侧脑室注射促性腺激素抑制激素/RF酰胺相关肽-3对卵巢摘除术后补充雌激素大鼠子宫腔液影响的蛋白质组学分析

赵雪莹; 程露阳; 司丽娜; 魏萌; 乔跃兵; 杨松鹤

doi:10.16098/j.issn.0529-1356.2021.04.001

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解剖学报 ›› 2021, Vol. 52 ›› Issue (4) : 499-505. DOI: 10.16098/j.issn.0529-1356.2021.04.001

神经生物学

侧脑室注射促性腺激素抑制激素/RF酰胺相关肽-3对卵巢摘除术后补充雌激素大鼠子宫腔液影响的蛋白质组学分析

赵雪莹¹ 司丽娜² 魏萌² 乔跃兵² 杨松鹤^2* 程露阳^1*

作者信息 +

Proteomic analysis of uterine luminal fluid in ovariectomized estrogen primed rats with lateral cerebral ventricle injection of gonadotropin-inhibitory hormone/RF amide-related peptide-3

ZHAO Xue-ying¹ SI Li-na² WEI Meng² QIAO Yue-bing² YANG Song-he^2* CHENG Lu-yang^1*

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摘要

目的运用生物信息学方法分析侧脑室注射促性腺激素抑制激素（GnIH）/RF酰胺相关肽-3（RFRP-3）对卵巢摘除术补充雌激素 (OEP) 大鼠子宫腔液蛋白的影响，探索GnIH/RFRP-3影响子宫的分子机制。方法本研究通过液相色谱-串联质谱（LC-MS/MS）技术结合生物信息学分析侧脑室微量注射GnIH对OEP模型大鼠（n=30）子宫腔液蛋白的影响。比较侧脑室注射GnIH/RFRP-3实验组（n=15，给予2 g/L RFRP-3，16 μl/kg）与生理盐水对照组（n=15，给予生理盐水16 μl/kg）6 h后子宫腔液的蛋白质组分，利用UniProt数据库筛选差异蛋白并对所得差异蛋白进行基因本体论（GO）富集分析和京都基因与基因百科全书（KEGG）通路分析。从STRING 数据库获得蛋白相互作用数据，利用Cytoscape 3.7.1软件构建蛋白相互作用网络（PPI）并筛选中心节点蛋白。结果应用LC-MS/MS技术所得质谱数据利用UniProt数据库进行搜索鉴定，筛选出差异蛋白419个，其中279个上调，138个下调；GO功能富集分析显示，差异蛋白主要富集于对有机物质的反应，对内源性刺激的反应，对含氧化合物的反应等生物过程；KEGG通路分析显示，差异蛋白在糖酵解、碳代谢、肌动蛋白骨架调节、甲状腺激素合成等代谢途径富集（P<0.05）；分析蛋白相互作用网络获得5个中心节点蛋白：白蛋白（Alb）、α-烯醇化酶1(Eno1）、过氧化物酶6（Prdx6)、组织型基质金属蛋白酶抑制剂1（Timp1）、Ras相关C3肉毒素底物1 (Rac1)。结论本实验结果提示，GnIH可能通过下丘脑-垂体生殖轴调节子宫腔液蛋白的分泌，并可能通过上调Alb、Eno1和Prdx6，下调Timp1和Rac1，调节子宫生理及病理过程。

Abstract

Objective To explore the roles and effects of gonadotropin inhibitory hormone gonadotropin-inhibitory hormone (GnIH)/RF amide-related peptide-3 (RFRP-3) on the uterus through the hypothalamic-pituitary reproductive axis. Methods Ovariectomized estrogen primed (OEP) rats model was divided into GnIH injection group and normal saline injection group with 15 rats in each group, 2 g/L GnIH (16 μl/kg) and normal saline (16 μl/kg) were injected into the lateral ventricle of rats in the 2 groups respectively. 6 hours after injection, the uterine fluid of rats was obtained. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to separate the differential proteins in uterine fluid and UniProt was used to identify. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed, and the proteinprotein interaction (PPI) network of the differentially expressed proteins was constructed using Cytoscape 3.7.1 software. Hub proteins were detected from PPI network. Results The result showed that the differentially expressed proteins separating by LC-MS/MS were 419 identified by UniProt. Among them, 279 were up-regulated and 138 were down-regulated. GO analysis showed that the differentially expressed proteins were mainly involved in response to organic substance, response to oxygen-containing compound, response to endogenous stimulus, response to stress. The top five enriched pathways obtained in the KEGG pathway analysis (P<0.05) were carbon metabolism, gap junction, long-term depression, regulation of actin cytoskeleton, biosynthesis of amino acids. Five hub proteins involved albumin（Alb），alpha-enolase 1(Eno1), peroxiredoxin-6 (Prdx6), tissue inhibitor of matrix metalloproteinases-1（Timp1）, Ras-related C3 botulinum toxin substrate 1 (Rac1) were obtained by analyzing PPI network. Conclusion The result of this study indicate that GnIH may regulate the secretion of uterine cavity fluid protein through hypothalamic-pituitary reproductive axis. And regulate the physiological and pathological process of uterus by up-regulating Alb, Eno1, Prdx6 and down-regulating Timp1 and Rac1.

导出引用

赵雪莹司丽娜魏萌乔跃兵杨松鹤程露阳. 侧脑室注射促性腺激素抑制激素/RF酰胺相关肽-3对卵巢摘除术后补充雌激素大鼠子宫腔液影响的蛋白质组学分析[J]. 解剖学报. 2021, 52(4): 499-505 https://doi.org/10.16098/j.issn.0529-1356.2021.04.001

ZHAO Xue-ying SI Li-na WEI Meng QIAO Yue-bing YANG Song-he CHENG Lu-yang. Proteomic analysis of uterine luminal fluid in ovariectomized estrogen primed rats with lateral cerebral ventricle injection of gonadotropin-inhibitory hormone/RF amide-related peptide-3[J]. Acta Anatomica Sinica. 2021, 52(4): 499-505 https://doi.org/10.16098/j.issn.0529-1356.2021.04.001

中图分类号： Q51

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