果蔬酵素对黑色素生成的影响
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摘要
研究3种果蔬酵素对体外培养小鼠黑色素瘤细胞B16的酪氨酸酶活性的作用及对胞内黑色素生成及活性氧簇(ROS)相对产生率的影响,评价3种果蔬酵素的美白功效。采用四甲基偶氮唑蓝(MTT)比色法测定酵素对细胞增殖率的影响,左旋多巴(L-DOPA)氧化法检测对酪氨酸酶活性的抑制作用,氢氧化钠裂解法检测对胞内黑色素含量的影响,流式细胞仪检测对偶氮二异丁脒盐酸盐(AAPH)诱导的胞内ROS产生量的影响,从抑制酪氨酸酶活、减少黑色素含量和抗氧化方面评价3种酵素的美白效果。结果表明, 3种酵素均有美白作用,相比于台湾酵素和日本酵素,宁夏酵素对酪氨酸酶活性的抑制效果更明显,能在一定程度上减少黑色素的生成并保护细胞免受氧化应激的损伤,提高其抗氧化能力。
The inhibition of tyrosinase activity, the regulation of melanogenesis and oxidative stress in the response of B16 murine melanoma cells in vitro to three kinds of fruit and vegetable enzymes and evaluate the whitening efficacy of three enzymes were studied. Through the Thiazolyl Blue Tetrazolium Bromide(MTT) assay, the effect of enzymes on cell proliferation rate was tested. The tyrosinase activity was detected by levodopa(L-DOPA) oxidation, the melanin was measured by NaOH-assay, and the reactive oxygen species(ROS) level in cell induced by 2, 2'-azobis [2-methylpropionamidine]dihydrochloride(AAPH) was detected by flow cytometry instrument to determine the whitening effect from the aspects of inhibiting the activity of tyrosinase, reduction of melanin content and antioxidant activity. The results show that all three enzymes have whitening effect. Compared with the enzymes in Taiwan and Japanese enzymes, Ningxia's enzymes have more pronounced inhibitory effect on tyrosinase activity, to some extent, it can reduce the production of melanin and protect cells against oxidation to improve its antioxidant capacity.
The inhibition of tyrosinase activity, the regulation of melanogenesis and oxidative stress in the response of B16 murine melanoma cells in vitro to three kinds of fruit and vegetable enzymes and evaluate the whitening efficacy of three enzymes were studied. Through the Thiazolyl Blue Tetrazolium Bromide(MTT) assay, the effect of enzymes on cell proliferation rate was tested. The tyrosinase activity was detected by levodopa(L-DOPA) oxidation, the melanin was measured by NaOH-assay, and the reactive oxygen species(ROS) level in cell induced by 2, 2'-azobis [2-methylpropionamidine]dihydrochloride(AAPH) was detected by flow cytometry instrument to determine the whitening effect from the aspects of inhibiting the activity of tyrosinase, reduction of melanin content and antioxidant activity. The results show that all three enzymes have whitening effect. Compared with the enzymes in Taiwan and Japanese enzymes, Ningxia's enzymes have more pronounced inhibitory effect on tyrosinase activity, to some extent, it can reduce the production of melanin and protect cells against oxidation to improve its antioxidant capacity.
引文
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[2]GILLBRO J M,OLSSON M J.The melanogenesis and mechanisms of skin-lightening agents-existing and new approaches[J].International Journal of Cosmetic Science,2011,33(3):210-221.
[3]刘加友,王振斌.微生物酵素食品研究进展[J].食品与发酵工业,2016,42(1):273-276.
[4]刘祺,陈朝银,赵声兰.酵素化妆品的研发与展望[J].药物生物技术,2016(5):462-466.
[5]刘朝圣,龚坚,申梦洁.湘西龙山百合对小鼠B-16黑色素瘤细胞黑色素含量、酪氨酸酶活性的影响[J].湖南中医药大学学报,2017,37(2):145-148.
[6]解士海.人表皮黑素细胞--角质形成细胞共培养及11种化合物对黑素沉着的影响研究[D].北京:中国协和医科大学,2006.
[7]步犁,程树军,马来记.黑色素形成分子调控及美白剂功效评价方法[J].华南国防医学杂志,2012,26(4):395-398.
[8]叶希韵,朱萍亚.黑色素的合成与美白产品的研究进展[J].华东师范大学学报(自然科学版),2016,2016(2):1-8.
[9]STARNER R J,MCCLELLAND L,ABDEL-MALEK Z,et al.PGE 2,is a UVR-inducible autocrine factor for human melanocytes that stimulates tyrosinase activation[J]Experimental Dermatology,2010,19(7):682.
[10]李玲,宋伟民,周华.4种皮肤美白剂对小鼠黑素瘤细胞的功效评价[J].环境与职业医学,2006,23(5):385-389.