红松松仁多糖对脂多糖诱导RAW264.7细胞炎症反应的抑制作用

目的:探讨红松松仁多糖PNP40c-1对脂多糖(Lipopolysaccharides,LPS)诱导RAW264.7细胞炎症反应的抑制作用及可能机制。方法:以LPS刺激RAW264.7细胞诱导炎症模型,分别采用MTT法、比色法、酶联免疫法、RT-PCR和Western Blot等方法,比较LPS诱导前后巨噬细胞的细胞活力、吞噬能力、一氧化氮(nitric oxide,NO)/一氧化氮合酶(nitric oxide synthase,iNOS)和细胞因子表达的变化;同时对Nrf2/HO-1信号通路中关键蛋白核因子E2相关因子2(nuclear factor(erythroid-derived 2)-like 2 protein,Nrf2)和血红素氧化酶-1(heme oxygenase-1,HO-1)的mRNA和蛋白表达水平进行研究,以探讨PNP40c-1的具体作用机制。结果:在LPS诱导RAW264.7细胞的炎症反应中,PNP40c-1以剂量依赖性显著提高巨噬细胞的吞噬能力(P<0.05),抑制LPS诱导的NO和iNOS过量表达;PNP40c-1还可通过Nrf2/HO-1信号通路调节炎症因子如肿瘤坏死因子(tumor necrosis factor-α,TNF-α),白介素-1β(interleukin-1β,IL-1β)、白介素-6(IL-6)的分泌,缓解炎症反应。结论:红松松仁多糖PNP40c-1对LPS诱导的炎症反应具有一定的抑制作用,其作用机制与调节Nrf2/HO-1信号通路有关。

Inhibitory Effects of Pinus koraiensis Pine Nut Polysaccharide on Inflammatory Response of RAW264.7 Cells Induced by Lipopolysaccharide

Objective:This study was to investigate the inhibitory effects of Pinus koraiensis pine nut polysaccharide(PNP40c-1)on the inflammatory response of RAW264.7 cells induced by lipopolysaccharide(LPS)and the possible mechanism. Methods:LPS was used to stimulate RAW264.7 macrophages to induce inflammation. MTT assay,colorimetric assay,enzyme-linked immunosorbent assay,RT-PCR and Western Blot were used to compare the cell viability,phagocytosis,nitric oxide/nitric oxide synthase and the expression of cytokines in macrophages before and after LPS-treatment. Furthermore,the mRNA and protein expression of Nrf2 and HO-1 in the Nrf2/HO-1 signaling pathway were also studied to explore the specific action mechanism of PNP40c-1.Results:In LPS-induced inflammatory response,PNP40c-1 significantly increased the phagocytosis(P<0.05)and inhibited the overexpression of NO and iNOS in RAW264.7 macrophages in a dose-dependent way. Moreover,PNP40c-1 also regulated the secretion of pro-inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)through Nrf2/HO-1 signaling pathway to relieve inflammation induced by LPS. Conclusion:Pinus koraiensis pine nut polysaccharide PNP40c-1 possesses inhibitory effects on LPS-induced inflammatory response and the action mechanism is related to the regulation of Nrf2/HO-1 signaling pathway.