vivo değerlendirilmesi. Ankara Üniv Vet Fak Derg, 56, 201-205.
27. Kumar, A., Majhi, R. K., Swain, N., Giri, S. C., Kar, S., Samanta, L., & Goswami, C. (2016). TRPV4 is endogenously expressed in vertebrate spermatozoa and regulates intracellular calcium in human sperm. Biochemical and biophysical research communications, 473(4), 781-788.
28. Lee, S. H., Richardson, R. L., Dashwood, R. H., & Baek, S. J. (2012). Capsaicin represses transcriptional activity of β-catenin in human colorectal cancer cells. The Journal of nutritional biochemistry, 23(6), 646-655.
29. Lee, Y. M., Kang, S. M., Lee, S. R., Kong, K. H., Lee, J. Y., Kim, E. J., & Chung, J. H. (2011). Inhibitory effects of TRPV1 blocker on UV-induced responses in the hairless mice. Archives of dermatological research, 303, 727-736.
30. Maccarrone, M., Barboni, B., Paradisi, A., Bernabò, N., Gasperi, V., Pistilli, M. G., & Mattioli, M. (2005). Characterization of the endocannabinoid system in boar spermatozoa and implications for sperm capacitation and acrosome reaction. Journal of Cell Science, 118(19), 4393-4404.
31. Majhi, R. K., Kumar, A., Giri, S. C., & Goswami, C. (2020). Differential expression and localization of thermosensitive Transient Receptor Potential Vanilloid (TRPV) channels in the mature sperm of white pekin duck (Anas platyrhynchos). bioRxiv, 2020-02.
32. Majhi, R. K., Kumar, A., Yadav, M., Swain, N., Kumari, S., Saha, A., & Goswami, C. (2013). Thermosensitive ion channel TRPV1 is endogenously expressed in the sperm of a fresh water teleost fish (Labeo rohita) and regulates sperm motility. Channels, 7(6), 483-492.
33. Manjunatha, H., & Srinivasan, K. (2006). Protective effect of dietary curcumin and capsaicin on induced oxidation of low‐density lipoprotein, iron‐induced hepatotoxicity and carrageenan‐induced inflammation in experimental rats. The FEBS journal, 273(19), 4528-4537.
34. Olğaç, K. T., & Akçay, E. (2021). Effects of Spermine and Spermidine supplemented extenders on post-thaw Spermatological Parameters in Stallion Semen Cryopreservation. Cryobiology, 100, 72-76.
35. Park, S. G., Yon, J. M., Lin, C., Gwon, L. W., Lee, J. G., Baek, I. J., & Nam, S. Y. (2017). Capsaicin attenuates spermatogenic cell death induced by scrotal hyperthermia through its antioxidative and anti‐apoptotic activities. Andrologia, 49(5), e12656.
36. Sabuncuoğlu, S., & Özgüneş, H. (2011). Kemoterapi, serbest radikaller ve oksidatif stres. Hacettepe University Journal of the Faculty of Pharmacy, (2), 137-150.
37. Salahshouri, S., Akbarian, F., Tavalaee, M., Seifati, S. M., & Nasr-Esfahani, M. H. (2022). Expression of TRPV1 as A Heat Sensitive Voltage-Dependent Ion Channel and Oxidative Stress in Sperm Samples of Infertile Men with Varicocele: A Case-Control Study. Cell Journal (Yakhteh), 24(6), 323.
38. Salmani, H., Nabi, M. M., Vaseghi-Dodaran, H., Rahman, M. B., Mohammadi-Sangcheshmeh, A., Shakeri, M., ... & Zhandi, M. (2013). Effect of glutathione in soybean lecithin-based semen extender on goat semen quality after freeze-thawing. Small ruminant research, 112(1-3), 123-127.
39. Sanocka, D., & Kurpisz, M. (2004). Reactive oxygen species and sperm cells. Reproductive biology and endocrinology, 2(1), 1-7.
40. Sarioglu‐Buke, A., Erdem, S., Gedikoglu, G., Bingol‐Kologlu, M., & Tanyel, F. C. (2001). Capsaicin effectively prevents apoptosis in the contralateral testis after ipsilateral testicular torsion. BJU international, 88(7), 787-789.
41. Szallasi, A., & Blumberg, P. M. (1999). Vanilloid (Capsaicin) receptors and mechanisms. Pharmacological reviews, 51(2), 159-212.
42. Szolcsányi, J. (2004). Forty years in capsaicin research for sensory pharmacology and physiology. Neuropeptides, 38(6), 377-384.
43. Üstüner, B., Nur, Z., Alcay, S., Toker, M. B., Sağırkaya, H., & Soylu, M. K. (2015). Effect of freezing rate on goat sperm morphology and DNA integrity. Turkish Journal of Veterinary & Animal Sciences, 39(1), 110-114.
44. Yeni, D. Güngör, Ş. Avdatek, F. Gülhan, M.F. Olğaç, K.T. İnanç, M.E. Denk, B. & Taşdemir, U. (2022). Investigation of Changes in Spermatozoon Characteristics, Chromatin Structure, and Antioxidant/Oxidant Parameters after Freeze-Thawing of Hesperidin (Vitamin P) Doses Added to Ram Semen. Life, 12, 1780.