Efectos de la pentoxifilina y la trehalosa sobre la calidad de los espermatozoides epididimarios de ratón durante el almacenamiento en estado líquido
Resumen
En este estudio se compararon los efectos de la pentoxifilina y la trehalosa sobre espermatozoides epididimarios de ratón durante el almacenamiento líquido. Se utilizaron veinte ratones machos adultos, y los espermatozoides epididimarios se distribuyeron en cuatro grupos: control, pentoxifilina 1 mM, pentoxifilina 3 mM y trehalosa 50 mM. Como medio basal se empleó un diluyente modificado a base de Tris. La motilidad y la capacitación evaluada mediante tinción con clortetraciclina se registraron a las 0, 24 y 48 horas; asimismo, el estado antioxidante total, el estado oxidante total y el índice de estrés oxidativo se determinaron a las 0 y 48 horas. Durante el almacenamiento, la motilidad disminuyó en todos los grupos. Sin embargo, solo se detectó una diferencia significativa a las 0 horas, con el valor más alto en el grupo trehalosa 50 mM. Inicialmente, los valores de estado antioxidante total fueron similares. Después de 48 h, la pentoxifilina 3 mM y la trehalosa 50 mM mostraron valores de estado antioxidante total más altos que el control. En estos mismos grupos, los menores valores de índice de estrés oxidativo observados a las 48 horas parecen estar relacionados principalmente con diferencias en los niveles de estado antioxidante total, dado que los valores de estado oxidante total no difirieron significativamente entre los tratamientos. Según los resultados de clortetraciclina, los grupos fueron similares a las 0 horas. A las 24 horas, los porcentajes de espermatozoides capacitados fueron mayores en todos los grupos tratados, mientras que a las 48 horas el valor más alto se observó con trehalosa 50 mM. En conjunto, estos hallazgos sugieren que el almacenamiento líquido a corto plazo influyó más claramente sobre el equilibrio redox y los cambios de membrana relacionados con la capacitación que sobre la conservación de la motilidad. Entre los aditivos evaluados, la trehalosa 50 mM mostró el perfil protector más consistente.
Descargas
Citas
Takeo T, Tsutsumi A, Omaru T, Fukumoto K, Haruguchi Y, Kondo T, Nakamuta Y, Takeshita Y, Matsunaga H, Tsuchiyama S, Sakoh K, Nakao S, Yoshimoto H, Shimizu N, Nakagata N. Establishment of a transport system for mouse epididymal sperm at refrigerated temperatures. Cryobiology. [Internet]. 2012; 65(3):163-168. doi: https://doi.org/f4dhv3 DOI: https://doi.org/10.1016/j.cryobiol.2012.06.002
Sankai T, Tsuchiya H, Ogonuki N. Short-term nonfrozen storage of mouse epididymal spermatozoa. Theriogenology. [Internet]. 2001; 55(8):1759-1768. doi: https://doi.org/dhhdjh DOI: https://doi.org/10.1016/S0093-691X(01)00518-0
Yoshimoto H, Takeo T, Nakagata N. Simple transportation of genetically engineered mice via cold storage techniques. In: Larson MA, editor. Transgenic Mouse. Methods Mol. Biol. New York, USA: Humana; 2020. p. 211-216. doi: https://doi.org/rkhb DOI: https://doi.org/10.1007/978-1-4939-9837-1_17
Yoshimoto H, Takeo T, Nakagata N. Dimethyl sulfoxide and quercetin prolong the survival, motility, and fertility of cold-stored mouse sperm for 10 days. Biol. Reprod. [Internet]. 2017; 97(6):883-891. doi: https://doi.org/gchtqw DOI: https://doi.org/10.1093/biolre/iox144
Takeo T, Fukumoto K, Kondo T, Haruguchi Y, Takeshita Y, Nakamuta Y, Tsuchiyama S, Yoshimoto H, Shimizu N, Li MW, Kinchen K, Vallelunga J, Lloyd KC, Nakagata N. Investigations of motility and fertilization potential in thawed cryopreserved mouse sperm from cold-stored epididymides. Cryobiology. [Internet]. 2014; 68(1):12-17. doi: https://doi.org/f5txmm DOI: https://doi.org/10.1016/j.cryobiol.2013.10.007
