基于两种液质联用技术研究西门塔尔种公牛精子活力与精浆肉碱的关系

doi:10.11843/j.issn.0366-6964.2019.11.021

摘要/Abstract

摘要： 旨在研究西门塔尔种公牛精子活力与精浆肉碱的关系。本研究共采集26份西门塔尔种公牛精液，根据精子活力进行分组，其中异常组12个样本，正常组14个样本。采用超高效液相色谱-四极杆-飞行时间质谱（UPLC-Q-TOF MS）技术对样品进行非靶测定，同时应用超高效液相色谱-三重四极杆-飞行时间质谱（UPLC-Q-Trap MS）技术进行广靶脂质测定，对于测定结果应用主成分分析（PCA）和正交偏最小二乘法-判别分析（OPLS-DA）进行统计分析。结果显示，异常组精浆代谢轮廓较正常组发生明显变化，将两种方法检测出的差异显著的肉碱进行分析，在精液活力异常组中，L-氯化棕榈酰肉碱、L-棕榈酰肉碱、硬脂酰肉碱、游离肉碱、乙酰肉碱、羟丁基-肉碱、3-羟基辛基肉碱、肉毒杆菌-肉碱、己烯基肉碱浓度显著高于活力正常组（P<0.05）。这些肉碱浓度与西门塔尔种公牛精液活力呈负相关，为研究牛精液品质的判定提供新的方法和思路。

Abstract: This experiment aimed to study the relationship between sperm motility and seminal plasma carnitine in Simmental bull. Twenty-six semen samples of Simmental bulls were collected, 12 of which were viewed as abnormal group, and 14 of which were viewed as normal group according to their sperm motility. Ultra-high Performance Liquid Chromatography-Quadrupole-Time-Of-Flight Mass Spectrometry (UPLC-Q-TOF MS) technique was used for non-target determination of samples, the Ultra-high Performance Liquid Chromatography-Quadrupole-Trap Mass Spectrometry (UPLC-Q-Trap MS) technology was used for broad-target lipid determination, principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used for statistical analysis. The results of multivariate statistics showed that the metabolic profile of seminal plasma in the abnormal group changed obviously compared with the normal group. The carnitine with significant difference detected by the two methods was analyzed, in the abnormal group, the concentration of decanoyl-L-carnitine, L-palmitoylcarnitine, stearoylcarnitine, free carnitine, acetyl-carnitine, hydroxybutyryl-carnitine, 3-hydroxyoctanoyl carnitine, myristoyl-carnitine and decenoyl-carnitine were significantly higher than those in the normal group (P<0.05). These carnitine concentrations were negatively correlated with the sperm motility of Simmental bulls, which would provide new methods and ideas for studying the determination of bovine semen quality.

中图分类号:

S823.3

罗芳, 刘继甜, 陶金忠. 基于两种液质联用技术研究西门塔尔种公牛精子活力与精浆肉碱的关系[J]. 畜牧兽医学报, 2019, 50(11): 2357-2364.

LUO Fang, LIU Jitian, TAO Jinzhong. Relationship between Sperm Motility and Seminal Plasma Carnitine in Simmental Bull Based on Two LC-MS Technology[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(11): 2357-2364.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2019.11.021

