衢枳壳黄酮组分抑制脂多糖诱导RAW264.7细胞炎症反应的作用机制研究

Anti-inflammatory Effect of the Flavonoids from Quzhiqiao (Fructus Auranti)on LPS-induced RAW264.7 Cells

目的:研究衢枳壳黄酮组分(the flavonoids from Quzhiqiao,TFQ)对脂多糖(lipopolysaccharides,LPS)诱导的RAW264. 7细胞的抗炎作用机制。方法:LPS(1 mg·L^-1)诱导RAW264. 7细胞建立细胞炎症模型。四甲基偶氮唑盐(MTT)法检测不同浓度TFQ对RAW264.7细胞活性的影响,RT-PCR法检测细胞炎性因子肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和白细胞介素1β(IL-1β) mRNA表达情况,Western blot法和免疫荧光法分别检测细胞核因子-κB(NF-κB p65)表达情况和入核情况。结果:TFQ在10~100 mg·L^-1时,对RAW264. 7细胞活性无影响。RT-PCR结果表明,TFQ能够降低LPS诱导的RAW264. 7细胞炎性因子TNF-α、IL-6和IL-1β表达,以TFQ 100 mg·L^-1作用显著(P <0. 05);Western blot法和免疫荧光结果表明,TFQ能够显著抑制p65蛋白表达与入核。结论:TFQ具有抑制LPS诱导的RAW264. 7细胞炎症反应的作用,其机制可能与抑制p65活化减少炎性因子表达有关。

Objective:To investigate the anti-inflammatory mechanism of the flavonoids from Quzhiqiao( TFQ) on LPS-induced RAW264.7 cells.Methods:RAW264.7 cells were induced to establish a cellular inflammatory model by LPS( 1 mg·L-1).The proliferation effect of TFQ with different concentrations on RAW264.7 cells was detected by using MTT assay.The mRNA expressions of TNF-α,IL-6 and IL-1β were detected by RT-PCR.Western blot and immunofluorescence were used to detect p65 expression and its nuclear translation.Results:TFQ had no effect on RAW264.7 cell viability at the dose 10 to 100 mg·L-1.The results of RT-PCR showed that TFQ could reduce the mRNA expressions of TNF-α,IL-6 and IL-1β on LPS-induced RAW264.7 cells,especially TFQ 100 mg·L-1( P < 0.05).The results of Western blot and immunofluorescence showed that TFQ could significantly inhibit the protein expression of p65 and promote its transport to nucleus.Conclusion:TFQ can inhibit the LPS-induced inflammatory response in RAW264.7 cells,which mechanism may be related to inhibit the activation of p65 to decrease the expression of inflammatory cytokines.