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目的:研究猪苓多糖对脂多糖(LPS)诱导的J774巨噬细胞炎症模型细胞因子的调节作用,并探讨其可能的抗炎作用机制。方法:J774细胞以2×105个/孔接种于6孔培养板,将细胞分为空白组、LPS模型组、LPS加猪苓多糖低剂量组(1 mg·L-1)、LPS加猪苓多糖中剂量组(10 mg·L-1)、LPS加猪苓多糖高剂量组(100 mg·L-1),每组6个复孔。J774给予10 mg·L-1LPS刺激3 h,同时猪苓多糖干预共刺激3 h后收集细胞,实时荧光定量PCR(RT-PCR)方法检测白细胞介素-1β(IL-1β),白细胞介素-10(IL-10),肿瘤坏死因子-α(TNF-α),干扰素-γ(IFN-γ)和白细胞介素-6(IL-6)mRNA的表达,Western blotting检测猪苓多糖对p38丝裂原活化蛋白激酶(p38),细胞外信号调节蛋白激酶42/44(ERK42/44),p65丝裂原活化蛋白激酶(p65)蛋白磷酸化表达的影响。结果:与空白组相比,LPS模型组细胞因子IL-1β,IL-10,TNF-α,IFN-γ和IL-6 mRNA表达量显著升高(P<0.01),炎症模型建立成功。给予猪苓多糖干预后,猪苓多糖可降低由LPS诱导导致的炎症因子的升高与LPS导致的p38,ERK42/44,p65磷酸化的表达(P<0.01,P<0.05)。结论:猪苓多糖可以降低LPS导致的炎症反应,可能是通过丝裂原活化蛋白激酶(MAPK)信号通路来降低炎症损伤。
AIM: To investigate the regulatory effect of Polyporus umbellatis polysaccharides on cytokines induced by lipopolysaccharide (LPS) in J774 macrophage inflammatory model and to explore its possible anti-inflammatory mechanism. Methods: J774 cells were inoculated into 6-well plates at 2 × 105 cells / well and divided into blank group, LPS model group, LPS plus low dose polysaccharide group (1 mg · L-1), LPS plus Polyporus umbellatus Polysaccharide medium dose group (10 mg · L-1), LPS plus Polyporus umbellatus polysaccharide high dose group (100 mg · L-1). J774 was stimulated with 10 mg · L-1 LPS for 3 h, and co-stimulated with Polyporus umbellatus polysaccharide for 3 h. The cells were collected and the expression of interleukin-1β (IL-1β), interleukin (IL-10), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and interleukin-6 (IL- p38 mitogen-activated protein kinase (p38), extracellular signal-regulated protein kinase 42/44 (ERK42 / 44), p65 mitogen-activated protein kinase (p65) Results: Compared with the blank group, the expression of IL-1β, IL-10, TNF-α, IFN-γ and IL-6 mRNA in LPS model group was significantly increased (P <0.01). Polyporus umbellatus polysaccharide could reduce the increase of inflammatory factors induced by LPS and the expression of p38, ERK42 / 44, p65 phosphorylation induced by LPS (P <0.01, P <0.05). Conclusion: Polyporus umbellatus polysaccharide can reduce the inflammatory response induced by LPS, which may be through mitogen-activated protein kinase (MAPK) signaling pathway to reduce the inflammatory injury.