ACTA VETERINARIA BRNO 审稿人-刊鹿论文编译

Glutathione as an antioxidant marker: determination of glutathione concentration in the breast muscles and liver of broilers supplemented with different selenium sources

Abstract:

The aim of the study was to determine the influence of different selenium sources on antioxidant properties. The glutathione (GSH) concentration and glutathione peroxidase (GPx) activity were measured in the breast muscles and liver of 60 one-day-old broiler chickens. Another goal was to compare these indices with the weights of individual tissues and the live weight of broilers. The broilers were divided into 4 groups according to the selenium source: group 1 (control), group 2 (selenized yeast), group 3 (selenomethionine), group 4 (sodium selenite). Treatment groups were supplemented with 0.2 mg of additional selenium/kg. No significant changes in the hepatic GSH concentrations (P > 0.05) were found in the experimental groups compared to control. Significantly higher GSH concentration (P < 0.05) was found in breast muscles of broilers in group 4 (sodium selenite) compared to control. However, no positive effect of selenium supplementation in the form of sodium selenite was observed. The differences in the GPx activity in breast muscles and liver between the experimental groups and the control group were not significant (P > 0.05). No significant differences were recorded in the experimental groups compared to control in relation to the GSH concentration and GPx activity measured in the tissues. A significantly positive correlation was noted between mean GPx activity in breast muscle and breast muscle weight (P < 0.01; r = 0.3790) and live weight (P < 0.05; r = 0.2690). Although changes in the GSH concentration and GPx activity were recorded in some experimental groups, the selected dose of additional selenium appeared to be too low to affect these concentrations and the antioxidant defence system. Antioxidant system, glutathione peroxidase, selenium supplementation, broiler chickens, tissue Glutathione is a tripeptide with a thiol group present at high concentrations in various cells and tissues (Shaik and Mehvar 2006; Cappiello et al. 2013). It is composed of glutamate, cysteine and glycine (γ-L-glutamyl-L-cysteinylglycine) and exists in millimolar concentrations (Kaplowitz et al. 1985). Glutathione is present in cells in its reduced form (GSH) and in oxidized forms – glutathione disulphide (GSSG) and glutathione mixed disulphides (Cappiello et al. 2013). The structure of glutathione, specifically its two forms, is related to its function and role in the organism (Meister 1988). Glutathione protects cells from oxidative stress directly by scavenging reactive oxygen species (ROS) as free radicals and peroxides that are formed in metabolism and by using glutathione-dependent enzymes such as glutathione peroxidase (GPx) and glutathione S-transferase (GST) (Meister 1983; Cappiello et al. 2013). For this reason, it is an extremely important endogenous antioxidant and plays a key role in the antioxidant defence system (Enkvetchakul et al. 1995). The imbalance of the antioxidant defence system in favour of prooxidants initiates oxidative stress and leads to the oxidation of different biological substrates – macromolecules (the reactive -OH· radical attacks lipids, DNA, proteins). Thus, oxidative stress may be the reason for processes such ACTA VET. BRNO 2019, 88: 157–163; https://doi.org/10.2754/avb201988020157 Address for correspondence: Mgr. Karolína Mikulková Large Animal Clinical Laboratory Faculty of Veterinary Medicine University of Veterinary and Pharmaceutical Sciences Brno Palackého tř. 1946/1, 612 42, Brno, Czech Republic Phone: +420 541 562 431 E-mail: mikulkovak@vfu.cz http://actavet.vfu.cz/ as lipid peroxidation, protein oxidation, membrane damage, carcinogenesis and other cell damage (Kaplowitz et al. 1985; Miller et al. 1993). The main function of glutathione is reduction of hydrogen peroxide (H2O2) and organic peroxides (lipid peroxides); these reactions are catalysed by GPx and GST (Meister 1983; Kaplowitz et al. 1985). The thiol group, present in reduced glutathione (GSH), acts as a free radical scavenger. During the reduction of H2O2 and lipid peroxides to H2O and alcohols, GSH is oxidized to GSSG and serves as an electron donor. Oxidized glutathione is converted back to GSH by glutathione reductase using reduced nicotinamide adenine dinucleotide phosphate (NADPH) (Cao et al. 2013). In healthy cells and tissues, the presence of the reduced form (GSH) predominates. Under pathophysiological conditions the GSH:GSSG ratio decreases. Oxidative stress that is caused by an imbalance between prooxidants and antioxidants can be the reason for an increase to the glutathione disulphide concentration. Thiol-disulphide exchange reactions are another type of conversion of intracellular GSH to GSSG catalysed by transhydrogenase (EC 1.8.4.2 – protein-disulphide reductase [glutathione]). These reactions are part of many metabolic and physiological functions such as protein synthesis and degradation and the activation and inactivation of enzymes (Meister 1983; Meister and Anderson 1983). According to several studies, the GSH/GSSG ratio may be considered the best indicator of oxidative stress (Tarin 1966; Tarin et al. 1998; Avanzo et al. 2001; Kaneko et al. 2001). Selenium is considered an essential trace element because of its irreplaceable role in the organism. It regulates the functions of the antioxidant defence system through the GSH and Se-containing antioxidant enzyme GPx (Mahmoud and Edens 2003; Horký 2014). The aim of this study was to evaluate the effect of additional selenium in the diet on the GSH concentration and GPx activity in the breast muscles and liver of broiler chickens. The breast muscle, liver and live weights of broilers were recorded and the correlation between weights and antioxidant properties were evaluated. Materials and Methods Animals and diets The study was carried out using a total of 60 one-day-old broiler chickens, following local Research Ethics Committee approval (84722/2014-MZE-17214). The chickens were divided into 4 groups – 1 control group and 3 treatment groups. Each group contained 15 chickens. All experimental groups were fed a basal diet (Table 1) and the 3 treatment groups were also supplemented with 0.2 mg of selenium/kg in three different form of additional selenium. The experimental groups were as follows: group 1 (control), group 2 (selenized yeast) supplemented with 118.9 g/t feed, group 3 (selenomethionine) supplemented with 8.70 g/t feed and group 4 (sodium selenite) supplemented with 6.67 g/t feed. The chickens were fed a Starter BR1 until 2 weeks of age, followed by a Grower BR2 until 5 weeks of age and a Finisher BR3 from 5 weeks of age (MTD Ústrašice, Czech Republic). Water and feed were provided ad libitum. After 6 weeks of feeding the chickens were stunned by electricity and slaughtered by cervical dislocation. Their body weight was recorded. Liver and breast muscle samples were immediately taken, weighed and stored at -70 °C. Sampling and analysis Homogenization of samples The tissues were washed in 0.9% NaCl solution and then homogenized. Disperser T10 basic ULTRA-TURRAX (IKA, Staufen, Germany) was used for homogenization. Tissues for GSH determination were homogenized in precipitation reagent at a ratio of 1:20 (supplied in the kit) and tissues for GPx determination in 50 mM sodium phosphate buffer (pH 7.4), where 1 ml of solution per 0.12 g of tissue was used. The samples were kept on ice during homogenization. After centrifugation at 3000 g for 10 min at 4 °C, the supernatant was collected and used for the assay. The tissues were stored at -70 °C and the GSH concentration and GPx activity were determined immediately after homogenization. Total glutathione determination Total glutathione concentrations were measured with a BIOXYTECH GSH-420 kit (OxisResearch, Portland, Oregon, USA) using the quantitative colorimetric method. This method is based on the formation of chromophoric thione and the chemical reaction proceeds in three steps. First, oxidized glutathione (GSSG) is reduced to the reduced form (GSH) by adding the reducing agent tris(2-carboxyethyl)phosphine (TCEP). The 158

