对香豆酸影响猪常温保存精液品质的研究

doi:10.16431/j.cnki.1671-7236.2021.08.025

摘要/Abstract

摘要： 本试验旨在探讨于猪精液基础稀释液（BTS）中添加不同浓度的对香豆酸（P-coumaric acid，PCA）对猪精液质量及抗氧化酶活性的影响。在猪常温稀释制剂中添加浓度为0、0.03、0.06、0.09、0.12、0.15 g/L的PCA，利用CASA全自动精子分析仪与荧光探针技术分别在1~5 d内检测精子运动学参数与精子功能完整性；利用抗氧化试剂盒在1、3和5 d检测精子活性氧（reactive oxygen species，ROS）和丙二醛（malondialdehyde，MDA）水平以及超氧化物歧化酶（superoxide dismutase，SOD）活性。试验结果表明，在精液保存的第5天，0.09 g/L的PCA处理组与空白组相比，精子活力显著提高（P<0.05），精子的顶体功能完整性、质膜功能完整性、线粒体功能完整性也显著升高（P<0.05），不仅如此，0.03、0.06、0.09和0.12 g/L PCA处理可显著降低精子ROS与MDA水平，提高SOD活性（P<0.05），其中0.09 g/L处理组相比其他组抗氧化活性效果最佳，而当添加浓度为0.15 g/L时，精子的抗氧化活性与0.09 g/L组相比显著降低（P<0.05）。综上，猪常温基础稀释液中添加PCA可改善精子常温保存品质，添加0.09 g/L效果最佳。

关键词: 猪精液; PCA; 常温保存; 精子动力学; 功能完整性; 抗氧化酶活性

Abstract: The purpose of this study was to investigate the effects of different concentrations of P-coumaric acid (PCA) added to Beltsville thawing solution (BTS) on boar semen quality and antioxidant enzyme activities.The concentrations of 0, 0.03, 0.06, 0.09, 0.12, 0.15 g/L PCA were added into boar semen dilution preserved at room temperature.The sperm kinetic parameters and sperm functional integrity were detected by CASA automatic sperm analyzer and fluorescence probe technology within 1-5 d.The levels of reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD) were determined by antioxidant kit on day 1, 3 and 5.The results showed that the sperm motility of the 0.09 g/L PCA group was significantly higher than that of the blank group on the 5th day of semen preservation (P<0.05), the sperm acrosome function integrity, plasma membrane function integrity and mitochondrial function integrity were also significantly higher than blank group (P<0.05).In addition, 0.03, 0.06, 0.09 and 0.12 g/L PCA treatment significantly reduced the levels of ROS and MDA, and increased the activity of SOD (P<0.05).The antioxidant activity of 0.09 g/L PCA, which in comparison with other treatment groups, was won the optimum antioxidant effect.However, when the adding concentration of PCA up to 0.15 g/L, the antioxidant activity of sperm was significantly decreased in comparison with 0.09 g/L (P<0.05).In summary, the addition of 0.09 g/L PCA to BTS could improve the quality of sperm, the best supplementation concentration of PCA was 0.09 g/L.

Key words: boar semen; PCA; room temperature preservation; sperm kinetic; functional integrity; antioxidant enzyme activity

中图分类号:

S814.1

蓝群, 谢颖瑜, 曹嘉程, 薛丽娥, 李家权, 赵芳露, 林瑞意, 方绍明, 肖天放. 对香豆酸影响猪常温保存精液品质的研究[J]. 中国畜牧兽医, 2021, 48(8): 2928-2936.

LAN Qun, XIE Yingyu, CAO Jiacheng, XUE Li'e, LI Jiaquan, ZHAO Fanglu, LIN Ruiyi, FANG Shaoming, XIAO Tianfang. Effect of P-coumaric Acid on the Quality of Boar Semen Preserved at Room Temperature[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 2928-2936.

