玉米赤霉烯酮对IPEC-J2细胞炎症相关因子mRNA表达和活性的影响  

Effect of ZEN on mRNA Expression and Activity of Inflammation Related Factors in IPEC-J2 Cells

作　　者：杨丽革 牟洋[1] 张薇[1] 李建平[1] 单安山[1] 

机构地区：[1]东北农业大学动物营养研究所,黑龙江哈尔滨150030

出　　处：《中国畜牧杂志》2017年第6期75-81,共7页Chinese Journal of Animal Science

基　　金：国家重点研发计划(2016YFD0501207);现代农业产业技术体系建设专项(CARS-36);动物普通疾病防治重点实验室开放课题

摘　　要：本试验旨在探讨玉米赤霉烯酮(Zearalenone,ZEN)对仔猪空肠上皮细胞(IPEC-J2)炎症相关因子m RNA表达和活性的影响。试验选取不同浓度(0~80μg/m L)ZEN处理IPEC-J2细胞12、24、48 h,CCK-8法检测细胞存活率并计算得出半数抑制浓度(IC50)。根据IC50选取用不同浓度ZEN(0、5、10、15μg/m L)处理细胞12、24、48 h,倒置显微镜下观察细胞形态,RT-PCR法测定炎症相关因子NF-κB、COX-2、PGES、IL-6、IL-8m RNA的表达量,ELISA方法检测炎症相关因子的活性。结果表明:ZEN以时间和剂量依赖性降低IPEC-J2细胞存活率,12、24、48 h ZEN的IC50分别为41.09、28.54、19.85μg/m L;随ZEN浓度升高和处理时间的延长,细胞形态被逐渐破坏;除COX-2的m RNA表达量和活性在12 h随ZEN浓度升高而升高外(P<0.05),炎症相关因子m RNA表达量和活性均随ZEN浓度的升高呈先升高后降低(P<0.05);ZEN浓度和处理时间对炎症相关因子m RNA表达量和活性的影响均有极显著的交互作用(P<0.001)。以上结果表明,ZEN能降低细胞存活率,破坏细胞形态,ZEN对细胞炎症相关因子m RNA表达和活性具有双向调节作用。The purpose of this study was to evaluate the effects of ZEN on mRNA expression of inflammation related factors in IPEC-J2. Cells were treated with different concentrations of ZEN （0-80 μg/mL） for 12, 24, 48 h. Cell viability was estimated by CCK-8 assay and the IC50 was calculated. After cells were treated with different concentrations of ZEN （0, 5, 10, 15 μg/mL） for 12, 24, 48 h, the changes of cells morphology were observed and the mRNA expression of NF-κB, PGES, COX-2, IL-6, IL-8 were detected by RT-PCR. The results showed as follows： the cell viability was reduced in a dose/time-dependent manner, IC50 of ZEN were 41.09, 28.54 and 19.85 μg/mL, respectively. With the increasing of ZEN concentrations and the extension of treatment time, the morphology of cells was gradually destroyed. In addition to the COX-2 mRNA expression increased with dose dependent of ZEN at 12h （P〈0.05）, the mRNA expression increased first then decreased with the concentration increased （P〈0.05）. When ZEN concentration was 5 μg/mL, the NF-κB in 12 h, PGES and IL-8 in 24 h, IL-6 and COX-2 in 48 h mRNA expression highest; when ZEN concentration was 10 μg/mL, the NF-KB, PGES and IL-8 in 12 h, COX-2 in 24 h, IL-6 in 48 h mRNA expression highest; when ZEN concentration was 15 μg/mL, all inflammation related factors mRNA expressed highest in 12 h. The interactions of ZEN concentration and treatment time on the expression of inflammatory cytokine mRNA was highly significant （P〈0.001）. The current findings indicated that ZEN could reduce cell viability and destroy cell morphology, ZEN had dual regulatory functions in expression of inflammation related factors.

关 键 词：玉米赤霉烯酮 IPEC-J2 炎症相关因子 

分 类 号：S828.5[农业科学—畜牧学]