芪冬活血饮对巨噬细胞Cav-1/NF-κB炎性反应信号通路的影响

Effects of Qidong Huoxue Decoction on Cav-1/NF-κB inflammatory signaling pathway in macrophages

目的:通过观察芪冬活血饮药物血清对脂多糖(LPS)致炎巨噬细胞株小窝蛋白(Cav-1)/细胞核因子-κB(NF-κB)炎性反应信号通路的影响,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。培养24h后,加入血清培养0.5h,后加入LPS培养12h,测定各组上清液肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-10(IL-10)水平,及细胞核因子-κB(NF-κB)p65、小窝蛋白-1(Cav-1)蛋白和m RNA表达情况。结果:LPS刺激各组TNF-α、IL-1β、IL-10水平增高,芪冬活血饮药物血清可降低TNF-α、IL-1β水平,升高IL-10水平,与其余各组比较差异均有统计学意义(P<0.01)。LPS刺激各组NF-κB p65、Cav-1蛋白及m RNA表达增加,芪冬活血饮药物血清可减少其表达,与其余各组比较差异均有统计学意义(P<0.01)。结论:芪冬活血饮药物血清可减轻LPS刺激的巨噬细胞炎性反应,其机制与抑制Cav-1/NF-κB p65炎性反应信号通路,降低促炎细胞因子TNF-α、IL-1β水平,升高抗炎细胞因子IL-10水平,纠正炎性反应失衡有关。

Objective： To observe the effect of serum containing Qidong Huoxue Decoction（QD） on Cav-1/NF-κB inflammatory signaling pathway of macrophages stimulated by lipopolysaccharide（LPS）, in order to explore the regulation mechanism of inflammatory reaction. Methods： The macrophages were divided into six groups as control group, LPS group, blank serum group, LPS combined with blank serum group, drug-serum group, and the LPS combined with drug-serum group. After culturing for 24 h, cells were cultured for 0.5 h with serum added, and then the cells were cultured for another 12 h with LPS added. Levels of TNF-α, IL-1β and IL-10 in cell supernatant, and the protein and m RNA expression of NF-κB p65 and Cav-1 of cells in each group were detected. Results： The LPS stimulation could increase the levels of TNF-α, IL-1β and IL-10 of cells. Serum containing QD could reduce the levels of IL-1β and TNF-α, and increase the level of IL-10, the differences were statistically significant compared with the rest groups（P0.01）. The protein and m RNA expression of NF-κB p65 and Cav-1 of cells in LPS stimulated groups increased, while serum containing QD could reduced the expression, and the differences were statistically significant compared with the rest groups（P0.01）. Conclusion： Serum containing QD could relieve the inflammatory responses of macrophages stimulated by LPS, and the mechanism related with the inhibition Cav-1/NF-κB p65 signaling pathway, reduction on TNF-α and IL-1β levels, promotion on IL-10 levels, and correction unbalanced inflammation.