绵羊子宫内膜容受前期与容受期的阴道微生物比较研究

doi:10.11843/j.issn.0366-6964.2025.02.020

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (2): 689-699.doi: 10.11843/j.issn.0366-6964.2025.02.020

绵羊子宫内膜容受前期与容受期的阴道微生物比较研究

楚翼健¹(), 崔久增¹, 李增开², 张磊¹, 褚婷婷¹, 黄艳平²^,*(), 宋宇轩¹^,*()

1. 西北农林科技大学动物科技学院, 杨凌 712100
2. 神木市畜牧业发展中心(神木市疾病预防控制中心), 神木 719300

收稿日期:2024-08-09 出版日期:2025-02-23 发布日期:2025-02-26
通讯作者: 黄艳平,宋宇轩 E-mail:yijianchu@outlook.com;huangyp121@126.com;syx98728@163.com
作者简介:楚翼健(1999-)，男，吉林通化人，硕士，主要从事动物生殖生理调控研究，E-mail: yijianchu@outlook.com
基金资助:
陕西省畜禽育种“两链’融合重点专项(2022GD-TSLD-46-0202)；高产优质奶绵羊快速扩繁与选育提高(SMXMKJ001)；陕西省中央引导地方科技发展资金项目(2024ZY-CGZY-20)；陕西省自然科学基础研究计划资助项目(2023-JC-QN-0181)

Comparative Study on Vaginal Microorganisms in Pre-endometrial Receptivity and Endometrial Receptivity of Sheep

CHU Yijian¹(), CUI Jiuzeng¹, LI Zengkai², ZHANG Lei¹, CHU Tingting¹, HUANG Yanping²^,*(), SONG Yuxuan¹^,*()

1. College of Animal Science and Technology, Northwest A & F University, Yangling 712100, China
2. Shenmu Animal Husbandry Development Center (Shenmu Center for Disease Control and Prevention), Shenmu 719300, China

Received:2024-08-09 Online:2025-02-23 Published:2025-02-26
Contact: HUANG Yanping, SONG Yuxuan E-mail:yijianchu@outlook.com;huangyp121@126.com;syx98728@163.com

摘要/Abstract

摘要：

旨在基于16S rRNA测序技术，解析绵羊子宫容受前期与容受期阴道微生物群落的差异变化，筛选绵羊子宫内膜容受期的标志菌群。本研究选取12只两岁龄、体重相近且健康的经产雌性湖羊为研究对象。试验分为两组，每组6只湖羊，利用同期发情技术，在绵羊配种后第5天(容受前期, pre-receptive endometrium组，PE组)与第15天(容受期，peceptive endometrium组，RE组)采集阴道分泌物及子宫内膜组织。利用RT-qPCR检测子宫内膜组织相关容受基因表达情况，并选择16S rRNA V3V4可变区对阴道分泌物进行测序分析，比较两组之间微生物组成差异。RT-qPCR结果显示，相比于配种后第5天，配种后第15天的子宫内膜组织中HOXA10、LIF和VEGFA容受基因显著升高(P < 0.01或P＜0.05)，表明配种后第15天的子宫内膜组织进入容受状态。Alpha多样性比较分析，容受前期组与容受期组的阴道微生物无显著差异(P>0.05)。Beta多样性比较分析，容受前期组与容受期组存在组间差异。物种相对丰度结果显示，在门分类水平上，Proteobacteria、Firmicutes等为优势菌门，占据70%以上。与容受前期组相比，容受期组Proteobacteria和Fusobacteriota菌门丰度下降; Firmicutes菌门丰度上升; 在属分类水平上，unidentified_Enterobacteriaceae、PorpHyromonas等为优势菌属。与容受前期组相比，容受期组unidentified_Enterobacteriaceae、Streptobacillus等菌属丰度下降；PorpHyromonas、Ureaplasma等菌属丰度增加。差异菌群比较结果显示，在门水平上，Fusobacteriota、Acidobacteriota等丰度存在显著差异(P＜0.05)。在属水平上，Occallatibacter、EdapHobacter、Roseiarcus丰度存在显著差异(P＜0.05)。功能预测分析表明，相比于容受前期组，容受期组孕酮介导的卵母细胞成熟、雌激素信号通路、抗原的处理和呈递等通路显著升高(P＜0.05)。绵羊子宫内膜容受前期与容受期组微生物Alpha多样性并无显著差异，Beta多样性表明两组间存在着组间差异。同时在多个分类水平上均有显著差异菌群，Facklamia、Turicibacter菌属、Firmicutes菌门可作为绵羊子宫内膜容受期的标志菌群。

