Reactive Oxygen Species Production and Antioxidant Enzyme Expression after Epstein-Barr Virus Lytic Cycle Induction in Raji Cell Line
Reactive Oxygen Species Production and Antioxidant Enzyme Expression after Epstein-Barr Virus Lytic Cycle Induction in Raji Cell Line
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DOI：
10.1007/s12011-011-9135-5
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发表时间：
2011-12-01
影响因子：
3.9
通讯作者：
Van Pelt, Jos
中科院分区：
文献类型：
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作者：
Gargouri, Bochra;Nasr, Rihab;Van Pelt, Jos
在之前的研究中，我们已经描述了在EB病毒裂解周期诱导过程中的氧化应激。观察到的高丙二醛水平和抗氧化酶活性的降低证明了氧化应激。我们推测，抗氧化酶活性降低的原因要么是活性氧自由基(ROS)的过量产生，要么是抗氧化酶基因表达的负调控。为了阐明这一情况，在Raji细胞系中用非应激剂量12-0-十四酰佛波醇-13-乙酸酯诱导EBV裂解周期。采用半定量逆转录聚合酶链式反应(RT-PCR)检测BZLF-1、超氧化物歧化酶和过氧化氢酶基因在6、12、24、36和48h的表达，化学发光法检测ROS的产生。为了确定ROS产生、BZLF-1和抗氧化基因表达是否相互关联，进行了一项研究。裂解周期的诱导导致超氧化物歧化酶和过氧化氢酶基因的表达增加，从24 h开始(p<；0.05)，48 h达到高峰(p<；0.05)。与未处理细胞相比，经TPA处理的Raji细胞在12h时ROS水平显著升高，在EBV裂解周期诱导后48h达到高峰。ROS的产生与BZLF-1、SOD0.05和CAT基因的表达呈正相关(r分别为0.913、0.978和0.955)。BZLF-1与抗氧化基因表达呈正相关(分别为r=0.961和r=0.987)。总之，观察到的SOD和CAT基因表达的增加消除了在裂解周期的诱导过程中各自基因受到抑制的假设。另一方面，观察到BZLF-1基因的表达和ROS产生之间的直接相关性表明该基因在氧化应激中发挥了作用。
In a previous study, we have described oxidative stress during Epstein-Barr virus lytic cycle induction. Oxidative stress was evidenced by the observed high MDA levels and the decreased activities of antioxidant enzymes. We hypothesised that the lower activities of the antioxidant enzymes decrease were the result of either the excessive production of reactive oxygen radical species (ROS) or a negative regulation of the antioxidant enzyme gene expressions. In an attempt to clarify this situation, EBV lytic cycle was induced in Raji cell line by a non-stressing dose of 12-0-tetradecanoylphorbol-13-acetate. BZLF-1, superoxide dismutase, and catalase gene expressions were then analysed using semi-quantitative RT-PCR, simultaneously at a kinetic of 6, 12, 24, 36, and 48 h. ROS production was evaluated by chemiluminescence. A study was conducted to establish whether ROS production, BZLF-1, and the expression of antioxidant genes were inter-correlated. Induction of the lytic cycle resulted in increased expressions of the genes of superoxide dismutase and catalase, which began at 24 h (p < 0.05) and reached a peak at 48 h (p < 0.05). Significant increases of the ROS levels were observed in TPA-treated Raji cell line at 12 h, as compared with untreated cells, reaching a peak at 48 h after EBV lytic cycle induction. ROS production correlates positively with BZLF-1, SOD, and CAT gene expressions (p < 0.05; r = 0.913, r = 0.978, and r = 0.955, respectively). A positive correlation was also observed between BZLF-1 and antioxidant gene expressions (p < 0.05; r = 0.961 and r = 0.987, respectively). In conclusion, the observed increases of the SOD and CAT gene expressions eliminate the hypothesis of a repression of the respective genes during the induction of the lytic cycle. On the other hand, the observed direct correlation between the BZLF-1 gene expression and the ROS production is indicative of a role of this gene in oxidative stress.