从光雷;刘青;孙伟;曾申明;
CONG Guanglei;LIU Qing;SUN Wei;ZENG Shenming;College of Animal Science and Technology, Yangzhou University;

• 1:扬州大学动物科学与技术学院

摘要(Abstract)：

人工授精能提高优良种公羊的利用率,提高养羊业的经济效益,也是良种选育和扩繁的必备技术。在此技术中,精液保存是最重要的环节,目前有液态和冷冻保存2种方法。精液在处理和保存过程中会不可避免地产生活性氧(ROS)家族和活性氮(RNS)家族,它们会对精子结构和遗传物质稳定性造成不可逆损伤。因此,在稀释液中添加适量抗氧化剂能够降低ROS和RNS含量,有效延长精子在体外保存时间。本文根据国内外最新进展,综述了抗氧化剂在绵羊精液保存中研究和应用现状,为新型稀释液的研究提供参考。
Artificial insemination can improve the utilization rate of high-quality rams and improve the economic benefits of the sheep-raising industry. It is also an indispensable technology for breeding and reproduction. In this technique, semen preservation is the most important step, which currently includes fresh storage and cryopreservation. During the process of treatment and preservation, the reactive oxygen species(ROS) and reactive nitrogen species(RNS) will be inevitably produced, which can cause irreversible damages to sperm structure and genetic material stability. Therefore, the semen extender supplemented with appropriate antioxidants can reduce the content of ROS and RNS and effectively prolong the preservation time of sperm in vitro. This article will comprehensively review the research and application status of antioxidants in sheep semen preservation according to the latest publications in this area to provide reference for new effective dilutors' design.

关键词(KeyWords)： 绵羊精液保存;ROS;RNS;抗氧化剂
Sheep;Semen preservation;ROS;RNS;Antioxidant

Abstract:

Keywords:

基金项目(Foundation): 江苏省重点研发计划(现代农业)项目(BE2018354);; 江苏省农业重大新品种创制项目(PZCZ201739);; 江苏省农业科技自主创新项目(CX(18)2003)

作者(Authors): 从光雷;刘青;孙伟;曾申明;
CONG Guanglei;LIU Qing;SUN Wei;ZENG Shenming;College of Animal Science and Technology, Yangzhou University;

