臺灣博碩士論文加值系統

近年來，酪胺酸酶抑制劑常被用於癌症和炎症治療，並在化妝品工業中開發作為增白或美白劑。酪胺酸酶抑制劑可以從天然的材料中提取，因此，本研究篩選17種中藥材，從中挑選出牡丹皮（Paeonia suffruticosa Andr.）做為進一步研究對象。牡丹皮於傳統中醫藥中用作為抗生素和抗糖尿病藥物，然而，關於該中藥材的酪胺酸酶抑制性質知之甚少。我們使用超聲波輔助提取法，溶劑使用水或95％乙醇進行提取。使用1,1-diphenyl-2-picrylhydrazyl (DPPH)清除試驗和測量總多酚及總黃酮含量，來研究兩種溶劑提取物的抗氧化能力。這些研究結果顯示兩種牡丹皮提取物皆具有良好的抗氧化性，但與水提取物相比，乙醇提取物顯示出更強的抗氧化活性（95.3％對94.1％）。乙醇提取物含有較高的酚類化合物含量（18.85 mg/g），其中的主要成分為牡丹酚(Paeonol)，皆對酪胺酸酶具有突出的抑制作用（IC50 = 0.85 mg/mL和1.10 mg/mL）。酵素抑制動力學分析表明牡丹皮的乙醇提取物和牡丹酚皆是非競爭性抑制劑，在它們存在時，酵素的Km值（0.51mM）保持不變，而酶反應的Vmax分別從24.33 mg/mL‧min降低到9.84 mg/mL‧min和3.40 mg/mL‧min，表示它們可以與酶或酶–基質複合物結合，以減少反應進行。我們的研究結果也發現，以紫外線誘導A2058人體黑色素瘤細胞是比以α-黑色素細胞刺激素誘導B16F10小鼠黑色素瘤細胞更接近實際使用的細胞美白效果的測試模型。近一步的研究結果顯示牡丹皮的乙醇提取物和牡丹酚皆能有效抑制細胞的黑色素合成和細胞內的酪胺酸酶表達。然而，以成本價格考量，牡丹皮的乙醇提取物是比牡丹酚較為經濟的美白替代品。

Recent years the development of tyrosinase inhibitors as a whitening agent is a hot research topic. It is typically extracted from natural resources. We screened 17 different Chinese herbs, and found that Paeonia suffruticosa Andr., commonly named as Cortex Moutan (CM), has the most potential to find new tyrosinase inhibitor. It traditionally used in Chinese medicine as an antibiotic and anti-diabetes medicine. However, little is known about the tyrosinase inhibitor properties of this plant. The plant was extracted with water or 95% ethyl alcohol using supersonic wave extraction. The antioxidant ability of different extracts was investigated using a DPPH scavenging assay and measuring the total polyphenol/flavones content. Subsequently, this investigation suggests that the P. suffruticosa Andr. extracts have good antioxidant properties. Furthermore, ethanol extracts show stronger oxidant activity compared with water extracts (95.3% vs. 94.1%). The alcohol extract contains phenoid compound content of 18.85 mg/g. The main ingredient of the alcohol extract is paeonol, both possess outstanding inhibition effects to the tyrosinase (IC50: 0.85 mg/mL and 1.10 mg/mL, respectfully). Kinetic analysis shows that the ethanol extracts of P. suffruticosa Andr. and paeonol are noncompetitive inhibitors. The Km value (0.51 mM) remained the same. However, the decrease in enzyme reaction Vmax (from 24.33 to 9.84/3.40 mg/mL‧min) suggested that the extract can bind with either the enzyme or the enzyme-substrate complex to diminish the enzyme reaction. The UV inducing A2058 human melanoma cells is a better testing model for skin whitening effectiveness than B16F10 cell line. Paeonol is effective in inhibiting melanin synthesis and cellular tyrosinase expression. However, in economic consideration, ethanol extract of P. suffruticosa Andr. is an appropriate alternative to Paeonol.

