槲皮黄酮对脂多糖诱导BV-2小胶质细胞损伤的保护机制研究

Protective mechanism of quercetin on the BV-2 Microglia injury induced by LPS

关键词:  槲皮黄酮  BV-2细胞  炎症反应  Traf6/p65信号通路

Key words:  quercetin  BV-2 microglial cells  inflammatory response  Traf6/p65 signaling pathway

摘要:

目的 探讨槲皮黄酮对脂多糖（LPS）诱导的BV-2小神经胶质细胞炎症反应的抑制作用及其可能机制。方法 体外培养BV-2细胞，采用LPS刺激细胞诱导炎症模型，将细胞分为正常对照组、模型组和槲皮黄酮低中高（10，20，40 μg·mL^-1）剂量组，CCK8法检测槲皮黄酮对BV-2细胞增殖情况的影响，ELISA法检测细胞上清中TNF-α和IL-6的表达量，免疫印迹实验检测iNOS、COX-2、Traf6、p65及p-p65蛋白的表达水平。结果 槲皮黄酮可显著促进BV-2细胞的增殖，并明显降低细胞的炎症反应。槲皮黄酮可明显抑制TNF-α和IL-6的释放，还可显著抑制炎症相关蛋白iNOS及COX-2的表达，并且还可以抑制Traf6/p65信号通路的活化。结论 槲皮黄酮可明显抑制LPS诱导的BV-2细胞炎症反应，抑制炎症相关蛋白的表达，其可能作用机制是通过抑制Traf6/p65信号通路的活化实现的。

Abstract:

OBJECTIVE To observe the protective effects of quercetin on the BV-2 microglia injury induced by LPS and study the possible mechanism. METHODS BV-2 cells were cultured in vitro and LPS was used to stimulate the inflammatory model of BV-2 cells. The cells were divided into five groups:blank control group, model group, quercetin treatment groups (10, 20, 40 μg·mL^-1). CCK-8 kit was used to determine the relative survival rate of BV-2 cells and ELISA kits were used to measure the expression levels of IL-6 and TNF-α. Western blot was used to analysis the expression levels of iNOS, COX-2, Traf6, p65 and p-p65. RESULTS The BV-2 cells injury model was induced by LPS. The BV-2 cells relative survival rates after treatment for different concentrations of quercetin were significantly increased compared to model group, while the expression levels of inflammatory cytokines IL-6 and TNF-α were significnatly decreased compared with model group. The results of Western blot showed that the expression levels of iNOS, COX-2, Traf6 and p-p65 were significantly decreased after treatment with quercetin. CONCLUSION Quercetin can significantly inhibit the inflammatory response of BV-2 microglial cells induced by LPS via suppressing the activation of Traf6/p65 signaling pathway.