DOI: 10.19556/j.0258-7033.20190520-01

参考文献(References)：

• [1]Azarabadi S, Abdollahi H, Torabi M, et al. ROS generation,oxidative burst and dynamic expression profiles of ROSscavenging enzymes of superoxide dismutase(SOD), catalase(CAT)and ascorbate peroxidase(APX)in response to Erwinia amylovora in pear(Pyrus communis L)[J]. Eur J Plant Pathol,2017, 147(2):279-294.
• [2]Tas Demir U, Tuncer P B, Büyükleblebi C S, et al. Effects of various antioxidants on cryopreserved bull sperm quality.[J].Kafkas Univ Vet Fak, 2014, 20(2):253-258.
• [3]Alomar M, Alzoabi M, Zarkawi M. Kinetics of hydrogen peroxide generated from live and dead ram spermatozoa and the effects of catalase and oxidase substrates addition.[J]. Czech J Anim Sci, 2016, 61(1):1-7.
• [4]徐志鹏.一氧化氮浓度对人精子活力的影响[J].中华男科学,2004, 10(3):204.
• [5]Kasimanickam R, Pelzer K D, Kasimanickam V, et al. Association of classical semen parameters, sperm DNA fragmentation index, lipid peroxidation and antioxidant enzymatic activity of semen in ram-lambs[J]. Theriogenology, 2006, 65(7):1407-1421.
• [6]Marti J I, Marti E, Cebrian-perez J A, et al. Survival rate and antioxidant enzyme activity of ram spermatozoa after dilution with different extenders or selection by a dextran swim-up procedure[J]. Theriogenology, 2003, 60(6):1025-1037.
• [7]Mohsen F, Saman F E, Sayyed Mprteza H, et al. Can permeable super oxide dismutase mimetic agents improve the quality of frozen-thawed ram semen?[J]. Cryobiology, 2013, 66(2):126-130.
• [8]Silva E C B, Cajueiro J F P, Silva S V, et al. Effect of antioxidants resveratrol and quercetin on in vitro evaluation of frozen ram sperm[J]. Theriogenology, 2012, 77(8):1722-1726.
• [9]Camara D R, Mello-Pinto M M C, Pinto L C, et al. Effects of reduced glutathione and catalase on the kinematics and membrane functionality of sperm during liquid storage of ram semen[J]. Small Ruminant Res, 2011, 100(1):44-49.
• [10]Camara D R, Silva S V, Almeida F C, et al. Effects of antioxidants and duration of pre-freezing equilibration on frozenthawed ram semen[J]. Theriogenology, 2011, 76(2):342-350.
• [11]AzawiOI,Hussei nEK.Effect of vitamins CorE supplementation to Tris diluent on the semen quality of Awassi rams preserved at 5?C[J]. Vet Res Forum, 2013, 4(3):157.
• [12]Kheradmand A, Babaei H, Abshenas J. Comparative evaluation of the effect of antioxidants on the chilled-stored ram semen[J].Iran J Vet Res, 2006, 7(4):40-45.
• [13]Maia M D S, Bicudo S D, Sicherle C C, et al. Lipid peroxidation and generation of hydrogen peroxide in frozen-thawed ram semen cryopreserved in extenders with antioxidants[J].Anim Reprod Sci, 2010, 122(1-2):118-123.
• [14]Mata-CampuzanoM,álvarez-Rodríguez, M, Tamayo-Canul J,et al. Refrigerated storage of ram sperm in presence of Trolox and GSH antioxidants:Effect of temperature, extender and storage time[J]. Anim Reprod Sci, 2014, 151(3-4):137-147.
• [15]Silva S V, Soares A T, Batista A M, et al. Vitamin E(Trolox)addition to Tris-egg yolk extender preserves ram spermatozoon structure and kinematics after cryopreservation[J]. Anim Reprod Sci, 2013, 137(1-2):37-44.
• [16]Maia M D S, Bicudo S D, Azevedo H C, et al. Motility and viability of ram sperm cryopreserved in a Tris-egg yolk extender supplemented with anti-oxidants[J]. Small Ruminant Res, 2009,85(2-3):85-90.
• [17]Choucair F, Saliba E, Jaoude I A, et al. Antioxidants modulation of sperm genome and epigenome damage:Fact or fad? Converging evidence from animal and human studies[J]. Middle East Fertility Society Journal, 2018, 23(2):85-90.
• [18]SánchezPartida LG, Setchell B P, Maxwell W M. Epididymal compounds and antioxidants in diluents for the frozen storage of ram spermatozoa[J]. Reprod Fert Develop, 1997, 9(7):689.
• [19]Rietjens I M C M, Boersma M G, Haan L D, et al. The prooxidant chemistry of the natural antioxidants vitamin C, vitamin E, carotenoids and flavonoids[J]. Environ Toxicol Phar, 2002,11(3):321-333.
