Interaction of porcine circovirus type 2 replication with intracellular redox status in vitro
Interaction of porcine circovirus type 2 replication with intracellular redox status in vitro
复制标题
猪圆环病毒2型复制与体外细胞内氧化还原状态的相互作用
DOI：
10.1179/1351000213y.0000000058
复制
发表时间：
2013-09-01
期刊：
影响因子：
3.8
通讯作者：
Huang, Kehe
中科院分区：
文献类型：
--
作者：
Chen, Xingxiang;Ren, Fei;Huang, Kehe
目的：氧化还原状态影响一些病毒的复制，但其对猪圆环病毒2型（PCV 2）的影响尚不清楚，PCV 2是新出现的猪疾病断奶后多系统衰竭综合征的主要病原体。方法：用5，5 ′-二硫代双（2-硝基苯甲酸）比色法测定细胞内谷胱甘肽（GSH）含量，并与细胞内还原型谷胱甘肽（GSH）浓度进行相关性分析。总超氧化物歧化酶（SOD）活性测定通过抑制羟胺氧化的黄嘌呤氧化酶系统。采用硫代巴比妥酸法测定丙二醛（MDA）含量。结果：PCV 2感染后48 h，GSH和SOD含量均显著降低，MDA含量显著升高;此外，PCV 2在PK 15细胞中的复制显着受损后，通过与抗氧化剂N-乙酰基-L-半胱氨酸（NAC），在GSH合成的前体治疗的细胞内GSH的升高。相反，PCV 2在PK 15细胞中的复制增强后，GSH水平通过H2 O2介导的氧化降低。结论：PCV 2感染PK 15细胞后，细胞内的氧化还原状态影响PCV 2的复制。
Objectives: Redox status influences replication of some viruses but its effect on porcine circovirus type 2 (PCV2), the primary causative agent of the emerging swine disease post-weaning multisystemic wasting syndrome is not known. The interaction of PCV2 replication with intracellular redox status in PK15 cells was examined in this study.Methods: Intracellular glutathione (GSH) was measured spectrophotometrically by reaction with 5, 5'-dithiobis (2-nitrobenzoic acid). Total superoxide dismutase activity (SOD) was assayed by inhibition of oxyamine oxidation by the xanthine oxidase system. Malondialdehyde (MDA) was assayed spectrophotometrically using the thiobarbituric acid reaction. Both quantification of PCV2 DNA by real-time polymerase chain reaction and indirect immunofluorescence of PCV2-infected cells were used to evaluate the replication of PCV2.Results: Both GSH and SOD decreased significantly at 48 hours after PCV2 infection, whereas MDA concentration increased significantly after 48 hour post-infection. Furthermore, PCV2 replication in PK15 cells was significantly impaired after the elevation of intracellular GSH through treatment with the antioxidant N-acetyl-L-cysteine (NAC), a precursor in GSH synthesis. In contrast, PCV2 replication in PK15 cells was enhanced after reduction of GSH levels through H2O2-mediated oxidation. In addition, NAC treatment blocked the increase of virus replication induced by H2O2.Conclusions: This study suggests that PCV2 infection induces oxidative stress and that intracellular redox status influences PCV2 replication in PK15 cells.