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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。
The effects of different concentrations of porcine circovirus type 2 (PCV2) on the level of oxidative stress induced by in vitro infection of 3D4 / 2 cells were investigated to establish the conditions for establishing the model of PCV2-induced 3D4 / 2 cell oxidative stress in vitro. Five different concentrations of PCV2 infection group (100,10-1,10-2,10-3,10-4) were applied to 3D4 / 2 cells for 2h, the virus liquid was discarded, and the medium containing 100mL / L fetal calf serum 1640 After the liquid culture, the cell supernatants or cells were collected at 4, 8, 12, 24 and 48 hours, and the indexes such as NO, ROS, GSH, GSSG, XOD, MPO and iNOS were measured. 10-1PCV2 infected 3D4 / 2 cells 24h, 48h significantly increased cell NO levels, 4h ~ 24h infection significantly increased cell ROS levels, reduce cell GSH levels and GSH / GSSG ratio; 10-1 PCV2 infected 3D4 / 2 cells 4h , 8h significantly increased cell GSSG levels; 10-1PCV2 infected 3D4 / 2 cells at 4,8,12,24 h were significantly increased cell XOD, MPO, iNOS activity, suggesting that 10-1PCV2 infected 3D4 / 2 cells changed to some extent The redox state of the cells induced oxidative stress in the cells, and no PCV2 nucleic acid was detected 48 h after virus infection. The results showed that 4h ~ 24h after PCV2 infection induced 3D4 / 2 cell oxidative stress, and 10-1PCV2 was chosen as the infection dose of oxidative stress model.