封面內頁
簽名頁
中文摘要 ................................................................................................... iii
英文摘要................................................................................................... v
誌謝................................................................................................... vi
圖目錄 ................................................................................................... ix
表目錄 ................................................................................................... x
前言................................................................................................... 1
第一章 文獻回顧 ................................................................................................... 3
第一節 皮膚色素代謝................................................................................................... 3
皮膚結構................................................................................................... 3
黑色素的形成與代謝 ................................................................................................... 5
抑制黑色素產生的基因調控 ................................................................................................... 9
減少黑色素被釋放到表皮細胞................................................................................................... 15
加速黑色素離開表皮層 ................................................................................................... 15
第二節 酵素抑制動力學 ................................................................................................... 16
第三節 中草藥美白研究 ................................................................................................... 22
第四節 現有之限制 ................................................................................................... 26
第二章 研究目標及架構 ................................................................................................... 32
第三章 材料及方法 ................................................................................................... 34
第一節 藥品與儀器設備 ................................................................................................... 34
第二節 儀器設備 ................................................................................................... 34
第三節 萃取方法 ................................................................................................... 35
第四節 DPPH抗氧化性試驗 ................................................................................................... 36
第五節 總酚含量分析 ................................................................................................... 36
第六節 抑制酪胺酸酶試驗 ................................................................................................... 37
第七節 高效能液相層析 ................................................................................................... 37
第八節 酵素動力學分析 ................................................................................................... 38
第九節 細胞培養 ................................................................................................... 38
第十節 MTT細胞毒性測試 ................................................................................................... 38
第十一節 細胞黑色素抑制試驗 ................................................................................................... 39
第十二節 細胞酪胺酸酶抑制試驗................................................................................................... 40
第四章 結果 ................................................................................................... 41
第一節 中藥萃取結果 ................................................................................................... 41
第二節 抗氧化性試驗 ................................................................................................... 43
第三節 抑制酪胺酸酶活性分析 ................................................................................................... 45
第四節 高效能液相層析 ................................................................................................... 49
第五節 酵素動力學分析 ................................................................................................... 51
第六節 細胞毒性試驗 ................................................................................................... 54
第七節 細胞黑色素量測定 ................................................................................................... 56
第八節 細胞酪胺酸酶抑制試驗 ................................................................................................... 58
第五章 討論 ................................................................................................... 60
參考文獻 ................................................................................................... 65
附件................................................................................................... 76
標準曲線 ................................................................................................... 76

1.曹智莞。2003。中草藥 植物精華 草本香料 最in的化妝保養品3大趨勢。市場情報。：14-6。
2.丁秀玉。1999。中藥牡丹皮、赤芍和津蒼朮化學成分及生物活性之研究：195。高雄醫學大學藥學研究所博士論文。高雄市，台灣。
3.王四切。嘉南藥理科技大學。2003。美容保健中草藥之開發與研究：1-8，嘉南藥理科技大學專題研究計畫，台南縣，台灣。
4.台大皮膚科部。2006。皮膚的構造及機能。實用皮膚醫學: 1-22。 孫啓璟。台北，台灣。台大醫學院。
5.李洪娟, 孙居锋, 代现平, 王术光 和张蕾。2009。桑白皮总黄酮超声波提取法研究。安徽農業科學。37(3)：1188-9。
6.張瑋玲。2005。中草藥美白活性研究 五倍子對b16黑色素細胞黑色素形成之抑制研究：93。台北醫學大學醫學研究所碩士論文。台北市，台灣。
7.楊武德, 王芳 and 丁寧。2008。HPLC法测定不同產地牡丹皮中丹皮酚含量。遼寧中醫藥大學學報。10(6)：187-8。
8.Alesiani, D., Cicconi, R., Mattei, M., Bei, R. and Canini, A. 2009. Inhibition of mek 1/2 kinase activity and stimulation of melanogenesis by 5, 7-dimethoxycoumarin treatment of melanoma cells. Int. J. Oncol. 34(6): 1727-35.
9.Ando, H., Ryu, A., Hashimoto, A., Oka, M. and Ichihashi, M. 1998. Linoleic acid and α-linolenic acid lightens ultraviolet-induced hyperpigmentation of the skin. Archives of Dermatological Research 290(7): 375-81.
