母猪妊娠早期血液中差异表达蛋白的鉴定

doi:10.11843/j.issn.0366-6964.2022.04.011

Abstract

Abstract: The study aimed to investigate the proteins expression in peripheral blood at early pregnancy, and to discover potential protein biomarkers for early pregnancy diagnosis in sows. In this study, the peripheral blood from 8 healthy, normal production performance and similar physical condition of pregnant and non-pregnant Landrace×Large White multiparous sows was collected on the 15th day after mating. Proteins were isolated, then iTRAQ technique and bioinformatics software were employed to screen differentially expressed proteins related to embryo attachment. ELISA was used to verify its expression and ROC curve was analyzed. A total of 24 differentially expressed proteins were screened in the peripheral blood of pregnant and non-pregnant sows(19 were up-regulated and 5 were down-regulated in pregnant sows), which mainly enriched in PI3K-Akt, p53, MAPK, Estrogen, mTOR and other important signaling pathways. ELISA test verified their expression trend were consistent with iTRAQ results, which proved the accuracy of iTRAQ results. ROC curve analysis showed that FLNC, ADIPOQ, HP, TERF2, FLNC-ADIPOQ and ADIPOQ-HP could be used as a potential biomarker for early pregnancy of sows, with good accuracy and specificity. These results suggest that the differential proteins FLNC, ADIPOQ, HP and TERF2 can be used as potential biomarkers for early pregnancy diagnosis in sows, and provide a new view for the study of the mechanism of embryos implantation in sows.

Key words: iTRAQ, proteomics, sows, early pregnancy, biomarkers

CLC Number:

S828.3

QI Mengfan, XIE Su, GAO Ruonan, SUN Yishan, SUN Xiaomei, HE Junfei, LU Huiwen, LU Shihao, CHEN Xin, LI Qingchun, HUANG Tao. Identification of Differentially Expressed Proteins in Blood of Sows at Early Pregnancy[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1109-1121.

