岗松总黄酮对脂多糖诱导RAW264.7细胞炎症的影响

Effect of Total Flavonoids from Baeckea frutescens on LPS-induced RAW264.7 Cell Inflammation

关键词:  岗松总黄酮  脂多糖  RAW264.7  NF-κB  抗炎

Key words:  total flavonoids from Baeckea frutescens  LPS  RAW264.7  NF-κB  anti-inflammation

摘要:

为探讨岗松总黄酮（GSH）的抗炎活性机制，通过NF-κB信号通路，研究脂多糖（LPS）诱导RAW264.7细胞炎症的保护作用。实验采用噻唑蓝（MTT）法检测GSH对RAW264.7细胞增殖活性的影响，采用酶联免疫法（ELISA）检测GSH对白介素（IL-1β、IL-8）及转换生长因子β （TGF-β）的影响，采用蛋白印迹法（WB）检测细胞中IκBα蛋白相对表达，采用实时定量基因扩增荧光检测法（QPCR）检测细胞中NF-κB p65的mRNA基因表达。研究结果表明， GSH处理细胞后，可促进RAW264.7细胞增殖（P<0.05），抑制IL-1β、IL-8、TGF-β含量（P<0.01），上调IκBα蛋白的相对表达（P<0.05或P<0.01），抑制NF-κB p65的mRNA基因表达（P<0.01）。推测岗松总黄酮抑制NF-κB信号通路可能是其抗炎活性作用机制之一。

Abstract:

In order to investigate the anti-inflammation activity mechanism of total flavonoids from Baeckea frutescens (GSH)，the protective effect of LPS-induced RAW264.7 cells was studied by NF-κB signaling pathway. MTT method was used to detect the effect of GSH on the proliferation activity of RAW264.7 cells. The effect of GSH on interleukin (IL-1β, IL-8) and transforming growth factor β (TGF-β) was detected by enzyme-linked immunosorbent assay (ELISA). The relative expression of IκBα protein in cells was detected by Western blotting (WB). The mRNA expression of NF-κB p65 in cells was detected by real-time quantitative gene amplification fluorescence assay (QPCR). The results show that after treated by GSH, the proliferation of RAW264.7 cells was promoted (P<0.05), the content of IL-1β、IL-8 and TGF-β were inhibited significantly (P<0.01), the IκBα protein expression was enhanced significantly (P<0.05 or P<0.01), the NF-κB p65 mRNA expression was inhibited significantly (P<0.01). It is speculated that the inhibition of NF-κB signaling pathway by total flavonoids may be one of the anti-inflammatory mechanisms.