10.Ando, H., Watabe, H., Valencia, J. C., Yasumoto, K.-I., Furumura, M., Funasaka, Y., Oka, M., Ichihashi, M. and Hearing, V. J. 2004. Fatty acids regulate pigmentation via proteasomal degradation of tyrosinase. J. Biol. Chem. 279(15): 15427-33.
11.Arung, E. T., Shimizu, K., Tanaka, H. and Kondo, R. 2010. 3-prenyl luteolin, a new prenylated flavone with melanin biosynthesis inhibitory activity from wood of artocarpus heterophyllus. Fitoterapia 81(6): 640-3.
12.Barsh, G., Gunn, T., He, L. I. N., Schlossman, S. and Duke-Cohan, J. 2000. Biochemical and genetic studies of pigment-type switching. Pigment Cell Res. 13: 48-53.
13.Bernard, Berthon and Bernard, P. 2000. Resveratrol: An original mechanism on tyrosinase inhibition. Int. J. Cosmet. Sci. 22(3): 219-26.
14.Bian, S., Doh, H. J., Zheng, J., Kim, J. S., Lee, C. H. and Kim, D. D. 2003. In vitro evaluation of patch formulations for topical delivery of gentisic acid in rats. Eur. J. Pharm. Sci. 18(2): 141-7.
15.Bonilla, C., Boxill, L. A., Mcdonald, S. A., Williams, T., Sylvester, N., Parra, E., Dios, S., Norton, H., Shriver, M. D. and Kittles, R. A. 2005. The 8818g allele of the agouti signaling protein (asip) gene is ancestral and is associated with darker skin color in african americans. Hum. Genet. 116(5): 402-6.
16.Cha, J. Y., Yang, H. J., Moon, H. I. and Cho, Y. S. 2012. Inhibitory effect and mechanism on melanogenesis from fermented herbals composition for medical or food uses. Food Research International 45(1): 225-31.
17.Chan, Y. Y., Kim, K. H. and Cheah, S. H. 2011. Inhibitory effects of sargassum polycystum on tyrosinase activity and melanin formation in b16f10 murine melanoma cells. J. Ethnopharmacol. 137(3): 1183-8.
18.Cheli, Y., Ohanna, M., Ballotti, R. and Bertolotto, C. 2010. Fifteen-year quest for microphthalmia-associated transcription factor target genes. Pigment Cell & Melanoma Research 23(1): 27-40.
19.Chen, L. G., Chang, W. L., Lee, C. J., Lee, L. T., Shih, C. M. and Wang, C. C. 2009. Melanogenesis inhibition by gallotannins from chinese galls in b16 mouse melanoma cells. Biol. Pharm. Bull. 32(8): 1447-52.
20.Chen, L. G., Chang, W. L., Lee, C. J., Lee, L. T., Shih, C. M. and Wang, C. C. 2009. Melanogenesis inhibition by gallotannins from chinese galls in b16 mouse melanoma cells. Biol. Pharm. Bull. 32(8): 6.
21.Chen, Q. X. and Kubo, I. 2002. Kinetics of mushroom tyrosinase inhibition by quercetin. J. Agric. Food Chem. 50(14): 4108-12.
22.Chen, W. S., Lin, L. D. and Tseng, C. F. 1988. Analysis of paeonol in moutan contex and it's pharmaceutical preparation by tlc and hplc. Annual Scientific Report of National Laboratories of Food and Drugs: 94-9 [In Chinese, English abstract].
23.Choi, H., Ahn, S., Lee, B. G., Chang, I. and Hwang, J. S. 2005. Inhibition of skin pigmentation by an extract of lepidium apetalum and its possible implication in il-6 mediated signaling. Pigment Cell Res. 18(6): 439-46.
24.Chu, H. L., Wang, B. S. and Duh, P. D. 2009. Effects of selected organo-sulfur compounds on melanin formation. Journal of Agricultural Food Chemistry 57(15): 7072-7.
