猪圆环病毒2型诱导3D4/2细胞氧化应激模型的建立

通过研究不同浓度猪圆环病毒2型(PCV2)对体外感染3D4/2细胞产生氧化应激水平的影响,来确定建立PCV2体外诱导3D4/2细胞氧化胁迫模型的条件。以5个不同浓度PCV2感染组(100、10-1、10-2、10-3、10-4)作用于3D4/2细胞2h,弃去病毒液,加入含100mL/L胎牛血清1640培养液后继续培养,于4、8、12、24、48h分别收集细胞上清液或细胞,测定NO、ROS、GSH、GSSG、XOD、MPO和iNOS等指标。10-1PCV2感染3D4/2细胞24h、48h后显著升高细胞NO水平,感染4h~24h显著升高细胞ROS水平,降低细胞GSH水平和GSH/GSSG比值;10-1 PCV2感染3D4/2细胞4h、8h显著升高细胞GSSG水平;10-1PCV2感染3D4/2细胞4、8、12、24h均能显著升高细胞XOD、MPO、iNOS活力,提示10-1PCV2感染3D4/2细胞一定程度上改变了细胞氧化还原状态,诱导了细胞产生氧化应激,而病毒感染后48h未检测到PCV2核酸。表明PCV2感染后4h~24h能诱发3D4/2细胞氧化应激,选择10-1PCV2作为氧化应激模型的感染剂量。

Establishment of Oxidative Stress Model of 3D4/2 Cells Induced by PCV2 in Vitro

The study was carried out to establish a oxidative stress model induced by PCV2 in vitro.The 3D4/2 cells were incubated with different concentrations of PCV2 to investigate the levels of oxidative stress in the cells.3D4/2 cells were incubated with PCV2 at dose of 100,10-1,10-2,10-3 or 10-4.The supernatant or cells were collected at different time to observe the levels of NO,ROS,GSH,GSSG,cell viability,and the activities of XOD,MPO,iNOS.The level of NO in 3D4/2 cells infected with 10-1 PCV2 was significantly increased after 24 h,48 h,and the levels of ROS,GSSG and the activities of XOD,MPO,iNOS were significantly upregulated post infection.These findings suggested that PCV2 infection changed the redox state of 3D4/2 cells and induced 3D4/2 cells to be in state of oxidative stress.PCV2 infection induced oxidative stress in 3D4/2 cells,and 10-1 PCV2 was selected as the infection dose of oxidative stress model,the infection time was determined to be 4 h-24 h.