猪圆环病毒2型体外诱导RAW264.7细胞氧化应激模型的建立

发布时间：2018-05-10 07:36

本文选题：猪圆环病毒型(PCV) + RAW.细胞　；参考：《南方农业学报》2017年01期

【摘要】：【目的】探讨猪圆环病毒2型(PCV2)感染量、感染时间与被感染RAW264.7细胞活性氧水平动态变化的相关性,为建立RAW264.7细胞氧化胁迫体外模型提供参考依据。【方法】PCV2原液调至5×106/m L,经10、102、103和104倍稀释(10-1、10-2、10-3和10-4稀释度)后,感染作用RAW264.7细胞2 h,弃病毒液,加入含5%FBS的DMEM培养液继续培养,分别于第4、8、12、24和48 h收集细胞上清液或细胞,测定一氧化氮(NO)、活性氧自由基(ROS)、还原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)、黄嘌呤氧化酶(XOD)、髓过氧化物酶(MPO)和诱生型一氧化氮合酶(i NOS)等指标。【结果】10-1 PCV2感染RAW264.7细胞后能有效升高细胞NO、ROS水平,降低细胞GSH水平和GSH/GSSG,升高细胞XOD、MPO和i NOS活性,表明以10-1 PCV2感染RAW264.7细胞一定程度上能改变细胞氧化还原状态,诱导细胞产生氧化应激。【结论】PCV2感染能诱导RAW264.7细胞发生氧化应激,并确立10-1 PCV2(5×105/m L)体外感染RAW264.7细胞4~24 h是建立RAW264.7细胞氧化胁迫模型的最佳条件。
[Abstract]:[objective] to investigate the relationship between the quantity and time of porcine circovirus type 2 (Porcine circovirus 2) infection and the level of reactive oxygen species (Ros) in infected RAW264.7 cells. In order to establish the model of oxidative stress of RAW264.7 cells in vitro, the model was established. [methods] the PCV2 solution was adjusted to 5 脳 10 ~ (-6) / mL, diluted with 10 ~ (-1) 10 ~ (-2) 10 ~ (-3) and 10 ~ (-4) dilution of 10 ~ (- 1) ~ 10 ~ (-2) 渭 m 路L ~ (-1), then infected with RAW264.7 cells for 2 h, the virus solution was discarded. The supernatants or cells were collected at 4h 81224 and 48 h, respectively. Nitric oxide (no), reactive oxygen free radical (Ros), reduced glutathione (GSH), oxidized glutathione (GSS), xanthine oxidase (XODO), myeloperoxidase (MPO) and inducible nitric oxide synthase (iNOS) were determined. [results] 10-1 PCV2 infected with RAW264.7. Retrocyte can effectively increase the level of NOROS in cells. The activity of XODMPO and I NOS was increased by decreasing the level of GSH and GSH / GSSG, indicating that the redox state of RAW264.7 cells infected with 10-1 PCV2 could be changed to some extent. [conclusion] PCV2 infection can induce oxidative stress in RAW264.7 cells, and it is the best condition to establish RAW264.7 cell oxidative stress model by 10 ~ (-1) PCV2(5 脳 10 ~ (5) / mL) infection of RAW264.7 cells for 4h in vitro.
【作者单位】：广西大学动物科学技术学院;
【基金】：国家自然科学基金项目(31260619) 高等学校博士学科点专项科研基金项目(20134501110004)
【分类号】：S852.651

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