Long-term exposure to high ammonia concentrations could severely impact chicken health. On the other hand, luteolin has been shown to protect against ammonia poisoning. Although phosphorylation is critically involved in toxicity induction, the specific role of phosphorylated proteins in ammonia poisoning remains unclear. Herein, we constructed an in vitro model to study chicken ammonia poisoning and also analyzed the protective effects of luteolin. Specifically, a combined series of organic techniques such as protein extraction, enzyme digestion, modified peptide enrichment, Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) analysis, and bioinformatics analysis were employed for a quantitative omics study of phosphorylation modification in three groups of samples. Our findings revealed thousands of Differentially Expressed Proteins (DEPs). The differentially expressed modified proteins were subjected to GO classification, KEGG pathway analysis, cluster analysis, and protein interaction analysis, revealing the detoxification mechanism encompassed mitochondrial maintenance, signal transduction, transcriptional regulation, and cytoskeleton regulation. In the process, mitochondria and Golgi apparatus were the key organelles. Furthermore, the AKT1/FOXO signaling pathway and Heat Shock Proteins (HSPs) were the key core modifiers of the proteins. We hope that our findings will provide a theoretical basis and experimental support for future research on luteolin’s detoxification mechanism against ammonia poisoning.

中文翻译：

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木犀草素介导的鸡脾淋巴细胞磷酸化组学变化：揭示针对氨诱导应激的解毒机制


长期暴露在高浓度的氨环境中可能会严重影响鸡的健康。另一方面，木犀草素已被证明可以防止氨中毒。尽管磷酸化与毒性诱导密切相关，但磷酸化蛋白在氨中毒中的具体作用仍不清楚。在此，我们构建了一个体外模型来研究鸡氨中毒，并分析了木犀草素的保护作用。具体来说，采用蛋白质提取、酶消化、修饰肽富集、液相色谱-串联质谱 （LC-MS/MS） 分析和生物信息学分析等一系列有机技术的组合技术对三组样品的磷酸化修饰进行定量组学研究。我们的研究结果揭示了数千种差异表达蛋白 （DEP）。对差异表达的修饰蛋白进行 GO 分类、 KEGG 通路分析、聚类分析和蛋白质相互作用分析，揭示线粒体维持、信号转导、转录调控和细胞骨架调控等解毒机制。在此过程中，线粒体和高尔基体是关键的细胞器。此外，AKT1/FOXO 信号通路和热休克蛋白 （HSP） 是蛋白质的关键核心修饰因子。我们希望我们的研究结果能为未来木犀草素对氨中毒解毒机制的研究提供理论依据和实验支持。