Effect of taxifolin on cold-shock damages in spermatozoa in rabbits

Abstract

Objective: To investigate the effect of taxifolin added to rabbit semen on freezing-induced cold-shock damages in spermatozoa.

Methods: Semen was collected from six adult New Zealand rabbits once a week by artificial vagina. The collected semen was pooled at 38 °C and divided into four equal volumes. They were diluted with 0, 50, 100 and 200 μM taxifolin-containing Tris + egg yolk extender at 38 °C and their temperatures were lowered to 4 °C. Following equilibration, semen drawn into 0.25 mL straws were frozen in an automatic semen freezing device and stored in liquid nitrogen container at -196 °C. Samples were thawed in 38 °C water for 25 s and the analyses of motility, kinematic parameters, morphological deformities, changes in membrane integrity, mitochondrial membrane potential, dead-live ratio, acrosomal damages and as well as oxidative stress analyses were performed in semen.

Results: Addition of 50 μM taxifolin significantly improved motility (total, progressive, rapid and static), high mitochondrial membrane potential and the ratios of spermatozoa with acrosomal damage compared to the control group. Compared to the control group, malondialdehyde (MDA) levels in the 50 and 100 μM taxifolin groups were significantly lower, while the MDA level was high and viable spermatozoa ratio was low in the 200 μM taxifolin group. There were no significant differences between the groups in terms of kinematic parameters, morphological deformities, membrane integrity and antioxidant levels.

Conclusions: The low dose of taxifolin (50 μM) has a positive effect and the high dose (200 μM) has a negative effect. Therefore, it is concluded that the addition of low-dose (50 μM) taxifolin to the extenders would be a useful additive in reducing cold-shock damage that occurs during freezing of rabbit semen.

Keywords

Taxifolin / Semen freezing / Cold-shock / Oxidative stress / Rabbit

Conflict of interest statement

The authors declare no conflict of interest.

Funding

This study was financially supported by Fırat University Scientific Research Projects Coordination Unit (Grant No: VF.21.02).

Authors' contributions

İrfan Yılmaz (this study was summarized from his master's thesis) was involved in conceptualization, investigation, formal analysis, resources, and writing original draft preparation. Gaffari Türk was involved in supervision, funding acquisition, statistical analyses, writing-reviewing and editing. Aslıhan Çakır Cihangiroğlu, Tutku Can Acısu, İbrahim Halil Güngör, Gözde Arkalı and Edanur Güler Ekmen were involved in investigation.

Publisher’ s Note

The Publisher of the Journal remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

[1] Nishijima K, Kitajima S, Matsuhisa F, Niimi M, Wang CC, Fan J. Strategies for highly efficient rabbit sperm cryopreservation. Animals (Basel) 2021; 11: 1220.

[2] De Lamirande E, Jiang H, Zini A, Kodama H, Gagnon C. Reactive oxygen species and sperm physiology. Rev Reprod 1997; 2: 48-54.

[3] Hafez ESE. Preservation and cryopreservation of gamets and embryos. In:Hafez ESE, Hafez B. (eds.) Reproduction in farm animals. 7th ed. Philadelphia: Lippincott Williams & Wilkins; 2000, p. 431-442.

[4] Donnelly ET, McClure N, Lewis SEM. Cryopreservation of human semen and prepared sperm. Effects on motility parameters and DNA integrity. Fertil Steril 2001; 76: 892-900.

[5] Curry MR. Cryopreservation of semen from domestic livestock. Rev Reprod 2000; 5: 46-52.

[6] Braun J, Hochi S, Oguri N, Sato K, Torres-Boggino F. Effect of different protein supplements on motility and plasma membrane integrity of frozen-thawed stallion spermatozoa. Cryobiology 1995; 32: 487-492.

[7] Zhu Z, Fan X, Lv Y, Zhang N, Fan C, Zhang P, et al. Vitamin E analogue improves rabbit sperm quality during the process of cryopreservation through its antioxidant action. PLoS One 2015; 10: e0145383.

[8] Cotelle N. Role of flavonoids in oxidative stress. Curr Top Med Chem 2001; 1: 569-590.

[9] Gupta MB, Bhall TN, Gupta CR, Mitra CR, Bhargava KP. Anti-inflammatory activity of taxifolin. Jpn J Pharmacol 1971; 21: 377-382.

[10] Çeribaşı S, Türk G, Özçelik M, Doğan G, Çeribaşı AO, İflazoğlu Mutlu S, et al. Negative effect of feeding with high energy diets on tests and metabolic blood parameters of male Japanese quails, and positive role of milk thistle seed. Theriogenology 2020; 144: 74-81.

[11] Attia YA, Hamed RS, Bovera F, Abd El AEHE, Al-Harthi MA, Shahba HA, et al. Semen quality, antioxidant status and reproductive performance of rabbits buck fed milk thistle seeds and rosemary leaves. Anim Reprod Sci 2017; 184: 178-186.

[12] Sevim S, Sarıözkan S. Effect of taxifolin on spermatological parameters of rabbit semen stored for a short time. J Vet Med Erciyes Univ 2020; 17: 164-172.

[13] Bucak MN, Keskin N, Ili P, Bodu M, Peker Akalın P, Öztürk AE, et al. Decreasing glycerol content by co-supplementation of trehalose and taxifolin hydrate in ram semen extender: Microscopic, oxidative stress, and gene expression analyzes. Cryobiology 2020; 96: 19-29.