Sieme H, Oldenhof H, Wolkers WF. Sperm membrane behaviour during cooling and cryopreservation. Reprod. Domest. Anim. [Internet]. 2015; 50(Suppl 3):20-26. doi: https://doi.org/f7xm23 DOI: https://doi.org/10.1111/rda.12594
Gautier C, Aurich C. “Fine feathers make fine birds” – The mammalian sperm plasma membrane lipid composition and effects on assisted reproduction. Anim Reprod. Sci. [Internet]. 2022; 246:106884. doi: https://doi.org/g9zp6f DOI: https://doi.org/10.1016/j.anireprosci.2021.106884
Aitken RJ, Drevet JR, Moazamian A, Gharagozloo P. Male infertility and oxidative stress: A focus on the underlying mechanisms. Antioxidants. [Internet]. 2022; 11(2):306. doi: https://doi.org/rkhc DOI: https://doi.org/10.3390/antiox11020306
Wang Y, Fu X, Li H. Mechanisms of oxidative stress-induced sperm dysfunction. Front. Endocrinol. [Internet]. 2025; 16:1520835. doi: https://doi.org/g9wfkd DOI: https://doi.org/10.3389/fendo.2025.1520835
Erel O. A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clin. Biochem. [Internet]. 2004; 37(4):277-285. doi: https://doi.org/frb5dh DOI: https://doi.org/10.1016/j.clinbiochem.2003.11.015
Erel O. A new automated colorimetric method for measuring total oxidant status. Clin Biochem. [Internet]. 2005; 38(12):1103-1111. doi: https://doi.org/dzjwc5 DOI: https://doi.org/10.1016/j.clinbiochem.2005.08.008
Ünal I, Uysal H. Seminal oxidative stress index can be used as a marker in the prediction of bull semen cryotolerance. Anim. Reprod. Sci. [Internet]. 2024; 270:107618. doi: https://doi.org/rkhd DOI: https://doi.org/10.1016/j.anireprosci.2024.107618
Rathi R, Colenbrander B, Bevers MM, Gadella BM. Evaluation of in vitro capacitation of stallion spermatozoa. Biol. Reprod. [Internet]. 2001; 65(2):462-470. doi: https://doi.org/bfxrrt DOI: https://doi.org/10.1095/biolreprod65.2.462
Terriou P, Hans E, Giorgetti C, Spach JL, Salzmann J, Urrutia V, Roulier R. Pentoxifylline initiates motility in spontaneously immotile epididymal and testicular spermatozoa and allows normal fertilization, pregnancy, and birth after intracytoplasmic sperm injection. J. Assist. Reprod. Genet. [Internet]. 2000; 17(4):194-199. doi: https://doi.org/cj48cd DOI: https://doi.org/10.1023/A:1009435732258
Griveau JF, Lobel B, Laurent MC, Michardière L, Le Lannou D. Interest of pentoxifylline in ICSI with frozen-thawed testicular spermatozoa from patients with non-obstructive azoospermia. Reprod. Biomed. Online. [Internet]. 2006; 12(1):14-18. doi: https://doi.org/db8tpt DOI: https://doi.org/10.1016/S1472-6483(10)60974-1
Guasti PN, Monteiro GA, Maziero RRD, Carmo MT, Dell’Aqua JA Jr, Crespilho AM, Rifai EA, Papa FO. Pentoxifylline effects on capacitation and fertility of stallion epididymal sperm. Anim. Reprod. Sci. [Internet]. 2017; 179:27-34. doi: https://doi.org/f9xwcg DOI: https://doi.org/10.1016/j.anireprosci.2017.01.013
Gholami D, Sharafi M, Esmaeili V, Nadri T, Alaei L, Riazi G, Shahverdi A. Beneficial effects of trehalose and gentiobiose on human sperm cryopreservation. PLoS One. [Internet]. 2023; 18(4):e0271210. doi: https://doi.org/rkhf DOI: https://doi.org/10.1371/journal.pone.0271210
Jia B, Allai L, Li C, Liang J, Lv C, Wu G, Quan G. A review on the functional roles of trehalose during cryopreservation of small ruminant semen. Front. Vet. Sci. [Internet]. 2024; 11:1467242. doi: https://doi.org/rkhg DOI: https://doi.org/10.3389/fvets.2024.1467242
Nazari P, Farshad A, Hosseini Y. Protective effects of trehalose and pentoxifylline on goat sperm exposed to chilling-freezing process. Biopreserv. Biobank. [Internet]. 2022; 20(6):540-550. doi: https://doi.org/rkhh DOI: https://doi.org/10.1089/bio.2021.0081
