后备母猪发情期和乏情期下丘脑-垂体-卵巢性腺轴miRNA-mRNA表达谱比较分析

doi:10.16431/j.cnki.1671-7236.2025.07.001

中国畜牧兽医 ›› 2025, Vol. 52 ›› Issue (7): 2965-2980.doi: 10.16431/j.cnki.1671-7236.2025.07.001

后备母猪发情期和乏情期下丘脑-垂体-卵巢性腺轴miRNA-mRNA表达谱比较分析

吕玲燕^1,2, 孙如玉¹, 林昌华^2,3, 张胜斌³, 覃秀珍³, 柏秀芳³, 吴永绍¹, 陈钊¹, 刘磊^1,2, 张冰¹, 蒋家霞^2,4, 张家庆⁵

1. 广西农业职业技术大学, 南宁 530007;
2. 广西大学动物科学技术学院, 南宁 530004;
3. 广西农垦永新畜牧集团西江有限公司, 贵港 537100;
4. 广西职业技术学院农业工程学院, 南宁 530226;
5. 河南省农业科学院畜牧研究所, 郑州 450002

收稿日期:2025-01-23 出版日期:2025-07-05 发布日期:2025-07-01
通讯作者: 蒋家霞, 张家庆 E-mail:279403392@qq.com;zjq8650612@163.com
作者简介:吕玲燕，E-mail：llyan159@163.com；孙如玉，E-mail：434783109@qq.com。
基金资助:
国家重点研发项目(2021YFD1301200)；广西农业职业技术大学校级课题(XKJ2336)；广西农业科技自筹经费项目(Z2022115)；河南省科技攻关项目(242102111030)；广西农业科技自筹经费项目(Z2023101)；2025年度广西高校中青年教师科研基础能力提升项目(2025KY1388)

Comparative Analysis of miRNA-mRNA Expression Profile in Hypothalamus-Pituitary-Ovary Gonad Axis During Estrus and Anestrus in Gilts

LYU Lingyan^1,2, SUN Ruyu¹, LIN Changhua^2,3, ZHANG Shengbin³, QIN Xiuzhen³, BAI Xiufang³, WU Yongshao¹, CHEN Zhao¹, LIU Lei^1,2, ZHANG Bing¹, JIANG Jiaxia^2,4, ZHANG Jiaqing⁵

1. Guangxi Vocational and Technical University of Agriculture, Nanning 530007, China;
2. College of Animal Science and Technology, Guangxi University, Nanning 530004, China;
3. Guangxi Nongken Yongxin Livestock Group Xijiang Co., Ltd., Guigang 537100, China;
4. College of Agricultural Engineering, Guangxi Vocational and Technical College, Nanning 530226, China;
5. Institute of Animal Husbandry, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China

Received:2025-01-23 Online:2025-07-05 Published:2025-07-01

摘要/Abstract

摘要： 【目的】探讨miRNA-mRNA互作网络在后备母猪发情调控中的关键作用，以期解释其在后备母猪发情活动中的遗传机制。【方法】以发情期和乏情期后备母猪的下丘脑、垂体、卵巢组织为研究对象，测定血清中促卵泡激素(FSH)、孕酮(P₄)、雌二醇(E₂)等生殖激素浓度，通过小RNA测序(sRNA-Seq)并利用生物信息学软件筛选出不同情期差异表达miRNA，通过R语言对差异表达miRNA靶基因进行GO功能及KEGG通路富集分析，并随机选取4个差异表达miRNAs进行实时荧光定量PCR验证。【结果】后备母猪血清中生殖激素浓度与母猪所处的生理周期相吻合；在发情期和乏情期母猪中共检测到742个已知的miRNAs和229个新的miRNAs。发情期和乏情期母猪下丘脑中有57个差异表达miRNAs，其中24个上调，33个下调；垂体中有71个差异表达miRNAs，其中44个上调，27个下调；卵巢中有140个差异表达miRNAs，其中63个上调，77个下调。KEGG通路富集分析发现，后备母猪下丘脑中差异表达miRNA靶基因主要参与癌症中的蛋白聚糖、NOD样受体信号通路、Toll样受体信号通路、细胞因子受体相互作用等；垂体中差异表达miRNA靶基因主要参与甘氨酸、丝氨酸、苏氨酸代谢过程，促性腺激素释放激素分泌，细胞黏附分子，新陈代谢信号通路等；卵巢中差异表达miRNA靶基因主要参与趋化因子信号通路、溶酶体、卵巢类固醇生成、胆固醇代谢、PPAR信号通路、ECM受体交互作用等。在与生殖相关的靶基因调控网络中筛选出可能由下丘脑-垂体-卵巢性腺轴介导调控后备母猪情期活动的关键miRNAs：miR-6240Z、ssc-miR-34a、ssc-miR-143-3P、ssc-miR-127、ssc-miR-21-5P、ssc-miR-381-3p。对后备母猪卵巢组织中4个差异表达miRNAs进行实时荧光定量PCR验证，其表达趋势与测序结果一致。【结论】本研究成功构建了发情期和乏情期后备母猪下丘脑-垂体-卵巢性腺轴miRNA表达谱，并对差异表达miRNA进行验证，筛选到与后备母猪情期活动显著相关的miRNA，为解析后备母猪发情机制提供了理论依据。