• [20]Amidi F, Pazhohan A, Nashtaei M S, et al. The role of antioxidants in sperm freezing:a review[J]. Cell Tissue Bank, 2016,17(4):745-756.
• [21]Abeyrathne E D N S, Lee H Y, Ahn D U. Sequential separation of lysozyme, ovomucin, ovotransferrin, and ovalbumin from egg white[J]. Poult Sci, 2014, 93(4):1001-1009.
• [22]Asadpour R, Jafari R, Tayefinasrabadi H. The effect of antioxidant supplementation in semen extenders on semen quality and lipid peroxidation of chilled bull spermatozoa[J].Iran J Vet Res, 2012, 13(3):246-249.
• [23]哈福,赵有璋.绵羊冷冻精液稀释液中添加复合维生素B提高冻精解冻后品质的研究[J].甘肃农业大学学报, 2003,38(1):17-19.
• [24]Uysal O, Bucak M N. Effects of oxidized glutathione, bovine serum albumin, cysteine and lycopene on the quality of frozenthawed ram semen[J]. Acta Vet Brno, 2007, 76(3):383-390.
• [25]?oyan K, Baspinar N, Bucak M N, et al. Effects of cysteine and ergothioneine on post-thawed Merino ram sperm and biochemical parameters[J]. Cryobiology, 2011, 63(1):1-6.
• [26]Sharafi M, Zhandi M, Sharif A A. Supplementation of soybean lecithin-based semen extender by antioxidants:complementary flowcytometric study on post-thawed ram spermatozoa[J]. Cell Tissue Bank, 2015, 16(2):261-269.
• [27]?oyan K, Baspinar N, Bucak M N, et al. Influence of methionine and dithioerythritol on sperm motility, lipid peroxidation and antioxidant capacities during liquid storage of ram semen[J]. Res Vet Sci, 2010, 89(3):426-431.
• [28]Bucak M N, Keskin N, Mehmet Ta?p?nar, et al. Raffinose and hypotaurine improve the post-thawed Merino ram sperm parameters[J]. Cryobiology, 2013, 67(1):34-39.
• [29]Mehr R A, Noori H. Effect of different levels of l-Glutamine and glycerol on freezing of ram spermatozoa[J]. Small Ruminant Res, 2013, 115(1-3):103-107.
• [30]Abozar N, Hossein Daghigh K, Hossein M, et al. Different concentrations of cysteamine and ergothioneine improve microscopic and oxidative parameters in ram semen frozen with a soybean lecithin extender[J]. Cryobiology, 2014, 69(1):68-73.
• [31]Saleh R A, Agarwal A. Oxidative stress and male infertility:from research bench to clinical practice[J]. Asian J Androl,2002, 23(6):737-752.
• [32]Banday M N, Lone F A, Rasool F, et al. Use of antioxidants reduce lipid peroxidation and improve quality of crossbred ram sperm during its cryopreservation[J]. Cryobiology, 2017, 74:25-30.
• [33]Sangeeta S, Arangasamy A, Kulkarni S, et al. Role of amino acids as additives on sperm motility, plasma membrane integrity and lipid peroxidation levels at pre-freeze and postthawed ram semen[J]. Anim Reprod Sci, 2015, 161:82-88.
• [34]Fukui Y, Kohno H, Togari T, et al. Fertility of Ewes inseminated intrauterinally with frozen semen using extender containing bovine serum albumin[J]. J Reprod Devlop, 2007, 53(4):959.
• [35]Matsuoka T, Imai H, Kohno H, et al. Effects of bovine serum albumin and trehalose in semen diluents for improvement of frozen-thawed ram spermatozoa[J]. J Reprod Devlop, 2006,52(5):675-683.
• [36]Baghshahi H, Riasi A, Mahdavi A H, et al. Antioxidant effects of clove bud(Syzygium aromaticum)extract used with different extenders on ram spermatozoa during cryopreservation[J].Cryobiology, 2014, 69(3):482-487.
• [37]Motlagh M K, Sharafi M, Zhandi M, et al. Antioxidant effect of rosemary(Rosmarinus officinalis L.)extract in soybean lecithin-based semen extender following freeze-thawing process of ram sperm[J]. Cryobiology, 2014, 69(2):217-222.
• [38]Gandomani V T, MahdaviA H, Rahmani H R, et al. Effects of different levels of clove bud(Syzygium aromaticum)on performance, intestinal microbial colonization, jejunal morphology, and immunocompetence of laying hens fed different n-6 to n-3 ratios[J]. Livest Sci, 2014, 167(9):236-248.
• [39]Najafi A, Najafi M, Zanganeh Z, et al. Cryopreservation of ram semen in extenders containing soybean lecithin as cryoprotectant and hyaluronic acid as antioxidant[J]. Reprod Domest Anim, 2015, 49(6):934-940.
