基于4D-DIA技术的伊犁马早期妊娠血清蛋白质组分析

doi:10.11843/j.issn.0366-6964.2024.08.022

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (8): 3493-3502.doi: 10.11843/j.issn.0366-6964.2024.08.022

基于4D-DIA技术的伊犁马早期妊娠血清蛋白质组分析

曾明敏¹(), 孟军¹^,², 曾亚琦¹^,², 王建文¹^,², 邓海峰³, 任万路¹, 薛宇恒¹, 尚婷婷¹, 高凤¹, 姚新奎¹^,²^,*()

1. 新疆农业大学动物科学学院/新疆农业大学马产业研究院，乌鲁木齐 830052
2. 新疆马繁育与运动生理重点实验室，乌鲁木齐 830052
3. 伊犁哈萨克自治州昭苏马场畜牧兽医科，昭苏 835600

收稿日期:2023-12-18 出版日期:2024-08-23 发布日期:2024-08-28
通讯作者: 姚新奎 E-mail:1536866543@qq.com;yxk61@126.com
作者简介:曾明敏(1998-)，女，新疆石河子人，硕士生，主要从事动物生产学研究，E-mail：1536866543@qq.com
基金资助:
新疆维吾尔自治区重大科技专项“国产马专门化品系高效繁育技术体系构建”(2022A02013-1);自治区创新环境(人才、基地)建设专项“马种质创新培育与高效健康养殖重点实验室平台建设”(PT2311);中央引导地方科技发展专项资金项目“专门化运动马培育技术服务支撑”(ZYYD2023C02)

Analysis of Serum Proteomics in Early Pregnancy of Yili Horses Based on 4D-DIA Technology

Mingmin ZENG¹(), Jun MENG¹^,², Yaqi ZENG¹^,², Jianwen WANG¹^,², Haifeng DENG³, Wanlu REN¹, Yuheng XUE¹, Tingting SHANG¹, Feng GAO¹, Xinkui YAO¹^,²^,*()

1. College of Animal Science, Xinjiang Agricultural University/Research Institute of Horse Industry, Xinjiang Agricultural University, Urumqi 830052, China
2. Xinjiang Key Laboratory of Equine Breeding and Exercise Physiology, Urumqi 830052, China
3. Animal Husbandry and Veterinary Department, Zhaosu Horse Farm, Yili Kazak Autonomous Prefecture, Zhaosu 835600, China

Received:2023-12-18 Online:2024-08-23 Published:2024-08-28
Contact: Xinkui YAO E-mail:1536866543@qq.com;yxk61@126.com

摘要/Abstract

摘要：

旨在利用蛋白质组学技术探究伊犁马妊娠早期血清蛋白质组特征，筛选差异蛋白质并分析其在妊娠中的作用。本研究选取年龄相同、膘情相近的健康伊犁马母马20匹，自然发情后进行人工授精。在母马排卵后第13天进行妊娠诊断，根据B超结果选择妊娠母马与未妊娠母马各3匹并采集其血清样品。利用4D-DIA技术对伊犁马母马血清样品进行蛋白定量定性分析，筛选差异蛋白质并进行GO、KEGG等生物信息学分析。结果显示，共鉴定出823个蛋白质，分子量范围在8~1 010 ku；根据差异倍数绝对值大于1.5倍、P＜0.05的阈值，筛选出53种差异显著性蛋白质，其中26个蛋白上调，27个蛋白下调。差异蛋白质主要参与细胞代谢、生物调节、代谢、对刺激的反应、细胞成分组织或生物发生等生物学过程；KEGG富集通路包含蛋白酶体、精氨酸生物合成、PI3K-Akt信号、ECM受体相互作用等与妊娠相关通路。根据GO、KEGG等生物学分析，初步筛选出6种差异显著性蛋白可能与早期妊娠存在关系，并用其绘制蛋白互作网络图，结果显示，CTSS、MMP1、SERPINA5等处在关键节点。研究初步筛选出6种妊娠差异蛋白，为今后筛选马属动物妊娠特异性蛋白及分子调控机制提供了基础资料。

关键词: 蛋白质组, 伊犁马, 妊娠, 血清

Abstract:

