傅里叶变换红外光谱衰减全反射法在测量身心应激后血浆中羰基化合物的适用性。
Applicability of FTIR-ATR Method to Measure Carbonyls in Blood Plasma after Physical and Mental Stress.
作者信息
Bujok Jolanta, Gąsior-Głogowska Marlena, Marszałek Michał, Trochanowska-Pauk Natalia, Zigo František, Pavľak Alexander, Komorowska Małgorzata, Walski Tomasz
机构信息
Department of Animal Physiology and Biostructure, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Norwida 31, 50-375 Wrocław, Poland.
Department of Biomedical Engineering, Faculty of Fundamental Problems of Technology, Wrocław University of Science and Technology, Wybrzeże Wyspiańskiego 27, 50-370 Wrocław, Poland.
出版信息
Biomed Res Int. 2019 Mar 26;2019:2181370. doi: 10.1155/2019/2181370. eCollection 2019.
INTRODUCTION
Oxidative stress is a state of imbalance between the production of reactive oxygen species and antioxidant defenses. It results in the oxidation of all cellular elements and, to a large extent, proteins, causing inter alia the formation of carbonyl groups in their structures. The study focused on assessment of changes in the plasma protein-bound carbonyls in police horses after combat training and after rest and the applicability of infrared spectroscopy with a Fourier transform, utilizing the attenuated total reflectance (FTIR-ATR) in detecting plasma protein oxidation.
METHODS
We evaluated the influence of both the different concentrations of hydrogen peroxide and combat training on protein carbonylation in horse blood plasma. The oxidation of plasma proteins was assessed using a spectrophotometric method based on the carbonyl groups derivatization with 2,4-dinitrophenylhydrazine (DNPH). The measured values were correlated with the carbonyl groups concentrations determined by means of the FTIR-ATR method.
RESULTS
The linear correlation between the DNPH and FTIR-ATR methods was shown. The concentration of plasma protein-bound carbonyls significantly deceased in police horses after one-day rest when compared to the values measured directly after the combat training (a drop by 23%, p<0.05 and 29%, p<0.01 measured by DNPH and FTIR-ATR methods, respectively). These results were consistent with the proteins phosphorylation analysis.
CONCLUSION
The FTIR-ATR method may be applied to measure the level of plasma proteins peroxidation.
引言
氧化应激是活性氧生成与抗氧化防御之间失衡的一种状态。它会导致所有细胞成分发生氧化，在很大程度上还会使蛋白质氧化，尤其会在其结构中形成羰基。本研究聚焦于评估警用马匹在战斗训练后及休息后的血浆蛋白结合羰基的变化，以及利用衰减全反射傅里叶变换红外光谱法（FTIR - ATR）检测血浆蛋白氧化的适用性。
方法
我们评估了不同浓度的过氧化氢和战斗训练对马血浆中蛋白质羰基化的影响。血浆蛋白的氧化通过基于用2,4 - 二硝基苯肼（DNPH）衍生化羰基的分光光度法进行评估。所测值与通过FTIR - ATR法测定的羰基浓度相关。
结果
显示了DNPH法与FTIR - ATR法之间的线性相关性。与战斗训练刚结束时直接测量的值相比，警用马匹在休息一天后血浆蛋白结合羰基的浓度显著下降（分别通过DNPH法和FTIR - ATR法测量，下降了23%，p < 0.05和29%，p < 0.01）。这些结果与蛋白质磷酸化分析一致。
结论
FTIR - ATR法可用于测量血浆蛋白过氧化水平。
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