Bucak MN, Keskin N, Taspinar M, Çoyan K, Baspinar N, Cenariu MC, Bilgili A, Öztürk C, Kursunlu AN. Raffinose and hypotaurine improve the post-thawed Merino ram sperm parameters. Cryobiology. [Internet]. 2013; 67(1):34-39. doi: https://doi.org/f439bz DOI: https://doi.org/10.1016/j.cryobiol.2013.04.007
Bi Y, Xu WM, Wong HY, Zhu H, Zhou ZM, Chan HC, Sha JH. NYD-SP27, a novel intrinsic decapacitation factor in sperm. Asian J. Androl. [Internet]. 2009; 11(2):229-239. doi: https://doi.org/cdttp2 DOI: https://doi.org/10.1038/aja.2009.6
Ward CR, Storey BT. Determination of the time course of capacitation in mouse spermatozoa using a chlortetracycline fluorescence assay. Dev. Biol. [Internet]. 1984; 104(2):287-296. doi: https://doi.org/fbwh7d DOI: https://doi.org/10.1016/0012-1606(84)90084-8
Silvestre MA, Yániz JL, Peña FJ, Santolaria P, Castelló-Ruiz M. Role of antioxidants in cooled liquid storage of mammal spermatozoa. Antioxidants. [Internet]. 2021; 10(7):1096. doi: https://doi.org/rkhj DOI: https://doi.org/10.3390/antiox10071096
Fleming SD, Thomson LK. The oxidative stress of human sperm cryopreservation. Antioxidants. [Internet]. 2025; 14(4):402. doi: https://doi.org/g9wfq5 DOI: https://doi.org/10.3390/antiox14040402
O’Flaherty C. Redox signaling regulation in human spermatozoa: A primary role of peroxiredoxins. Asian J. Androl. [Internet]. 2025; 27(5):556-563. doi: https://doi.org/rkhk DOI: https://doi.org/10.4103/aja2024126
Pavuluri H, Bakhtiary Z, Panner-Selvam MK, Hellstrom WJG. Oxidative stress-associated male infertility: Current diagnostic and therapeutic approaches. Medicina. [Internet]. 2024; 60(6):1008. doi: https://doi.org/rkhm DOI: https://doi.org/10.3390/medicina60061008
Shapovalova N, Buch T, Bollwein H, Vom-Berg J, Malama E. Beyond swimming: Emerging parameters for predicting the fertility of mouse spermatozoa. Lab. Anim. [Internet]. 2025; 54(12):343-354. doi: https://doi.org/rkhn DOI: https://doi.org/10.1038/s41684-025-01647-9
Aboagla EME, Terada T. Trehalose-enhanced fluidity of the goat sperm membrane and its protection during freezing. Biol. Reprod. [Internet]. 2003; 69(4):1245-1250. doi: https://doi.org/c6c8p4 DOI: https://doi.org/10.1095/biolreprod.103.017889
Hu JH, Zan LS, Zhao XL, Li QW, Jiang ZL, Li YK, Li X. Effects of trehalose supplementation on semen quality and oxidative stress variables in frozen-thawed bovine semen. J. Anim. Sci. [Internet]. 2010; 88(5):1657-1662. doi: https://doi.org/d6sg54 DOI: https://doi.org/10.2527/jas.2009-2335
Sariözkan S, Bucak MN, Cantürk F, Özdamar S, Yay A, Tuncer PB, Özcan S, Sorgucu N, Caner Y. The effects of different sugars on motility, morphology and DNA damage during the liquid storage of rat epididymal sperm at 4°C. Cryobiology. [Internet]. 2012; 65(2):93-97. doi: https://doi.org/f367z4 DOI: https://doi.org/10.1016/j.cryobiol.2012.05.007
Takei GL. Molecular mechanisms of mammalian sperm capacitation, and its regulation by sodium-dependent secondary active transporters. Reprod. Med. Biol. [Internet]. 2024; 23(1):e12614. doi: https://doi.org/rkhp DOI: https://doi.org/10.1002/rmb2.12614
Satish M, Kumari S, Deeksha W, Abhishek S, Nitin K, Adiga SK, Hegde P, Dasappa JP, Kalthur G, Rajakumara E. Structure-based redesigning of pentoxifylline analogs against selective phosphodiesterases to modulate sperm functional competence for assisted reproductive technologies. Sci. Rep. [Internet]. 2021; 11(1):12293. doi: https://doi.org/gkhvhw DOI: https://doi.org/10.1038/s41598-021-91636-y
Benko F, Mohammadi-Sangcheshmeh A, Duracka M, Lukác N, Tvrdá E. In vitro versus cryo-induced capacitation of bovine spermatozoa, part 1: Structural, functional, and oxidative similarities and differences. PLoS One. [Internet]. 2022; 17(10):e0276683. doi: https://doi.org/rkhq DOI: https://doi.org/10.1371/journal.pone.0276683
