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目的初步探讨土荆皮乙酸(PLAB)对脂多糖(LPS)诱导RAW264.7细胞抗炎作用及影响M1表型偏移的分子机制。方法 LPS诱导RAW264.7细胞建立体外炎症模型,给予0.5μmol/L PLAB和1μmol/L过氧化物酶体增殖物激活受体γ(PPARγ)阻滞剂GW9662处理。流式细胞术检测细胞周期变化,实时定量PCR检测PPARγ和M1型巨噬细胞标志物白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)的mRNA表达,Western blot法检测核因子κB(NF-κB)信号通路相关信号分子水平。结果 PLAB能够明显降低LPS诱导RAW264.7细胞的IL-1β、TNF-α的mRNA水平,上调PPARγ的mRNA水平。下调NF-κB p65、p NF-κB p65、IKKα、IKKβ、p IKKα/β、IκBα、p IκBα的蛋白水平,使RAW264.7细胞阻滞在G0和G2期。GW9662可以抵抗PLAB的抗炎作用。结论 PLAB抑制LPS诱导RAW264.7细胞炎症反应并抑制巨噬细胞向M1表型偏移,与影响细胞周期分布、调控NF-κB/PPARγ通路有关。
OBJECTIVE: To investigate the anti-inflammatory effect and the molecular mechanism of PLAB on lipopolysaccharide (LPS) -induced RAW264.7 cells. Methods RAW264.7 cells were induced by lipopolysaccharide (LPS) in vitro, and treated with 0.5 μmol / L PLAB and 1 μmol / L of PPARγ blocker GW9662. The changes of cell cycle were detected by flow cytometry. The mRNA expressions of IL-1β and TNF-α of PPARγ and M1 macrophage markers were detected by real-time quantitative PCR. Factor κB (NF-κB) signaling pathway-related signaling molecule levels. Results PLAB could significantly decrease the mRNA level of IL-1β and TNF-α in LPS-induced RAW264.7 cells and up-regulate the mRNA level of PPARγ. The protein levels of NF-κB p65, IKKα, IKKβ, p IKKα / β, IκBα, and p IκBα were down-regulated in RAW264.7 cells in the G0 and G2 phases. GW9662 can resist the anti-inflammatory effect of PLAB. Conclusion PLAB can inhibit LPS-induced inflammatory response of RAW264.7 cells and inhibit the migration of macrophages to M1 phenotype, which is related to cell cycle distribution and NF-κB / PPARγ pathway.