25.Cone, R. D. and Mountjoy, K. G. 1993. Molecular genetics of the acth and melanocyte-stimulating hormone receptors. Trends Endocrinol. Metab. 4(7): 242-7.
26.Cooray, S. N. and Clark, A. J. L. 2011. Melanocortin receptors and their accessory proteins. Mol. Cell. Endocrinol. 331(2): 215-21.
27.Costin, G. E. and Hearing, V. J. 2007. Human skin pigmentation: Melanocytes modulate skin color in response to stress. Federation of American Societies for Experimental Biology Journal 21(4): 976-94.
28.Costin, G. E., Trif, M., Nichita, N., Dwek, R. A. and Petrescu, S. M. 2002. Ph-sensitive liposomes are efficient carriers for endoplasmic reticulum-targeted drugs in mouse melanoma cells. Biochem. Biophys. Res. Commun. 293(3): 918-23.
29.De La Lande, A., Maddaluno, J., Parisel, O., Darden, T. and Piquemal, J. 2010. Study of the docking of competitive inhibitors at a model of tyrosinase active site: Insights from joint broken-symmetry/spin-flip dft computations and elf topological analysis. Interdisciplinary Sciences: Computational Life Sciences 2(1): 3-11.
30.Dessinioti, C., Stratigos, A. J., Rigopoulos, D. and Katsambas, A. D. 2009. A review of genetic disorders of hypopigmentation: Lessons learned from the biology of melanocytes. Exp. Dermatol. 18(9): 741-9.
31.Fais, A., Corda, M., Era, B., Fadda, M. B., Matos, M. J., Quezada, E., Santana, L., Picciau, C., Podda, G. and Delogu, G. 2009. Tyrosinase inhibitor activity of coumarin-resveratrol hybrids. Molecules 14(7): 2514-20.
32.Ferioli, V., Rustichelli, C., Pavesi, G. and Gamberini, G. 2001. New combined treatment of hypermelanosis: Analytical studies on efficacy and stability improvement. Int. J. Cosmet. Sci. 23(6): 333-40.
33.Funasaka, Y., Komoto, M. and Ichihashi, M. 2000. Depigmenting effect of α-tocopheryl ferulate on normal human melanocytes. Pigment Cell Res. 13: 170-4.
34.Granata, A., Monzani, E., Bubacco, L. and Casella, L. 2006. Mechanistic insight into the activity of tyrosinase from variable-temperature studies in an aqueous/organic solvent. Chemistry – A European Journal 12(9): 2504-14.
35.Hakozaki, T., Minwalla, L., Zhuang, J., Chhoa, M., Matsubara, A., Miyamoto, K., Greatens, A., Hillebrand, G., Bissett, D. and Boissy, R. 2002. The effect of niacinamide on reducing cutaneous pigmentation and suppression of melanosome transfer. Br. J. Dermatol. 147(1): 20-31.
36.Hirobe, T. 2005. Role of keratinocyte-derived factors involved in regulating the proliferation and differentiation of mammalian epidermal melanocytes. Pigment Cell Res. 18(1): 2-12.
37.Hwang, J. H. and Lee, B. M. 2007. Inhibitory effects of plant extracts on tyrosinase, l-dopa oxidation, and melanin synthesis. Journal of Toxicology and Environmental Health, Part A 70(5): 393 - 407.
38.Hyun, S. K., Lee, W. H., Jeong Da, M., Kim, Y. and Choi, J. S. 2008. Inhibitory effects of kurarinol, kuraridinol, and trifolirhizin from sophora flavescens on tyrosinase and melanin synthesis. Biol. Pharm. Bull. 31(1): 154-8.
39.Jimenez, M., Chazarra, S., Escribano, J., Cabanes, J. and Garcia-Carmona, F. 2001. Competitive inhibition of mushroom tyrosinase by 4-substituted benzaldehydes. J. Agric. Food Chem. 49(8): 4060-3.
40.Karolien Van Den Bossche, J.-M. N. J. L. 2006. The quest for the mechanism of melanin transfer. Traffic 7(7): 769-78.
