基于蛋白质组学探讨绿原酸抗玉米赤霉烯酮致卵泡颗粒细胞死亡的作用机制

doi:10.11843/j.issn.0366-6964.2025.12.048

摘要/Abstract

摘要： 玉米赤霉烯酮(zearalenone，ZEA)是一种常见的霉菌毒素，对人和动物有严重的生殖毒性。本团队前期研究发现绿原酸(chlorogenic acid)可缓解ZEA诱导的卵巢颗粒细胞毒性，然而其作用机制并不清楚。本试验旨在通过蛋白质组学技术解析绿原酸拮抗ZEA诱导小鼠卵泡颗粒细胞死亡的分子机制。将原代分离培养的小鼠卵巢颗粒细胞分为两组：ZEA组和绿原酸干预组，处理24 h后，采用定量蛋白质组学技术，结合生物信息学分析和RT-qPCR验证，解析绿原酸的调控网络。结果表明，绿原酸干预组与ZEA处理组相比，共鉴定出32个差异表达蛋白(differentially expressed proteins，DEPs)，其中15个显著上调，17个显著下调。通过GO功能注释和KEGG通路富集分析，发现DEPs主要参与有丝分裂调控、细胞骨架重构等生物学过程，并显著富集于凋亡、黏着斑、Toll样受体信号通路和NF-κB信号通路等关键调控途径。进一步通过RT-qPCR验证关键调控因子RIPK1的表达变化，发现ZEA处理显著抑制RIPK1表达(P<0.001)，而绿原酸干预可部分恢复其表达水平(P<0.01)。绿原酸通过调控细胞周期进程和凋亡、维持细胞结构稳态等多靶点多通路协同作用，从而拮抗ZEA诱导的生殖细胞毒性。此外，RIPK1作为RIPK1作为细胞生存和死亡的关键调控开关可能发挥关键作用。本研究从蛋白层面揭示绿原酸拮抗ZEA生殖毒性的作用机制，为开发基于绿原酸的ZEA解毒剂或功能性饲料添加剂提供了新的分子靶点和理论依据。

关键词: 玉米赤霉烯酮, 绿原酸, 卵泡颗粒细胞, 蛋白质组学

Abstract: Zearalenone (ZEA) is a common mycotoxin that can cause severe reproductive toxicity. In our previous study, we found that chlorogenic acid could alleviate ZEA-induced ovarian granulosa cell toxicity, however, the mechanism of action is not clear. The aim of this study was to investigate the molecular mechanism by which chlorogenic acid antagonizes ZEA-induced follicular granulosa cell death in mice using proteomics techniques. Primary isolated and cultured mouse ovarian granulosa cells were divided into two groups: the ZEA group and the chlorogenic acid intervention group. After 24 h of treatment, the regulatory network of chlorogenic acid was systematically analyzed using quantitative proteomics technology combined with bioinformatics analysis and RT-qPCR validation. The results showed that a total of 32 differentially expressed proteins (DEPs) were identified in the chlorogenic acid-treated group compared with the ZEA-treated group, of which 15 were significantly up-regulated and 17 were significantly down-regulated. Through GO functional annotation and KEGG pathway enrichment analysis, the DEPs were mainly involved in biological processes such as mitotic regulation and cytoskeletal remodeling, and were significantly enriched in key regulatory pathways such as apoptosis, focal adhesion, Toll-like receptor signaling pathway and NF-κB signaling pathway. The changes in the expression of the key regulator RIPK1 were further verified by RT-qPCR and it was found that ZEA treatment significantly inhibited the expression of RIPK1, whereas chlorogenic acid intervention partially restored its expression level. Chlorogenic acid acts synergistically through multi-targets and multi-pathways such as regulation of cell cycle progression and apoptosis and maintenance of cellular structural homeostasis to antagonize ZEA-induced reproductive toxicity. In addition, RIPK1 may play a critical role as a key regulatory switch for cell survival and death. The present study reveals the mechanism of action of chlorogenic acid in antagonizing reproductive toxicity of ZEA at the protein level, providing new molecular targets and theoretical basis for the development of chlorogenic acid-based ZEA detoxifying agents or functional feed additives.

Key words: zearalenone, chlorogenic acid, ovarian granulosa cells, proteomics

中图分类号:

扆妍妍, 孙盼盼, 孙娜, 李宏全. 基于蛋白质组学探讨绿原酸抗玉米赤霉烯酮致卵泡颗粒细胞死亡的作用机制[J]. 畜牧兽医学报, 2025, 56(12): 6477-6486.

YI Yanyan, SUN Panpan, SUN Na, LI Hongquan. Proteomic Profiling Unveils the Protective Mechanism of Chlorogenic Acid against Zearalenone-induced Ovarian Granulosa Cell Death[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6477-6486.

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