[14] Özer Kaya Ş, Güngör H, Dayan Cinkara S, Acısu TC, Koca RH, Akarsu SA, et al. Effect of different doses of hydrated C60 fullerene nanoparticles on ram semen during cool storage. Turk J Vet Anim Sci 2021; 45: 139-147.

[15] Garner DL, Johnson LA. Viability assessment of mammalian sperm using SYBR-14 and propidium iodide. Biol Reprod 1995; 53: 276-284.

[16] Inanc ME, Gungor S, Ozturk C, Korkma F, Baştan I, Çil B. Cholesterol-loaded cyclodextrin plus trehalose improves quality of frozen-thawed ram sperm. Vet Med Czech 2019; 64: 118-124.

[17] Placer ZA, Cushman LL, Johnson BC. Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Anal Biochem 1966; 16: 359-364.

[18] Sedlak J, Lindsay RH. Estimation of total, protein-bound and nonprotein sulfhydryl groups in tissue with Ellman's reagent. Anal Biochem 1968; 25: 192-205.

[19] Lawrence RA, Burk RF. Glutathione peroxidase activity in selenium deficient rat liver. Biochem Biophys Res Commun 2012; 425: 503-509.

[20] Goth LA. Simple method for determination of serum catalase activity and revision of reference range. Clin Chim Acta 1991; 196: 143-151.

[21] Weitze KF, Petzoldt R. Preservation of semen. Anim Reprod Sci 1992; 28: 229-235.

[22] Salamon S, Maxwell WMC. Frozen storage of ram semen I. Processing, freezing, thawing and fertility after cervical insemination. Anim Reprod Sci 1995; 37: 185-249.

[23] Ford WCL. Regulation of sperm function by reactive oxygen species. Hum Reprod Update 2004; 10: 387-399.

[24] Amidi F, Pazhohan A, Shabani Nashtaei M, Khodarahmian M, Nekoonam S. The role of antioxidants in sperm freezing: A review. Cell Tissue Bank 2016; 17: 745-756.

[25] Wang Y, Zu Y, Long J, Fu Y, Li S, Zhang D, et al. Enzymatic water extraction of taxifolin from wood sawdust of Larix gmelini (Rupr.) Rupr. and evaluation of its antioxidant activity. Food Chem 2011; 126: 1178-1185.

[26] Aruoma GI, Spencer JPE, Warren QD, Jenner P, Butler J, Halliwell B. Characterization of food using commercial garlicand ginger preparations. Food Chem 1997; 60: 149-156.

[27] Yang CL, Lin YS, Liu KF, Peng WH, Hsu CM. Hepatoprotective mechanisms of taxifolin on carbon tetrachloride-induced acute liver injury in mice. Nutrients 2019; 11: 655.

[28] Zribi N, Chakroun NF, Ben Abdallah F, Elleuch H, Sellami A, Gargouri J. Effect of freezing-thawing process and quercetin on human sperm survival and DNA integrity. Cryobiology 2012; 65: 326-331.

[29] Kawasaki Y, Sakurai D, Yoshihara T, Tsuchida M, Harakawa S, Suzuki H. Effect of quercetin on the motility of cryopreserved canine spermatozoa. Cryobiology 2020; 96: 50-54.

[30] Gibb Z, Butler TJ, Morris LHA, Maxwell WMC, Grupen CG. Quercetin improves the postthaw characteristics of cryopreserved sex-sorted and nonsorted stallion sperm. Theriogenology 2013; 79: 1001-1009.

[31] Silva ECB, Cajueiro JFP, Silva SV, Soares PC, Guerra MMP. Effect of antioxidants resveratrol and quercetin on in vitro evaluation of frozen ram sperm. Theriogenology 2012; 77: 1722-1726.

[32] Gottlieb C, Svanborg K, Bygdeman M. Adenosine triphosphate (ATP) in human spermatozoa. Andrologia 1991; 23: 421-425.

[33] Johinke D, de Graaf SP, Bathgate R. Investigation of in vitro parameters and in vivo fertility of rabbit spermatozoa after chilled storage. Anim Reprod Sci 2014; 147: 135-143.

[34] El-Khawagah ARM, Kandiel MMM, Samir H. Effect of quercetin supplementation in extender on sperm kinematics, extracellular enzymes release, and oxidative stress of Egyptian buffalo bulls frozen-thawed semen. Front Vet Sci 2020; 7: 604460.

[35] Cheraghi E, Sajadi SMS, Soleimani Mehranjani M. The effect of quercetin on the quality of sperm parameters in frozen-thawed semen of patients with asthenospermia. Andrologia 2021; 53: e14167.

[36] 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 2017; 97: 883-891.

[37] Rakha BA, Qurrat-Ul-Ain, Ansari MS, Akhter S, Akhter A, Awan MA. Effect of quercetin on oxidative stress, mitochondrial activity and quality of Indian red jungle fowl (Gallus gallus murghi) sperm. Biopreserv Biobank 2020; 18: 311-320.

[38] Barbagallo F, La Vignera S, Cannarella R, Aversa A, Calogero AE, Condorelli RA. Evaluation of sperm mitochondrial function: A key organelle for sperm motility. J Clin Med 2020; 9: 363.

[39] Seifi-Jamadi A, Ahmad E, Ansari M, Kohram H. Antioxidant effect of quercetin in an extender containing DMA or glycerol on freezing capacity of goat semen. Cryobiology 2017; 75: 15-20.