Boşgelmez I İpek, Güvendik Gülin, Dilbaz Nesrin, Esen Metin
Department of Toxicology, Faculty of Pharmacy, Erciyes University, Kayseri 38280, Türkiye.
Department of Toxicology, Faculty of Pharmacy, Istanbul Health & Technology University, İstanbul 34275, Türkiye.
Int J Mol Sci. 2025 Aug 23;26(17):8199. doi: 10.3390/ijms26178199.
Alcohol Use Disorder (AUD) poses global health challenges, and causes hematological alterations such as macrocytosis and oxidative stress. Disruption of protein structures by alcohol and/or its metabolites may exacerbate AUDs; proteomics can elucidate the underlying biological mechanisms. This study examined the proteins differentially expressed in the cytosol and membrane fractions of erythrocytes obtained from 30 male patients with AUD, comparing them to samples from 15 age- and BMI-matched social drinkers (SDs) and 15 non-drinkers (control). The analysis aimed to identify the molecular differences related to alcohol consumption. The AUD patient subgrouping was based on mean corpuscular volume (MCV), with 16 individuals classified as having a normal MCV and 14 having a high MCV. Proteins were separated via two-dimensional(2D)-gel electrophoresis, digested with trypsin, and identified via Matrix-Assisted Laser Desorption/Ionization Time-of-Flight (TOF) mass spectrometry (MALDI-TOF/TOF). Additionally, levels of malondialdehyde and 4-hydroxyalkenals (MDA + HAE), reduced glutathione (GSH), oxidized glutathione (GSSG), serum carbohydrate-deficient transferrin (%CDT), disialotransferrin (%DST), and sialic acid (SA) were analyzed. The results showed increased MDA + HAE and decreased total thiols in AUD patients, with GSSG elevated and the GSH/GSSG ratio reduced in the AUD MCV-high subgroup. Serum %CDT, %DST, and SA were significantly higher in AUD. Compared to the control profiles, the AUD group exhibited differential protein expression. Few proteins, such as bisphosphoglycerate mutase, were downregulated in AUD versus control and SD, as well as in the MCV-high AUD subgroup. Conversely, endoplasmin and gelsolin were upregulated in AUD relative to control. Cytoskeletal proteins, including spectrin-alpha chain, actin cytoplasmic 2, were overexpressed in the AUD group and MCV-high AUD subgroup. Several proteins, such as 14-3-3 isoforms, alpha-synuclein, translation initiation factors, heat shock proteins, and others, were upregulated in the MCV-high AUD subgroup. Under-expressed proteins in this subgroup include band 3 anion transport protein, bisphosphoglycerate mutase, tropomyosin alpha-3 chain, uroporphyrinogen decarboxylase, and WD repeat-containing protein 1. Our findings highlight the specific changes in protein expression associated with oxidative stress, cytoskeletal alterations, and metabolic dysregulation, specifically in AUD patients with an elevated MCV. Understanding these mechanisms is crucial for developing targeted interventions and identifying biomarkers of alcohol-induced cellular damage. The complex interplay between oxidative stress, membrane composition, and cellular function illustrates how chronic alcohol exposure affects cellular physiology.
酒精使用障碍（AUD）给全球健康带来挑战，并导致血液学改变，如大细胞性贫血和氧化应激。酒精及其代谢产物对蛋白质结构的破坏可能会加剧酒精使用障碍；蛋白质组学可以阐明其潜在的生物学机制。本研究检测了30例男性酒精使用障碍患者红细胞胞质和膜部分中差异表达的蛋白质，并将其与15例年龄和体重指数匹配的社交饮酒者（SD）及15例非饮酒者（对照组）的样本进行比较。该分析旨在确定与饮酒相关的分子差异。酒精使用障碍患者亚组是根据平均红细胞体积（MCV）划分的，16例个体MCV正常，14例MCV高。蛋白质通过二维（2D）凝胶电泳分离，用胰蛋白酶消化，并通过基质辅助激光解吸/电离飞行时间（TOF）质谱（MALDI-TOF/TOF）进行鉴定。此外，还分析了丙二醛和4-羟基烯醛（MDA+HAE）、还原型谷胱甘肽（GSH）、氧化型谷胱甘肽（GSSG）、血清缺糖转铁蛋白（%CDT）、双唾液酸转铁蛋白（%DST）和唾液酸（SA）的水平。结果显示，酒精使用障碍患者的MDA+HAE增加，总硫醇减少，酒精使用障碍MCV高亚组的GSSG升高，GSH/GSSG比值降低。酒精使用障碍患者的血清%CDT、%DST和SA显著更高。与对照组相比，酒精使用障碍组表现出差异蛋白质表达。与对照组和社交饮酒者相比，以及在酒精使用障碍MCV高亚组中，少数蛋白质如二磷酸甘油酸变位酶表达下调。相反，相对于对照组，内质蛋白和凝溶胶蛋白在酒精使用障碍患者中上调。包括血影蛋白α链、肌动蛋白胞质2在内的细胞骨架蛋白在酒精使用障碍组和酒精使用障碍MCV高亚组中过表达。该亚组中表达不足的蛋白质包括带3阴离子转运蛋白、二磷酸甘油酸变位酶、原肌球蛋白α-3链、尿卟啉原脱羧酶和含WD重复序列蛋白1。我们的研究结果突出了与氧化应激、细胞骨架改变和代谢失调相关的蛋白质表达的特定变化，特别是在MCV升高的酒精使用障碍患者中。了解这些机制对于制定有针对性的干预措施和识别酒精诱导细胞损伤的生物标志物至关重要。氧化应激、膜组成和细胞功能之间的复杂相互作用说明了慢性酒精暴露如何影响细胞生理学。
