β-连环蛋白在脂多糖诱导巨噬细胞炎性因子表达中的作用  

Effect of β-catenin on lipopolysaccharide-induced pro-inflammatory cytokines expression in macrophages

作　　者：何永辉[1] 陈佳[2] 谈健[1] 胡建鹏[1] 崔飞伦[1] 

机构地区：[1]江苏大学附属人民医院泌尿外科,镇江212002 [2]第二军医大学附属长征医院心内科

出　　处：《中华实验外科杂志》2015年第3期576-578,共3页Chinese Journal of Experimental Surgery

摘　　要：目的 探讨β-连环蛋白（β-catenin）缺陷对脂多糖（LPS）诱导巨噬细胞炎性因子表达的影响及其机制.方法 分离培养野生型及β-catenin缺陷小鼠腹腔巨噬细胞,分为4组（n=5）,β-catenin=^＋/＋、β-catenin^-/-、β-catenin=^＋/＋LPS、β-catenin^-/-＋LPS组.Western blot检测巨噬细胞β-catenin表达及p65磷酸化水平的变化,反转录-聚合酶链反应（RT-PCR）定量分析肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1β和IL-6等炎性因子的表达;酶联免疫吸附试验（ELISA）检测细胞上清中TNF-α、IL-1β和IL-6的浓度.结果 β-catenin=^＋/＋ LPS组β-catenin蛋白表达（0.717±0.101）与β-catenin=^＋/＋组（0.524±0.091）比较增加,两组差异有统计学意义（P＜0.05）;β-catenin^-/-＋LPS组与β-catenin^＋/＋＋ LPS组比较TNF-α、IL-1β和IL-6 mRNA及细胞上清中TNF-α、IL-1β和IL-6的浓度都较β-catenin^＋/＋＋ LPS组明显增高,两组差异有统计学意义（P＜0.05）;β-catenin^-/-＋LPS组p65的磷酸化水平（0.9646±0.1510）比β-catenin^＋/＋＋ LPS组（0.728 8±0.093 0）增加,两组差异有统计学意义（P＜0.05）.结论 β-catenin缺陷使LPS诱导巨噬细胞活化增加,促进炎性因子的分泌,其机制可能与β-catenin促进p65磷酸化,增加核因子-κB （NF-κB）的转录活性有关.Objective To investigate the effect of β-catenin on the pro-inflammatory cytokines expression in murine peritoneal macrophage sstimulated with lipopolysaccharide （LPS） and the molecular mechanisms.Methods The peritoneal macrophages obtained from the β-catenin knockout mice and wild-type mice were divided into four groups （n =5 each）：β-catenin^＋/＋ group,β-catenin^-/-group,β-catenin^＋/＋ ＋LPS group and β-catenin^-/-＋ LPS group.The protein level of β-catenin and the phosphorylated level of p65 in macrophages were determined by Western blotting.The tumor necrosis factor-α （TNF-α）,interleukin （IL）-1 b and IL-6 mRNA levels were detected by reverse transcriptase-polymerase chain reaction （RT-PCR）.The concentrations of TNF-α,IL-1β and IL-6 in the supernatants were measured by an enzyme-linked immunosorbent assay （ELISA）.Results Compared with the β-catenin^＋/＋ group （0.524 ± 0.091）,β-catenin expression in β-catenin^＋/＋ ＋ LPS group （0.717 ± 0.101） was increased （P 〈 0.05） after LPS stimulation.The mRNA levels of TNF-α,IL-1β and IL-6 and the concentrations of TNF-α,IL-1β and IL-6 in the supernatants were significantly increased in β-catenin^-/-＋ LPS group as compared with β-catenin ^＋/＋ group （P 〈 0.05）.P65 phosphorylation was increased markedly in β-catenin^-/-＋ LPS group as compared with β-catenin^＋/＋ group （0.964 6 ±0.151 0 vs.0.728 8 ±0.093 0,P 〈0.05）.Conclusion LPS increased β-catenin in murine peritoneal macrophages and β-catenin knockdown promotes macrophage activation partly through increased phosphorylation of p65.

关 键 词：Β-连环蛋白 P65 巨噬细胞 

分 类 号：R392[医药卫生—免疫学]