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Melanin Biosynthesis Inhibition Effects of Ginsenoside Rb2 Isolated from Panax ginseng Berry
Melanin Biosynthesis Inhibition Effects of Ginsenoside Rb2 Isolated from Panax ginseng Berry
Journal of Microbiology and Biotechnology. 2015. Dec, 25(12): 2011-2015
Copyright © 2015, The Korean Society For Microbiology And Biotechnology
  • Received : May 22, 2015
  • Accepted : October 04, 2015
  • Published : December 28, 2015
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About the Authors
Dae Young Lee
Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong 27709, Republic of Korea
Yong Tae Jeong
HK BIO, Business Incubator, Daegu Haany University, Gyeongsan, Gyeongbuk 38578, Republic of Korea
Sang Chul Jeong
Freshwater Bioresources Utilization Division, Nakdonggang National Institute of Biological Resources, SangJu-si 37242, Republic of Korea
Mi Kyoung Lee
Cancer Preventive Material Development Research Center, College of Oriental Medicine, Kyung Hee University, Seoul 02447, Republic of Korea
Jin Woo Min
Research & Development Center, GFC Co., Ltd., Suwon 16690, Republic of Korea
Jae Won Lee
Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong 27709, Republic of Korea
Geum Soog Kim
Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong 27709, Republic of Korea
Seung Eun Lee
Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong 27709, Republic of Korea
Young Sup Ahn
Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong 27709, Republic of Korea
Hee Cheol Kang
Research & Development Center, GFC Co., Ltd., Suwon 16690, Republic of Korea
Jin Hee Kim
College of Herbal Bio-industry, Daegu Haany University, Gyeongsan 42158, Republic of Korea
jinheekim@dhu.ac.kr

Abstract
Ginsenoside Rb2 (Gin-Rb2) was purified from the fruit extract of Panax ginseng . Its chemical structure was measured by spectroscopic analysis, including HR-FAB-MS, 1 H-NMR, and IR spectroscopy. Gin-Rb2 decreased potent melanogenesis in melan-a cells, with 23.4% at 80 μM without cytotoxicity. Gin-Rb2 also decreased tyrosinase and MITF protein expression in melan-a cells. Furthermore, Gin-Rb2 presented inhibition of the body pigmentation in the zebrafish in vivo system and reduced melanin contents and tyrosinase activity. These results show that Gin-Rb2 isolated from P. ginseng may be an effective skin-whitening agent via the in vitro and in vivo systems.
Keywords
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( A ) The structure of ginsenoside Rb2. ( B ) Effect of cell cytotoxicity was measured with triplicate experiments. The cells were cultured with 0-160 μM of Gin-Rb2 for 3 days, and cell cytotoxicity was determined by a CCK-8 cell counting kit. ( C ) Inhibition of melanin synthesis was measured with triplicate experiment. The cells were cultured with 0-80 μM of Gin-Rb2 for 3 days. Each value is expressed the mean ± SD of triplicate determinations. * p < 0.05, ** p < 0.01 versus control group.
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Melan-a cells were treated with 0-80 μM of Gin-Rb2 for 24 h. Total cell lysates were then prepared and subjected to western blotting using tyrosinase and MITF.
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Synchronized embryos were treated with the indicated concentrations of PTU and ginsenoside Rb2. Gin-Rb2 was dissolved in DMSO (0.1%) and then added to the embryo medium. ( A ) The change of phenotype on zebrafish was assessed using a microscope. ( B ) Total melanin content and tyrosinase activity were quantified using a spectrometer. Results shown are the mean of three independent experiments ± SD. * p < 0.05, *** p < 0.001 versus control group.
Acknowledgements
This work was carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ01035103)” Rural Development Administration, Republic of Korea.
