白藜芦醇对LPS诱导小鼠腹腔巨噬细胞NO和炎性因子生成的调控

Effects of Resveratrol on Production of NO and Pro - inflammatory Factors From Endotoxin Activated Murine Peritoneal Macrophages

目的探讨白藜芦醇（Res）对内毒素（LPS）诱导小鼠腹腔巨噬细胞核因子-κB（NF—κB）活化及炎性细胞因子（TNF—α、IL—1β、IL-6）基因表达的调节，为Res的临床运用提供理论依据。方法分别用LPS或Res＋LPS处理体外培养的小鼠巨噬细胞，采用电子顺磁共振（EPR）自旋捕集技术直接检测巨噬细胞产生的NO，电泳迁移率改变分析法（EMSA）检测细胞中NF—κB活性，逆转录一聚合酶链反应（RT—PCR）和酶联免疫吸附法（ELISA）检测细胞中TNF—α、IL—1β、IL-6 mRNA和蛋白的表达。结果表明LPS组NO含量、NF—κB活性和TNF—α、IL-1β、IL-6含量在刺激后2—12h明显高于正常对照组（P〈0．01），而Res＋LPS组NO含量、NF—κB活性和TNF—α、IL-1β、IL-6含量均显著低于LPS组（P〈0．01）。结论提示LPS可诱导巨噬细胞NF—KB活化，导致TNF-α、IL-1β、IL-6基因表达增强，而Res能抑制NF—κB活化而调节TNF—α、IL-1β、IL-6基因的表达。

Objective To investigate activation of nuclear factor -κB （NF -κB） and the expression of inflammatory cytokines in lipopolysaccharide - induced murine peritoneal macrophages stimulated with resveratrol （Res） Methods Peritoneal macrophages cultured in vitro were given with LPS or Res ＋ LPS. The NO produced from activated peritoneal macrophages was detected directly by electron paramagnetic resonance （EPR） spin trapping. The NF-κB activity of peritoneal macrophages was tested by electrophoretic mobility shift assay （EMSA） . The expression of TNF -α、IL-1β, IL- 6 in peritoneal macrophages were measured by reverse transcription polymerase chain reaction techniques （ RT - PCR ） and enzyme - linked immuno sorbent assay （ELISA ） . Results The results showed that the activity of NF - κB and the level of TNF -α、IL-1β, IL - 6 significantly increased from 2 tO i2 hour after LPS stimulation （ P 〈 0.01） Compared with LPS - stimulated group, both NF - κB activity and concentration of TNF -α、IL-1β, IL - 6 were significantly lowered in Res ＋ LPS group （ P 〈 0.01 } . Conclusion The results suggested LPS might activate NF - κB in peritoneal macrophages and induce the increase of transcription and expression of TNF -α、IL-1β, IL - 6 genes; Res could inhibit the activation of NF- κB and regulate the expression of TNF -α、IL-1β and IL - 6 genes in peritoneal macrophages.