聚乙烯吡咯烷酮对公猪精液冷冻的影响

doi:10.16431/j.cnki.1671-7236.2020.08.024

中国畜牧兽医 ›› 2020, Vol. 47 ›› Issue (8): 2553-2560.doi: 10.16431/j.cnki.1671-7236.2020.08.024

聚乙烯吡咯烷酮对公猪精液冷冻的影响

菅芯蕊^1,2, 钱芙蓉¹, 王珺^1,2, 李晓霞⁴, 张绍萱³, 张志彬¹, 于永生¹, 高凯², 刁云飞⁴, 张树敏¹, 李兆华¹

1. 吉林省农业科学院畜牧科学分院, 公主岭 136100;
2. 延边大学农学院, 延吉 133002;
3. 吉林大学动物科学学院, 长春 130062;
4. 中国农业科学院特产研究所, 长春 130062

收稿日期:2019-12-20 出版日期:2020-08-20 发布日期:2020-08-15
通讯作者: 李兆华 E-mail:379493818@qq.com
作者简介:菅芯蕊(1994-),女,山西忻州人,硕士生,研究方向:动物遗传育种与繁殖,E-mail:jxrsxau@163.com
基金资助:
吉林省科技发展计划项目"新吉林黑猪扩繁与产业示范化"（20170307017NY）；长春市国际科技合作计划"猪精子抗冻性分子标记的研究"（12ZX73）；吉林省农业科技创新工程项目"抗氧化剂与冷冻保护剂对冷冻解冻后猪精子质量及配种成绩影响的研究"（C92072003）

Effect of Polyvinylpyrrolidone on Cryopreservation of Boar Semen

JIAN Xinrui^1,2, QIAN Furong¹, WANG Jun^1,2, LI Xiaoxia⁴, ZHANG Shaoxuan³, ZHANG Zhibin¹, YU Yongsheng¹, GAO Kai², DIAO Yunfei⁴, ZHANG Shumin¹, LI Zhaohua¹

1. Branch of Animal Husbandry, Jilin Academy of Agricultural Science, Gongzhuling 136100, China;
2. Agricultural College of Yanbian University, Yanji 133002, China;
3. College of Animal Science, Jilin University, Changchun 130062, China;
4. Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun 130062, China

Received:2019-12-20 Online:2020-08-20 Published:2020-08-15

摘要/Abstract

摘要： 试验旨在评价聚乙烯吡咯烷酮（polyvinylpyrrolidone，PVP）对公猪精液冷冻的影响。试验分为5组，分别为对照组（不添加PVP）和PVP处理组（在冷冻基础液中分别加入0.25%、0.50%、1.00%、2.00% PVP）。采用手握法采集松辽黑猪精液，用5种冷冻基础液稀释，在25 ℃平衡1 h，17 ℃平衡2 h，4 ℃平衡3 h后灌装于0.5 mL细管中，在液氮上方3 cm处熏蒸10 min，保存在液氮罐中30 d后进行检测。样本解冻后分别检测精子活力、质膜完整性、顶体完整性、线粒体活性、DNA完整性、过氧化氢酶（catalase，CAT）活性、超氧化物歧化酶（superoxide dismutase，SOD）活性、谷胱甘肽过氧化物酶（glutathioneperoxidase，GSH-Px）活性、活性氧簇（reactive oxygen species，ROS）水平及丙二醛（malondialdehyde，MDA）水平。结果显示，与对照组相比，冷冻基础液中添加0.50% PVP显著提高冻融后精子的活力、质膜完整性、顶体完整性、线粒体活性、CAT活性、SOD活性、GSH-Px活性（P<0.05），显著降低精子ROS和MDA水平（P<0.05）；与对照组相比，添加PVP有利于提高DNA完整性，但差异不显著（P>0.05）。因此，猪精液冷冻基础液中添加PVP可改善冻融后精子质量，添加0.50%效果最佳。

