菜籽饼提取物抑制黑色素生成作用
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摘要
为了研究菜籽饼提取物对黑色素合成的抑制作用,为菜籽饼提取物作为美白化妆品的天然添加剂提供实验依据。采用ABTS法、DPPH法检测菜籽饼提取物清除自由基的能力,测定A-375黑色素细胞中酪氨酸酶活性和黑色素含量,并利用各自的试剂盒检测A-375细胞中抗氧化相关因子(MDA, SOD, GSH)。研究结果表明:菜籽饼提取物具有清除DPPH和ABTS的能力,对DPPH和ABTS的半数抑制率分别为304μg/mL和305.8μg/mL。菜籽饼提取物具有显著的抑制A-375黑色素的作用,药物作用48 h后,其IC50为113.3μg/m L,同时能够显著的降低黑色素的含量和酪氨酸酶的活性。与空白对照组(17.66 mol/ng)相比,菜籽饼提取物能显著降低A-375细胞内MDA含量,其抑制率为9.21 mol/ng,且能够显著提高SOD的含量,当浓度为125μg/m L时,SOD的含量较空白对照组提高了36.54 U/mL,与此同时,菜籽饼提取物亦能增加GSH的含量,当浓度为62.5μg/mL时,细胞中GSH的含量为6.43 mol/ng,其含量显著高于空白对照组。因此,菜籽饼提取物在降低黑色素含量、抑制酪氨酸酶活性的同时,也能够通过显著的抗氧化能力和清除自由基的能力,抑制A-375细胞中黑色素的生成。
In order to study the inhibitory effect of rapeseed cake extract on the synthesis of melanin, and to provide experimental basis for rapeseed extracts as natural additive for whitening cosmetics, the ABTS method and DPPH method were used to detect the ability of free radical scavenging by rapeseed extract. Besides, the tyrosinase activity and melanin content of A-375 melanoma cells were detected, as well as the effect of antioxidant related factor(MDA,SOD, GSH) in cell was also detected with each kit. The results showed that rapeseed cake extract had the scavenging ability of DPPH and ABTS, and their IC50 to DPPH and ABTs were 304 μg/mL and 305.8 μg/m L, respectively.Rapeseed cake extract had significant inhibition effect on A-375 melanoma. After treated 48 h by medicine, the IC50 of rapeseed cake was 113.3 μg/m L, and it could significantly reduce the melanin content and tyrosinase activity. Compared with control group(17.66 mol/ng) rapeseed cake extract could significantly reduce the content of MDA in A-375 cell,of which the inhibition rate was 9.21 mol/ng Rapeseed cake extract could significantly improve the content of SOD.When the concentration was 125 μg/m L, the content of SOD increased 36.54 U/m L than the control group. At the same time, rapeseed extract could also increase the content of GSH. When the concentration was 62.5 μg/m L, the content of GSH in cell was 6.43 mol/ng, which was significantly higher than in the control group. Therefore, rapeseed extracts had the ability of reducing melanin content and tyrosinase inhibition activity, at the same time, it could inhibit the formation of melanin in A-375 cells through its antioxidant ability and free radical scavenging capacity.
In order to study the inhibitory effect of rapeseed cake extract on the synthesis of melanin, and to provide experimental basis for rapeseed extracts as natural additive for whitening cosmetics, the ABTS method and DPPH method were used to detect the ability of free radical scavenging by rapeseed extract. Besides, the tyrosinase activity and melanin content of A-375 melanoma cells were detected, as well as the effect of antioxidant related factor(MDA,SOD, GSH) in cell was also detected with each kit. The results showed that rapeseed cake extract had the scavenging ability of DPPH and ABTS, and their IC50 to DPPH and ABTs were 304 μg/mL and 305.8 μg/m L, respectively.Rapeseed cake extract had significant inhibition effect on A-375 melanoma. After treated 48 h by medicine, the IC50 of rapeseed cake was 113.3 μg/m L, and it could significantly reduce the melanin content and tyrosinase activity. Compared with control group(17.66 mol/ng) rapeseed cake extract could significantly reduce the content of MDA in A-375 cell,of which the inhibition rate was 9.21 mol/ng Rapeseed cake extract could significantly improve the content of SOD.When the concentration was 125 μg/m L, the content of SOD increased 36.54 U/m L than the control group. At the same time, rapeseed extract could also increase the content of GSH. When the concentration was 62.5 μg/m L, the content of GSH in cell was 6.43 mol/ng, which was significantly higher than in the control group. Therefore, rapeseed extracts had the ability of reducing melanin content and tyrosinase inhibition activity, at the same time, it could inhibit the formation of melanin in A-375 cells through its antioxidant ability and free radical scavenging capacity.
