Similar articles

Abstract: Resveratrol (3,5,4'-trihydroxystilbene, RSV) has been widely used in mammalian cells, but whether it can be used during freezing boar semen is still unknown. The effects of RSV treatment during boar semen freezing on its anti-freezing ability, apoptosis, and possible apoptotic pathways were observed in this study. Sperm motility, mitochondrial membrane potential (ΔΨ_m), adenosine triphosphate (ATP) content, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL)-positive apoptotic state, and messenger RNA (mRNA) expression levels of apoptotic genes involved in different apoptotic pathways after freezing with or without RSV treatment were tested. The results showed that: (1) Compared with fresh sperm, the motility, normal acrosome rate, and plasma membrane integrity rate of frozen boar sperm decreased significantly (P<0.05), and RSV did not significantly increase the sperm motility (0.44 vs. 0.40, P>0.05), but it did significantly improve the normal acrosome rate (57.65% vs. 47.00%, P<0.05) and plasma membrane integrity rate (46.67% vs. 38.85%, P<0.05). (2) After freezing, most boar sperm showed low mitochondrial ΔΨ_m. RSV treatment could increase the rate of high mitochondrial ΔΨ_m of boar sperm. (3) RSV treatment significantly decreased reactive oxygen species (ROS) levels (58.65% vs. 88.41%, P<0.05) and increased the ATP content (0.49 μmol/L vs. 0.25 μmol/L, P<0.05) of boar sperm during freezing. (4) The apoptotic rate of the freezing group (80.41%) with TUNEL detection increased significantly compared to the fresh group (9.70%, P<0.05), and RSV treatment greatly decreased the apoptotic rate (68.32%, P<0.05). (5) Real-time polymerase chain reaction (RT-PCR) showed that not only the genes from the death receptor-mediated apoptotic pathway (tumor necrosis factor-α (TNF-α), Fas ligand (FasL), and Caspase-8), but also the genes from the mitochondria-mediated apoptotic pathway (manganese superoxide dismutase (MnSOD), B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and Caspase-9) were both significantly changed after freezing. RSV treatment during freezing greatly changed their expression levels. Although RSV treatment during boar semen freezing did not significantly increase motility after thawing, it still played an efficient antioxidant role, which could enhance the mitochondrial function and decrease the apoptotic level induced by both the death receptor- and mitochondria-mediated apoptotic pathways.

猪精液冷冻过程中白藜芦醇处理对其凋亡和凋亡途径的影响

[1]Aitken RJ, Koppers AJ, 2011. Apoptosis and DNA damage in human spermatozoa. Asian J Androl, 13(1):36-42.

[2]Awda BJ, Mackenzie-Bell M, Buhr MM, 2009. Reactive oxygen species and boar sperm function. Biol Reprod, 81(3):553-561.

[3]Branco CS, Garcez ME, Pasqualotto FF, et al., 2010. Resveratrol and ascorbic acid prevent DNA damage induced by cryopreservation in human semen. Cryobiology, 60(2):235-237.

[4]Bucak MN, Ataman MB, Başpınar N, et al., 2015. Lycopene and resveratrol improve post-thaw bull sperm parameters: sperm motility, mitochondrial activity and DNA integrity. Andrologia, 47(5):545-552.

[5]Collodel G, Federico MG, Geminiani M, et al., 2011. Effect of trans-resveratrol on induced oxidative stress in human sperm and in rat germinal cells. Reprod Toxicol, 31(2):239-246.

[6]Dai JJ, Wu CF, Zhang DF, et al., 2009. Some factors affecting freezing of boar semen in 5 mL maxi-straws. Asian Australas J Anim Sci, 22(4):507-515.

[7]Dai JJ, Wu CF, Muneri CW, et al., 2015. Changes in mitochondrial function in porcine vitrified MII-stage oocytes and their impacts on apoptosis and developmental ability. Cryobiology, 71(2):291-298.

[8]Dai JJ, Niu YF, Wu CF, et al., 2016. Both death receptor and mitochondria mediated apoptotic pathways participated the occurrence of apoptosis in porcine vitrified MII stage oocytes. Cryo Letters, 37(2):129-136.

[9]de Oliveira MR, Nabavi SF, Manayi A, et al., 2016. Resveratrol and the mitochondria: from triggering the intrinsic apoptotic pathway to inducing mitochondrial biogenesis, a mechanistic view. Biochim Biophys Acta Gen Subj, 1860(4):727-745.

[10]Gadea J, Sellés E, Marco MA, et al., 2004. Decrease in glutathione content in boar sperm after cryopreservation: effect of the addition of reduced glutathione to the freezing and thawing extenders. Theriogenology, 62(3-4):690-701.

