首页|花生红衣中反式白藜芦醇对HEK293T细胞抗氧化的影响

花生红衣中反式白藜芦醇对HEK293T细胞抗氧化的影响

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目的:分析花生红衣中的白藜芦醇成分,探究其主要成分反式白藜芦醇(RES)对人胚胎肾细胞293(HEK293T)抗氧化的影响及其潜在的分子机制.方法:采用超声波辅助酶法提取花生红衣中的白藜芦醇,采用高效液相色谱-质谱联用(HPLC-MS)技术分析花生红衣中白藜芦醇粗提液的组成成分.通过测定细胞活力,检测RES对HEK293T细胞的最高毒性.通过荧光素酶报告基因试验及免疫印迹试验检测其对Keap1-Nrf2-ARE抗氧化信号通路的影响,以DPPH自由基清除率评价其体外抗氧化能力.结果:HPLC-MS结果表明,花生红衣中含有白藜芦醇苷、白皮杉醇和RES.由于白藜芦醇在自然界中主要以反式白藜芦醇形式存在,因此选择RES进行后续试验.细胞存活力检测结果表明,RES对HEK293T细胞的最高无毒浓度为50 μmol/L,体外抗氧化作用呈浓度依赖性.荧光素酶报告基因分析表明,RES显著诱导了 ARE介导的转录激活.免疫印迹结果显示,RES能诱导Nrf2介导的3个抗氧化蛋白表达[血红素氧合酶1(HO-1)、醌氧化还原酶1(NQO1)和谷氨酸半胱氨酸连接酶(GCLM)]表达.此外,DPPH自由基清除率测定结果表明,RES能有效清除DPPH自由基,具有体外抗氧化能力.结论:花生红衣中白藜芦醇的主要成分RES对HEK293T细胞有抗氧化作用,可通过Keap1-Nrf2-ARE信号通路激活潜在的抗氧化活性.
Effects of Trans-resveratrol in Peanut Red Coat on Antioxidant in HEK293T Cells
Objective:This experiment analyzed the resveratrol composition in peanut red coat to investigate the antioxi-dant effects of its main component trans-resveratrol(RES)on human embryonic kidney cell 293(HEK293T)and its po-tential molecular mechanism.Methods:The resveratrol in peanut red coat was extracted by ultrasonic-assisted enzymatic method,and the constituents of the crude extract of resveratrol in peanut red coat were analyzed by high performance liquid chromatography-mass spectrometry(HPLC-MS).The highest toxicity of RES on HEK293T cells was detected by cell viability assay,its effects of Keap1-Nrf2-ARE antioxidant signaling pathway was detected by luciferase reporter gene assay and Western blot assay,and its in vitro antioxidant capacity was determined by DPPH free radical clearance.Re-sults:The results of HPLC-MS showed that resveratrol glycosides,paclitaxel and RES were contained in peanut red coat.Since resveratrol mainly exists in the form of trans-resveratrol in nature,RES was selected for the subsequent experi-ments.The results of cell viability assay showed that the highest non-toxic concentration of RES on HEK293T cells was 50 μmol/L,and the in vitro antioxidant effects was concentration-dependent.Western blot results showed that RES in-duced Nrf2-mediated expression of three target proteins heme oxygenase 1(HO-1),quinone oxidoreductase 1(NQO1)and glutamate cysteine ligase(GCLM).In addition,the results of the DPPH radical scavenging assay showed that RES could effectively scavenge DPPH radicals and had in vitro antioxidant capacity.Conclusion:The major components of resveratrol in peanut red coat have antioxidant effects on HEK293T cells and can activate a potential antioxidant activity through the Keap1-Nrf2-ARE signaling pathway.

peanut red coatresveratrolHEK293Thigh performance liquid chromatography-mass spectrometry(HPLC-MS)Keap1-Nrf2-ARE signaling pathwayantioxidant

孙畅、周春田、岳玉兰、吕呈蔚、李云飞、黄威、李铁柱、胡济美

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吉林省农业科学院农产品加工研究所 长春 130033

包头轻工职业技术学院 内蒙古包头 014035

中国国际工程咨询有限公司 北京 100048

花生红衣 白藜芦醇 HEK293T 高效液相色谱-质谱联用 Keap1-Nrf2-ARE信号通路 抗氧化

吉林省科技厅重点研发项目吉林省科技厅重点研发项目吉林省预算内基本建设资金项目(2022)吉林省农业科技创新工程项目吉林省农业科技创新工程项目

20210203182SF20200403063SF2022c043-12CXGC2021TD006CXGC2021RCY028

2024

10.16429/j.1009-7848.2024.05.011

中国食品学报
中国食品科学技术学会

中国食品学报

CSTPCD北大核心EI
影响因子：1.079
ISSN：1009-7848
年,卷(期)：2024.24(5)
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