41.Kim, D. S., Lee, S., Lee, H. K., Park, S. H., Ryoo, I. J., Yoo, I. D., Kwon, S. B., Baek, K. J., Na, J. I. and Park, K. C. 2008. The hypopigmentary action of ki-063 (a new tyrosinase inhibitor) combined with terrein J. Pharm. Pharmacol. 60(3): 343-8.
42.Kim, D. S., Park, S. H., Kwon, S. B., Park, E. S., Huh, C. H., Youn, S. W. and Park, K. C. 2006. Sphingosylphosphorylcholine-induced erk activation inhibits melanin synthesis in human melanocytes. Pigment Cell Res. 19(2): 146-53.
43.Kim, D. S., Park, S. H. and Park, K. C. 2004. Transforming growth factor-[beta]1 decreases melanin synthesis via delayed extracellular signal-regulated kinase activation. The International Journal of Biochemistry & Cell Biology 36(8): 1482-91.
44.Kim, S. J., Son, K. H., Chang, H. W., Kang, S. S. and Kim, H. P. 2003. Tyrosinase inhibitory prenylated flavonoids from sophora flavescens. Biological Pharmaceutical Bulletin 26(9): 1348-50.
45.Kim, S. J., Son, K. H., Chang, H. W., Kang, S. S. and Kim, H. P. 2003. Tyrosinase inhibitory prenylated flavonoids from sophora flavescens. Biol. Pharm. Bull. 26(9): 1348-50.
46.Kim, Y. J. and Uyama, H. 2005. Tyrosinase inhibitors from natural and synthetic sources: Structure, inhibition mechanism and perspective for the future. Cell. Mol. Life Sci. 62(15): 1707-23.
47.Kubo, I., Chen, Q. X. and Nihei, K. 2003. Molecular design of antibrowning agents: Antioxidative tyrosinase inhibitors. Food Chem. 81(2): 241-7.
48.Kubo, I., Chen, Q. X., Nihei, K., Calderón, J. S. and Céspedes, C. L. 2003. Tyrosinase inhibition kinetics of anisic acid. Z Naturforsch [C] 58(9-10): 713-8.
49.Kubo, I. and Kinst-Hori, I. 1999. Flavonols from saffron flower: Tyrosinase inhibitory activity and inhibition mechanism. J. Agric. Food Chem. 47(10): 4121-5.
50.Kumaran, A. and Karunakaran, R. J. 2007. Activity-guided isolation and identification of free radical-scavenging components from an aqueous extract of coleus aromaticus. Food Chem. 100(1): 356-61.
51.Laidler, K. J. and Bunting, P. S. 1973. The chemical kinetics of enzyme action, Clarendon Press Oxford.
52.Lam, R. Y., Lin, Z. X., Sviderskaya, E. and Cheng, C. H. 2010. Application of a combined sulphorhodamine b and melanin assay to the evaluation of chinese medicines and their constituent compounds for hyperpigmentation treatment. J. Ethnopharmacol. 132(1): 274-9.
53.Lee, M. H., Kim, H. J., Ha, D. J., Paik, J. H. and Kim, H. Y. 2002. Therapeutic effect of topical application of linoleic acid and lincomycin in combination with betamethasone valerate in melasma patients. J. Korean Med. Sci. 17(4): 518-23.
54.Liang, C. H., Chou, T. H. and Ding, H. Y. 2010. Inhibition of melanogensis by a novel origanoside from origanum vulgare. J. Dermatol. Sci. 57(3): 170-7.
55.Lim, J. Y., Ishiguro, K. and Kubo, I. 1999. Tyrosinase inhibitory p-coumaric acid from ginseng leaves. Phytother. Res. 13(5): 371-5.
56.Maeda, K., Tomita, Y., Naganuma, M. and Tagaml, H. 1996. Phospholipases induce melanogenesis in organ-cultured skin. Photochem. Photobiol. 64(1): 220-3.
57.Maresca, V., Flori, E., Cardinali, G., Briganti, S., Lombardi, D., Mileo, A. M., Paggi, M. G. and Picardo, M. 2006. Ferritin light chain down-modulation generates depigmentation in human metastatic melanoma cells by influencing tyrosinase maturation. J. Cell. Physiol. 206(3): 843-8.