https://www.xmsyxb.com/CN/Y2019/V50/I11/2357

参考文献 32

[1]	BROOKS D E.Carnitine in the male reproductive tract and its relation to the metabolism of the epididymis and spermatozoa[J].Carnit Biosynth Metab Funct,1980:219-235.
[2]	BORNMAN M S,DU TOIT D,OTTO B,et al.Seminal carnitine,epididymal function and spermatozoal motility[J].S Afr Med J, 1989,75(1):20-21.
[3]	DE ROSA D,BOGGIA B,AMALFI B,et al.Correlation between seminal carnitine and functional spermatozoal characteristics in men with semen dysfunction of various origins[J].Drugs R D,2005,6(1):1-9.
[4]	MATALLIOTAKIS I,KOUMANTAKI Y,EVAGELIOU A,et al.L-carnitine levels in the seminal plasma of fertile and infertile men:correlation with sperm quality[J].Int J Fertil Womens Med,2000,45(3):236-240.
[5]	陆鹏荣.精加力复合肉碱联合左卡尼汀口服液治疗精子DNA损伤的临床分析[J].系统医学,2018,3(12):27-29.
	LU P R.Clinical analysis of sperm DNA damage treated by compound carnitine plus L-carnitine oral liquid[J].Systems Medicine,2018,3(12):27-29.(in Chinese)
[6]	陆金春.精液生化指标的全自动检测及临床应用[J].中华男科学杂志,2018,24(4):291-296.
	LU J C.Automatic detection and clinical application of semen biochemical markers[J].National Journal of Andrology, 2018, 24(4):291-296.(in Chinese)
[7]	孔祥军,姜睿.中药治疗少弱精子症的分子机制研究进展[J].中国男科学杂志,2018,32(1):68-72.
	KONG X J,JIANG R.Research progress on the molecular mechanism of traditional Chinese medicine in the treatment of oligoasthenospermia[J].Chinese Journal of Andrology,2018,32(1):68-72.(in Chinese)
[8]	赵宪林,昝林森,胡建宏,等.维生素E对秦川牛冷冻精液的抗氧化保护作用[J].畜牧兽医学报,2010,41(1):117-122.
	ZHAO X L,ZAN L S,HU J H,et al.The antioxidation protective effects of vitamin E on frozen-thawed semen in Qinchuan cattle[J].Acta Veterinaria et Zootechnica Sinica,2010,41(1):117-122.(in Chinese)
[9]	孙鹏.精加力复合肉碱对精子DNA完整性的影响及其在IVF-ET治疗中的临床疗效[J].临床医药文献电子杂志,2018,5(8):151.
	SUN P.Effects of spermatogenic plus carnitine on sperm DNA integrity and its clinical efficacy in IVF-ET treatment[J].Journal of Clinical Medical Literature,2018,5(8):151.(in Chinese)
[10]	李克,李伟,黄宇烽,等.精浆中游离L-肉碱水平与附属性腺生化指标的相关性研究[J].中华男科学杂志,2007, 13(6):507-510.
	LI K,LI W,HUANG Y F,et al.Correlation of free L-carnitine level with accessory gland markers and its clinical significance[J].National Journal of Andrology,2007,13(6):507-510.(in Chinese)
[11]	ZHOU B,XIAO J F,TULI L,et al.LC-MS-based metabolomics[J].Mol Biosyst,2012,8(2):470-481.
[12]	葛庆华,康修远,马欢,等.LC-MS/MS生物样品分析方法的应用与评价[J].中国医药工业杂志,2019,50(1):93-98.
	GE Q H,KANG X Y,MA H,et al.Application and evaluation of some validated LC-MS/MS bioanalytical methods used in routine sample analysis[J].Chinese Journal of Pharmaceuticals,2019,50(1):93-98.(in Chinese)
[13]	郭延生,陶金忠.基于LC-Q/TOF-MS代谢组学方法筛选奶牛妊娠识别阶段乳样生物标志物的研究[J].畜牧兽医学报,2018, 49(8):1633-1641.
	GUO Y S,TAO J Z.Selection of milk biomarkers of pregnancy recognition in dairy cows based on LC-Q/TOF-MS metabolomics[J].Acta Veterinaria et Zootechnica Sinica,2018,49(8):1633-1641.(in Chinese)
[14]	罗正中,白利鹏,黄逸馨,等.酵母硒对分娩应激状态下围产期奶牛代谢的影响[J].西北农林科技大学学报:自然科学版,2019,47(2):7-15.
	LUO Z Z,BAI L P,HUANG Y X,et al.Effect of selenium yeast on plasma metabolism of transition dairy cow in parturition stress status[J].Journal of Northwest A&F University:Natural Science Edition,2019,47(2):7-15.(in Chinese)
[15]	DAL BOSCO C,PANERO S,NAVARRA M A,et al.Screening and assessment of low-molecular-weight biomarkers of milk from cow and water buffalo:an alternative approach for the rapid identification of adulterated water buffalo mozzarellas[J].J Agric Food Chem,2018,66(21):5410-5417.
[16]	柳建明,姜辉,洪锴,等.男性不育患者精浆中游离睾酮、游离左旋肉碱水平与精液参数的相关性分析[J].第二军医大学学报, 2010,31(7):767-769.
	LIU J M,JIANG H,HONG K,et al.Correlation analysis of seminal plasma free testosterone and L-carnitine with semen parameters[J].Academic Journal of Second Military Medical University,2010,31(7):767-769.(in Chinese)