引文
Bao J.L.,Ding R.B.,Zou L.D.,Zhang C.,Wang K.,Liu F.,Li P.,Chen M.W.,Wan J.B.,Su H.X.,Wang Y.T.,and He C.W.,2016,Forsythiae fructus inhibits B16 melanoma growth involving MAPKs/Nrf2/HO-1 mediated anti-oxidation and antiinflammation,The American Journal of Chinese Medicine,44(5):1-19
DudonnéS.,Vitrac X.,Coutière P.,Woillez M.,and Mérillon J.M.,2009,Comparative study of antioxidant properties and total phenolic content of 30 plant extracts of industrial interest using DPPH,ABTS,FRAP,SOD,and ORAC assays,Journal of Agricultural and Food Chemistry,57(5):1768-1774
Fu R.,Zhang Y.T.,Guo Y.R.,Peng T.,and Chen F.,2016,Hepatoprotection using Brassica rapa var.rapa L.seeds and its bioactive compound,sinapine thiocyanate,for CCl4-induced liver injury,Journal of Functional Foods,22:73-81
Jia A.Q.,Sun Y.,and Wang J.X.,2001,Adcance in studies of melanins inhibting mechanism and development of skin lightenors,Riyong Huaxue Gongye(China Surfactant Detergent and Cosmetics),31(1):41-44(贾爱群,孙洋,王建新,2001,美白剂的发展现状及其黑色素抑制机理的研究进展,日用化学工业,31(1):41-44)
Li J.X.,Yang Z.Q.,and Wang X.Z.,2006,Biological function of reactive oxygen free radicals in animals,Dongwu Yixue Jinzhan(Progress in Veterinary Medicine),27(10):33-36(李建喜,杨志强,王学智,2006,活性氧自由基在动物机体内的生物学作用,动物医学进展,27(10):33-36)
Li L.L.,Xing R.,Deng Y.Y.,Xie L.F.,and Wang L.,2016,New developments of tyrosinase inhibitors,Shipin Gongye(The Food Industry),37(8):235-239(李莉莉,邢蕊,邓阳阳,谢乐芳,王力,2016,酪氨酸酶抑制剂的研究新进展,食品工业,37(8):235-239)
Lin J.W.,Chiang H.M.,Lin Y.C.,and Wen K.C.,2008,Natural products with skin-whitening effects,Journal of food and drug analysis,16(2):l-10
Li P.Y.,Huo L.N.,Su W.,Lu R.M.,Deng C.C.,Liu L.Q.,Deng Y.K.,Guo N.N.,Lu C.S.,and He C.L.,2011,Free radicalscavenging capacity,antioxidant activity and phenolic content of Pouzolzia zeylanica,Journal of the Serbian Chemical Society,76(5):709-717
Li Y.W.,Xu A.E.,Wei X.D.,Zhang D.M.,and Liu J.F.,2005,Effects of glossy privet fruit on the proliferation melanin compound and c-kit gene expression in normal human melanocytes in vitro,Zhongguo Zhongxiyi Jiehe Pifuxing Bingxue Zazhi(Chinese Journal of Integrated Traditional and Western Medicine Dermatology),4(3):150-152(李永伟,许爱娥,尉晓冬,张迪敏,刘继峰,2005,女贞子对黑素细胞的黑素合成,细胞增殖和c-kit基因表达的影响,中国中西医结合皮肤性病学杂志,4(3):150-152)
Munoz-Munoz J.L.,García-Molina F.,Varón R.,Tudela J.,GarcíaCánovas F.,and Rodríguez-López J.N.,2009,Generation of hydrogen peroxide in the melanin biosynthesis pathway,Biochimica et biophysica acta,Proteins and Proteomics,1794(7):1017-1029
Shi F.,Jia X.,Zhao C.,and Chen Y.,2010,Antioxidant activities of various extracts from Artemisisa selengensis Turcz(Lu Hao),Molecules,15(7):4934-4946
Sies H.,1997,Oxidative stress:oxidants and antioxidants,Exper imental Physiology,82(2):291-295
Wang K.H.,Lin R.D.,Hsu F.L.,Huang Y.H.,Chang H.C.,Huang C.Y.,and Lee M.H.,2006,Cosmetic application of selected traditional Chinese herbal medicines,Journal of Ethnopharmacology,106(3):353-359
