食品科学 ›› 2017, Vol. 38 ›› Issue (19): 182-188.doi: 10.7506/spkx1002-6630-201719029

• 营养卫生 • 上一篇    下一篇

蒲公英糖蛋白体外抗炎作用及对NF-κB信号通路的调控

夏 炎1,管晓辉1,崔艳艳1,高 超1,陈 玲1,董 然2,沈明浩1,*   

  1. 1.吉林农业大学食品科学与工程学院,吉林 长春 130118; 2.长春科技学院,长白山生态资源开发工程研究中心,吉林 长春 130600
  • 出版日期:2017-10-15 发布日期:2017-09-29
  • 基金资助:
    吉林省科技厅科技支撑项目(20100259)

Inhibitory Effect of Taraxacum Glycoprotein on Lipopolysaccharide-Induced Inflammation via Regulating NF-κB Pathway

XIA Yan1, GUAN Xiaohui1, CUI Yanyan1, GAO Chao1, CHEN Ling1, DONG Ran2, SHEN Minghao1,*   

  1. 1. College of Food Science and Engineering, Jilin Agricultural University, Changchun 130118, China; 2. Changbai Mountain Ecological Resources Development Engineering Research Center, Changchun Science and Technology College, Changchun 130600, China
  • Online:2017-10-15 Published:2017-09-29

摘要: 探讨蒲公英糖蛋白(glycoprotein from Taraxacum,TG)对由脂多糖引起的RAW264.7细胞炎症的抗炎效 果,并阐明其活性的基本分子机制。利用脂多糖刺激RAW264.7细胞,建立体外炎症模型,采用噻唑蓝比色法检 测TG对RAW264.7细胞的增殖毒性,Griess试剂法检测了一氧化氮(nitric oxide,NO)的分泌情况,反转录聚合酶 链式反应检测炎症细胞因子——白细胞介素-6(interleukin 6,IL-6)和肿瘤坏死因子-α(tumor necrosis factor-α, TNF-α)mRNA以及诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)mRNA的表达水平,酶联免疫 吸附测定法测定IL-6和TNF-α的分泌量,Western blot检测P-IκB-α蛋白的表达水平,用以研究TG对核转录因子-κB (nuclear factor-kappa B,NF-κB)信号转导通路的抑制作用。结果表明:TG能够显著甚至极显著地抑制NO的分 泌,IL-6、TNF-α、iNOS的mRNA的表达,IL-6和TNF-α的分泌(P<0.05、P<0.01)。TG高度显著上调了IκB-α的 蛋白表达(P<0.001),并显著下调了P-IκB-α的蛋白表达(P<0.05),且与TG质量浓度成正比。其中在TG质量 浓度为250、500、1 000 μg/mL时对TNF-α分泌量的抑制率分别为28.6%、65.4%、89.3%,对IL-6分泌量的抑制率分 别为32.3%、54.1%、85.7%。TG间接抑制了NF-κB信号转导途径,有显著的体外抗炎效果且抗炎效果与TG的质量 浓度呈剂量依赖性。

关键词: 蒲公英, 糖蛋白, 炎症因子, 核转录因子-κB

Abstract: To investigate the inhibitory effect of Taraxacum glycoprotein on lipopolysaccharide-induced (LPS-induced) inflammation in RAW264.7 cells and elucidate the underlying molecular mechanism, an in vitro model of inflammation in RAW264.7 macrophages stimulated by LPS was established. The proliferation of RAW264.7 cells were detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) method and the secretion of nitric oxide (NO) was detected by Griess reagent method. The inhibitory effect of Taraxacum glycoprotein on the mRNA expression of LPS-induced inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and inducible nitric oxide synthase (iNOS) in RAW264.7 cells was examined using reverse transcription-polymerase chain reaction, and the effect of Taraxacum glycoprotein on the secretion levels of LPS-induced inflammatory cytokines such as TNF-α and IL-6 was detected by enzyme-linked immunosorbent assay as well as the effect on its upstream signaling proteins including P-IκB-α and IκB-α by Western blot. Results: Taraxacum glycoprotein could significantly inhibit the secretion of NO, the mRNA expression of IL-6, TNF-α and iNOS and the secretion of IL-6 and TNF-α (P < 0.05, P < 0.01, respectively). Taraxacum glycoprotein resulted in a significant up-regulation of IκB-α protein expression (P < 0.001) and a significant down-regulation of P-IκB-α in a concentration-dependent manner (P < 0.05). The percentage inhibition of TNF-α secretion was 28.6%, 65.4% and 89.3%, and the percentage inhibition of IL-6 secretion was 32.3%, 54.1% and 85.7%, respectively, at concentrations of 250, 500 and 1 000 μg/mL. Conclusion: The nuclear factor-kappa B signal transduction pathway is indirectly inhibited by dandelion glycoprotein, and it has a significant anti-inflammatory effect in vitro in a dose-dependent manner.

Key words: Taraxacum F.H.Wigg., glycoprotein, inflammatory factor, nuclear factor-kappa B

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