群体感应分子3-氧代癸酰基高丝氨酸内酯(3-oxo-C10-HSL)抑制脂多糖诱导的RAW264.7巨噬细胞炎症反应

Quorum sensing molecule N-3-oxodecanoyl-L-homoserine lactone (3-oxo-C-HSL) inhibits lipopolysaccharide-induced inflammatory responses of RAW264.7 macrophages

目的探讨群体感应分子3-氧代癸酰基高丝氨酸内酯(3-oxo-C10-HSL)对脂多糖(LPS)诱导的RAW264.7巨噬细胞炎症反应的调控作用。方法RAW264.7巨噬细胞分为实验组、对照组与空白组。实验组用不同浓度的3-oxo-C10-HSL与LPS共处理,对照组加入DMSO与LPS,空白组加入DMSO与PBS。刺激12 h后,实时定量PCR检测细胞白细胞介素6(IL-6)、IL-1β、肿瘤坏死因子α(TNF-α)、单核细胞趋化蛋白1(MCP-1)的mRNA表达,ELISA检测上清液IL-6、TNF-α水平。以25μmol/L 3-oxo-C10-HSL与100 ng/mL LPS共刺激细胞15、30、60 min,Western blot法检测核因子κBp65(NF-κBp65)的磷酸化情况。结果3-oxo-C10-HSL能剂量依赖性地降低LPS诱导的RAW264.7巨噬细胞的IL-6、IL-1β、TNF-α、MCP-1的mRNA水平以及IL-6、TNF-α蛋白水平。3-oxo-C10-HSL能抑制LPS诱导的NF-κBp65磷酸化。结论3-oxo-C10-HSL能通过抑制NF-κB信号通路活化减轻LPS诱导的RAW264.7巨噬细胞炎症反应。

Objective To explore the regulatory effect of quorum sensing molecule N-3-oxodecanoyl-L-homoserine lactone(3-oxo-C10-HSL)on lipopolysaccharide(LPS)-induced inflammation in RAW264.7 macrophages.MethodsRAW264.7 macrophages were divided into experimental group,control group and blank group.The experimental group was treated with different concentrations of 3-oxo-C10-HSL and LPS;the control group was treated with DMSO and LPS;and the blank group was treated with DMSO and PBS.Cells and supernatants were collected after 12 hours of stimulation.The mRNA expression of interleukin-6(IL-6),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and monocyte chemoattractant protein-1(MCP-1)were detected by real-time quantitative PCR,and the protein levels of IL-6 and TNF-αin supernatant were detected by ELSIA.Further,25μmol/L 3-oxo-C10-HSL and 100 ng/mL LPS were used to stimulate the cells for 15,30 and 60 minutes,and the phosphorylation of nuclear factorκBp65(NF-κBp65)was detected by Western blot analysis.Results The 3-oxo-C10-HSL could decrease the mRNA levels of IL-6,IL-1β,TNF-α,MCP-1 and the protein levels of IL-6 and TNF-αin LPS-treated RAW264.7 macrophages in a dose-dependent manner.In addition,3-oxo-C10-HSL could inhibit the phosphorylation of NF-κBp65 induced by LPS.Conclusion 3-oxo-C10-HSL can alleviate LPS-induced inflammatory responses in RAW264.7 macrophages by inhibiting activation of NF-κB signaling pathway.