Author Listing： Karolína Mikulková;Josef Illek;Zdenka Bezděková;Ivana Šimková

Volume： 88

Pages： 157-163

DOI： 10.2754/AVB201988020157

Language： English

Journal： Acta Veterinaria Brno

ACTA VETERINARIA BRNO

ACTA VET BRNO

影响因子：0.6

是否综述期刊：否

是否OA：是

是否预警：不在预警名单内

发行时间：1969

ISSN：0001-7213

发刊频率：Quarterly

收录数据库：SCIE/Scopus收录

出版国家/地区：CZECH REPUBLIC

出版社：University of Veterinary and Pharmaceutical Sciences

期刊介绍

ACTA VETERINARIA BRNO is a scientific journal of the University of Veterinary and Pharmaceutical Sciences in Brno, Czech Republic.The scientific journal Acta Veterinaria Brno is dedicated to the publication of original research findings and clinical observations in veterinary and biomedical sciences. Original scientific research articles reporting new and substantial contribution to veterinary science and original methods that have not been submitted for publication elsewhere are considered for publication. A written statement to this effect should accompany the manuscript, along with approval for publication by the author´s head of department. The authors bear full responsibility for the contents of their contribution. Book reviews are published, too.

《布尔诺兽医学报》（Acta Veterinaria Brno）是捷克共和国布尔诺兽医和制药科学大学的科学期刊，专门发表兽医和生物医学科学的原创研究成果和临床观察。报告对兽医科学有新的和实质性贡献的原始科学研究文章以及尚未在其他地方提交发表的原始方法将被考虑发表。手稿应附有一份书面声明，沿着附有作者所在部门负责人的出版批准。作者对其稿件内容承担全部责任。书评也出版了。

年发文量	53
国人发稿量	-
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自引率	33.3%
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平均审稿周期	较慢,18-36周
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