Khosravifarsani Meysam, Monfared Ali Shabestani, Pouramir Mahdi, Zabihi Ebrahim
M.Sc. of Radiobiology and Radiation Protection, MPhil, Cellular and Molecular Biology Research Center, Faculty of Medicine, Babol University of Medical Sciences, Babol, Iran.
Ph.D. of Medical Physics, Professor, Cellular and Molecular Biology Research Center, Faculty of Medicine, Babol University of Medical Sciences, Babol, Iran.
Electron Physician. 2016 Sep 20;8(9):2970-2976. doi: 10.19082/2970. eCollection 2016 Sep.
Human serum albumin (HSA) is a critical protein in human blood plasma, which can be highly damaged by oxidative stress. The aim of this study was to analyze modifications of this protein after oxidation using a Fenton system.
In this 2015 experiment, different ratios of Fenton reagent (Fe2+/HO) was incubated with one concentration of human serum albumin (1mg/ml). Hence, HSA was incubated 30 min with various combinations of a Fenton system and quantified oxidation products such as carbonyl groups, fragmentations, degradations, and oxidized free thiol group using reliable techniques. Image and data analysis were carried out using ImageJ software and Excel (version 2007), respectively.
An SDS-PAGE profile showed no cross link and aggregation. However, protein band intensity has decreased to 50% in the highest ratio of HO/Fe. Carbonylation assay indicated carbonyl/protein (molc/molp) ratio increased linearly in lower ratios and the values plateau at higher levels of HO/Fe 2+. The only free sulfhydryl group on HSA was oxidized in all ratios of the Fenton system.
To sum, the structure of HSA has been changed following treatment with Hydroxyl Radical as the main product of Fenton reaction. These data confirm the antioxidant activity of HSA.
人血清白蛋白（HSA）是人体血浆中的一种关键蛋白质，极易受到氧化应激的损伤。本研究旨在分析使用芬顿体系氧化后该蛋白质的修饰情况。
在2015年的这项实验中，将不同比例的芬顿试剂（Fe2+/HO）与一种浓度的人血清白蛋白（1mg/ml）一起孵育。因此，将HSA与芬顿体系的各种组合孵育30分钟，并使用可靠技术对羰基、片段化、降解和氧化游离巯基等氧化产物进行定量分析。分别使用ImageJ软件和Excel（2007版）进行图像和数据分析。
SDS-PAGE图谱显示没有交联和聚集。然而，在HO/Fe的最高比例下，蛋白条带强度下降至50%。羰基化分析表明，在较低比例下，羰基/蛋白质（molc/molp）比值呈线性增加，在较高水平的HO/Fe 2+时达到平稳。在芬顿体系的所有比例下，HSA上唯一的游离巯基都被氧化。
总之，作为芬顿反应主要产物的羟基自由基处理后，HSA的结构发生了变化。这些数据证实了HSA的抗氧化活性。
