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

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和细胞保护作用

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

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 via 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-1) 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 (p < 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 (i.e., 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.

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