【摘要】 为了探究柔嫩艾美耳球虫(E.tenella)子孢子感染对鸡胚肾(CEK)细胞免疫反应及抗氧化程度的影响，试验在成功构建E.tenella感染CEK细胞模型的基础上，将CEK细胞分为空白组和感染组，采用酶联免疫吸附试验(ELISA)和实时荧光定量PCR方法分别检测子孢子感染后第3,9,12,24小时时细胞上清液中与E.tenella感染相关的4种免疫因子白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、白细胞介素-1β(IL-1β)及肿瘤坏死因子-α(TNF-α)的浓度及其在CEK细胞中mRNA相对表达量，同时检测细胞上清液中4种氧化指标过氧化氢酶(CAT)、超氧化物歧化酶(SOD)活性及一氧化氮(NO)和活性氧(ROS)浓度。结果表明：子孢子感染后第3,9小时时，感染组IL-6、IL-8及IL-1β mRNA相对表达量及其浓度均极显著高于空白组(P<0.01),且随着感染时间的延长呈下降趋势；感染后第12,24小时时，感染组TNF-α mRNA相对表达量和其浓度均极显著高于空白组(P<0.01),且在感染子孢子后呈上升趋势。此外，感染组CAT和SOD活性在子孢子感染后第3,9,12小时时极显著低于空白组(P<0.01),并呈下降趋势；在感染后第3,9小时时，感染组NO和ROS浓度显著或极显著高于空白组(P<0.05或P<0.01),这些氧化指标的变化主要发生在感染后的9 h或12 h内。说明E.tenella的感染会显著影响CEK细胞的氧化动态平衡与免疫细胞因子的表达水平。更多还原

【Abstract】 In order to investigate the effects of Eimeria tenella(E. tenella) sporozoite infection on the immune response and antioxidant level of chicken embryo kidney(CEK) cells, in the experiment, based on the successful construction of an E. tenella-infected CEK cell model, CEK cells were divided into blank group and infection group; enzyme-linked immunosorbent assay(ELISA) and real-time quantitative PCR technology were used to detect the concentrations of four immune factors in cell supernatants, interleukin-6(IL-6), interleukin-8(IL-8), interleukin-1β(IL-1β), and tumor necrosis factor-α(TNF-α), and their relative mRNA expression levels in CEK cells at 3, 9, 12, and 24 h after sporozoite infection respectively, which were related to E. tenella infection; at the same time, four antioxidant indexes catalase(CAT), superoxide dismutase(SOD), nitric oxide(NO) and reactive oxygen species(ROS) were detected in cell supernatants. The results showed that at 3 and 9 h after sporozoite infection, the relative mRNA expression and protein concentration of IL-6, IL-8 and IL-1β in the infection group were significantly higher than those in the blank group(P<0.01), which showed a downward trend with the extension of infection time. At 12 and 24 h after infection, the relative expression and protein concentration of TNF-α mRNA in the infection group were significantly higher than those in the blank group(P<0.01), which showed an upward trend after sporozoite infection. In addition, the activities of CAT and SOD in the infection group were significantly lower than those in the blank group at 3, 9 and 12 h after sporozoite infection(P<0.01), which showed a downward trend. At 3 and 9 h after infection, the concentrations of NO and ROS in the infection group were significantly or extremely significantly higher than those in the blank group(P<0.05 or P<0.01). The changes in these antioxidant indexes mainly occurred within 9 h or 12 h after infection. These results suggested that E. tenella infection significantly affected the oxidative homeostasis and immune cytokine expression levels of CEK cells.更多还原