58.Martínez Esparza, M., Ferrer, C., Castells, M. T., García Borrón, J. C. and Zuasti, A. 2001. Transforming growth factor [beta]1 mediates hypopigmentation of b16 mouse melanoma cells by inhibition of melanin formation and melanosome maturation. The International Journal of Biochemistry & Cell Biology 33(10): 971-83.
59.Minwalla, L., Zhao, Y., Cornelius, J., Babcock, G. F., Wickett, R. R., Le Poole, I. C. and Boissy, R. E. 2001. Inhibition of melanosome transfer from melanocytes to keratinocytes by lectins and neoglycoproteins in an in vitro model system. Pigment Cell Res. 14(3): 185-94.
60.Nair, X., Parab, P., Suhr, L. and Tramposch, K. M. 1993. Combination of 4-hydroxyanisole and all-trans retinoic acid produces synergistic skin depigmentation in swine. Journal of Investigative Dermatology 101(2): 145-9.
61.Nakajima, M., Shinoda, I., Mikogami, T., Iwamoto, H., Hashimoto, S. I., Miyauchi, H., Fukuwatari, Y. and Hayasawa, H. 1997. Β-lactoglobulin suppresses melanogenesis in cultured human melanocytes. Pigment Cell Res. 10(6): 410-3.
62.Nanda, V., Parwaz, M. A. and Handa, S. 2006. Linear hypopigmentation after triamcinolone injection: A rare complication of a common procedure. Aesthetic Plast. Surg. 30(1): 118-9.
63.No, J. K., Kim, Y. J., Lee, J. S. and Chung, H. Y. 2006. Inhibition of melanogenic activity by 4,4'-dihydroxybiphenyl in melanoma cells. Biol. Pharm. Bull. 29(1): 14-6.
64.Nordlund, J. J. 2007. The melanocyte and the epidermal melanin unit: An expanded concept. Dermatol. Clin. 25(3): 271-81.
65.Ohguchi, K., Akao, Y. and Nozawa, Y. 2005. Involvement of calpain in melanogenesis of mouse b16 melanoma cells. Mol. Cell. Biochem. 275(1): 103-7.
66.Olivares, C., Jiménez-Cervantes, C., Lozano, J. A., Solano, F. and García-Borrón, J. C. 2001. The 5,6-dihydroxyindole-2-carboxylic acid (dhica) oxidase activity of human tyrosinase. Biochem. J. 354(1): 131-9.
67.Ong, E. S. 2004. Extraction methods and chemical standardization of botanicals and herbal preparations. Journal of Chromatography B 812(1–2): 23-33.
68.Paniwnyk, L., Beaufoy, E., Lorimer, J. P. and Mason, T. J. 2001. The extraction of rutin from flower buds of sophora japonica. Ultrason. Sonochem. 8(3): 299-301.
69.Parvez, S., Kang, M., Chung, H. S. and Bae, H. 2007. Naturally occurring tyrosinase inhibitors: Mechanism and applications in skin health, cosmetics and agriculture industries. Phytother. Res. 21(9): 805-16.
70.Parvez, S., Kang, M., Chung, H. S., Cho, C., Hong, M. C., Shin, M. K. and Bae, H. 2006. Survey and mechanism of skin depigmenting and lightening agents. Phytother. Res. 20(11): 921-34.
71.Peng, L. H., Liu, S., Xu, S. Y., Chen, L., Shan, Y. H., Wei, W., Liang, W. Q. and Gao, J. Q. 2013. Inhibitory effects of salidroside and paeonol on tyrosinase activity and melanin synthesis in mouse b16f10 melanoma cells and ultraviolet b-induced pigmentation in guinea pig skin. Phytomedicine 20(12): 1082-7.
72.Rendon, M. I. and Gaviria, J. I. 2005. Review of skin‐lightening agents. Dermatol. Surg. 31(s1): 886-90.
