食品科学 ›› 2023, Vol. 44 ›› Issue (17): 67-78.doi: 10.7506/spkx1002-6630-20220928-307

• 营养卫生 • 上一篇    下一篇

高白鲑鱼油通过抑制核因子κB、信号传导转录激活因子1和琥珀酸盐/缺氧诱导因子-1α信号通路的抗炎机制

刘宏旭,宋国库,王珍,夏效东,秦宁波   

  1. (大连工业大学食品学院,辽宁 大连 116034)
  • 出版日期:2023-09-15 发布日期:2023-09-29
  • 基金资助:
    国家自然科学基金青年科学基金项目(32101963);辽宁省教育厅高校基本科研项目(面上项目)(LJKMZ20220890)

Anti-inflammatory Mechanism of Coregonus peled Oil via the Inhibition of Nuclear Factor Kappa-B (NF-κB), Signal Transducer and Activator of Transcription 1 (STAT1), and Succinate/Hypoxia-Inducible Factor-1α (HIF-1α) Signaling Pathways

LIU Hongxu, SONG Guoku, WANG Zhen, XIA Xiaodong, QIN Ningbo   

  1. (College of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, China)
  • Online:2023-09-15 Published:2023-09-29

摘要: 目的:探讨高白鲑鱼油的化学成分和抗炎作用机制。方法:采用匀浆萃取超声法从高白鲑中提取鱼油,并对其主要化学成分进行检测。采用气相色谱-质谱联用仪对高白鲑鱼油进行脂肪酸成分分析。构建巨噬细胞RAW264.7炎症细胞模型,用脂多糖(lipopolysaccharide,LPS)刺激后,通过MTT法检测细胞毒性,采用液相色谱串联质谱联用仪对琥珀酸盐含量进行测定;采用试剂盒法测定上清液中一氧化氮(nitric oxide,NO)含量和一氧化氮合酶(inducible nitric oxide synthase,iNOS)活力;采用酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)测定细胞上清液中白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)质量浓度。采用实时荧光定量聚合酶链式反应法测定细胞上清液中CD68、CD86、Arg1、CD206、YM1、HIF-1α基因的相对表达量,并通过蛋白免疫印迹法测定其蛋白的相对表达量。结果表明:高白鲑鱼油的主要化学成分为棕榈油酸、油酸、二十碳五烯酸和二十二碳六烯酸。其可以显著抑制NO、iNOS、IL-6、TNF-α、IL-1β的产生及相关基因的表达。鱼油可通过抑制LPS诱导RAW264.7细胞中的核因子-κB(nuclear factor kappa-B,NF-κB)抑制剂α的降解和减少p65核易位来抑制NF-κB的激活。此外,高白鲑鱼油可通过抑制信号传导转录激活因子1(signal transducer and activator of transcription 1,STAT1)的磷酸化,将巨噬细胞从M1型极化为M2型。另外,通过测定琥珀酸盐的含量、缺氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)基因和蛋白的表达量发现,高白鲑鱼油可以通过抑制琥珀酸盐的积累来抑制IL-1β的产生。结论:高白鲑鱼油可能通过抑制NF-κB、STAT1和琥珀酸盐/HIF-1α信号通路抑制炎症。

关键词: 高白鲑;鱼油;炎症;极化;琥珀酸盐/缺氧诱导因子-1α

Abstract: Objective: To investigate the chemical composition and anti-inflammatory mechanism of Coregonus peled oil (CPO). Methods: CPO was extracted by ultrasonic homogenization and its major chemical components were detected. In addition, its fatty acid (FA) composition was analyzed by gas chromatography-mass spectrometry (GC-MS). Macrophage RAW264.7 cells were cultured as an inflammation model and stimulated with lipopolysaccharides (LPS). Cytotoxicity was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The accumulation of succinate was detected by liquid chromatography tandem mass spectrometry (LC-MS/MS). The concentration of nitric oxide (NO) and the activity of inducible nitric oxide synthase (iNOS) in the supernatant were measured by commercial kits. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of interleukin-6 (IL-6), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the supernatant. The expression levels of CD68, CD86, Arg1, CD206, YM1 and HIF-1α were detected by quantitative real-time polymerase chain reaction (qPCR), and their protein expression levels were determined by Western blotting. Results: The major chemical constituents of CPO were palmitoleic acid, oleic acid, eicosapntemacnioc acid (EPA), and docosahexaenoic acid (DHA). CPO significantly inhibited the production of NO, iNOS, IL-6, TNF-α, and IL-1β as well as their gene expression levels. CPO blocked NF-κB activation by inhibiting the degradation of inhibitor α of NF-κB (IκBα) and decreasing p65 nuclear translocation in LPS-induced RAW264.7 cells. In addition, CPO promoted macrophage polarization from M1 to M2 types by inhibiting the phosphorylation of signal transducer and activator of transcription 1 (STAT1). The content of succinate and the expression levels of hypoxia-inducible factor-1α (HIF-1α) gene and protein indicated that CPO could inhibit the production of IL-1β by inhibiting the accumulation of succinate. Conclusion: The anti-inflammatory mechanism of CPO may be due to the inhibition of the NF-κB, STAT1, and succinate/HIF-1α signaling pathways.

Key words: Coregonus peled; fish oil; inflammation; polarization; succinate/hypoxia-inducible factor-1α

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