关键词: miRNA-mRNA; 后备母猪; 发情期; 乏情期; 下丘脑-垂体-卵巢性腺轴

Abstract: 【Objective】 The purpose of this study was to explore the key role of miRNA-mRNA interaction network in the regulation of estrus in gilts for explaining the genetic mechanism of miRNA-mRNA interaction in the estrus activity of gilts. 【Method】 The hypothalamus,pituitary and ovary of gilts in estrus and anestrus were selected as the study objects.The concentrations of follicle-stimulating hormone (FSH),progesterone (P₄),estradiol (E₂), and other reproductive hormones in serum were measured.After sRNA-Seq analysis,the differentially expressed miRNA were screened out by bioinformatics software,GO function and KEGG pathway enrichment analysis of differentially expressed miRNA target genes were performed using R language,and 4 differentially expressed miRNAs were verified by Real-time quantitative PCR. 【Result】 The concentration of reproductive hormone in serum was consistent with the physiological cycle of gilts.A total of 742 known miRNAs and 229 novel miRNAs in gilts were detected during estrus and anestrus,57 differentially expressed miRNAs (24 up-regulated and 33 down-regulated) were found in hypothalamus of gilts during estrus and anestrus,71 differentially expressed miRNAs (44 up-regulated and 27 down-regulated) in pituitary were found,and 140 differentially expressed miRNAs (63 up-regulated and 77 down-regulated) in ovary were found. KEGG pathway enrichment analysis showed that differentially expressed miRNA target genes in hypothalamus of gilts were mainly involved in proteoglycan in cancer,NOD-like receptor signaling pathway,Toll-like receptor signaling pathway,and cytokine receptor interaction.The differentially expressed miRNA target genes in pituitary were mainly involved in glycine, serine and threonine metabolism,GnRH secretion,cell adhesion molecules,metabolic signaling pathway.The differentially expressed miRNA target genes in ovary were mainly involved in chemokine signaling pathway,lysosome,ovarian sterogenesis, cholesterol metabolism,PPAR signaling pathway,and ECM-receptor interaction.In the target gene regulatory network related to reproduction,the key miRNA that might be mediated by hypothalamus-pituitary-ovary gonad axis to regulate the estrus activity of gilts were screened:miR-6240Z,ssc-miR-34a,ssc-miR-143-3P,ssc-miR-127,ssc-miR-21-5P,and ssc-miR-381-3p.Four differentially expressed miRNAs in ovary of gilts were verified by Real-time quantitative PCR,and the expression trends were consistent with the sequencing results. 【Conclusion】 In this study,miRNA expression profiles of hypothalamus-pituitary-ovary gonad axis of gilts in estrus and anestrus were successfully constructed,and the differentially expressed miRNA was verified.miRNAs that were significantly related to the estrus activity of gilts were screened,which provided theoretical basis for the analysis of the estrus mechanism of gilts.

Key words: miRNA-mRNA; gilts; estrus; anestrus; hypothalamus-pituitary-ovary gonad axis

中图分类号:

S814.8

吕玲燕, 孙如玉, 林昌华, 张胜斌, 覃秀珍, 柏秀芳, 吴永绍, 陈钊, 刘磊, 张冰, 蒋家霞, 张家庆. 后备母猪发情期和乏情期下丘脑-垂体-卵巢性腺轴miRNA-mRNA表达谱比较分析[J]. 中国畜牧兽医, 2025, 52(7): 2965-2980.

LYU Lingyan, SUN Ruyu, LIN Changhua, ZHANG Shengbin, QIN Xiuzhen, BAI Xiufang, WU Yongshao, CHEN Zhao, LIU Lei, ZHANG Bing, JIANG Jiaxia, ZHANG Jiaqing. Comparative Analysis of miRNA-mRNA Expression Profile in Hypothalamus-Pituitary-Ovary Gonad Axis During Estrus and Anestrus in Gilts[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 2965-2980.

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后备母猪发情期和乏情期下丘脑-垂体-卵巢性腺轴miRNA-mRNA表达谱比较分析

Comparative Analysis of miRNA-mRNA Expression Profile in Hypothalamus-Pituitary-Ovary Gonad Axis During Estrus and Anestrus in Gilts

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