参考文献

[1] 戈新,张宝珣,王建华,等.商品稀释剂pH值、渗透压测定及其对猪常温精液保存效果的比较[J].畜牧与兽医,2011,43(2):37-40. GE X,ZHANG B X,WANG J H,et al.Determination of pH value and osmotic pressure of different commercial diluents and their effects on liquid preservation of boar semen[J].Animal Husbandry and Veterinary Medicine,2011,43(2):37-40.(in Chinese)
[2] 赵媛.常见稀释粉对猪精液品质的影响试验[J].中国畜禽种业,2015,11(1):60-61. ZHAO Y.Effect of common diluents on boar semen quality[J].The Chinese Livestock and Poultry Breeding,2015,11(1):60-61.(in Chinese)
[3] TESI M,SABATINI C,VANNOZZI I,et al.Variables affecting semen quality and its relation to fertility in the dog:A retrospective study[J].Theriogenology,2018,118:34-39.
[4] BARRANCO I,PADILLA L,TVARIJONAVICIUTE A,et al.Levels of activity of superoxide dismutase in seminal plasma do not predict fertility of pig AI-semen doses[J].Theriogenology,2019,140:18-24.
[5] LI J,BARRANCO I,TVARIJONAVICIUTE A,et al.Seminal plasma antioxidants are directly involved in boar sperm cryotolerance[J].Theriogenology,2018,107:27-35.
[6] 杨威.三种抗氧化剂对猪精子冷冻保存效果的影响[D].杨凌:西北农林科技大学,2015. YANG W.Effects of three antioxidants on cryopreservation of boar sperm[D].Yangling:Northwest A&F University,2015.(in Chinese)
[7] BUCCI D,SPINACI M,YESTE M,et al.Combined effects of resveratrol and epigallocatechin-3-gallate on post thaw boar sperm and IVF parameters[J].Theriogenology,2018,117:16-25.
[8] WANG S,SUN M,WANG N,et al.Effects of L-glutamine on boar sperm quality during liquid storage at 17 degrees[J].Animal Reproduction Science,2018,191:76-84.
[9] ZHANG X G,LI H,WANG L,et al.The effects of different levels of superoxide dismutase in Modena on boar semen quality during liquid preservation at 17 degrees C[J].Animal Science Journal,2017,88(1):55-62.
[10] MOON P D,HAN N R,LEE J S,et al.p-coumaric acid,an active ingredient of Panax ginseng,ameliolates atopic dermatitis-like skin lesions through inhibition of thymic stromal lymphopoietin in mice[J].Journal of Ginseng Research,2021,45(1):176-182.
[11] BOEING T,COSTA P,VENZON L,et al.Gastric healing effect of p-coumaric acid isolated from Baccharis dracunculifolia DC on animal model[J].Naunyn-Schmiedebergs Archives of Pharmacology,2021,394(1):49-57.
[12] PRAGASAM S J,MURUNIKARA V,SABINA E P,et al.Antiperoxidative potential of p-coumaric acid,a common dietary phenol,in adjuvant-induced arthritis in rats[J].Zhong Xi Yi Jie He Xue Bao,2012,10(8):932-938.
[13] WEIKE S Y,LI Q,REN Z,et al.Effects of kojic acid on boar sperm quality and anti-bacterial activity during liquid preservation at 17C[J].Theriogenology,2019,140:124-135.
[14] LI Q,SHAOYONG W,LI Y,et al.Effects of oligomeric proanthocyanidins on quality of boar semen during liquid preservation at 17 degrees C[J].Animal Reproduction Science,2018,198:47-56.
[15] FENG T Y,REN F,FANG Q,et al.Effects of sulfanilamide on boar sperm quality,bacterial composition,and fertility during liquid storage at 17 degrees[J].Animal Science Journal,2019,90(9):1161-1169.
[16] SUN L,FAN X,ZENG Y,et al.Resveratrol protects boar sperm in vitro via its antioxidant capacity[J].Zygote,2020,2:1-8.
[17] ZOU T,LIU X,DING S,et al.Evaluation of sperm mitochondrial function using rh123/PI dual fluorescent staining in asthenospermia and oligoasthenozoospermia[J].Journal of Biomedical Research,2010,24(5):404-410.
[18] UCKUN F M,LIU X P,D'CRUZ O J.Human sperm immobilizing activity of aminophenyl arsenic acid and its N-substituted quinazoline,pyrimidine,and purine derivatives:Protective effect of glutathione[J].Reproductive Toxicology,2002,16(1):57-64.
[19] GADEA J,SELLES E,MARCO M A,et al.Decrease in glutathione content in boar sperm after cryopreservation.Effect of the addition of reduced glutathione to the freezing and thawing extenders[J].Theriogenology,2004,62(3-4):690-701.
