LPS刺激对鸡HD11细胞RIPK2基因甲基化的影响及调控  

Effect and Regulation of LPS Stimulation on the Methylation of RIPK2 Gene in Chicken HD11 Cells

作　　者：李欢 马武生[1] 孙长花[1] LI Huan;MA Wu-sheng;SUN Chang-hua(Yangzhou Polytechnic College,Yangzhou 225009,China)

机构地区：[1]扬州职业大学,江苏扬州225009

出　　处：《扬州职业大学学报》2023年第1期22-27,共6页Journal of Yangzhou Polytechnic College

基　　金：江苏省高校“青蓝工程”资助项目(2022JS-29);扬州职业大学“中青年学术带头人”培养项目(yzd2020-08);扬州市“英才培育计划”资助项目(2020YZYC-098)。

摘　　要：为分析LPS刺激前后对RIPK2基因甲基化的影响,试验采用凝胶电泳、亚硫酸氢盐、双荧光素酶报告系统和qRT-PCR方法分析LPS刺激前后RIPK2基因的甲基化模式及甲基化相关试剂对鸡RIPK2启动子活性及表达水平影响。结果表明:LPS刺激后鸡RIPK2基因甲基化水平从53.8%下降至15.4%;5-氮杂胞苷显著性提高鸡RIPK2启动子活性,促进基因表达;而CpG甲基转移酶M. SssI显著性抑制鸡RIPK2启动子活性,抑制基因表达。研究结果对提高家禽免疫和控制过度性炎症反应具有十分重要的现实意义。To analyze the effect of LPS stimulation on the methylation level of chicken RIPK2 gene,gel electrophoresis,bisulfite,dual luciferase reporter system,and qRT-PCR are used to analyze the methylation pattern of RIPK2 gene before and after LPS stimulation,as well as the effects of methylation-related reagents on chicken RIPK2 promoter activity and its expression level.Results show that the methylation level of RIPK2 gene is decreased from 53.8%to 15.4%after LPS stimulation.Moreover,5-azacytidine can significantly activate the promoter activity of chicken RIPK2 and increase gene expression level.However,CpG methyltransferase M.SssI significantly inhibits the promoter activity of chicken RIPK2 and reduces gene expression level.These results are of great practical significance to improving poultry immunity and controlling excessive inflammatory response.

关 键 词：鸡RIPK2 LPS 5-氮杂胞苷 M.SssI 基因表达 

分 类 号：S858.31[农业科学—临床兽医学]