关键词: 公猪; 精液; 聚乙烯吡咯烷酮(PVP); 冷冻保存; 精子质量

Abstract: The purpose of this study was to investigate the effect of polyvinylpyrrolidone (PVP) on boar semen freezing.There were five groups in the experiment:Control group (not add PVP) and PVP treatment groups (add 0.25%,0.50%,1.00%,and 2.00% PVP to the frozen base fluid).Collecting the semen of Songliao Black pig by hand,diluted with five frozen base solutions,equilibrated for 1 h at 25 ℃,2 h at 17 ℃,3 h at 4 ℃,then filled in 0.5 mL straw,fumigated 3 cm above liquid nitrogen for 10 min,stored in a liquid nitrogen tank for 30 d and tested.Sperm viability,plasma membrane integrity,acrosome integrity,mitochondrial activity,DNA integrity,catalase (CAT) activity,superoxide dismutase (SOD) activity,glutathione peroxidase (GSH-Px) activity,reactive oxygen species (ROS) level and malondialdehyde (MDA) level were tested after thawing.The results showed that compared with the control group,adding 0.50% PVP to the frozen base fluid could significantly improve the sperm viability,plasma membrane integrity,acrosome integrity,mitochondrial activity,CAT activity,SOD activity,GSH-Px activity (P<0.05);and significantly reduced the sperm ROS and MDA level (P<0.05).Adding PVP was beneficial to improve DNA integrity,but had no significant effect on DNA integrity compared with the control group (P>0.05).In conclusion,adding PVP to boar semen in the frozen base solution could improve the quality of sperm after freezing and thawing,the best concentration was 0.50%.

Key words: boar; semen; polyvinylpyrrolidone (PVP); cryopreservation; semen quality

中图分类号:

S823.8⁺3

菅芯蕊, 钱芙蓉, 王珺, 李晓霞, 张绍萱, 张志彬, 于永生, 高凯, 刁云飞, 张树敏, 李兆华. 聚乙烯吡咯烷酮对公猪精液冷冻的影响[J]. 中国畜牧兽医, 2020, 47(8): 2553-2560.

JIAN Xinrui, QIAN Furong, WANG Jun, LI Xiaoxia, ZHANG Shaoxuan, ZHANG Zhibin, YU Yongsheng, GAO Kai, DIAO Yunfei, ZHANG Shumin, LI Zhaohua. Effect of Polyvinylpyrrolidone on Cryopreservation of Boar Semen[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(8): 2553-2560.

