食品科学 ›› 2024, Vol. 45 ›› Issue (17): 96-104.doi: 10.7506/spkx1002-6630-20240122-196

• 营养卫生 • 上一篇    下一篇

基于RAW264.7细胞模型的槲皮万寿菊素与叶黄素协同改善急性肺损伤的作用机制

冯丹琦, 王向红, 吴迪, 连运河, 程鑫颖, 刘卫华, 米思   

  1. (1.河北农业大学食品科技学院,河北 保定 071000;2.晨光生物科技集团股份有限公司,河北 邯郸 057250;3.河北晨光检测技术服务有限公司,河北 邯郸 057250)
  • 出版日期:2024-09-15 发布日期:2024-09-09
  • 基金资助:
    “十四五”国家重点研发计划重点专项(2022YFE0124900);国家自然科学基金青年科学基金项目(32202013); 河北省自然科学基金生物农业联合基金培育项目(C2022204222)

Synergistic Ameliorative Effect of Quercetagetin and Lutein on Acute Lung Injury in RAW264.7 Cells

FENG Danqi, WANG Xianghong, WU Di, LIAN Yunhe, CHENG Xinying, LIU Weihua, MI Si   

  1. (1. College of Food Science and Technology, Hebei Agricultural University, Baoding 071000, China; 2. Chenguang Biotechnology Group Co. Ltd., Handan 057250, China; 3. Hebei Chenguang Detection Technology Services Co. Ltd., Handan 057250, China)
  • Online:2024-09-15 Published:2024-09-09

摘要: 为解析槲皮万寿菊素、槲皮素与叶黄素单独处理以及联合处理对急性肺损伤的作用机制,以脂多糖诱导构建RAW264.7细胞炎症模型,以一氧化氮(nitric oxide,NO)相对含量为评价指标,采用联合指数法确定槲皮万寿菊素与叶黄素以及槲皮素与叶黄素的最佳复配比例;分析比较槲皮万寿菊素、槲皮素与叶黄素单独及联合处理对RAW264.7细胞中炎症因子(肿瘤坏死因子-α、白细胞介素(interleukin,IL)-1β、IL-6)含量、丙二醛含量、超氧化物歧化酶活性以及谷胱甘肽过氧化物酶活性的影响;采用免疫印迹法测定核因子κB(nuclear factor kappa-B,NF-κB)信号通路中p65、p50以及沉默信息调节因子1(silent information regulator 1,SIRT1)、核因子E2相关因子2(nuclear factor‑erythroid 2 related factor 2,Nrf2)和NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)的相对表达量。结果表明,槲皮万寿菊素与叶黄素高剂量3∶1(30 μg/mL+10 μg/mL)复配能够最大程度降低RAW264.7细胞中的NO相对含量。二者单独及联合作用均能通过降低炎症因子、丙二醛含量,提高超氧化物歧化酶和谷胱甘肽还原酶活性,下调NF-κB p65、p50以及NLRP3表达水平并上调SIRT1、Nrf2蛋白相对表达量发挥改善急性肺损伤的作用,且联合处理效果优于单独处理组。

关键词: 万寿菊;槲皮万寿菊素;叶黄素;槲皮素;RAW264.7细胞;急性肺损伤

Abstract: This study aimed to elucidate the ameliorative effect and mechanism of quercetagetin, quercetin and lutein, alone and in combination, on acute lung injury. An lipopolysaccharide-induced inflammation model was established based on RAW264.7 cells. The optimal mixture ratios between quercetagetin or quercetin and lutein were determined based on the content of nitric oxide (NO) in RAW264.7 cells using combination index (CI) analysis. The individual and combined effects of quercetagetin, quercetin and lutein on the contents of malondialdehyde (MDA), inflammatory cytokines (including tumor necrosis factor α (TNF-α), interleukin (IL)-1β and IL-6) and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in LPS-stimulated RAW264.7 cells were investigated. In addition, the relative expression levels of p65, p50, silent information regulator 1 (SIRT1), nuclear factor‑erythroid 2 related factor 2 (Nrf2), and NOD-like receptor thermal protein domain associated protein 3 (NLRP3) were evaluated by Western blot. The results showed that the lowest NO level in RAW264.7 cells was observed by treatment with a mixture of high-dose quercetagetin and lutein of 3:1 (30 μg/mL + 10 μg/mL). Quercetagetin, lutein and their combination all decreased the contents of MDA and inflammatory cytokines, increased the activities of SOD and GSH-Px, down-regulated the expression of nuclear factor kappa-B (NF-κB) p65, p50 and NLRP3, and up-regulated the expression of SIRT1 and Nrf2, the combination being more effective than either treatment alone.

Key words: marigold (Tagetes erecta L.); quercetagetin; lutein; quercetin; RAW264.7 cells; acute lung injury

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