芪冬活血饮对巨噬细胞TLR4/NF-κB炎性信号通路的影响

Effects of Qidong Huoxue Decoction on TLR4/NF-κB inflammation signaling pathway in macrophages

目的:观察芪冬活血饮药物血清对脂多糖致炎RAW264.7巨噬细胞TLR4/NF-κB炎性信号通路的影响,探讨其调控炎性反应的机制。方法:将巨噬细胞分为空白对照组、LPS组、空白血清组、空白血清加LPS组、药物血清组、药物血清加LPS组6组。细胞培养24h后,加入血清培养0.5h,然后加入LPS培养12h,测定细胞上清液细胞因子及TLR4、NF-κBp65蛋白及m NRA表达。结果:LPS刺激后TNF-α、IL-1β、IL-10水平升高,芪冬活血饮药物血清可降低TNF-α、IL-1β水平,升高IL-10水平,与空白血清加LPS组比较差异均有统计学意义(P<0.01)。LPS刺激后各组NF-κBp65、TLR4蛋白及m RNA表达增加,芪冬活血饮药物血清可减少NF-κBp65、TLR4蛋白及m RNA表达(P<0.05)。结论:芪冬活血饮药物血清可减轻LPS诱导的巨噬细胞炎性反应,其机制与抑制TLR4/NF-κBp65炎性信号通路及下游炎性因子有关。

Objective： To observe the effect of Qidong Huoxue Decoction（QD）-serum on TLR4/NF-kappa B inflammation signaling pathway in RAW264.7 cells, and explore its mechanism. Methods： RAW264.7 cells were divided into six groups： control group, LPS group, blank serum group, blank serum plus LPS group, drug serum group, drug serum plus LPS group. After culturing for 24 h, cells were added the QD-serum for 0.5h, then adding LPS for 12 h. The cytokines and the protein and m RNA expression of TLR4 and NF-κBp65 in cell supernatant were detected. Results： The levels of TNF-α, IL-1β, IL-10 were increased in the LPS stimulated groups. The QD-serum group reduced the levels of IL-1β and TNF-α, and increased the level of IL-10. There were statistical differences among the QD-serum group and blank serum plus PLS group（P〈0.01）. The protein and m RNA expression of NF-κBp65 and TLR4 in LPS stimulated groups increased, while QD-serum reduced the expression. Conclusion： QD-serum could relieve the inflammatory responses of macrophages stimulated by LPS, which might be related to the inhibition of TLR4/NF-κBp65 signaling pathway and downstream inflammatory factors.