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田七总皂苷对PCV-2诱导RAW264.7细胞氧化应激的调节作用

发表时间:2016-11-15  浏览量:1255  下载量:344
全部作者: 尹 丹,郝祝兵,韦英益,杨 剑,胡庭俊
作者单位: 广西大学动物科学技术学院
摘 要: 本试验以PCV-2体外感染小鼠巨噬细胞系RAW264.7细胞,探讨田七总皂苷(Pannax notoginseng saponins,PNS)对PCV-2感染RAW264.7细胞氧化应激的调节作用。采用维生素C(vitamin C,Vc,200 µmol/L)作为对照药,用50,100,200 mg/L PNS作用于PCV-2感染的RAW264.7细胞,MTT法检测细胞存活率,Griess法检测NO水平,DCFH-DA荧光探针检测细胞总活性氧(reactive oxygen species,ROS)水平,邻苯二甲醛(OPT)荧光法检测细胞氧化还原状态,化学发光法检测细胞黄嘌呤氧化酶(xanthine oxidase,XOD)、髓过氧化物酶(myeloperoxidase,MPO)、诱生型一氧化氮合酶(inducible nitric oxide synthase,iNOS)活力,观察4,8,12,24 h PNS对PCV-2感染RAW264.7细胞NO、ROS、谷胱甘肽(glutathione,GSH)、氧化型谷胱甘肽oxidized glutathione,(GSSG)水平,以及对XOD、MPO、iNOS活力的影响。结果显示,PCV-2感染RAW264.7细胞后显著升高细胞分泌NO水平(P<0.05),显著升高细胞ROS水平、GSSG水平、MPO活力、XOD活力及iNOS活力(P<0.05),显著降低细胞 GSH 水平及GSH/GSSG比值(P<0.05);加入PNS处理后,200 mg/L的PNS在4,12 h显著降低细胞分泌NO水平;在4,8,12,24 h不同程度地降低ROS水平、GSSG水平、MPO活力、XOD活力及iNOS活力、升高细胞GSH水平及GSH/GSSG比值。结果表明,PNS能调节PCV-2诱导RAW264.7细胞的氧化应激,但其详细分子作用机制有待进一步研究。
关 键 词: 中兽医学;田七总皂苷;PCV-2;RAW264.7细胞;氧化应激
Title: Effect of Pannax notoginseng saponins on oxidative stress induced by PCV-2 infection in RAW264.7 cells
Author: YIN Dan, HAO Zhubing, WEI Yingyi, YANG Jian, HU Tingjun
Organization: College of Animal Science and Technology, Guangxi University
Abstract: The objective of the current study is to investigate the effects of Pannax notoginseng saponins (PNS) on oxidative stress induced by PCV-2 infection in RAW264.7 cells. Vitamin C (Vc, 200 μmol/L) was used as controlled drugs, PCV-2 infected RAW264.7 cells were treated with PNS (50, 100, 200 mg/L) or Vc, to observe NO, reactive oxygen species (ROS), glutathione (GSH), oxidized glutathione (GSSG), myeloperoxidase (MPO), xanthine oxidase (XOD), inducible nitric oxide synthase (iNOS) production, cell viability and ratio of GSH to GSSG at 4, 8, 12, 24 h. MTT method was used to measure the cell viability. The level of NO was determined by Griess regent. ROS production was evaluated with DCFH-DA fluorescence probe. GSH and GSSG levels were determined with fluorescence method in OPT. MPO, XOD, iNOS activity were measured by chemiluminescence assay. The results showed that PCV-2 infection significantly increased the production of NO, ROS and GSSG (P<0.05), and significantly raised activity of MPO, XOD, iNOS (P<0.05) and significantly decreased the level of GSH and ratio of GSH to GSSG (P<0.05) in PCV-2 infected cells. Treatment with PNS at 200 mg/L significantly reduced the secretion of NO at 4 h and 12 h after PCV-2 infection, the levels of ROS, GSSG and the activity of MPO, XOD, iNOS were significantly decreased at 4, 8, 12 and 24 h. And the ratio of GSH to GSSG was significantly increased in infected RAW264.7 cells. The results showed that PCV-2 infection resulted in cell oxidative stress and PNS acted against PCV-2 infection by scavenging ROS in RAW264.7 cells and improving the enzyme activity, but the detailed molecular mechanism needed further research.
Key words: traditional Chinese veterinary science; Pannax notoginseng saponins; PCV-2; RAW264.7 cell; oxidative stress
发表期数: 2016年11月第21期
引用格式: 尹 丹,郝祝兵,韦英益,等. 田七总皂苷对PCV-2诱导RAW264.7细胞氧化应激的调节作用[J]. 中国科技论文在线精品论文,2016,9(21):2265-2272.
 
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