[17]	MONGIOI L,CALOGERO A E,VICARI E,et al.The role of carnitine in male infertility[J].Andrology,2016,4(5):800-807.
[18]	尹彪,刘红杰,赵明,等.精浆中锌、果糖和肉碱含量与精液参数的关系[J].中华男科学杂志,2013,19(11):1051-1053.
	YIN B,LIU H J,ZHAO M,et al.Relationship between the contents of zinc,fructose and carnitine in seminal plasma and semen parameters[J].National Journal of Andrology,2013,19(11):1051-1053.(in Chinese)
[19]	刘丽,张学红,陆彪,等.线粒体脂肪酸β-氧化障碍的研究新进展[J].中国优生与遗传杂志,2014,22(5):4-7.
	LIU L,ZHANG X H,LU B,et al.Novel progress in mitochondrial fatty acid β-oxidation disorders[J].Chinese Journal of Birth Health & Heredity,2014,22(5):4-7.(in Chinese)
[20]	AGARWAL A,SAID T M.Carnitines and male infertility[J].J Urol,2004,8(4):376-384.
[21]	JEULIN C,LEWIN L M.Role of free L-carnitine and acetyl-L-carnitine in post-gonadal maturation of mammalian spermatozoa[J]. Hum Reprod Update,1996,2(2):87-102.
[22]	康宁,马洁桦,周鑫,等.L-肉碱对糖尿病大鼠生精细胞凋亡及附睾精子数量和活动率的影响[J].中华男科学杂志,2011,17(5):422-426.
	KANG N,MA J H,ZHOU X,et al.Effects of L-carnitine on the apoptosis of spermatogenic cells and epididymal sperm count and motility in rats with diabetes mellitus[J].National Journal of Andrology,2011,17(5):422-426.(in Chinese)
[23]	MATALLIOTAKIS I,KOUMANTAKI Y,EVAGELIOU A,et al.L-carnitine levels in the seminal plasma of fertile and infertile men:correlation with sperm quality[J].Int J Fertil Womens Med,2000,45(3):236-240.
[24]	ZÖPFGEN A,PRIEM F,SUDHOFF F,et al.Relationship between semen quality and the seminal plasma components carnitine,alpha-glucosidase,fructose,citrate and granulocyte elastase in infertile men compared with a normal population[J]. Hum Reprod, 2000,15(4):840-845.
[25]	余颜,阳方,董良,等.左卡尼汀对梗阻性无精子症患者睾丸内精子质量的影响[J].中国性科学,2017,26(2):94-96.
	YU Y,YANG F,DONG L,et al.Effect of levocarnitine on testicular sperm quality in patients with obstructive azoospermia[J]. Chinese Journal of Human Sexuality,2017,26(2):94-96.(in Chinese)
[26]	阴继宁.观察L-肉碱联合维生素E和C治疗特发性少弱精子症的疗效[J].中国继续医学教育,2016,8(13):174-175.
	YIN J N.Observation curative effect on L-carnitine combine with vitamin E and C in treatment of idiopathic asthenospermia[J]. China Continuing Medical Education,2016,8(13):174-175.(in Chinese)
[27]	高国政,常轲祎,王芳.短期应用复方左旋肉碱在单精子卵胞浆注射治疗男性不育中的作用[J].中国实用医刊,2017, 44(6):68-70.
	GAO G Z,CHANG K Y,WANG F.Short term application of compound L-carnitine in the treatment of male infertility by intracytoplasmic sperm injection[J].Chinese Journal of Practical Medicine,2017,44(6):68-70.(in Chinese)
[28]	NADA E A,EL TAIEB M A,IBRAHIM H M,et al.Efficacy of tamoxifen and L-carnitine on sperm ultrastructure and seminal oxidative stress in patients with idiopathic oligoasthenoteratozoospermia[J].Andrologia,2015,47(7):801-810.
[29]	吴正沐,陆湘,王永卫,等.短期应用左卡尼汀在卵细胞胞质内单精子注射治疗少弱精子症中的作用[J].中华男科学杂志,2012,18(3):253-256.
	WU Z M,LU X,WANG Y W,et al.Short-term medication of L-carnitine before intracytoplasmic sperm injection for infertile men with oligoasthenozoospermia[J].National Journal of Andrology,2012,18(3):253-256.(in Chinese)
[30]	牛玉森.左旋肉碱治疗男性不育症有效性的Meta分析[J].兰州大学学报:医学版,2014,40(2):41-46.
	NIU Y S.A Meta analysis of the efficacy of L-carnitine on male infertility[J].Journal of Lanzhou University:Medical Sciences,2014,40(2):41-46.(in Chinese)
[31]	卢彦欣,杨瑞,姚旭旺,等.左旋肉碱治疗小鼠少弱精症机理[J].中国兽医学报,2018,38(8):1598-1602.
	LU Y X,YANG R,YAO X W,et al.L-carnitine improves the therapeutic effect of male mouse with cyclophosphamide-induced asthenospermia[J].Chinese Journal of Veterinary Science,2018,38(8):1598-1602.(in Chinese)
[32]	单妹,王燕桃,朱才兴,等.左旋肉碱和维生素对公猪精液品质的影响[J].猪业科学,2010,27(9):98-100.
	SHAN M,WANG Y T,ZHU C X,et al.Effects of L-carnitine and vitamin on boar semen quality[J].Swine Industry Science,2010,27(9):98-100.(in Chinese)