The purpose of this article was to explore the serum proteomic characteristics of early pregnancy in Yili horses using proteomic techniques, screen for differential proteins, and analyze their roles in pregnancy. In this study, 20 healthy Yili mares with the same age and similar weight were selected for artificial insemination after natural estrus. Pregnancy diagnosis was performed on the 13th day after ovulation in the mare. Three pregnant mares and three non pregnant mares were selected based on ultrasound results, and their serum samples were collected. Using 4D-DIA technology for protein quantitative and qualitative analysis of serum samples from Yili mares, screening differential proteins, and conducting bioinformatics analysis such as GO and KEGG. The results showed that a total of 823 proteins were identified, with a molecular weight range of 8-1 010 ku; Based on the threshold of absolute multiple of differences greater than 1.5 times and P < 0.05, 53 proteins with significant differences were screened, of which 26 proteins were upregulated and 27 proteins were downregulated. Differential proteins mainly participated in biological processes such as cellular metabolism, biological regulation, metabolism, response to stimuli, cellular component organization, or biogenesis; The KEGG enrichment pathways included proteasomes, arginine biosynthesis, PI3K Akt signaling, ECM receptor interactions, and other pregnancy related pathways. Based on biological analysis such as GO and KEGG, six significantly different proteins were preliminarily screened for possible relationship with early pregnancy, and protein interaction network diagrams were drawn using them. The results showed that CTSS, MMP1, SERPINA5, and others were at key nodes. Six pregnancy specific proteins have been preliminarily screened in this study, providing basic data for screening pregnancy specific proteins and molecular regulatory mechanisms in equine animals in the future.

Key words: proteome, Yili horse, pregnancy, serum

中图分类号:

S821.2

曾明敏, 孟军, 曾亚琦, 王建文, 邓海峰, 任万路, 薛宇恒, 尚婷婷, 高凤, 姚新奎. 基于4D-DIA技术的伊犁马早期妊娠血清蛋白质组分析[J]. 畜牧兽医学报, 2024, 55(8): 3493-3502.

Mingmin ZENG, Jun MENG, Yaqi ZENG, Jianwen WANG, Haifeng DENG, Wanlu REN, Yuheng XUE, Tingting SHANG, Feng GAO, Xinkui YAO. Analysis of Serum Proteomics in Early Pregnancy of Yili Horses Based on 4D-DIA Technology[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3493-3502.

图/表 7

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蛋白ID Accession ID	蛋白质名称 Protein name	基因 Gene	上调/下调 Up/Down	差异倍数 log ₂(FC)值	P值 P value
F6ZRF6	SERPIN家族A成员7	SERPINA7	↑	0.696 498	9.15×10^-9
F6PPF8	钙粘蛋白17	CDH17	↑	0.714 742	3.54×10^-6
F7D2I6	蛋白质精氨酸甲基转移酶5	PRMT5	↑	1.452 295	0.000 116
F7AWQ9	白细胞介素1受体辅助蛋白	IL1RAP	↑	0.605 578	0.000 243
F6VUW2	组织蛋白酶S	CTSS	↑	0.589 026	0.000 274
F7C373	热休克蛋白家族A(Hsp70)成员9	HSPA9	↑	1.169 765	0.001 492
F6Q4Q1	异构核核糖核蛋白K	HNRNPK	↑	1.199 253	0.001 955
A0A5S7NAP8	氯化物通道附件2	CLCA2	↓	-0.745 096	1.62×10^-11
A0A3Q2HKG3	乳转铁蛋白	LTF	↓	-0.668 901	9.00×10^-6
F6V881	玻连蛋白	VTN	↓	-0.671 550	4.58×10^-5
F7C0D2	脂多糖结合蛋白	LBP	↓	-0.649 453	0.000 555
A0A3Q2HNU8	SERPIN家族A成员5	SERPINA5	↓	-1.558 677	0.000 592
A0A3Q2HN87	硫运载蛋白2	LCN2	↓	-0.808 099	0.002 139
F7C349	血清淀粉样蛋白P组分	APCS	↓	-1.685 178	0.002 163

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基于4D-DIA技术的伊犁马早期妊娠血清蛋白质组分析

Analysis of Serum Proteomics in Early Pregnancy of Yili Horses Based on 4D-DIA Technology

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