73.Sanchez-Laorden, B. L., Sanchez-Mas, J., Martinez-Alonso, E., Martinez-Menarguez, J. A., Garcia-Borron, J. C. and Jimenez-Cervantes, C. 2006. Dimerization of the human melanocortin 1 receptor: Functional consequences and dominant-negative effects. J. Invest. Dermatol. 126(1): 172-81.
74.Sapkota, K., Park, S. E., Kim, J. E., Kim, S., Choi, H. S., Chun, H. S. and Kim, S. J. 2010. Antioxidant and antimelanogenic properties of chestnut flower extract. Biosci. Biotechnol. Biochem. 74(8): 1527-33.
75.Schiöth, H. B., Phillips, S. R., Rudzish, R., Birch-Machin, M. A., Wikberg, J. E. S. and Rees, J. L. 1999. Loss of function mutations of the human melanocortin 1 receptor are common and are associated with red hair. Biochem. Biophys. Res. Commun. 260(2): 488-91.
76.Segel, I. H. 1975. Enzyme kinetics : Behavior and analysis of rapid equilibrium and steady-state enzime systems, New York : John Wiley & Sons.
77.Seiberg, M., Paine, C., Sharlow, E., Andrade-Gordon, P., Costanzo, M., Eisinger, M. and Shapiro, S. S. 2000. Inhibition of melanosome transfer results in skin lightening. J. Invest. Dermatol. 115(2): 162-7.
78.Seo, D.H., Jung, J.H., Lee, J.E., Jeon, E.J., Kim, W. and Park, C.-S. 2012. Biotechnological production of arbutins (α- and β-arbutins), skin-lightening agents, and their derivatives. Appl. Microbiol. Biotechnol. 95(6): 1417-25.
79.Shimizu, K., Kondo And, R. and Sakai, K. 2000. Inhibition of tyrosinase by flavonoids, stilbenes and related 4-substituted resorcinols: Structure-activity investigations. Planta Med. 66(01): 11-5.
80.Shin, N. H., Ryu, S. Y., Choi, E. J., Kang, S. H., Chang, I. M., Min, K. R. and Kim, Y. 1998. Oxyresveratrol as the potent inhibitor on dopa oxidase activity of mushroom tyrosinase. Biochem. Biophys. Res. Commun. 243(3): 801-3.
81.Slominski, A., Plonka, P. M., Pisarchik, A., Smart, J. L., Tolle, V., Wortsman, J. and Low, M. J. 2005. Preservation of eumelanin hair pigmentation in proopiomelanocortin-deficient mice on a nonagouti (a/a) genetic background. Endocrinology 146(3): 1245-53.
82.Solano, F., Briganti, S., Picardo, M. and Ghanem, G. 2006. Hypopigmenting agents: An updated review on biological, chemical and clinical aspects. Pigment Cell Res. 19(6): 550-71.
83.Song, T. Y., Chen, C. H., Yang, N. C. and Fu, C. S. 2009. The correlation of in vitro mushroom tyrosinase activity with cellular tyrosinase activity and melanin formation in melanoma cells a2058. Journal of Food and Drug Analysis 17(3).
84.Sprong, H., Degroote, S., Claessens, T., Van Drunen, J., Oorschot, V., Westerink, B. H. C., Hirabayashi, Y., Klumperman, J., Van Der Sluijs, P. and Van Meer, G. 2001. Glycosphingolipids are required for sorting melanosomal proteins in the golgi complex. The Journal of Cell Biology 155(3): 369-80.
85.Star, R. A., Rajora, N., Huang, J., Stock, R. C., Catania, A. and Lipton, J. M. 1995. Evidence of autocrine modulation of macrophage nitric oxide synthase by alpha-melanocyte-stimulating hormone. Proceedings of the National Academy of Sciences 92(17): 8016-20.
86.Steingrímsson, E., Copeland, N. G. and Jenkins, N. A. 2004. Melanocytes and the microphthalmia transcription factor network. Annu. Rev. Genet. 38(1): 365-411.
