动物营养学报 >

2018 , Vol. 30 >Issue 9: 3517 - 3523

DOI: https://doi.org/10.3969/j.issn.1006-267x.2018.09.021

反刍动物营养 Ruminant nutrition

二乙烯三胺/一氧化氮聚合物诱导的奶牛外周血单个核细胞氧化损伤模型的建立

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  • 内蒙古农业大学动物科学学院, 呼和浩特 010018
郑亚光(1990-),男,内蒙古包头人,博士研究生,从事动物矿物质与维生素营养研究。E-mail:zhengyaguangg@163.com

收稿日期: 2018-02-09

  网络出版日期: 2018-09-20

基金资助

国家自然科学基金（31672463）

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Establishment of Oxidative Damage Model of Dairy Cow Peripheral Blood Mononuclear Cells Induced by Diethylenetriamine/Nitric Oxide Adduct

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  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China

Received date: 2018-02-09

  Online published: 2018-09-20

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摘要

本试验旨在利用二乙烯三胺/一氧化氮聚合物（DETA/NO）作为一氧化氮（NO）供体，以细胞存活率和抗氧化指标作为判断依据，确定建立奶牛外周血单个核细胞（PBMC）的氧化损伤模型的适宜条件。试验分2部分进行。试验1采用单因子完全随机试验设计，以不同浓度[0（对照）、50、100、200、300、500 μmol/L]的DETA/NO作为刺激源，在37℃下分别作用细胞2、4、6、8、12 h，通过测定细胞存活率，初步确定适宜的DETA/NO作用时间。试验2根据试验1得出的DETA/NO适宜作用时间，以不同浓度[0（对照）、50、100、200、300、500 μmol/L]的DETA/NO作为刺激源，依据抗氧化指标和炎症因子含量进一步筛选出适宜的DETA/NO作用浓度。结果显示：200 μmol/L DETA/NO作用细胞4 h，PBMC存活率降低至72.3%，超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶活性均较对照组显著降低（P<0.05），白细胞介素-1、白细胞介素-6、肿瘤坏死因子-α、丙二醛和NO含量均较对照组显著上升（P<0.05）。结果提示，在37℃下，DETA/NO诱导PBMC建立PBMC氧化损伤模型的适宜作用浓度和作用时间分别为200 μmol/L和4 h。

关键词： 奶牛外周血单个核细胞; 二乙烯三胺/一氧化氮聚合物; 氧化损伤

本文引用格式

郑亚光, 齐敬宇, 张博綦, 史彬林, 闫素梅 . 二乙烯三胺/一氧化氮聚合物诱导的奶牛外周血单个核细胞氧化损伤模型的建立[J]. 动物营养学报, 2018 , 30(9) : 3517 -3523 . DOI: 10.3969/j.issn.1006-267x.2018.09.021

Abstract

This study was conducted to investigate the suitable condition for oxidative damage model of dairy cow peripheral blood mononuclear cells (PBMC) induced by diethylenetriamine/nitric oxide adduct (DETA/NO) which was a nitric oxide (NO) donor. The oxidative damage model of PBMC was established by detecting the cell survival rate and antioxidant parameters. The test was divided into 2 parts. Test 1 was conducted as a single factor randomized arrangement, PBMC was exposed in DETA/NO with different concentrations[0 (control), 50, 100, 200, 300 and 500 μmol/L] for 2, 4, 6, 8 and 12 hours at 37℃, respectively. The suitable action time of DETA/NO was determined by detecting the cell survival rate. Based on the results of suitable action time of DETA/NO in test 1, the cells exposed in DETA/NO with different concentrations[0 (control), 50, 100, 200, 300 and 500 μmol/L] in test 2, the suitable action concentration of DETA/NO was further screened by detecting the antioxidant parameters and inflammatory cytokine contents. The results showed that the PBMC survival rate decreased to 72.3%, the activities of superoxide dismutase, catalase and glutathione peroxidase decreased significantly (P<0.05), and the contents of interleukin-1, interleukin-6, tumor necrosis factor-α, malondialdehyde and NO increased significantly (P<0.05) after treated with 200 μmol/L DETA/NO for 4 hours. It is concluded that the suitable action concentration and action time at 37℃ of DETA/NO for establishing the oxidative damage model of dairy cow PBMC are 200 μmol/L and 4 hours, respectively.

Key words： peripheral blood mononuclear cell; diethylenetriamine/nitric oxide adduct; oxidative damage

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