白藜芦醇-葡萄多酚复方口服液的抗氧化活性和美白作用评价
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摘要
本研究研制了一款白藜芦醇-葡萄多酚复方美白口服液,评价其抗氧化活性和美白作用,并对机理进行初步探讨。抗氧化活性实验结果表明,美白口服液具有较强的抗氧化活性,不仅能还原铁离子,还对DPPH+、ABTS+、过氧化自由基均有显著的清除作用。对DPPH+、ABTS+的IC50分别为10.32±1.20μg/m L、87.06±9.52μg/m L;对过氧自由基的清除、吸收能力分别为1649.00±2.78μmol equiv. vit. C/g、4251.30±566.71μmol of trolox equivalents (TE)/g。以人肝癌细胞Hep G2为细胞模型,美白口服液的胞内抗氧化活性可达5570.10±312.09μmol of quercetin equivalents (QE)/100 g。选择与人体黑色素细胞生理上较为接近的小鼠黑色素瘤B16细胞为研究模型,进行体外美白作用评价。结果表明,在一定添加量下,美白液能抑制B16细胞增殖、胞酪氨酸酶活性和蛋白表达量,从而减少黑色素的生成。这款复方美白口服液具有良好的抗氧化性与美白作用,将在功能食品、保健品行业有较好的应用前景。
This study developed a resveratrol-grape polyphenol composite whitening oral solution, evaluated its antioxidant activity and whitening effect, and discussed preliminarily the underlying mechanism. The results of the antioxidant activity experiments showed that the whitening oral solution exhibited strong antioxidant activity: not only reduced iron ions, but also exerted significant scavenging effects on the DPPH+ and ABTS+ free radicals and peroxides. The IC50 values for DPPH+ and ABTS+ radical scavenging activities were 10.32±1.20 μg/m L and 87.06±9.52 μg/m L, respectively. The peroxyl radical scavenging and absorption capacities were 1649.00±2.78 μmol Vit. C equivalents/g and 4251.30±566.71 μmol Trolox equivalents(TE)/g, respectively. In human hepatoma cell line Hep G2 model, the intracellular antioxidant activity of the whitening oral solution reached 5570.10±312.09 μmol quercetin equivalents(QE)/100 g. The mouse melanoma B16 cells, which are physiologically relevant to the human melanocytes, were selected as the research models to evaluate in vitro the whitening effect. The results showed that at a certain dose, the whitening oral solution could inhibit the proliferation of B16 cells, cellular tyrosinase activity and protein expression of B16 cells, thereby reducing the production of melanin. This composite whitening oral solution exhibited good antioxidant activity and whitening effect, thus, will have great prospects for the functional foods and healthcare products industry.
This study developed a resveratrol-grape polyphenol composite whitening oral solution, evaluated its antioxidant activity and whitening effect, and discussed preliminarily the underlying mechanism. The results of the antioxidant activity experiments showed that the whitening oral solution exhibited strong antioxidant activity: not only reduced iron ions, but also exerted significant scavenging effects on the DPPH+ and ABTS+ free radicals and peroxides. The IC50 values for DPPH+ and ABTS+ radical scavenging activities were 10.32±1.20 μg/m L and 87.06±9.52 μg/m L, respectively. The peroxyl radical scavenging and absorption capacities were 1649.00±2.78 μmol Vit. C equivalents/g and 4251.30±566.71 μmol Trolox equivalents(TE)/g, respectively. In human hepatoma cell line Hep G2 model, the intracellular antioxidant activity of the whitening oral solution reached 5570.10±312.09 μmol quercetin equivalents(QE)/100 g. The mouse melanoma B16 cells, which are physiologically relevant to the human melanocytes, were selected as the research models to evaluate in vitro the whitening effect. The results showed that at a certain dose, the whitening oral solution could inhibit the proliferation of B16 cells, cellular tyrosinase activity and protein expression of B16 cells, thereby reducing the production of melanin. This composite whitening oral solution exhibited good antioxidant activity and whitening effect, thus, will have great prospects for the functional foods and healthcare products industry.
