RUMINANT AND HERBIVORE NUTRITION

Establishment of Oxidative Damage Model of Bovine Mammary Epithelial Cells Induced by Diethylenetriamine/Nitric Oxide Adduct

  • GUO Yongmei ,
  • ZHANG Boqi ,
  • SHI Huiyu ,
  • YAN Sumei ,
  • SHI Binlin ,
  • GUO Xiaoyu
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  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China

Received date: 2016-02-04

  Online published: 2016-08-17

Abstract

Due to exuberant metabolism, dairy cows produce a great deal of nitric oxide (NO)during lactation, which leads to oxidative stress, and then results in cell toxicity. This experiment was conducted to investigate the suitable condition for oxidative damage model of bovine mammary epithelial cells (BMEC) induced by diethylenetriamine/nitric oxide adduct (DETA/NO) and establish an oxidative damage model. The oxidative damage model of BMEC was established using DETA/NO as the stimulation source. BMEC was exposed in DETA/NO with different medium concentrations (0, 10, 30, 100, 500 and 1 000 μmol/L) and for different times (2, 4, 6, 8, 12 and 24 h). According to the analysis of cell survival rate and antioxidative parameters, inflammatory cytokines and nitric oxide (NO) contents, as well as inducible nitric oxide synthase (iNOS) activity, the suitable treatment time and concentration of DETA/NO were screened. The results showed that the survival rate decreased to 74.27%, and the activitise of superoxide dismutase, catalase and glutathione peroxidase also significantly reduced (P<0.05) after that BMEC was cultured with 1 000 μmol/L DETA/NO for 6 h. However, the activity of iNOS and the contents of NO, interleukin-1, interleukin-6, tumor necrosis factor-α and malondialdehyde showed opposite changes and increased significantly (P<0.05). In summary, the treatment of 1 000 μmol/L DETA/NO for 6 h is suitable for establishment of oxidative damage model.

Cite this article

GUO Yongmei , ZHANG Boqi , SHI Huiyu , YAN Sumei , SHI Binlin , GUO Xiaoyu . Establishment of Oxidative Damage Model of Bovine Mammary Epithelial Cells Induced by Diethylenetriamine/Nitric Oxide Adduct[J]. Chinese Journal of Animal Nutrition, 2016 , 28(8) : 2378 -2384 . DOI: 10.3969/j.issn.1006-267x.2016.08.008

References

[1] 刘建新,王艳明.奶牛氧化应激研究进展及其影响因素[J].饲料工业,2010(增刊):9-14.
[2] 王振云,周璇,李惠侠,等.茶多酚对氧化应激所致奶牛乳腺上皮细胞损伤的保护作用[J].南京农业大学学报,2012,35(3):101-106.
[3] MACMICKING J,XIE Q W,NATHAN C.Nitric oxide and macrophage function[J].Annual Review of Immunology,1997,15:323-350.
[4] SANG J R,JIANG M Y,LIN F,et al.Nitric oxide reduces hydrogen peroxide accumulation involved in water stress-induced subcellular anti-oxidant defense in maize plants[J].Journal of Integrative Plant Biology,2008,50(2):231-243.  
[5] KEEFER L K,NIMS R W,DAVIES K M,et al."NONOates"(1-substituted diazen-1-ium-1,2-diolates) as nitric oxide donors:convenient nitric oxide dosage forms[J].Methods in Enzymology,1996,268:281-293.
[6] 王新朋,闫素梅,齐利枝,等.β-羟丁酸浓度对奶牛乳腺上皮细胞内乳蛋白合成相关基因表达量的影响[J].动物营养学报,2014,26(12):3836-3842.
[7] PETKOV S G,MARKS H,KLEIN T,et al.In vitro culture and characterization of putative porcine embryonic germ cells derived from domestic breeds and Yucatan mini pig embryos at days 20-24 of gestation[J].Stem Cell Research,2011,6(3):226-237.  
[8] BIAN K,MURAD F.Nitric oxide (NO)-biogeneration,regulation,and relevance to human diseases[J].Frontiers in Bioscience,2002,8:d264-d278.
[9] 章丹丹,刘俊,MURAD F,等.诱导型一氧化氮合酶与疾病[J].现代生物医学进展,2007,7(10):1571-1573,1577.
[10] 孙婧陶,李兆华,张宝修,等.过氧化氢诱导延边奶山羊乳腺上皮细胞氧化损伤模型的建立[J].江苏农业科学,2013,41(10):149-152.
[11] 齐晓龙,赵芹,张亚男,等.过氧化氢诱导产蛋鸡原代肝细胞氧化应激模型的建立[J].中国畜牧杂志,2013,49(11):49-52.
[12] 金鹿,闫素梅,史彬林,等.过氧化氢诱导的奶牛乳腺上皮细胞氧化损伤模型的建立[J].动物营养学报,2014,26(12):3651-3658.
[13] 袁施彬.仔猪氧化应激及硒的抗应激效应和机理的研究[D].博士学位论文.雅安:四川农业大学,2007.
[14] 李忠鹏.奶牛肢蹄病患牛体内矿物元素和氧化应激指标的变化[D].硕士学位论文.泰安:山东农业大学,2014.
[15] 程广东.连翘酯苷A对鸡脾和血中炎症细胞因子及抗氧化性能影响[D].博士学位论文.哈尔滨:东北农业大学,2014.
[16] 范精华,刘康,刘保林.NO在炎症及免疫应答中的调节作用[J].中外医疗,2009,28(25):163,166.
[17] BIAN K,KE Y,KAMISAKI Y,et al.Proteomic modification by nitric oxide[J].Journal of Pharmacological Sciences,2006,101(4):271-279.  
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