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

摘要/Abstract

摘要： 为了研究母猪妊娠早期外周血液中蛋白质的表达情况，寻找潜在的母猪妊娠早期生物标志物，本试验采集了8头健康、生产表现正常且体况相近的配种后15 d妊娠与非妊娠的长大二元经产母猪外周血液，分离蛋白，利用iTRAQ技术及生物信息学方法筛选与胚胎附植相关的差异表达蛋白，用ELISA验证其表达情况，并进行ROC曲线分析。结果显示，筛选到配种后15 d妊娠与非妊娠母猪外周血中的24个差异表达蛋白(妊娠母猪中19个上调，5个下调)，主要富集的信号通路包括PI3K-Akt、p53、MAPK、Estrogen和mTOR等重要信号通路。ELISA结果验证其表达趋势与iTRAQ检测结果一致。ROC曲线分析发现，配种后15 d血液中的FLNC、ADIPOQ、HP和TERF2以及FLNC-ADIPOQ、ADIPOQ-HP联合使用均可作为母猪早期妊娠潜在的生物标志物，且准确性与特异性较好。本研究鉴定了配种后15 d母猪血液中差异表达蛋白，且发现FLNC、ADIPOQ、HP和TERF2可作为母猪妊娠早期的潜在生物标志物，并为母猪胚胎附植机制研究提供新视角。

关键词: iTRAQ, 蛋白组学, 猪, 早期妊娠, 生物标志物

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

中图分类号:

S828.3

祁梦凡, 谢苏, 高若男, 孙义姗, 孙晓梅, 和军飞, 鲁慧文, 卢世豪, 陈鑫, 李清春, 黄涛. 母猪妊娠早期血液中差异表达蛋白的鉴定[J]. 畜牧兽医学报, 2022, 53(4): 1109-1121.

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.

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