柔嫩艾美耳球虫子孢子感染对CEK细胞免疫因子表达及抗氧化能力的影响  

Effects of Eimeria tenella sporozoite infection on the expression of immune factors and antioxidant capacity of CEK cells

作　　者：汤建强 于海亮 董理月 唐美慧 AREEJ Arif 范作堃 张涛[1] 张跟喜[1] 谢恺舟[1] 赵振华[2] 戴国俊[1] TANG Jianqiang;YU Hailiang;DONG Liyue;TANG Meihui;AREEJ Arif;FAN Zuokun;ZHANG Tao;ZHANG Genxi;XIE Kaizhou;ZHAO Zhenhua;DAI Guojun(College of Animal Science and Technology/Ministry of Education Joint International Research Laboratory of Agriculture and Agri-Product Safety,Yangzhou University,Yangzhou 225009,China;Poultry Institute,Chinese Academy of Agricultural Sciences,Yangzhou 225125,China)

机构地区：[1]扬州大学动物科学与技术学院/教育部农业与农产品安全国际合作联合实验室,江苏扬州225009 [2]中国农业科学院家禽研究所,江苏扬州225125

出　　处：《黑龙江畜牧兽医》2024年第21期62-68,126,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家现代农业产业技术体系专项(CARS-41-G23);江苏现代农业产业技术体系建设专项(JATS[2023]447);江苏省高校优势学科建设工程项目(PAPD)。

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-1βmRNA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-αmRNA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。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-1βin 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-αmRNA 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 expressio

关 键 词：柔嫩艾美耳球虫 子孢子 CEK细胞 细胞因子 基因表达 

分 类 号：S852.723[农业科学—基础兽医学]