[11]Garcez ME, dos Santos Branco C, Lara LV, et al., 2010. Effects of resveratrol supplementation on cryopreservation medium of human semen. Fertil Steril, 94(6):2118-2121.

[12]Green DR, Reed JC, 1998. Mitochondria and apoptosis. Science, 281(5381):1309-1312.

[13]Gürler H, Malama E, Heppelmann M, et al., 2016. Effects of cryopreservation on sperm viability, synthesis of reactive oxygen species, and DNA damage of bovine sperm. Theriogenology, 86(2):562-571.

[14]Hsuuw YD, Chan WH, Yu JS, 2013. Ochratoxin a inhibits mouse embryonic development by activating a mitochondrion-dependent apoptotic signaling pathway. Int J Mol Sci, 14(1):935-953.

[15]Jeulin C, Soufir JC, Weber P, et al., 1989. Catalase activity in human spermatozoa and seminal plasma. Gamete Res, 24(2):185-196.

[16]Kefer JC, Agarwal A, Sabanegh E, 2009. Role of antioxidants in the treatment of male infertility. Int J Urol, 16(5):449-457.

[17]Li ZL, Lin QL, Liu RJ, et al., 2010. Protective effects of ascorbate and catalase on human spermatozoa during cryopreservation. J Androl, 31(5):437-444.

[18]Luño V, Gil L, Olaciregui M, et al., 2014. Rosmarinic acid improves function and in vitro fertilising ability of boar sperm after cryopreservation. Cryobiology, 69(1):157-162.

[19]Martinez-Soto JC, de DiosHourcade J, Gutiérrez-Adán A, et al., 2010. Effect of genistein supplementation of thawing medium on characteristics of frozen human spermatozoa. Asian J Androl, 12(3):431-441.

[20]Mukherjee A, Malik H, Saha AP, et al., 2014. Resveratrol treatment during goat oocytes maturation enhances developmental competence of parthenogenetic and hand-made cloned blastocysts by modulating intracellular glutathione level and embryonic gene expression. J Assist Reprod Genet, 31(2):229-239.

[21]Ou XH, Li S, Wang ZB, et al., 2012. Maternal insulin resistance causes oxidative stress and mitochondrial dysfunction in mouse oocytes. Hum Reprod, 27(7):2130-2145.

[22]Pasqualotto FF, Gupta S, Borges E Jr, et al., 2006. Impact of agriculture on semen quality and oxidative stress amongst infertile couples. Fertil Steril, 86(3):S445-S446.

[23]Perchec G, Jeulin C, Cosson J, et al., 1995. Relationship between sperm ATP content and motility of carp spermatozoa. J Cell Sci, 108(2):747-753.

[24]Qiu Y, Wang LG, Jia YF, et al., 2011. Effects of the crude extract of Polygala tenuifolia Willd on human sperm in vitro. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 12(6):448-454.

[25]Schäfer S, Holzmann A, 2000. The use of transmigration and Spermac™ stain to evaluate epididymal cat spermatozoa. Anim Reprod Sci, 59(3-4):201-211.

[26]Scott FL, Stec B, Pop C, et al., 2009. The Fas-FADD death domain complex structure unravels signalling by receptor clustering. Nature, 457(7232):1019-1022.

[27]Shimizu K, Miyagi S, Miyazawa K, et al., 2016. Resveratrol prevents warm ischemia-reperfusion injury in liver grafts from non-heart-beating donor rats. Transplan Proc, 48(4):1221-1225.

[28]Silva ECB, Cajueiro JFP, Silva SV, et al., 2012. Effect of antioxidants resveratrol and quercetin on in vitro evaluation of frozen ram sperm. Theriogenology, 77(8):1722-1726.

[29]Tao ZH, Li C, Xu XF, et al., 2019. Scavenging activity and mechanism study of ferulic acid against reactive carbonyl species acrolein. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 20(11):868-876.

[30]Toni LS, Garcia AM, Jeffrey DA, et al., 2018. Optimization of phenol-chloroform RNA extraction. MethodsX, 5:599-608.

[31]Trzcińska M, Bryła M, Gajda B, et al., 2015. Fertility of boar semen cryopreserved in extender supplemented with butylated hydroxytoluene. Theriogenology, 83(3):307-313.

[32]Zeng CJ, Tang KY, He L, et al., 2014. Effects of glycerol on apoptotic signaling pathways during boar spermatozoa cryopreservation. Cryobiology, 68(3):395-404.