87.Sugimoto, K., Nishimura, T., Nomura, K. and Kuriki, T. 2004. Inhibitory effects of alpha-arbutin on melanin synthesis in cultured human melanoma cells and a three-dimensional human skin model. Biol. Pharm. Bull. 27(4): 510-4.
88.Tadokoro, T., Rouzaud, F., Itami, S., Hearing, V. J. and Yoshikawa, K. 2003. The inhibitory effect of androgen and sex-hormone-binding globulin on the intracellular camp level and tyrosinase activity of normal human melanocytes. Pigment Cell Res. 16(3): 190-7.
89.Tang, Y. P., Li, Y. F., Hu, J. and Lou, F. C. 2002. Isolation and identification of antioxidants from sophora japonica. Journal of Asian Natural Products Research 4(2): 123-8.
90.Thompson, R. C., Morris, S., Krull, I. and Swartz, M. 2001. Striving to validate nutraceuticals. LC-GC North America 19(11): 1142-9.
91.Van Den Bossche, K., Naeyaert, J. M. and Lambert, J. 2006. The quest for the mechanism of melanin transfer. Traffic 7(7): 769-78.
92.Wang, K. H., Lin, R. D., Hsu, F. L., Huang, Y. H., Chang, H. C., Huang, C. Y. and Lee, M. H. 2006. Cosmetic applications of selected traditional chinese herbal medicines. J. Ethnopharmacol. 106(3): 353-9.
93.Winder, A. J. and Harris, H. 1991. New assays for the tyrosine hydroxylase and dopa oxidase activities of tyrosinase. Eur. J. Biochem. 198(2): 317-26.
94.Woosley, J. T. and Muldoon, T. G. 1977. Comparison of the accuracy of the scatchard, lineweaver-burk and direct linear plots for the analysis of steroid-protein interactions. J. Steroid Biochem. 8(6): 625-9.
95.Xia, Y., Wikberg, J. E. S. and Chhajlani, V. 1995. Expression of melanocortin 1 receptor in periaqueductal gray matter. Neuroreport 6(16): 2193-6.
96.Yamaguchi, Y., Itami, S., Watabe, H., Yasumoto, K. I., Abdel Malek, Z. A., Kubo, T., Rouzaud, F., Tanemura, A., Yoshikawa, K. and Hearing, V. J. 2004. Mesenchymal–epithelial interactions in the skin. The Journal of Cell Biology 165(2): 275-85.
97.Yang, Y., Zhang, T., Xiao, L. and Chen, R.-Y. 2010. Two novel flavanes from the leaves of morus alba l. Journal of Asian Natural Products Research 12(3): 194-8.
98.Ye, Y., Chu, J. H., Wang, H., Xu, H., Chou, G. X., Leung, A. K. M., Fong, W. F. and Yu, Z. L. 2010. Involvement of p38 mapk signaling pathway in the anti-melanogenic effect of san-bai-tang, a chinese herbal formula, in b16 cells. J. Ethnopharmacol. 132(2): 533-5.
99.Yoon, N. Y., Eom, T. K., Kim, M. M. and Kim, S. K. 2009. Inhibitory effect of phlorotannins isolated from ecklonia cava on mushroom tyrosinase activity and melanin formation in mouse b16f10 melanoma cells. J. Agric. Food Chem. 57(10): 4124-9.
100.Zhang, L., Ding, Z., Xu, P., Wang, Y., Gu, Z., Qian, Z., Shi, G. and Zhang, K. 2011. Methyl lucidenate f isolated from the ethanol-soluble-acidic components of ganoderma lucidum is a novel tyrosinase inhibitor. Biotechnology and Bioprocess Engineering 16(3): 457-61.
101.Zheng, Z. P., Tan, H. Y. and Wang, M. F. 2012. Tyrosinase inhibition constituents from the roots of morus australis. Fitoterapia 83(6): 1008-13.
102.Zhong, S., Wu, Y., S.M., A., Zhao, J., Wang, K., Yang, S., Jae-Ho, Y. and Zhu, X. 2006. Depigmentation of melanocytes by the treatment of extracts from traditional chinese herbs: A cell culture assay. Biol. Pharm. Bull. 29(9): 1947-51.