关键词: 绵羊, 容受前期, 容受期, 16S rRNA

Abstract:

Based on 16S rRNA sequencing technology, this study aimed to analyze the differential changes of vaginal microbial communities in sheep during pre- and endometrial receptivity, and screen the marker flora of endometrial receptivity in sheep. In this experiment, 12 healthy multiparous female Hu sheep aged two years old with similar body weight were selected as the research objects. The experiment was divided into 2 groups, 6 sheep in each group. The vaginal secretions and endometrial tissues were collected on the 5th day (pre-receptive endometrium group, PE group) and the 15th day (peceptive endometrium group, RE group) after mating. RT-qPCR was used to detect the expression of endometrial tissue-related receptivity genes, and the 16S rRNA V3V4 variable region was selected for sequencing analysis of vaginal secretions to compare the differences in microbial composition between the 2 groups. RT-qPCR results showed that the expression of HOXA10, LIF and VEGFA genes in the endometrium on the 15 th day after mating was significantly higher than that on the 5 th day after mating (P < 0.01 or P < 0.05), indicating that the endometrium on the 15 th day after mating entered the receptive state. Alpha diversity analysis showed that there was no significant difference in vaginal microorganisms between the pre-receptive group and the receptive group (P>0.05). Comparative analysis of Beta diversity showed that there were differences between the pre-tolerance group and the tolerance group. The results of relative abundance of species showed that Proteobacteria and Firmicutes were the dominant phyla at the level of phylum classification, accounting for more than 70%. Compared with the pre-tolerance group, the abundance of Proteobacteria and Fusobacteriota in the tolerance group decreased, the abundance of Firmicutes increased; at the genus level, unidentified _ Enterobacteriaceae and PorpHyromonas were the dominant genera. Compared with the pre-tolerance group, the abundance of unidentified _ Enterobacteriaceae, Streptobacillus and other bacteria in the tolerance group decreased; the abundance of PorpHyromonas and Ureaplasma increased. The results of differential flora comparison showed that there were significant differences in the abundance of Fusobacteriota and Acidobacteriota at the phylum level (P < 0.05). At the genus level, there were significant differences in the abundance of Occallatibacter, EdapHobacter and Roseiarcus (P < 0.05). Functional prediction analysis showed that compared with the pre-receptive group, progesterone-mediated oocyte maturation, estrogen signaling pathway, antigen processing and presentation were significantly increased in the receptive group (P < 0.05). There was no significant difference in microbial Alpha diversity between the pre-receptive and receptive groups of sheep endometrium, and Beta diversity indicated that there were differences between the two groups. At the same time, there were significant differences in flora at multiple classification levels. Facklamia, Turicibacter and Firmicutes could be used as the marker flora of endometrial receptivity in sheep.

Key words: sheep, pre-tolerance, tolerance, 16S rRNA

中图分类号:

S826.3

楚翼健, 崔久增, 李增开, 张磊, 褚婷婷, 黄艳平, 宋宇轩. 绵羊子宫内膜容受前期与容受期的阴道微生物比较研究[J]. 畜牧兽医学报, 2025, 56(2): 689-699.

CHU Yijian, CUI Jiuzeng, LI Zengkai, ZHANG Lei, CHU Tingting, HUANG Yanping, SONG Yuxuan. Comparative Study on Vaginal Microorganisms in Pre-endometrial Receptivity and Endometrial Receptivity of Sheep[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(2): 689-699.

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基因名称 Gene name	(5′→3′)引物序列(Forward) Primer sequences (Forward)	(3′→5′)引物序列(Reverse) Primer sequences (Reverse)
LIF	AACTCAACAGCAGTGCCAACAG	CGTGGAAGGGCGGGAAGTC
VEGFA	TGGCAGAAGGAGGGCAGAAAC	AGATGTCCACCAGGGTCTCAATG
HOXA10	CGAGGCTGGCGGTGGTG	GGCGGACGGCAGGTAGAC
β-ACTIN	GCAAGCGTGGCATCCTAACC	GCTCGTTGTAGAAGGTGTGGTG

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绵羊子宫内膜容受前期与容受期的阴道微生物比较研究

Comparative Study on Vaginal Microorganisms in Pre-endometrial Receptivity and Endometrial Receptivity of Sheep

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