引文
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[19]赵建,刘璇,文镜.荧光分光光度计测定保健食品总抗氧化能力[J].食品科学,2010,22:301-305ZHAO Jian,LIU Xuan,WEN Jing.Determination of total antioxidant capacity of health foods by fluorescence spectrophotometer[J].Food Science,2010,22:301-305
[20]杨涛,吴辉辉,徐青,等.抗氧化性能评价ORAC法及最新研究进展[J].食品工业科技,2009,7:352-355YANG Tao,WU Hui-hui,XU Qing,et al.ORAC method for antioxidant performance evaluation and recent research progress[J].Science and Technology of Food Industry,2009,7:352-355
[21]李谣,陈金龙,夏春燕,等.基于Hep G2细胞模型的香菇柄粉多酚抗氧化及抗增殖活性[J].食品科学,2016,11:190-196LI Yao,CHEN Jin-long,XIA Chun-yan,et al.Antioxidant and antiproliferative activity of polyphenols from mushroom handle powder based on HepG2 cell model[J].Food Science,2016,11:190-196
[22]Wolfe K L,Xinmei K,Xiangjiu H,et al.Cellular antioxidant activity of common fruits[J].Journal of Agricultural&Food Chemistry,2008,56(18):8418-8426
[23]陈贞纯,贾玲燕,毛祖法,等.茶叶提取物对B16小鼠黑色素瘤细胞的生物学效应及机理研究[J].茶叶科学,2014,5:465-472CHEN Zhen-chun,JIA Ling-yan,MAO Zu-fa,et al.Biological effects and mechanism of tea extract on B16mouse melanoma cells[J].Tea Science,2014,5:465-472
[24]Crowley L C,Marfell B J,Waterhouse N J.Analyzing cell death by nuclear staining with Hoechst 33342[J].Cold Spring Harbor Protocols,2016,9:778-781
[25]黄海潮,郑公铭,张小红,等.龙眼果核中没食子酸的体外美白作用及机制探索[J].中国实验方剂学杂志,2015,19:121-124HUANG Hai-chao,ZHENG Gong-ming,ZHANGXiao-hong,et al.In vitro whitening effect and mechanism of gallic acid in longan fruit nucleus[J].Chinese Journal of Experimental Traditional Medical Formulae,2015,19:121-124
[26]Sato K,Takahashi H,Iraha R,et al.Down-regulation of tyrosinase expression by acetylsalicylic acid in murine B16melanoma[J].Biological&Pharmaceutical Bulletin,2008,31(1):33-37
[27]王立峰,鞠兴荣.3种薏米的多酚提取物清除过氧化氢自由基抗氧化能力评价[J].食品科学,2012,19:39-44WANG Li-feng,JU Xing-rong.Evaluation of antioxidant capacity of hydrogen peroxide free radicals extracted from polyphenols extracted from three kinds of glutinous rice[J].Food Science,2012,19:39-44
[2]Fenoll L G,Penalver M J,Rodriguez-Lopez J N,et al.Tyrosinase kinetics:Discrimination between two models to explain the oxidation mechanism of monophenol and diphenol substrates[J].International Journal of Biochemistry&Cell Biology,2004,36(2):235-246
[3]邱凌.天然植物活性物质对酪氨酸酶作用机理的研究[D].厦门:厦门大学,2009QIU Ling.Study on the mechanism of action of natural plant active substances on tyrosinase[D].Xiamen:Xiamen University,2009
[4]王一帆,赖家珍,龙晓英,等.中药美白机制及功效评价进展[J].广东药学院学报,2014,4:525-529WANG Yi-fan,LAI Jia-zhen,LONG Xiao-ying,et al.Progress in whitening mechanism and efficacy evaluation of traditional Chinese medicine[J].Journal of Guangdong College of Pharmacy,2014,4:525-529
[5]Liu R H.Dietary bioactive compounds and their health implications[J].Journal of Food Science,2013,781:A18-A25
[6]Dejian H,Boxin O,Maureen H W,et al.High-throughput assay of oxygen radical absorbance capacity(ORAC)using a multichannel liquid handling system coupled with a microplate fluorescence reader in 96-well format[J].Journal of Agricultural&Food Chemistry,2002,50(16):4437
[7]Ming W Z,Rui F Z,Fang X Z.Phenolic profiles and antioxidant activity of black rice bran of different commercially available varieties[J].Journal of Agricultural&Food Chemistry,2010,58(13):7580-7587
[8]Wolfe K L,Xinmei K,Xiangjiu H,et al.Cellular antioxidant activity of common fruits[J].Journal of Agricultural&Food Chemistry,2008,56(18):8418-8426
[9]Chang T S,Lin V C.Melanogenesis inhibitory activity of two generic drugs:Cinnarizine and trazodone in mouse B16melanoma cells[J].International Journal of Molecular Sciences,2011,12(12):8787-8796
[10]Xin M,Yuan-Yuan W,Xiao M,et al.Antagonism of phenanthrene cytotoxicity for human embryo lung fibroblast cell line HFL-I by green tea polyphenols[J].Environmental Pollution,2011,159(1):164-168
