Fayyad-Kazan Mohammad, Bourgoin-Voillard Sandrine, Rachidi Walid, Seve Michel
Department of Natural and Applied Sciences, School of Arts and Sciences, The American University of Iraq - Baghdad, Baghdad, Iraq.
Université Grenoble Alpes, CNRS, U5525, Grenoble INP, TIMC, EPSP, Grenoble, France.
Methods Mol Biol. 2025;2879:83-92. doi: 10.1007/7651_2024_555.
Autophagy refers to the natural cellular process by which cells degrade and recycle their own damaged or dysfunctional cellular components. It is an essential mechanism for maintaining cellular homeostasis removing toxic substances and providing energy during times of stress or nutrient deprivation. When autophagy is dysregulated or impaired, it can have detrimental effects on cell function and overall health. Studying autophagy in skin exposed to pollutants can provide valuable insights into the cellular mechanisms underlying pollutant-induced skin damage. Proteomic methods, which involve the large-scale analysis of proteins, can be employed to investigate the changes in protein expression associated with biological processes including autophagy. Here, we thus describe a method where LC-MS/MS was applied to identify the deregulated proteins in pollutant exposed-skin. Using bioinformatics and statistical analysis, we extracted the qualitative and quantitative information for proteins involved in autophagy. These deregulated proteins were then validated by immunohistochemistry (IHC). These methods help to understand how the pollutants affect the autophagy process.
自噬是指细胞自然降解和循环利用自身受损或功能失调的细胞成分的过程。它是维持细胞内稳态、清除有毒物质以及在应激或营养缺乏时提供能量的重要机制。当自噬失调或受损时,会对细胞功能和整体健康产生有害影响。研究暴露于污染物的皮肤中的自噬,可以为污染物诱导的皮肤损伤的细胞机制提供有价值的见解。蛋白质组学方法涉及对蛋白质进行大规模分析,可用于研究与包括自噬在内的生物过程相关的蛋白质表达变化。在此,我们描述了一种应用液相色谱-串联质谱(LC-MS/MS)来鉴定暴露于污染物的皮肤中失调蛋白质的方法。通过生物信息学和统计分析,我们提取了参与自噬的蛋白质的定性和定量信息。然后通过免疫组织化学(IHC)对这些失调的蛋白质进行验证。这些方法有助于了解污染物如何影响自噬过程。