参考文献

[1] YANG C H,WU T W,CHENG F P,et al.Effects of different cryoprotectants and freezing methods on post-thaw boar semen quality[J].Reproductive Bio-logy,2016,16(1):41-46.
[2] 席利萌,罗军,杨地坤,等.渗透性冷冻保护剂对山羊精子的冷冻保护效果[J].西北农林科技大学学报(自然科学版),2015,43(8):27-32. XI L M,LUO J,YANG D K,et al.Cryoprotective effect of osmotic cryoprotectant on goat sperm[J].Journal of Northwest A&F University (Natural Science Edition),2015,43(8):27-32.(in Chinese)
[3] 张美娜,郭富强,宋国欣,等.冷冻保护剂对家畜精液冷冻保存的研究进展[J].黑龙江畜牧兽医,2019,21:37-41. ZHANG M N,GUO F Q,SONG G X,et al.Research progress on cryopreservatives for cryopreservation of livestock semen[J].Heilongjiang Animal Science and Veterinary Medicine,2019,21:37-41.(in Chinese)
[4] ZENG C J,TANG K Y,HE L,et al.Effects of glycerol on apoptotic signaling pathways during boar spermatozoa cryopreservation[J].Cryobiology,2014,68(3):395-404.
[5] 刘宏,王刚第,李青旺,等.二甲基甲酰胺对猪精液冷冻保存效果的影响[J].中国畜牧兽医,2010,37(3):137-141. LIU H,WANG G D,LI Q W,et al.Effects of dimethylformamide on cryopreservation of boar semen[J].China Animal Husbandry & Veterinary Medicine,2010,37(3):137-141.(in Chinese)
[6] 马丽,李青旺,吴民耀.二甲基亚砜对猪精液冷冻保存效果研究[J].动物医学进展,2014,35(5):56-61. MA L,LI Q W,WU M Y.Effect of dimethyl sulfoxide on cryopreservation of boar semen[J].Progress in Veterinary Medicine,2014,35(5):56-61.(in Chinese)
[7] 毋状元,罗永明,董红,等.不同卵黄成分、大豆卵磷脂及β-环糊精胆固醇对马精子冷冻效果的影响[J].中国畜牧兽医,2016,43(9):2435-2440. WU Z Y,LUO Y M,DONG H,et al.Effect of different yolk composition,soybean lecithin and cholesterol-loaded cyclodextrin on equine sperm cryopreservation[J].China Animal Husbandry & Veterinary Medicine,2016,43(9):2435-2440.(in Chinese)
[8] RAKHA B A,ANSARI M,AKHTER S,et al.Cryopreservation of Indian red jungle fowl (Gallus gallus murghi) semen[J].Animal Reproduction Science,2016,174:45-55.
[9] 朱振舒.聚乙烯吡咯烷酮纳米药物传输系统研究[D].南京:南京大学,2011. ZHU Z S.Study on the nanodrug delivery system of polyvinylpyrrolidone[D].Nanjing:Nanjing University,2011.(in Chinese)
[10] FAHY G M,WOWK B,WU J,et al.Improved vitrifification solutions based on the predictability of vitrifification solution toxicity[J].Cryobiology,2004,48:22-35.
[11] HERRERA J A,QUINTANA J A,LOPEZ M A,et al.Individual cryopreservation with dimethyl sulfoxide and polyvinylpyrrolidone of ejaculates and pooled semen of three avian species[J].Archives of Andrology,2005,51(5):353-360.
[12] BUSHRA A R,MUHAMMAD S A,SHAMIM A,et al.Cryopreservation of Indian red jungle fowl (Gallus gallus murghi) semen with polyvinylpyrrolidone[J].Cryobiology,2017,78:27-33.
[13] SCHÄFER J,WABERSKI D,JUNG M,et al.Impact of holding and equilibration time on post-thaw quality of shipped boar semen[J].Animal Reproduction Science,2017,187:109-115.
[14] 马婷芳,史铁钧.聚乙烯吡咯烷酮的性能、合成及应用[J].应用化工,2002,31(3):16-19. MA T F,SHI T J.Properties,synthesis and application of polyvinylpyrrolidone[J].Applied Chemical Industry,2002,31(3):16-19.(in Chinese)
[15] 陈东峰,李伟风,赵海峰,等.聚乙烯吡咯烷酮、聚乙烯醇对公猪精液常温保存的影响[J].养猪,2012,4:41-42. CHEN D F,LI W F,ZHAO H F,et al.Effect of PVP and PVA on the normal temperature preservation of boar semen[J].Swine Production,2012,4:41-42.(in Chinese)
[16] LUO V,GIL L,OLACIREGUI M,et al.Antioxidant effect of lemon balm (Melissa officinalis) and mate tea (Ilex paraguensys) on quality,lipid peroxidation and DNA oxidation of cryopreserved boar epididymal spermatozoa[J].Andrologia,2015,47(9):1004-1011.
[17] SIEME H,OLDENHOF H,WOLKERS W F.Mode of action of cryoprotectants for sperm preservation[J].Animal Reproduction Science,2016,169:2-5.
[18] YESTE M,RODRÍGUEZ-GIL,JOAN E,et al.Artificial insemination with frozen-thawed boar sperm[J].Molecular Reproduction and Development,2017,84(9):802-813.
[19] ZHAO J,FU B,PENG W,et al.Melatonin protect the development of preimplantation mouse embryos from sodium fluoride-induced oxidative injury[J].Environmental Toxicology and Pharmacology,2017,54:133-141.
[20] JENG H A C,LIN W Y,CHAO M,et al.Semen quality and sperm DNA damage associa-revised-final-finalted with oxidative stress in relation to exposure to polycyclic aromatic hydroc-arbons[J].Journal of Environmental Science and Health,2018,53(14):1221-1228.
[21] MURATORI M,TAMBURRINO L,MARCHIANI S,et al.Investigation on the origin of sperm DNA fragmentation:Role of apoptosis,immaturity and oxidative stress[J].Molecular Medicine,2015,21(1):109-122.
[22] CHAMPROUX A,DAMON-SOUBEYRAND C,GOUBELY C,et al.Nuclear integrity but not topology of mouse sperm chromosome is affected by oxidative DNA damage[J].Genes,2018,9(10):501-517.
[23] TREULEN F,URIBE P,BOGUEN R,et al.Mitochondrial permeability transition increases reactive oxygen species production and induces DNA fragmentation in human spermatozoa[J].Human Reproduction,2015,30(4):767-776.

