基于多组学与网络药理学探究淫羊藿对后备母猪发情的作用

doi:10.11843/j.issn.0366-6964.2024.04.024

摘要/Abstract

摘要： 旨在揭示淫羊藿对后备母猪发情的作用及其机理。本试验选择210~220日龄，体重(99.547±1.987)kg，发育成熟且符合配种条件的后备待配二元母猪32头，随机分为对照组和试验组，每组16头母猪，每个重复1头母猪。对照组饲喂基础日粮，试验组在饲喂基础日粮的基础上补充淫羊藿粗提物50 mg·d^-1。试验饲喂28 d。结果表明，试验组母猪发情提前，血清FSH、LH和E₂显著增加(P＜0.05)。卵巢转录组结果表明，检测出477个上调差异mRNAs，754个下调差异mRNAs。GO富集分析发现，差异的mRNA主要富集在运输囊泡、脂肪细胞分化、节律行为、发情周期等过程；KEGG富集分析发现，差异的mRNA主要富集在紧密连接、碳水化合物代谢、刺猬信号通路、GnRH分泌和PI3K-AKT信号通路等信号通路。卵巢代谢组结果表明共有1 616个代谢物上调，1 254个代谢物下调。KEGG富集分析表明，差异代谢物主要富集在α-亚麻酸代谢、ATP转运蛋白、亚油酸代谢、β-丙氨酸代谢、氨基苯甲酸盐降解、生物素代谢等通路。网络药理学分析结果表明淫羊藿作用卵巢的潜在作用靶点共161个。PPI互作网络得到degree前10的蛋白作为核心靶点，根据得分依次为TP53、SRC、AKT1、CCND1、TNF、ESR1、EP300、ERBB2、JAK2、PARP1。GO分析结果表明，淫羊藿作用卵巢的靶点主要参与了蛋白磷酸化、MAPK级联反应的正调控、生物节律、基因表达的正调控、细胞凋亡过程的负调控、细胞内钙离子浓度的正调控、RNA聚合酶II启动子转录的正调控等生物学过程。KEGG分析结果显示，靶点蛋白信号通路富集在PI3K-Akt信号通路、细胞周期、细胞衰老、HIF-1信号通路、孕激素介导的卵母细胞成熟等途径。以上结果从活体、代谢、转录和分子层面揭示了淫羊藿对后备母猪发情的影响和作用途径，本研究发现饲喂淫羊藿能够改变后备母猪卵巢转录与代谢模式，显著提高FSH、LH和E₂水平，进而促进母猪发情，淫羊藿的主要成分能够与TP53、SRC、AKT1、CCND1、TNF、ESR1、EP300、ERBB2、JAK2、PARP1结合发挥调控作用。本研究为淫羊藿应用提高后备母猪发情利用率提供理论依据。

关键词: 淫羊藿, 后备母猪, 发情, 转录组, 代谢组, 网络药理学

Abstract: The aim of this study was to reveal the effect and mechanism of epimedium on the estrus of gilts. A total of 32 gilts aged 210-220 days old, weighing (99.547±1.987)kg, mature and meeting breeding conditions were selected for the experiment. They were randomly divided into a control group and an experimental group, with 16 sows in each group and one sow in each replicate. Individuals in the control group was fed with a basic diet, while individuals in the experimental group was supplemented with 50 mg·d^-1 crude extract of epimedium in addition to the basic diet. Experimental feeding for 28 days. The results showed that gilts in the experimental group experienced early estrus, and serum FSH, LH, and E₂ significantly increased (P < 0.05). The ovarian transcriptome results showed that 477 upregulated differential mRNAs and 754 downregulated differential mRNAs were detected. GO enrichment analysis revealed that differential mRNAs was mainly enriched in processes such as transport vesicles, adipocyte differentiation, rhythmic behavior, and estrus cycle; KEGG enrichment analysis revealed that differential mRNAs was mainly enriched in tight junctions, carbohydrate metabolism, hedgehog signaling pathways, GnRH secretion, and PI3K-AKT signaling pathways. The results of ovarian metabolomics showed a total of 1 616 metabolites upregulated and 1 254 metabolites downregulated. KEGG enrichment analysis showed that differential metabolites were mainly enriched in α-linoleic acid metabolism, ATP transporters, linoleic acid metabolism β-pathways such as alanine metabolism, aminobenzoate degradation, and biotin metabolism. The results of network pharmacology analysis indicated that there were a total of 161 potential targets for the action of epimedium on the ovaries. The PPI interaction network obtained the top 10 protein grades as core targets, which were TP53, SRC, AKT1, CCND1, TNF, ESR1, EP300, ERBB2, JAK2, and PARP1 according to their scores. The GO analysis results indicated that the targets of epimedium on the ovaries were mainly involved in biological processes such as protein phosphorylation, positive regulation of MAPK cascade reaction, biological rhythm, positive regulation of gene expression, negative regulation of cell apoptosis process, positive regulation of intracellular calcium ion concentration, and positive regulation of RNA polymerase II promoter transcription. The KEGG analysis results showed that the target protein signaling pathway was enriched in PI3K-AKT signaling pathway, cell cycle, cell aging, HIF-1 signaling pathway, progesterone mediated oocyte maturation, and other pathways. The above results reveal the effects and pathways of epimedium on the estrus of gilts from the perspectives of in vivo, metabolism, transcription, and molecular level. This study found that feeding epimedium can change the transcription and metabolism patterns of sows′ ovaries, significantly increase FSH, LH, and E₂ levels, and promote estrus in gilts. The main components of epimedium can bind with TP53, SRC, AKT1, CCND1, TNF, ESR1, EP300, ERBB2, JAK2, and PARP1 to exert regulatory effects. This study provides a theoretical basis for the application of epimedium to improve the utilization rate of estrus in gilts.

Key words: epimedium, gilts, estrus, transcriptome, metabolic group, network pharmacology

中图分类号:

S828.3

徐俊杰, 张璐通, 王津洁, 陈晓晨, 何伟先, 蔡传江, 褚瑰燕, 杨公社. 基于多组学与网络药理学探究淫羊藿对后备母猪发情的作用[J]. 畜牧兽医学报, 2024, 55(4): 1615-1628.

XU Junjie, ZHANG Lutong, WANG Jinjie, CHEN Xiaochen, HE Weixian, CAI Chuanjiang, CHU Guiyan, YANG Gongshe. Exploring the Effect of Epimedium on Estrus of Gilts Based on Multiomics and Network Pharmacology[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1615-1628.

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