[20] TZAMELI I.The evolving role of mitochondria in metabolism[J].Trends in Endocrinology and Metabolism,2012,23(9):417-419.
[21] TVRDA E,KNAZICKA Z,BARDOS L,et al.Impact of oxidative stress on male fertility——A review[J].Acta Veterinaria Hungarica,2011,59(4):465-484.
[22] AITKEN R J,DREVET J R.The Importance of oxidative stress in determining the functionality of mammalian spermatozoa:A two-edged sword[J].Antioxidants (Basel),2020,9(2):111.
[23] AGARWAL A,VIRK G,ONG C,et al.Effect of oxidative stress on male reproduction[J].World Journal of Mens Health,2014,32(1):1-17.
[24] BANASZEWSKA D,BIESIADA-DRZAZGA B,ANDRASZEK K.Frequency of cytoplasmic droplets depends on the breed and age of insemination boars[J].Folia Biologica (Krakow),2015,63(1):9-18.
[25] LOPEZ R A,RIJSSELAERE T,BEEK J,et al.Boar seminal plasma components and their relation with semen quality[J].Systerms Biology Reproductive Medicine,2013,59(1):5-12.
[26] AITKEN R J.Free radicals,lipid peroxidation and sperm function[J].Reproduction,Fertility,and Development,1995,7:659-668.
[27] ARMSTRONG J S,RAJASEKARAN M,CHAMULITRAT W,et al.Characterization of reactive oxygen species induced effects on human spermatozoa movement and energy metabolism[J].Free Radical Biology and Medicine,1999,26:869-880.
[28] KUMAR R,VENKATESH S,KUMAR M,et al.Oxidative stress and sperm mitochondrial DNA mutation in idiopathic oligoasthenozoospermic men[J].Indian Journal of Biochemistry and Biophysics,2009,46(2):172-177.
[29] VENKATESH S,DEECARAMAN M,KUMAR R,et al.Role of reactive oxygen species in the pathogenesis of mitochondrial DNA (mtDNA) mutations in male infertility[J]. Indian Journal of Biochemistry and Biophysics,2009,129:127-137.
[30] PEZO F,ZAMBRANO F,URIBE P,et al.Oxidative and nitrosative stress in frozen-thawed pig spermatozoa.II:Effect of the addition of saccharides to freezing medium on sperm function[J].Cryobiology,2020,97:5-11.
[31] 管西芹,毛近隆,唐迎雪,等.对香豆酸的药理作用研究进展[J].中草药,2018,49(17):4162-4170. GUAN X Q,MAO J L,TANG Y X,et al.Research progress on pharmacological action of p-coumaric acid[J]. Chinese Traditional and Herbal Drugs,2018,49(17):4162-4170.(in Chinese)
[32] WEI D,ZHANG X L,WANG Y Z,et al.Lipid peroxidation levels,total oxidant status and superoxide dismutase in serum,saliva and gingival crevicular fluid in chronic periodontitis patients before and after periodontal therapy[J].Australian Dental Journal,2010,55(1):70-78.
[33] AHMADIFAR E,KALHOR N,DAWOOD M,et al.The blood and mRNA levels of antioxidant-related factors in common carp (Cyprinus carpio) fed p-Coumaric acid[J].Fish Physiology and Biochemistry,2021,47(1):59-68.
[34] LODOVICI M,RAIMONDI L,GUGLIELMI F,et al.Protection against ultraviolet B-induced oxidative DNA damage in rabbit corneal-derived cells (SIRC) by 4-coumaric acid[J].Toxicology,2003,184(2-3):141-147.
[35] SABITHA R,NISHI K,GUNASEKARAN V P,et al.p-Coumaric acid attenuates alcohol exposed hepatic injury through MAPKs,apoptosis and Nrf2 signaling in experimental models[J].Chemico-Biological Interactions,2020,321:109044.
[36] KHEIRY M,DIANAT M,BADAVI M,et al.p-coumaric acid attenuates lipopolysaccharide-induced lung inflammation in rats by scavenging ROS production:An in vivo and in vitro study[J].Inflammation,2019,42(6):1939-1950.
[37] KHLIFAOUI M,BATTUT I,BRUYAS J F,et al.Effects of glutamine on post-thaw motility of stallion spermatozoa:An approach of the mechanism of action at spermatozoa level[J].Theriogenology,2005,63(1):138-149.
[38] DEVASAGAYAM T P,TILAK J C,BOLOOR K K,et al.Free radicals and antioxidants in human health:Current status and future prospects[J]. Journal of The Association of Physicians India,2004,52:794-804.
[39] HU J H,ZAN L S,ZHAO X L,et al.Effects of trehalose supplementation on semen quality and oxidative stress variables in frozen-thawed bovine semen[J].Journal of Animal Science,2010,88(5):1657-1662.