References 35

[1]	CHOE C,PARK J W,KIM E S,et al.Proteomic analysis of differentially expressed proteins in bovine endometrium with endometritis[J]. Korean J Physiol Pharmacol, 2010,14(4):205-212.
[2]	MULLEN M P,ELIA G,HILLIARD M,et al.Proteomic characterization of histotroph during the preimplantation phase of the estrous cycle in cattle[J].J Proteome Res, 2012,11(5):3004-3018.
[3]	KOCH J M,RAMADOSS J,MAGNESS R R.Proteomic profile of uterine luminal fluid from early pregnant ewes[J].J Proteome Res, 2010,9(8):3878-3885.
[4]	WADSACK C,TABANO S,MAIER A,et al.Intrauterine growth restriction is associated with alterations in placental lipoprotein receptors and maternal lipoprotein composition[J].Am J Physiol Endocrinol Metab, 2007,292(2):E476-E484.
[5]	王煜.microRNA在流产及早孕蜕膜组织中的表达及miR-199的功能鉴定[D].呼和浩特:内蒙古大学,2015.WANG Y.Indntification of microRNA in miscarriage and preganancy decidua and miR-199 functional study[D].Hohhot:Inner Mongolia University,2015.(in Chinese)
[6]	张诗萌,佟翠,李鹏,等.早孕因子及其临床应用研究进展[J].黑龙江畜牧兽医,2019(5):41-43.ZHANG S M,TONG C,LI P,et al.Research progress of early pregnancy factor and its clinical application[J].Heilongjiang Animal Science and Veterinary Medicine, 2019(5):41-43.(in Chinese)
[7]	黄喜英,丘彦.早期妊娠血清β-HCG、E2、P的监测意义[J].中国医药科学,2013,3(7):47-49.HUANG X Y,QIU Y.The value of monitoring serum levels of human chorionic gonadotrophin,estradiol and progesterone during early pregnancy[J].China Medicine and Pharmacy, 2013,3(7):47-49.(in Chinese)
[8]	薄小辉.妊娠相关糖蛋白在奶牛早期妊娠诊断上的应用研究[D].北京:中国农业科学院,2017.BO X H.The application of pregnancy-associated glycoproteins in early pregnancy diagnosis of dairy cows[D].Beijing:Chinese Academy of Agricultural Sciences,2017.(in Chinese)
[9]	程蕾,王定发,向敏,等.奶牛早期妊娠阶段外周血中IFITM1 和RSAD2 基因的表达[J].华中农业大学学报,2015,34(3): 89-96.CHENG L,WANG D F,XIANG M,et al.Transcriptional analysis of IFITM1 and RSAD2 in maternal peripheral blood during early pregnancy of dairy cows[J].Journal of Huazhong Agricultural University, 2015,34(3):89-96.(in Chinese)
[10]	陈骊珠,张墨,马巍,等.蛋白质组学技术在产前诊断生物标志物筛查中的研究进展[J].中华全科医学,2018,16(3): 461-464.CHEN L Z,ZHANG M,MA W,et al.The main advances in the application of proteomics in prenatal diagnosis[J].Chinese Journal of General Practice, 2018,16(3):461-464.(in Chinese)
[11]	石娟,李勇勇,周丽萍,等.基于iTRAQ技术研究冻藏条件对中华管鞭虾肌肉蛋白质的影响[J].中国食品学报,2021,21(11): 120-132.SHI J,LI Y Y,ZHOU L P,et al.The effect of frozen storage conditions on muscular protein of Solenocera melantho based on iTRAQ technology[J].Journal of Chinese Institute of Food Science and Technology, 2021,21(11):120-132.(in Chinese)
[12]	彭梦玲,胡文业,李乃馨,等.组学技术分析肉鸡胚胎发育过程中肝脏蛋白表达的变化[J].畜牧兽医学报,2020,51(2):252-259.PENG M L,HU W Y,LI N X,et al.The changes of hepatic proteins during chicken embryonic development based on proteomics analysis[J]. Acta Veterinaria et Zootechnica Sinica, 2020,51(2):252-259.(in Chinese)
[13]	陈敏,张玲.iTRAQ技术在胃癌蛋白质组学研究中的应用进展[J].内蒙古医学杂志,2021,53(9):1075-1077,1080.CHEN M,ZHANG L.Application advances of iTRAQ in the study of gastric cancer proteomics[J].Inner Mongolia Medical Journal, 2021,53(9):1075-1077,1080.(in Chinese)
[14]	FRANTZI M,VLAHOU A.Ten years of proteomics in bladder cancer:progress and future directions[J].Bladder Cancer, 2017, 3(1):1-18.
[15]	傅强,邓小芳,GAEL A,等.高通量技术筛选主动脉夹层生物标志物的研究[J].临床心血管病杂志,2021,37(10):952-958.FU Q,DENG X F,GAEL A,et al.Biomarkers of aortic dissection screened by high-throughput technology[J].Journal of Clinical Cardiology (China), 2021,37(10):952-958.(in Chinese)
[16]	常程.基于iTRAQ技术的急性心肌梗死差异表达蛋白质的筛选及血清淀粉样蛋白A1在心肌缺血性损伤作用机制的研究[D].沈阳:中国医科大学,2021.CHANG C.Screening of differentially expressed proteins in acute myocardial infarction based on iTRAQ technology and study on the mechanism of serum amyloid A1 in myocardial ischemic injury[D].Shenyang:China Medical University,2021.(in Chinese)
[17]	李梦柔,韦明邦,张健,等.基于iTRAQ技术挖掘猪肾组织高原低氧适应性关键蛋白[J].畜牧兽医学报,2021,52(5): 1238-1246.LI M R,WEI M B,ZHANG J,et al.Mining key proteins of high altitude hypoxic adaptability in porcine kidney tissue based on iTRAQ technology[J].Acta Veterinaria et Zootechnica Sinica, 2021,52(5):1238-1246.(in Chinese)
