猪圆环病毒2型诱导小鼠体内免疫细胞氧化胁迫模型的建立

doi:10.16431/j.cnki.1671-7236.2017.02.015

摘要/Abstract

摘要：

试验旨在建立猪圆环病毒2型（PCV2）诱导的小鼠体内免疫细胞氧化胁迫模型。筛选PCV2感染剂量及感染时间，采用腹腔注射、滴鼻和灌胃3种途径联合方式于第1、2、3 天或第1、3、5、7天给予昆明系小鼠感染PCV2病毒原液或10^－1 PCV2病毒液，分别于感染后7、14、21 d剖杀小鼠，测定活性氧（ROS）、总谷胱甘肽（T-GSH）、还原型谷胱甘肽（GSH）、氧化型谷胱甘肽（GSSG）水平及黄嘌呤氧化酶（XOD）、髓过氧化物酶（MPO）、诱导型一氧化氮合酶（iNOS）活性，探讨PCV2感染时间与活性氧水平变化的关系，建立免疫细胞氧化胁迫动物模型。结果显示，第1、2、3天每天经3种途径联合感染PCV2病毒原液，1 mL/只，为后续试验感染最佳方案。PCV2感染小鼠3个时间点细胞内ROS水平较空白对照组均极显著升高（P < 0.01），感染后7、14 d GSH水平显著降低（P < 0.05）；感染后7 d GSSG水平极显著高于空白对照组（P < 0.01）；感染后7 d T-GSH水平显著低于空白对照组（P < 0.05），感染后14 d T-GSH水平极显著低于空白对照组（P < 0.01）。感染后7 d脾脏XOD、MPO、iNOS活性与空白对照组相比均存在显著差异（P < 0.05）。结果表明，PCV2成功感染小鼠，试验感染方案为第1、2、3 天每天经3种途径联合感染PCV2病毒原液，1 mL/只，且用PCV2病毒原液感染7 d是建立小鼠体内免疫细胞氧化胁迫模型的最佳条件。

关键词: 猪圆环病毒2型; 小鼠; 免疫细胞; 氧化胁迫模型

Abstract:

The aim of this experiment was to establish the oxidative stress model of immune cells in mice induced by PCV2. Optimal infection titer and time of PCV2 were selected. On the 1st,2nd and 3rd day or 1st, 3rd,5th and 7th day,Kunming mice were treated with 10⁰ PCV2 or 10^-1 PCV2 by 3 ways (intraperitoneal injection, intranasal administration and intragastrical administration). 7, 14, 21 d post administration, we killed the mice. Reactive oxygen species (ROS),total glutathione (T-GSH),reduced glutathione (GSH),oxidized glutathione (GSSG) levels and xanthine oxidase (XOD), myeloperoxidase (MPO) and inducible nitric oxide synthase (iNOS) activity were determined to investigate the relationship between the infection time point and the change of ROS level after PCV2 infection. Thus to establish mice immune cells oxidative stress model. The results showed that infection with 10⁰ PCV2 virus in 3 ways every day both at day 1, 2, 3 and 1 mL per mouse was the best program for the follow-up experiment. The intracellular level of ROS of mice infected with PCV2 was extremely significantly increased both at 7, 14 and 21 d (P < 0.01);7 and 14 d post infection, GSH level of PCV2 infected mice had significant difference compared with that of control group (P < 0.05). 7 d after infection, GSSG level was extremely significantly higher than that of control group (P < 0.01); 7 d after infection, T-GSH level was significantly lower than that of control group (P < 0.05), T-GSH was extremely significantly lower than that of control group at 14 d post infection (P < 0.01). XOD, MPO and iNOS activity between PCV2 infection group and blank control group were significantly different at 7 d post infection (P < 0.05). It suggested that PCV2 successfully infected mice, experimental infection programme was Kunming mice were treated with 10⁰ PCV2 by three ways (intraperitoneal injection, intranasal administration and intragastrical administration) both at the 1st,2nd and 3rd day of the experiment.The best condition to establish the oxidative stress model of immune cells in mice was using 10⁰ PCV2 to infect for 7 days.

Key words: PCV2; mice; immune cells; oxidative stress model

中图分类号:

S852.65

杨剑, 尹丹, 罗文涓, 李璐, 文露婷, 郝祝兵, 韦英益, 曾芸, 胡庭俊. 猪圆环病毒2型诱导小鼠体内免疫细胞氧化胁迫模型的建立[J]. 《中国畜牧兽医》, 2017, 44(2): 402-410.

YANG Jian, YIN Dan, LUO Wen-juan, LI Lu, WEN Lu-ting, HAO Zhu-bing, WEI Ying-yi, ZENG Yun, HU Ting-jun. Establishment of Model for Oxidative Stress in Mice Immune Cells Induced by PCV2 in vivo[J]. , 2017, 44(2): 402-410.

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