聚乙烯吡咯烷酮对公猪精液冷冻的影响

Effect of Polyvinylpyrrolidone on Cryopreservation of Boar Semen

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	吕春荣, 梁家充, 邵庆勇, 吴国权, 张玉梅, 权国波. 丙酮酸钠对山羊冷冻精子质量的影响[J]. 中国畜牧兽医, 2023, 50(6): 2363-2369.
[2]	刘锦钰, 张杨, 马旭梅, 周千, 周扬, 张春香, 石磊, 任有蛇. 谷胱甘肽对绵羊精液低温保存效果的影响[J]. 中国畜牧兽医, 2023, 50(6): 2388-2394.
[3]	宋承磊, 张哲, 韦怡林, 杨松柏, 豆雅晴, 李晨雷, 刘蓥珂, 李新建, 杨峰, 李秀领, 王克君, 乔瑞敏, 韩雪蕾. 不同品种、季节和月龄对公猪精液品质的影响[J]. 中国畜牧兽医, 2023, 50(5): 1938-1946.
[4]	杜朝晖, 葛梓豪, 游君怡, 刘晓玮, 梁国栋, 孙世铎, 胡建宏, 李晓. 丁酸梭菌和丁酸钠对高脂饮食诱导雄性小鼠生殖损伤的影响[J]. 中国畜牧兽医, 2023, 50(4): 1603-1616.
[5]	张笑科, 廖伟莉, 李婷婷, 李瑶, 李加琪, 黄翔, 张豪. 采精员和猪舍环境参数对杜洛克公猪精液品质的影响[J]. 中国畜牧兽医, 2023, 50(1): 220-229.
[6]	王赫川, 张涵, 李天峰, 王骞, 张群, 郭铭晖, 孙超, 周彦芝, 李远, 李雁冰, 魏国生, 李井春. 不同浓度牛磺酸对猪精液负压条件常温保存效果的影响[J]. 中国畜牧兽医, 2022, 49(9): 3484-3491.
[7]	刘刚, 朱芳贤, 章晓炜, 顾峰, 刘莉君, 苏锐, 孟飞, 许海涛, 孙飞舟. 湖羊胚胎和精液超低温保存效果的评价[J]. 中国畜牧兽医, 2022, 49(9): 3492-3499.
[8]	袁崇善, 王军, 吕文发. 多不饱和脂肪酸对动物精液品质影响的研究进展[J]. 中国畜牧兽医, 2022, 49(4): 1422-1429.
[9]	梁家充, 吕春荣, 洪琼花, 吴国权, 权国波. 云南半细毛羊精子冷冻前后转录组表达差异分析[J]. 中国畜牧兽医, 2022, 49(3): 973-981.
[10]	罗兰, 崔超, 董瑞兰, 刘浩域, 柳小斌, 于光辉. 不同浓度芦丁和冷冻速率对猪精子冷冻保存效果的研究[J]. 中国畜牧兽医, 2022, 49(3): 982-991.
[11]	陈嘉康, 吕春荣, 苗永旺, 权国波. 脂质组学技术在家畜精液质量评价和体外保存研究中的应用研究进展[J]. 中国畜牧兽医, 2022, 49(12): 4697-4706.
[12]	薛丽娥, 曹嘉程, 陈德军, 饶勇勇, 林瑞意, 肖天放, 方绍明. 鱼类抗冻蛋白在动物精液、细胞、胚胎低温保存中的研究进展[J]. 中国畜牧兽医, 2022, 49(12): 4715-4724.
[13]	李梦杰, 赵伟刚, 郭肖兰, 顾士钢, 赵全民, 徐超. 冷冻稀释液中添加大豆卵磷脂对梅花鹿冻融精子质量的影响[J]. 中国畜牧兽医, 2022, 49(11): 4327-4334.
[14]	张笑科, 廖伟莉, 李瑶, 吴煜伟, 庄汝柏, 贾百威, 袁晓龙, 李加琪, 黄翔, 张豪. 品种、季节、胎次、采精月龄及采精间隔对猪精液品质的影响[J]. 中国畜牧兽医, 2022, 49(10): 3879-3890.
[15]	陈璇, 吕艳秋, 韩越, 徐一超, 金一. 海藻糖替代部分甘油对延边黄牛冻融精子质量的影响[J]. 中国畜牧兽医, 2022, 49(1): 256-264.