[18]	赵畅,张江,白云龙,等.基于iTRAQ技术的卵巢静止奶牛血清差异蛋白分析[J].畜牧兽医学报,2019,50(5):972-982.ZHAO C,ZHANG J,BAI Y L,et al.Analysis of serum differential proteins in cows with inactive ovaries based on iTRAQ technology[J]. Acta Veterinaria et Zootechnica Sinica, 2019,50(5):972-982.(in Chinese)
[19]	SHARPE-TIMMS K L,RICKE E A,PIVA M,et al.Differential expression and localization of de-novo synthesized endometriotic haptoglobin in endometrium and endometriotic lesions[J].Hum Reprod, 2000,15(10):2180-2185.
[20]	BERKOVA N,LEMAY A,DRESSER D W,et al.Haptoglobin is present in human endometrium and shows elevated levels in the decidua during pregnancy[J].Mol Hum Reprod, 2001,7(8):747-754.
[21]	JALALI B M,BOGACKI M,DIETRICH M,et al.Proteomic analysis of porcine endometrial tissue during peri-implantation period reveals altered protein abundance[J].J Proteomics, 2015,125:76-88.
[22]	VANNUCCHI C I,MIRANDOLA R M,OLIVEIRA C M.Acute-phase protein profile during gestation and diestrous:proposal for an early pregnancy test in bitches[J].Anim Repord Sci, 2002,74(1-2):87-99.
[23]	CHO J K,KIM Y H,PARK I Y,et al.Polymorphism of haptoglobin in patients with premature rupture of membrane[J].Yonsei Med J, 2009,50(1):132-136.
[24]	GARCÍA-VÁZQUEZ F A,MOROS-NICOLÁS C,LÓPEZ- AU＇U BEDA R,et al.Evidence of haptoglobin in the porcine female genital tract during oestrous cycle and its effect on in vitro embryo production[J].Sci Rep, 2021,11(1):12041.
[25]	NARYZHNY S N,LEGINA O K.Haptoglobin as a biomarker[J].Biochem Mosc Suppl B Biomed Chem, 2021,15(3):184-198.
[26]	PALIN M F,BORDIGNON V V,MURPHY B D.Adiponectin and the control of female reproductive functions[J].Vitam Horm, 2012, 90:239-287.
[27]	TAKEMURA Y,OSUGA Y,YAMAUCHI T,et al.Expression of adiponectin receptors and its possible implication in the human endometrium[J].Endocrinology, 2006,147(7):3203-3210.
[28]	LORD E,LEDOUX S,MURPHY B D,et al.Expression of adiponectin and its receptors in swine[J].J Anim Sci, 2005,83(3): 565-578.
[29]	张小亚.PCOS小鼠子宫内膜、卵巢和胚胎中脂联素及其受体水平的变化以及对受孕率的影响[D].广州:暨南大学,2018.ZHANG X Y.Changes of adiponectin and its receptor levels in endometrium,ovary and embryo and their effects on conception rates in mice with PCOS[D].Guangzhou:Jinan University,2018.(in Chinese)
[30]	刘彩,朱君花,郝莉娜,等.重组脂联素对自然流产模型小鼠子宫蜕膜组织中FOXO3蛋白表达的影响[J].临床和实验医学杂志,2021,20(19):2048-2051.LIU C,ZHU J H,HAO L N,et al.Effect of recombinant adiponectin on FOXO3 protein expression in decidua tissues of spontaneous abortion model mice[J].Journal of Clinical and Experimental Medicine, 2021,20(19):2048-2051.(in Chinese)
[31]	卫雅娴,伊诺,周明书,等.重组脂联素对自然流产模型小鼠妊娠结局及Th17/Treg平衡的影响[J].临床和实验医学杂志, 2020,19(19):2017-2021.WEI Y X,YI N,ZHOU M S,et al.Effect of recombinant adiponectin on pregnancy outcome and Th17/Treg balance in spontaneous abortion mice model[J].Journal of Clinical and Experimental Medicine, 2020,19(19):2017-2021.(in Chinese)
[32]	SALKER M S,STEEL J H,HOSSEINZADEH Z,et al.Activation of SGK1 in endometrial epithelial cells in response to PI3K/AKT inhibition impairs embryo implantation[J].Cell Physiol Biochem, 2016,39(5):2077-2087.
[33]	LIU X Q,GAO R F,CHEN X M,et al.Possible roles of mmu-miR-141 in the endometrium of mice in early pregnancy following embryo implantation[J].PLoS One, 2013,8(6):e67382.
[34]	刘丽媛,何俊琳,刘学庆,等.PI3k/Akt信号通路及MMP-2、MMP-9在正常妊娠及自然流产绒毛组织中的表达[J].重庆医科大学学报,2014,39(11):1511-1515.LIU L Y,HE J L,LIU X Q,et al.Expression of PI3K/AKt signal pathway and MMP-2,MMP-9 in villi tissues of normal pregnancy and spontaneous abortion[J].Journal of Chongqing Medical University, 2014,39(11):1511-1515.(in Chinese)
[35]	SHEN J Y,ZHOU C L,ZHU S E,et al.Comparative transcriptome analysis reveals early pregnancy-specific genes expressed in peripheral blood of pregnant sows[J].PLoS One, 2014,9(12):e114036.

Identification of Differentially Expressed Proteins in Blood of Sows at Early Pregnancy

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