Wang P.,Li S.X.,Han F.S.,Wang Y.G.,and Shi H.J.,2013,Inhibition effect of grape seed extract on melanin formation in B16murine melanoma cells,Riyong Huaxue Gongye(China Surfactant Detergent and Cosmetics),43(4):280-284(王鹏,李素霞,韩福森,王勇刚,施惠娟,2013,葡萄籽提取物对B16细胞内黑色素合成的抑制作用,日用化学工业,43(4):280-284)
Wang Y.,Zhu B.,Jiang J.J.,and Wang Y.P.,2015,An review on feeding value and quality improvement of rapeseed meal,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),13(4):929-936(王月,朱宝,蒋金金,王幼平,2015,菜籽饼粕饲用价值及其品质改良的研究进展,分子植物育种,13(4):929-936)
Wilson M.J.,and Spaziani E.,1973,Testosterone regulation of pigmentation in scrotal epidermis of the rat,Cell and Tissue Research,140:451-458
Zhang H.Y.,Xing F.L.,Luo G.H.,Ye S.B.,Zhang Z.X.,Deng S.C.,and Hao J.Y.,2017,The inhibitory effects of grape seed proanthocyanidins on melanogenesis in mouse melanoma B16cell,Zhongyao Yaoli Yu Linchuang(Pharmacology and Clinics of Chinese Materia Medica),33(1):63-66(张慧瑛,邢芙玲,罗光宏,叶生宝,张泽星,邓世川,郝军元,2017,葡萄籽原花青素抑制小鼠B16-F0黑色素瘤细胞黑色素生成的影响,中药药理与临床,33(1):63-66)
Zhang J.,Chen Z.H.,Ye S.,Si Y.X.,and Li C.Y.,2012,Study on inhibition effect of Sargassum fusiforme polysaccharide extracts on activity of tyrosinase,Shipin Gongye Keji(Science and Technology of Food Industry),33(20):116-120(张捷,陈忠浩,叶森,斯越秀,李彩燕,2012,羊栖菜多糖提取物对酪氨酸酶的抑制作用研究,食品工业科技,33(20):116-120)
Zhu L.,Lu Y.,Yu W.G.,Zhao X.,and Lu Y.H.,2016,Anti-photoageing and anti-melanogenesis activities of chrysin,Pharmaceutical Biology,54(11):2692-2700
DudonnéS.,Vitrac X.,Coutière P.,Woillez M.,and Mérillon J.M.,2009,Comparative study of antioxidant properties and total phenolic content of 30 plant extracts of industrial interest using DPPH,ABTS,FRAP,SOD,and ORAC assays,Journal of Agricultural and Food Chemistry,57(5):1768-1774
Fu R.,Zhang Y.T.,Guo Y.R.,Peng T.,and Chen F.,2016,Hepatoprotection using Brassica rapa var.rapa L.seeds and its bioactive compound,sinapine thiocyanate,for CCl4-induced liver injury,Journal of Functional Foods,22:73-81
Jia A.Q.,Sun Y.,and Wang J.X.,2001,Adcance in studies of melanins inhibting mechanism and development of skin lightenors,Riyong Huaxue Gongye(China Surfactant Detergent and Cosmetics),31(1):41-44(贾爱群,孙洋,王建新,2001,美白剂的发展现状及其黑色素抑制机理的研究进展,日用化学工业,31(1):41-44)
Li J.X.,Yang Z.Q.,and Wang X.Z.,2006,Biological function of reactive oxygen free radicals in animals,Dongwu Yixue Jinzhan(Progress in Veterinary Medicine),27(10):33-36(李建喜,杨志强,王学智,2006,活性氧自由基在动物机体内的生物学作用,动物医学进展,27(10):33-36)
Li L.L.,Xing R.,Deng Y.Y.,Xie L.F.,and Wang L.,2016,New developments of tyrosinase inhibitors,Shipin Gongye(The Food Industry),37(8):235-239(李莉莉,邢蕊,邓阳阳,谢乐芳,王力,2016,酪氨酸酶抑制剂的研究新进展,食品工业,37(8):235-239)
Lin J.W.,Chiang H.M.,Lin Y.C.,and Wen K.C.,2008,Natural products with skin-whitening effects,Journal of food and drug analysis,16(2):l-10
