广陈皮中黄酮类化合物提取物对t-BHP诱导HepG2细胞氧化损伤保护作用机制
作者:
作者单位:

华中农业大学食品科学技术学院/环境食品学教育部重点实验室/ 果蔬加工与品质调控湖北省重点实验室,武汉 430070

作者简介:

陶叶杏,E-mail: yexing.tao@webmail.hzau.edu.cn

通讯作者:

徐晓云,E-mail: xuxiaoyun@mail.hzau.edu.cn

中图分类号:

R285.5

基金项目:

国家自然科学基金项目(32172140);中央高校基本科研业务费专项(2662020SPPY008)


Protective mechanism of flavonoids extracts from Pericarpium Citri Reticulatae ‘Chachi’ on t-BHP-induced oxidative damage in HepG2 cells
Author:
Affiliation:

College of Food Science and Technology/Key Laboratory of Environment Correlative Dietology,Ministry of Education/Hubei Key Laboratory of Fruit & Vegetable Processing & Quality Control,Huazhong Agricultural University,Wuhan 430070,China

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    摘要:

    为了研究陈化对于广陈皮抗氧化活性的影响,以新鲜干燥果皮、贮藏1 a和10 a广陈皮(Pericarpium Citri Reticulatae ‘Chachi’,PCR-C)为研究对象,提取纯化得到广陈皮黄酮类化合物提取物(PCR-CF),以叔丁基过氧化氢(t-BHP)诱导HepG2 细胞作为氧化应激损伤模型,评价陈皮的抗氧化活性,同时利用qRT-PCR和Western blot技术进一步探究其抗氧化活性作用分子机制。结果显示,广陈皮陈化过程中,橙皮苷含量呈现波动,但总体上是下降趋势;而多甲氧基黄酮及其总含量呈增加趋势,其中PCR-C10F中含量最高,PMFs总含量达到(98.66±0.56) mg/g。而且,不同贮藏年份PCR-CF可明显提高SOD、GSH水平和降低MDA含量,且不同年份间差异显著,PCR-C10F抗氧化活性最强,SOD、GSH、MDA水平分别达到(139.38±17.38) U/mg和(117.81±3.22) μmol/g、(0.39±0.03) nmol/mg。相关性分析表明,川陈皮素的积累是PCR-C10F维持氧化还原平衡的主要活性成分。此外,PCR-C10F显著上调Nrf2 mRNA以及其在胞核蛋白中的表达;从而激活抗氧化酶的mRNA和蛋白表达水平,HO-1的mRNA表达水平和蛋白表达水平显著降低。研究表明,陈皮在陈化过程中,多甲氧基黄酮含量呈现增加趋势,抗氧化活性不断增强,PCR-C10F能够通过调控Nrf2-ARE抗氧化信号通路起到氧化应激抵御保护作用。

    Abstract:

    The flavonoids extracts of Pericarpium Citri Reticulatae ‘Chachi’ (PCR-CF) were extracted and purified from fresh-dried tangerine peel and Pericarpium Citri Reticulatae ‘Chachi’ (PCR-C) stored for 1 year and 10 years to study the effects of aging process on the antioxidant activities of Pericarpium Citri Reticulatae ‘Chachi’. The changes of contents of flavonoids were analyzed with HPLC. The antioxidant activities of PCR-CF were evaluated by using tert-butyl hydroperoxide (t-BHP)-induced oxidative damage in human hepatoma cells (HepG2 cells) as the model. The molecular mechanism of antioxidant activity was further explored via real-time fluorescence quantitative PCR (qRT-PCR) and Western blot. The results showed that the content of hesperidin fluctuated,but the overall trend was decreasing during the aging process of dried tangerine peel. The contents of polymethoxyflavonoids(PMFs) and total PMFs showed an increasing trend,among which the content of PCR-C10F was the highest,and the content of total PMFs reached (98.66±0.56) mg/g. Moreover,PCR-CF from different storage years significantly increased the levels of SOD and GSH and reduced the contents of MDA There was significant difference between different storage periods. The antioxidant activity of PCR-C10F was the strongest,in which the level of SOD,GSH and MDA reached (139.38±17.38) U/mg,(117.81±3.22) μmol/g and (0.39±0.03) nmol/mg,respectively. The results of correlation analyses showed that the accumulation of nobiletin was the main active component of PCR-C10F to maintain redox balance. PCR-C10F significantly up-regulated the mRNA expression levels of Nrf2 (P<0.001) and increased the expression levels of Nrf2 (P<0.05) in nuclear protein. The mRNA (P<0.001) and protein (P<0.01) expression levels of antioxidant enzymes were significantly activated,and the mRNA (P<0.001) and protein (P<0.05) expression levels of HO-1 was significantly decreased. It is indicated that the contents of PMFs in PCR-CF showed increasing trends and the antioxidant activities enhanced during aging process. PCR-C10F can protect against oxidative stress by regulating the Nrf2-ARE antioxidant signaling pathway. It will provide a theoretical basis for rational storage and quality control of PCR-C.

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陶叶杏,余倩,刘瑞婷,张曦文,吴婷,潘思轶,徐晓云.广陈皮中黄酮类化合物提取物对t-BHP诱导HepG2细胞氧化损伤保护作用机制[J].华中农业大学学报,2022,41(5):161-168

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  • 收稿日期:2022-05-22
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  • 在线发布日期: 2022-10-11
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