脂多糖诱导仔猪小肠上皮细胞炎性损伤模型的建立  

Establishment of Inflammatory Injury Model of Intestinal Porcine Epithelial Cells Induced by Lipopolysaccharide

作　　者：赵磊 刘云卓 王毅 孙会[2] 张爱忠[1] 李沐阳 Zhao Lei;Liu Yunzhuo;Wang Yi;Sun Hui;Zhang Aizhong;Li Muyang(College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University,Daqing 163319;College of Animal Science and Technology,Jilin Agricultural University)

机构地区：[1]黑龙江八一农垦大学动物科技学院,大庆163319 [2]吉林农业大学动物科学技术学院

出　　处：《黑龙江八一农垦大学学报》2024年第3期50-57,共8页journal of heilongjiang bayi agricultural university

基　　金：国家重点研发计划(2017YFD0500506);黑龙江八一农垦大学学成、引进人才科研启动计划(NO.XYB202109)。

摘　　要：LPS诱导IPEC-J2细胞炎性损伤模拟仔猪体内早期断奶应激,构建IPEC-J2细胞炎性损伤模型。首先采用0、0.1、1、10、100μg·mL^(-1)LPS处理IPEC-J2细胞,MTS法检测LPS处理IPEC-J2细胞0、4、8、12、16、24 h后的增殖率,初步确定炎性损伤模型构建的适宜浓度为10μg·mL^(-1),适宜时间为16 h。为了进一步确定LPS的最佳浓度,采用0~100μg·mL^(-1)LPS处理IPEC-J2细胞16 h,探究不同浓度LPS对IPEC-J2细胞的影响。结果表明:0.1~100μg·mL^(-1)LPS均可以不同程度的提高IPECJ2细胞IL^(-1)β、IL-6、IL-8和TNF-α含量及基因表达量,下调ZO-1、Occludin、Claudin-1基因表达量,降低IL-10、IgM、IgA和IgG含量,其中10μg·mL^(-1)的作用效果最佳;0.1~100μg·mL^(-1)LPS均可以不同程度的提高IPEC-J2细胞中ROS含量和细胞凋亡率,其中10μg·mL^(-1)的作用效果最佳。由此可见,LPS诱导细胞构建炎性损伤模型的适宜条件为10μg·mL^(-1)和16 h。LPS-induced inflammatory injury in IPEC-J2 cells simulated early weaning stress in piglets in vivo,and built an inflammatory injury model of IPEC-J2 cells.Firstly,IPEC-J2 cells were treated with LPS of 0,0.1,1,10 and 100μg·mL^(-1).MTS method was used to detect the proliferation rate of IPEC-J2 cells treated with LPS for 0,4,8,12,16 and 24 h.It was preliminarily determined that the appropriate concentration for the establishment of inflammatory injury model was 10μg·mL^(-1),and the appropriate time was 16 h.In order to further determine the optimal concentration of LPS,IPEC-J2 cells were treated with LPS at concentrations of 0,0.1,1,10 and 100μg·mL^(-1)for 16 hours to explore the effects of different concentrations of LPS on IPEC-J2 cells.The results showed that 0.1~100μg·mL^(-1)LPS increased the contents or genes of IL^(-1)β,IL-6,IL-8 and TNF-α,down-regulate the expression of ZO-1,occludin,and claudin-1 genes,and decreased the contents of IL-10,IgM,IgA and IgG in IPEC-J2 cells to varying degrees.The result of 10μg·mL^(-1)LPS played a better effect.0.1~100μg·mL^(-1)LPS increased the ROS content and apoptosis rate in IPEC-J2 cells to varying degrees,the result of 10μg·mL^(-1)LPS played a better effect.On the basis of not affecting subsequent experiments,the result of 10μg·mL^(-1)played a better effect.It was concluded that the appropriate conditions for LPS induced cells to construct an inflammatory injury model were 10μg·mL^(-1)and 16 h.

关 键 词：仔猪 IPEC-J2细胞 脂多糖 炎性损伤模型 

分 类 号：S816[农业科学—饲料科学]