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猪苓多糖对LPS诱导的J774炎症模型的抗炎作用及其机制 被引量:19

Anti-inflammatory Effect and Mechanism of Polyporus Polysaccharide on LPS-stimulated J774 Cells
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摘要 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。 Objective: To study the effect of cytokines of polyporus polysaccharide (PPS) on lipopolysaccharide (LPS) -stimulated J774, and to explore its anti-inflammatory mechanism. Method: J774 cells were inoculated in 6-well plates with the density of 2 × 10^5 per well. The experiment was divided into blank group, model group, LPS plus low-dose PPS group ( 1 mg ·L^-1 ) , LPS plus middle-dose PPS group ( 10 mg ·L^-1 ) and LPS plus high-dose PPS group (100 mg ·L^-1) were set up in this study. Each group had six wells. J774 cells were polarized to inflammatory macrophage by treating with 10 mg ·L^-1 LPS for 3 h. Meanwhile, LPS-stimulated J774 cells were treated with PPS of 1, 10, 100 mg ·L^-1 for 3 h. The expressions of interleukin-1β (IL-1β), interleukin-lO (IL-10), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and interleukin-6 (IL-6) mRNA were detected by real-time quantitative PCR (RT-PCR). And the expressions of p38 mitogen-activated protein kinase (p38) , extracellular signal-regulated kinase (ERK42/44) , p65 mitogen-activated protein kinase (p65) protein were determined by using Western blotting. Result: Compared to blank group, the percentages of IL-1β, IL-10, TNF-α, IFN-γ and IL-6 mRNA of model group were significantly higher (P 〈 0.01 ). It meant the inflammation model was successfully established. After treated with PPS, the expressions of IL-1β, IL-10, TNF-α, IFN-γ and IL-6 mRNA were reduced. And the percentages of p38, ERK42/44, p65 protein were decreased (P 〈 0.01, P 〈 0.05). Conclusion: PPS could reduce inflammation in LPS-stimulated J774 cells. It might be achived through the mitogen activated protein kinase (MAPK) pathway to reduce inflammation damage.
出处 《中国实验方剂学杂志》 CAS CSCD 北大核心 2015年第3期156-159,共4页 Chinese Journal of Experimental Traditional Medical Formulae
基金 广东省科技计划项目(2010B030700059)
关键词 猪苓多糖 炎症模型 细胞因子 丝裂原活化蛋白激酶 protein kinasepolyporus polysaccharide inflammation model cytokines mitogen activated
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参考文献11

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