连翘脂素对LPS诱导RAW264.7细胞炎症反应的影响

为研究连翘脂素的抗炎效应及其抗炎机制,以地塞米松作为阳性对照,建立脂多糖(LPS)诱导小鼠巨噬细胞RAW264.7炎症模型,检测炎症因子的释放及相关蛋白和mRNA的表达,以期提高对连翘脂素抗炎作用的全面认识并为连翘脂素临床开发提供有力的科学依据。实验采用Griess法检测细胞上清液中NO含量,ELISA法检测TNF-α和IL-6的含量,Westernblot法检测iNOS、COX-2蛋白的表达,RT-qPCR法检测iNOS、COX-2mRNA的表达。与LPS组比较,连翘脂素组和地塞米松组可以明显降低LPS诱导的RAW264.7细胞释放NO、TNF-α和IL-6的量,并呈现浓度依赖关系。Westrenblot和RT-qPCR结果显示连翘脂素能抑制LPS诱导的iNOS、COX-2的蛋白表达以及mRNA的表达,并呈浓度依赖关系。实验研究表明连翘脂素能够明显抑制LPS诱导的RAW264.7细胞炎症因子的释放,iNOS、COX-2蛋白及mRNA的表达从而抑制炎症反应。

Effect of Phillygenin on inflammatory response in LPS-induced RAW 264.7 cells

To study the anti-inflammatory effect and anti-inflammatory mechanism of Phillygenin,dexamethasone as a positive control,established a lipopolysaccharide (LPS)-induced RAW264.7 inflammation model,to detect the release of inflammatory factors and the expression of related proteins and mRNA,in order to improve the comprehensive understanding of the anti-inflammatory effect of Phillygenin and provide a strong scientific basis for the clinical development of Phillygenin.The NO production was determined by assaying nitrite in culture supernatants with the Griess reagent.The levels of TNF-α and IL-6 in culture media were measured with ELISA kits.Western blot assay was performed to illustrate the inhibitory effects of Phillygenin on iNOS and COX-2.RT-qPCR was detected for mRNA expression of iNOS and COX-2.Treatment with Phillygenin or Dexamethasone dose-dependently inhibited the production expression of NO,TNF-α and IL-6 in RAW264.7 macrophages.Western blot and RT-qPCR assay suggested that the mechanism of the anti-inflammatory effect was associated with the inhibition of iNOS and COX-2.In conclusion,Phillygenin exhibited obvious anti-inflammatory effect and its mechanism may be related to its regulation on inflammatory cytokines,and inhibition of the expression of iNOS and COX-2.