[11]Komoike Y,Matsuoka M.Endoplasmic reticulum stress-mediated neuronal apoptosis by acrylamide exposure[J].Toxicology&Applied Pharmacology,2016,310:68-77
[12]李莹,王英,杨丙晔,等.Z-十八碳-9-烯-丙磺酰胺对小鼠B16细胞黑素生成的抑制作用[J].中国皮肤性病学杂志,2018,10:1120-1123LI Ying,WANG Ying,YANG Bing-ye,et al.Inhibition of Z-octadec-9-ene-propanesulfonamide on melanogenesis in mouse B16 cells[J].Chinese Journal of Dermatology and Venereology,2018,10:1120-1123
[13]Ou B,Huang D,Hampsch-Woodill M,et al.Analysis of antioxidant activities of common vegetables employing oxygen radical absorbance capacity(ORAC)and ferric reducing antioxidant power(FRAP)assays:A comparative study[J].J Agric Food Chem.,2002,50(11):3122-3128
[14]Porto C D,Calligaris S,Celotti E,et al.Antiradical properties of commercial cognacs assessed by the DPPH·test[J].Journal of Agricultural&Food Chemistry,2000,48(9):4241-4245
[15]Standley L,Winterton P,Marnewick J L,et al.Influence of processing stages on antimutagenic and antioxidant potentials of rooibos tea[J].Journal of Agricultural&Food Chemistry,2001,49(1):114-117
[16]李小燕,何婉莺,彭颖,等.琯溪蜜柚精油的抗氧化及其对B16细胞中酪氨酸酶的抑制效果[J].食品工业科技,2018,5:86-89LI Xiao-yan,HE Wan-ying,PENG Ying,et al.Antioxidation of Guanxi honey pomelo essential oil and its inhibitory effect on tyrosinase in B16 cells[J].Science and Technology of Food Industry,2018,5:86-89
[17]Labrinea E P,Georgiou C A.Stopped-flow method for assessment of p H and timing effect on the ABTS total antioxidant capacity assay[J].Analytica Chimica Acta,2004,526(1):63-68
[18]王立峰,鞠兴荣.3种薏米的多酚提取物清除过氧化氢自由基抗氧化能力评价[J].食品科学,2012,19:39-44WANG Li-feng,JU Xing-rong.Evaluation of antioxidant capacity of hydrogen peroxide free radicals extracted from polyphenols extracted from three kinds of glutinous rice[J].Food Science,2012,19:39-44
[19]赵建,刘璇,文镜.荧光分光光度计测定保健食品总抗氧化能力[J].食品科学,2010,22:301-305ZHAO Jian,LIU Xuan,WEN Jing.Determination of total antioxidant capacity of health foods by fluorescence spectrophotometer[J].Food Science,2010,22:301-305
[20]杨涛,吴辉辉,徐青,等.抗氧化性能评价ORAC法及最新研究进展[J].食品工业科技,2009,7:352-355YANG Tao,WU Hui-hui,XU Qing,et al.ORAC method for antioxidant performance evaluation and recent research progress[J].Science and Technology of Food Industry,2009,7:352-355
[21]李谣,陈金龙,夏春燕,等.基于Hep G2细胞模型的香菇柄粉多酚抗氧化及抗增殖活性[J].食品科学,2016,11:190-196LI Yao,CHEN Jin-long,XIA Chun-yan,et al.Antioxidant and antiproliferative activity of polyphenols from mushroom handle powder based on HepG2 cell model[J].Food Science,2016,11:190-196
[22]Wolfe K L,Xinmei K,Xiangjiu H,et al.Cellular antioxidant activity of common fruits[J].Journal of Agricultural&Food Chemistry,2008,56(18):8418-8426
[23]陈贞纯,贾玲燕,毛祖法,等.茶叶提取物对B16小鼠黑色素瘤细胞的生物学效应及机理研究[J].茶叶科学,2014,5:465-472CHEN Zhen-chun,JIA Ling-yan,MAO Zu-fa,et al.Biological effects and mechanism of tea extract on B16mouse melanoma cells[J].Tea Science,2014,5:465-472
[24]Crowley L C,Marfell B J,Waterhouse N J.Analyzing cell death by nuclear staining with Hoechst 33342[J].Cold Spring Harbor Protocols,2016,9:778-781
[25]黄海潮,郑公铭,张小红,等.龙眼果核中没食子酸的体外美白作用及机制探索[J].中国实验方剂学杂志,2015,19:121-124HUANG Hai-chao,ZHENG Gong-ming,ZHANGXiao-hong,et al.In vitro whitening effect and mechanism of gallic acid in longan fruit nucleus[J].Chinese Journal of Experimental Traditional Medical Formulae,2015,19:121-124
[26]Sato K,Takahashi H,Iraha R,et al.Down-regulation of tyrosinase expression by acetylsalicylic acid in murine B16melanoma[J].Biological&Pharmaceutical Bulletin,2008,31(1):33-37
[27]王立峰,鞠兴荣.3种薏米的多酚提取物清除过氧化氢自由基抗氧化能力评价[J].食品科学,2012,19:39-44WANG Li-feng,JU Xing-rong.Evaluation of antioxidant capacity of hydrogen peroxide free radicals extracted from polyphenols extracted from three kinds of glutinous rice[J].Food Science,2012,19:39-44