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香草醛、4-甲基愈创木酚和4-乙基愈创木酚对AAPH诱导的HepG2细胞异常氧化应激的定量分析及细胞保护作用。

Quantification and cytoprotection by vanillin, 4-methylguaiacol and 4-ethylguaiacol against AAPH-induced abnormal oxidative stress in HepG2 cells.

作者信息

Zhao Dongrui, Shi Dongmei, Sun Jinyuan, Li Hehe, Zhao Mouming, Sun Baoguo

机构信息

School of Food Science and Engineering, South China University of Technology Guangzhou 510640 P. R. China.

Beijing Laboratory for Food Quality and Safety, Beijing Technology and Business University Beijing 100048 P. R. China

出版信息

RSC Adv. 2018 Oct 16;8(62):35474-35484. doi: 10.1039/c8ra06505e. eCollection 2018 Oct 15.

Abstract

Vanillin, 4-methylguaiacol, and 4-ethylguaiacol, three phenolic compounds in Gujinggong (GJG) Chinese baijiu (Chinese liquor), were quantified by liquid-liquid extraction (LLE) combined with gas chromatography-mass spectrometry (GC-MS) and evaluated for their possible cytoprotective effects by AAPH-induced HepG2 cell model. To confirm whether vanillin, 4-methylguaiacol, and 4-ethylguaiacol protected HepG2 cells against AAPH-induced abnormal oxidative stress motivating the Keap1-Nrf2 pathway, the gene and protein expression of Nrf2, Keap1, SOD, CAT, and GPx from the Keap1-Nrf2 pathway were measured with real-time PCR and western blot. Three levels of treatment doses (1000, 500, and 100 mg L) were applied. Results showed that vanillin, 4-methylguaiacol, and 4-ethylguaiacol exhibited potent cytoprotective effect in a dose-dependent manner, greatly alleviating or reversing the increased oxidative stress induced by AAPH through up-regulating the mRNA and protein expression levels of Nrf2, SOD, CAT, and GPx, and thereby, significantly improving the intracellular antioxidant defense system in HepG2 cells ( < 0.05). Based on these findings, it was confirmed that vanillin, 4-methylguaiacol, and 4-ethylguaiacol, natural components of Chinese baijiu, were able to modulate the expression of Nrf2 and its downstream antioxidative enzymes (, SOD, CAT, and GPx) against AAPH-induced abnormal oxidative stress. Further, this study lays the foundation for better illustrating the health benefits of Chinese baijiu.

摘要

采用液液萃取(LLE)结合气相色谱 - 质谱联用(GC - MS)技术对古井贡酒(GJG)白酒中的三种酚类化合物香草醛、4 - 甲基愈创木酚和4 - 乙基愈创木酚进行定量分析,并通过AAPH诱导的HepG2细胞模型评估它们可能的细胞保护作用。为了证实香草醛、4 - 甲基愈创木酚和4 - 乙基愈创木酚是否能保护HepG2细胞免受AAPH诱导的异常氧化应激并激活Keap1 - Nrf2通路,采用实时荧光定量PCR和蛋白质免疫印迹法检测Keap1 - Nrf2通路中Nrf2、Keap1、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GPx)的基因和蛋白表达。设置了三个处理剂量水平(1000、500和100 mg/L)。结果表明,香草醛、4 - 甲基愈创木酚和4 - 乙基愈创木酚呈剂量依赖性地表现出强大的细胞保护作用,通过上调Nrf2、SOD、CAT和GPx的mRNA和蛋白表达水平,极大地减轻或逆转了AAPH诱导的氧化应激增加,从而显著改善了HepG2细胞内的抗氧化防御系统(P < 0.05)。基于这些发现,证实了白酒中的天然成分香草醛、4 - 甲基愈创木酚和4 - 乙基愈创木酚能够调节Nrf2及其下游抗氧化酶(SOD、CAT和GPx)的表达,以对抗AAPH诱导的异常氧化应激。此外,本研究为更好地阐明白酒对健康的益处奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0a3a/9087904/771d81cbff63/c8ra06505e-f1.jpg

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