• [40]Mata-Campuzano M, Soleilhavoup C, Tsikis G, et al. Motility of liquid stored ram spermatozoa is altered by dilution rate independent of seminal plasma concentration[J]. Anim Reprod Sci, 2015, 162(11):31-36.
• [41]Allai L, Druart X,?zturk M, et al. Protective effects of Opuntia ficus-indica extract on ram sperm quality, lipid peroxidation and DNA fragmentation during liquid storage[J]. Anim Reprod Sci,2016, 175:1-9.
• [42]Sarlós P, Molnár A, Kókai M, et al. Comparative evaluation of the effect of antioxidants in the conservation of ram semen[J].Acta Vet Hung, 2002, 50(2):235-245.
• [43]Moradi A R, Malekinejad H, Farrokhiardabili F, et al. Royal Jelly improves the sperm parameters of ram semen during liquid storage and serves as an antioxidant source[J]. Small Ruminant Res, 2013, 113(2-3):346-352.
• [44]Jerez-Ebensperger R, Gil L, Gonzales N, et al. The combined use of honey, garlic(Allium Sstivum L.)and skimmed milk as an extender for chilling sheep semen[J]. Cryoletters, 2015,36(4):243.
• [45]Seifi-Jamadi A, Kohram H, Zareh-Shahne A, et al. Effect of various concentrations of butylated hydroxyanisole and butylated hydroxytoluene on freezing capacity of Turkman stallion sperm[J]. Anim Reprod Sci, 2016, 170:108-113.
• [46]Farshad A, Khalili B, Jafaroghli M. Effects of butylated hydroxytoluene on freezability of ram spermatozoa.[J]. Asian Austr J Anim Sci, 2010, 23(10):1276-1281.
• [47]?palekováE,Makarevich A.V, Kubovi?ováE, et al. Effect of caffeine on functions of cooling-stored ram sperm in vitro[J].Acta Vet Brno, 2014, 83(1):19-25.
• [48]Kubovi?ováE, Makarevich A V,?paleková, E, et al. Motility and fertilizing ability of frozen-thawed ram sperm from two sheep breeds[J]. Slovak J Anim Sci, 2011:134-139.
• [49]Meque L C, Gil L, Mualuzanga D, et al.Effect of different concentrations of two types of gelatine in soya milk extender for storage of liquid ram semen[J]. Reprod Domest Anim, 2005,40(4):345-410.
• [50]Yániz J, MartíJ I, Silvestre M A, et al. Effects of solid storage of sheep spermatozoa at 15 degrees C on their survival and penetrating capacity[J]. Theriogenology, 2005, 64:1844-1851.
• [51]Lai J P, Jiang Y, He X W, et al. Separation and determination of astaxanthin from microalgal and yeast samples by molecularly imprinted microspheres[J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2004, 804(1):25-30.
• [52]Peker Akalin P, Bucak M N, GüNG R, et al. Influence of lycopene and cysteamine on sperm and oxidative stress parameters during liquid storage of ram semen at 5℃[J]. Small Ruminant Res, 2016, 137:117-123.
• [53]Souza H M, Arruda Lúcia C P, Monteiro M M, et al. The Effect of Canthaxanthin on the Quality of Frozen Ram Spermatozoa[J]. Biopreserv Biobank, 2016, 15(3):220-227.
• [54]Reiter R J, Tan D X, Osuna C, et al. Actions of melatonin in the reduction of oxidative stress[J]. J Biomed Sci, 2000, 7(6):444-458.
• [55]Ashrafi I, Kohram H, Ardabili F F. Antioxidative effects of melatonin on kinetics, microscopic and oxidative parameters of cryopreserved bull spermatozoa[J]. Anim Reprod Sci, 2013,139(1-4):25-30.
• [56]Li X X, Yang X G, Lu Y Q, et al. Protective effects of melatonin against oxidative stress in flow cytometry-sorted buffalo sperm[J]. Reprod Domest Anim, 2012, 47(2):299-307.
• [57]Martín-Hidalgo D, Barón F J, Bragado M J, et al. The effect of melatonin on the quality of extended boar semen after long-term storage at 17℃[J]. Theriogenology, 2011, 75(8):1550-1560.
• [58]Succu S, Berlinguer F, Pasciu V, et al. Melatonin protects ram spermatozoa from cryopreservation injuries in a dosedependent manner[J]. J Pineal Res, 2011, 50(3):310-318.
• [59]Upreti G C, Jensen K, Munday R, et al. Studies on aromatic amino acid oxidase activity in ram spermatozoa:Role of pyruvate as an antioxidant[J]. Anim Reprod Sci, 1998, 51(4):275.