槲皮素对LPS诱导小鼠RAW264.7细胞炎症的保护作用

目的研究槲皮素对脂多糖(LPS)诱导小鼠RAW264.7细胞炎症的保护作用。方法 CCK-8法检测细胞活力,Griess法检测NO释放,RT-PCR法检测TNF-α、IL-6、IL-1β、iNOS、COX-2、MCP-1、TLR2、TLR4、MyD88 mRNA表达,Western blot法检测iNOS、COX-2、TLR4、IκBα、p-IκBα、p-NF-κB p65蛋白表达。结果不同浓度(0~50μmol/L)槲皮素对细胞活力无明显影响(P>0.05)。与LPS组比较,槲皮素组(25、50μmol/L)显著抑制NO释放(P<0.05);槲皮素组(5、15、25μmol/L)显著降低TNF-α、IL-6、IL-1β、iNOS、COX-2、MCP-1、TLR4、MyD88 mRNA表达和iNOS、COX-2蛋白表达(P<0.05,P<0.01),并呈浓度依赖性;槲皮素组(25μmol/L)显著下调TLR4、p-IκBα、p-NF-κB p65蛋白表达(P<0.05),显著上调IκBα蛋白表达(P<0.05)。结论槲皮素可抑制LPS诱导小鼠RAW264.7细胞炎症,其机制可能与调节TLR4/NF-κB信号通路有关。

Protective effects of quercetin on the inflammation of mice RAW264.7 cells induced by LPS

AIM To study the protective effects of quercetin on inflammation of mice RAW264.7 cells induced by lipopolysaccharide(LPS).METHODS Cell viability was detected by CCK-8 method, NO release was detected by Griess method, TNF-α, IL-6, IL-1β, iNOS, COX-2, MCP-1, TLR2, TLR4 and MyD88 mRNA expressions were detected by RT-PCR, iNOS, COX-2, TLR4, IκBα, p-IκBα and p-NF-κB p65 protein expressions were detected by Western blot.RESULTS Different concentrations(0-50 μmol/L) of quercetin demonstrated no obvious effects on cell activity(P>0.05). Compared with LPS group, quercetin groups(25, 50 μmol/L) shared significantly inhibited NO release(P<0.05);quercetin groups(5, 15, 25 μmol/L) displayed markedly decreased TNF-α, IL-6, IL-1β, iNOS, COX-2, MCP-1, TLR4 and MyD88 mRNA expressions, and iNOS and COX-2 protein expressions(P<0.05, P<0.01) in a concentration-dependent manner;quercetin group(25 μmol/L) demonstrated obviously down-regulated TLR4, p-IκBα and p-NF-κB p65 protein expressions(P<0.05), and obviously up-regulated IκBα protein expression(P<0.05).CONCLUSION Quercetin can inhibit the inflammation of mice RAW264.7 cells induced by LPS, whose mechanism may contribute to the regulation of TLR4/NF-κB signaling pathway.