Li P.Y.,Huo L.N.,Su W.,Lu R.M.,Deng C.C.,Liu L.Q.,Deng Y.K.,Guo N.N.,Lu C.S.,and He C.L.,2011,Free radicalscavenging capacity,antioxidant activity and phenolic content of Pouzolzia zeylanica,Journal of the Serbian Chemical Society,76(5):709-717
Li Y.W.,Xu A.E.,Wei X.D.,Zhang D.M.,and Liu J.F.,2005,Effects of glossy privet fruit on the proliferation melanin compound and c-kit gene expression in normal human melanocytes in vitro,Zhongguo Zhongxiyi Jiehe Pifuxing Bingxue Zazhi(Chinese Journal of Integrated Traditional and Western Medicine Dermatology),4(3):150-152(李永伟,许爱娥,尉晓冬,张迪敏,刘继峰,2005,女贞子对黑素细胞的黑素合成,细胞增殖和c-kit基因表达的影响,中国中西医结合皮肤性病学杂志,4(3):150-152)
Munoz-Munoz J.L.,García-Molina F.,Varón R.,Tudela J.,GarcíaCánovas F.,and Rodríguez-López J.N.,2009,Generation of hydrogen peroxide in the melanin biosynthesis pathway,Biochimica et biophysica acta,Proteins and Proteomics,1794(7):1017-1029
Shi F.,Jia X.,Zhao C.,and Chen Y.,2010,Antioxidant activities of various extracts from Artemisisa selengensis Turcz(Lu Hao),Molecules,15(7):4934-4946
Sies H.,1997,Oxidative stress:oxidants and antioxidants,Exper imental Physiology,82(2):291-295
Wang K.H.,Lin R.D.,Hsu F.L.,Huang Y.H.,Chang H.C.,Huang C.Y.,and Lee M.H.,2006,Cosmetic application of selected traditional Chinese herbal medicines,Journal of Ethnopharmacology,106(3):353-359
Wang P.,Li S.X.,Han F.S.,Wang Y.G.,and Shi H.J.,2013,Inhibition effect of grape seed extract on melanin formation in B16murine melanoma cells,Riyong Huaxue Gongye(China Surfactant Detergent and Cosmetics),43(4):280-284(王鹏,李素霞,韩福森,王勇刚,施惠娟,2013,葡萄籽提取物对B16细胞内黑色素合成的抑制作用,日用化学工业,43(4):280-284)
Wang Y.,Zhu B.,Jiang J.J.,and Wang Y.P.,2015,An review on feeding value and quality improvement of rapeseed meal,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),13(4):929-936(王月,朱宝,蒋金金,王幼平,2015,菜籽饼粕饲用价值及其品质改良的研究进展,分子植物育种,13(4):929-936)
Wilson M.J.,and Spaziani E.,1973,Testosterone regulation of pigmentation in scrotal epidermis of the rat,Cell and Tissue Research,140:451-458
Zhang H.Y.,Xing F.L.,Luo G.H.,Ye S.B.,Zhang Z.X.,Deng S.C.,and Hao J.Y.,2017,The inhibitory effects of grape seed proanthocyanidins on melanogenesis in mouse melanoma B16cell,Zhongyao Yaoli Yu Linchuang(Pharmacology and Clinics of Chinese Materia Medica),33(1):63-66(张慧瑛,邢芙玲,罗光宏,叶生宝,张泽星,邓世川,郝军元,2017,葡萄籽原花青素抑制小鼠B16-F0黑色素瘤细胞黑色素生成的影响,中药药理与临床,33(1):63-66)
Zhang J.,Chen Z.H.,Ye S.,Si Y.X.,and Li C.Y.,2012,Study on inhibition effect of Sargassum fusiforme polysaccharide extracts on activity of tyrosinase,Shipin Gongye Keji(Science and Technology of Food Industry),33(20):116-120(张捷,陈忠浩,叶森,斯越秀,李彩燕,2012,羊栖菜多糖提取物对酪氨酸酶的抑制作用研究,食品工业科技,33(20):116-120)
Zhu L.,Lu Y.,Yu W.G.,Zhao X.,and Lu Y.H.,2016,Anti-photoageing and anti-melanogenesis activities of chrysin,Pharmaceutical Biology,54(11):2692-2700
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