食品科学 ›› 2022, Vol. 43 ›› Issue (23): 141-148.doi: 10.7506/spkx1002-6630-20211230-348

• 营养卫生 • 上一篇    下一篇

羊栖菜多酚通过核转录因子-κB/丝裂原活化蛋白激酶通路缓解脂多糖诱导的RAW264.7细胞炎症反应

黄平,洪静霞,米杰,张攀学,李超,杨文鸽   

  1. (宁波大学食品与药学学院,浙江省动物蛋白食品精深加工技术重点实验室,浙江 宁波 315211)
  • 出版日期:2022-12-15 发布日期:2022-12-28
  • 基金资助:
    “十三五”国家重点研发计划重点专项(2018YFD0901105);宁波市自然科学基金项目(202003N4128)

Polyphenols Extracted from Hizikia fusiformis Relieves Lipopolysaccharide-Induced Inflammation in RAW264.7 Cells via the Nuclear Factor-κB/Mitogen-Activated Protein Kinase Signaling Pathways

HUANG Ping, HONG Jingxia, MI Jie, ZHANG Panxue, LI Chao, YANG Wenge   

  1. (Key Laboratory of Animal Protein Food Deep Processing Technology of Zhejiang Province, College of Food and Pharmaceutical Sciences, Ningbo University, Ningbo 315211, China)
  • Online:2022-12-15 Published:2022-12-28

摘要: 目的:研究羊栖菜多酚对脂多糖(lipopolysaccharide,LPS)诱导小鼠单核巨噬细胞白血病细胞RAW264.7细胞炎症反应的影响。方法:噻唑蓝法测定细胞活力;Griess法测定一氧化氮(NO)水平;实时荧光定量聚合酶链式反应测定白细胞介素(interleukin,IL)-6、IL-1β、肿瘤坏死因子(tumor necrosis factor,TNF)-α、环氧合酶2(cyclooxygenase-2,COX-2)和诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)的基因相对表达量;流式细胞术测定细胞吞噬能力;蛋白免疫印迹法测定信号通路丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPKs)和核转录因子(nuclear factor,NF)-κB信号通路关键蛋白表达水平。结果:羊栖菜多酚对RAW264.7细胞的安全质量浓度范围为0~160 μg/mL。与LPS组相比,羊栖菜多酚剂量依赖性降低巨噬细胞吞噬能力并抑制NO的生成。同时,羊栖菜多酚下调促炎细胞因子(IL-1β、IL-6、TNF-α)和炎症诱导酶(iNOS、COX-2)的mRNA水平,且作用效果与给药剂量及LPS刺激时间相关。这些炎性介质的表达与羊栖菜多酚抑制p38 MAPK和NF-κB p65的激活有关。结论:羊栖菜多酚可通过减弱p38 MAPK和NF-κB p65信号通路的激活水平,抑制下游炎症介质的转录表达,从而缓解LPS诱导的巨噬细胞炎症反应。

关键词: 羊栖菜多酚;RAW264.7细胞;抗炎;核转录因子-κB信号通路;丝裂原活化蛋白激酶信号通路

Abstract: Objective: To study the effect of Hizikia fusiformis polyphenols (HFPs) on lipopolysaccharide (LPS)-induced inflammatory response in RAW264.7 cells. Methods: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method was used to examine the cellular viability. Nitric oxide (NO) was measured by the Griess method. Quantitative real-time polymerase chain reaction (qPCR) was applied to analyze the gene expression of interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF)-α, cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). Flow cytometry was used to determine macrophage phagocytosis. The expression of the key proteins involved in the mitogen-activated protein kinases (MAPKs) and nuclear factor (NF)-κB signaling pathways was investigated by Western blot. Results: HFPs at concentrations of 0–160 μg/mL had no cytotoxic effect on RAW264.7 cells. Compared with the LPS-induced group, HFPs dose-dependently inhibited the phagocytic capacity of macrophage and NO production. Besides, the mRNA levels of several inflammatory mediators, including IL-1β, IL-6, TNF-α, iNOS and COX-2 were down-regulated, and this effect was dependent on the dose of HFPs and LPCS stimulation time. The expression of these inflammatory mediators may be related to the inhibition of the activation of the activation of p38 MAPK and NF-κB p65 by HFPs. Conclusion: HFPs alleviates LPS-induced inflammation in RAW264.7 cells by down-regulating the activation of the p38 MAPK and NF-κB p65 signaling pathways and inhibiting the transcription and expression of downstream inflammatory mediators.

Key words: Hizikia fusiformis polyphenols; RAW264.7 cells; anti-inflammation; nuclear factor-κB signaling pathway; mitogen-activated protein kinases singling pathway

中图分类号: