硒对脂多糖诱导的奶牛乳腺上皮细胞氧化损伤的保护作用

发布时间：2018-04-01 10:07

本文选题：硒代蛋氨酸　切入点：脂多糖　出处：《动物营养学报》2017年09期

【摘要】：本试验旨在研究硒(Se)对脂多糖(LPS)诱导的奶牛乳腺上皮细胞(BMEC)氧化损伤的保护作用及其机制。将贴壁生长的第3代BMEC随机分为8组,每组6个重复,每个重复1个培养孔。对照(CON)组采用基础培养液,不添加Se和LPS,培养30h;LPS组和6个Se保护组在基础培养液中分别添加不同水平的Se(0、10、20、50、100、150和200nmol/L),培养24h后,加入1μg/mL LPS作为外源刺激作用6h。结果表明:1)与CON组相比,LPS组BMEC的相对增殖率显著下降(P0.05),谷胱甘肽过氧化物酶(GPx)、硫氧还蛋白还原酶(TrxR)、总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)活性和总抗氧化能力(T-AOC)均显著下降(P0.05),GPx1和TrxR1的基因和蛋白表达量、硒蛋白P(SelP)含量也显著下调(P0.05);而LPS组的一氧化氮(NO)含量,诱导型一氧化氮合酶(iNOS)活性及其基因和蛋白表达量,炎症因子肿瘤坏死因子-α(TNF-α)、白介素-1(IL-1)和白介素-6(IL-6)含量及其基因表达量,活性氧(ROS)活性,丙二醛(MDA)含量均显著升高(P0.05),丝裂原活化蛋白激酶(MAPK)信号通路相关因子p38丝裂原活化蛋白激酶(p38 MAPK)、c-Jun氨基端激酶(JNK)、细胞外信号调节激酶1/2(ERK1/2)的基因表达量呈相似变化。2)与LPS组相比,Se保护组随Se添加水平的增加,相对增殖率,T-SOD、CAT、GPx、TrxR活性,T-AOC,GPx1、TrxR1基因和蛋白表达量均呈先升高后下降趋势;而NO含量,iNOS活性及其基因和蛋白表达量,炎症因子TNF-α、IL-1、IL-6含量及其基因表达量,MAPK信号通路相关因子ERK1/2、JNK、p38 MAPK的基因表达量,ROS活性,MDA含量呈先降低后升高的趋势;以20~100nmol/L Se保护效果较好,综合来看50nmol/L Se保护效果最好。结果提示,Se可提高BMEC的抗氧化功能,对LPS引起的细胞氧化损伤具有保护作用,其机制是Se增强TrxR活性从而抑制MAPK信号通路的激活,最终减少NO的大量释放,但过高水平的Se会对细胞造成损伤。培养液中20~100nmol/L Se的保护作用较好,尤其以50nmol/L Se效果最好。
[Abstract]:The aim of this study was to investigate the protective effect of selenoside on oxidative damage induced by lipopolysaccharide (LPS) in dairy cow mammary epithelial cells (BMECs) and its mechanism. The third passage BMEC of adherent growth was randomly divided into 8 groups with 6 replicates in each group. The control group was treated with basic culture medium without se and LPS.The 30 h LPS group and 6 se protection group were cultured with different levels of Seo 01020D 50100150 and 200nmol / L 1, respectively, and cultured for 24 h. Addition of 1 渭 g/mL LPS as exogenous stimulator for 6 h. The results showed that the relative proliferation rate of BMEC in CON group was significantly lower than that in CON group (P 0.05), glutathione peroxidase (Glutathione peroxidase), thioredoxin reductase (TRA), total superoxide dismutase (T-SOD), catalase. The activity of CAT and total antioxidant capacity (T-AOC) significantly decreased the gene and protein expression of P0.05, GPx1 and TrxR1. In LPS group, the content of nitric oxide (no), the activity of inducible nitric oxide synthase (iNOS), its gene and protein expression, the content of inflammatory factor tumor necrosis factor- 伪 (TNF- 伪), interleukin-1 (IL-1) and interleukin-6IL-6 (IL-6) and its gene expression were also significantly down-regulated. Reactive oxygen species (Ros) activity, The content of malondialdehyde (MDA) MDAs increased significantly (P 0.05). The expression of p38 mitogen-activated protein kinase (p38 MAPK) signal pathway associated with c-Jun amino terminal kinase (JNKK) and extracellular signal-regulated kinase 1 / 2 ERK1 / 2 were similar. Compared with the LPS group, the se protection group increased with the increase of se addition level. The relative proliferative rate and the activity of T-SOD CATX TXR gene and protein increased at first and then decreased, while the no content and the activity of iNOS and its gene and protein expression showed a tendency to increase first, and then to decrease, while the activity of iNOS and the expression of gene and protein in the activity of T-AOCU GPx1 + TrxR1 showed a tendency to increase first and then to decrease. The content of IL-6 and its gene expression in inflammatory factor TNF- 伪 and its gene expression; the gene expression of ERK1 / 2 MAPK and Ros activity were decreased first and then increased, and the protective effect of 20~100nmol/L se was better. The results suggest that se can enhance the antioxidant function of BMEC and protect cells from oxidative damage induced by LPS. The mechanism is that se enhances the activity of TrxR and inhibits the activation of MAPK signaling pathway. In the end, the release of no was reduced, but too high level of se could damage the cells. The protective effect of 20~100nmol/L se in culture medium was better, especially 50nmol/L se was the best.
【作者单位】：内蒙古农业大学动物科学学院;
【基金】：国家自然科学基金(31560650)
【分类号】：S823.5

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