References
Bae HM , Cho OS , Kim SJ , Im BO , Cho SH , Lee S 2012 Inhibitory effects of ginsenoside re isolated from ginseng berry on histamine and cytokine release in human mast cells and human alveolar epithelial cells. J. Ginseng Res. 36 369 - 374    DOI : 10.5142/jgr.2012.36.4.369
Bennett DC , Cooper PJ , Hart IR 1987 A line of non-tumorigenic mouse melanocytes, syngeneic with the B16 melanoma and requiring a tumour promoter for growth. Int. J. Cancer 39 414 - 418    DOI : 10.1002/ijc.2910390324
Bentley NJ , Eisen T , Goding CR 1994 Melanocyte-specific expression of the human tyrosinase promoter: activation by the microphthalmia gene product and role of the initiator. Mol. Cell. Biol. 14 7996 - 8006    DOI : 10.1128/MCB.14.12.7996
Briganti S , Camera E , Picardo M 2003 Chemical and instrumental approaches to treat hyperpigmentation. Pigment Cell Res. 16 101 - 110    DOI : 10.1034/j.1600-0749.2003.00029.x
Choi TY , Kim JH , Ko DH , Kim CH , Hwang JS , Ahn S 2007 Zebrafish as a new model for phenotype-based screening of melanogenic regulatory compounds. Pigment Cell Res. 20 120 - 127    DOI : 10.1111/j.1600-0749.2007.00365.x
Elsalini OA , Rohr KB 2003 Phenylthiourea disrupts thyroid function in developing zebrafish. Dev. Genes Evol. 212 593 - 598
Fujimoto N , Watanabe H , Nakatani T , Roy G , Ito A 1998 Induction of thyroid tumours in (C57BL/6N x C3H/N)F1 mice by oral administration of kojic acid. Food Chem. Toxicol. 36 697 - 703    DOI : 10.1016/S0278-6915(98)00030-1
Gaggioli C , Busca R , Abbe P , Ortonne JP , Ballotti R 2003 Microphthalmia-associated transcription factor (MITF) is required but is not sufficient to induce the expression of melanogenic genes. Pigment Cell Res. 16 374 - 382    DOI : 10.1034/j.1600-0749.2003.00057.x
Huo YS 1984 Anti-senility action of saponin inPanax ginsengfruit in 327 cases. Chin. J. Modern Dev. Trad. Med. 4
Im SJ , Kim KN , Yun YG , Lee JC , Mun YJ , Kim JH , Woo WH 2003 Effect of Radix Ginseng and Radix Trichosanthis on the melanogenesis. Biol. Pharm. Bull. 26 849 - 853    DOI : 10.1248/bpb.26.849
Kim KN , Yang HM , Kang SM , Ahn GN , Roh SW , Lee W 2015 Whitening effect of octaphlorethol a isolated fromIshige foliaceain anin vivozebrafish model. J. Microbiol. Biotechnol. 25 448 - 451    DOI : 10.4014/jmb.1409.09033
Kim S , Jung SH , Cho CW 2008 Physicochemical studies of a newly synthesized molecule, 6-methyl-3-phenethyl-3,4- dihydro-1H-quinazoline-2-thione (JSH18) for topical formulations. Arch. Pharm. Res. 31 1363 - 1368    DOI : 10.1007/s12272-001-2118-x
Kong YH , Jo YO , Cho CW , Son D , Park S , Rho J , Choi SY 2008 Inhibitory effects of cinnamic acid on melanin biosynthesis in skin. Biol. Pharm. Bull. 31 946 - 948    DOI : 10.1248/bpb.31.946
Lee DY , Cha BJ , Lee YS , Kim GS , Noh HJ , Kim SY 2015 The potential of minor ginsenosides isolated from the leaves ofPanax ginsengas inhibitors of melanogenesis. Int. J. Mol. Sci. 16 1677 - 1690    DOI : 10.3390/ijms16011677
Lee HJ , Kim SR , Kim JC , Kang CM , Lee YS , Jo SK 2006 In vivoradioprotective effect ofPanax ginsengC.A. Meyer and identification of active ginsenosides. Phytother. Res. 20 392 - 395    DOI : 10.1002/ptr.1867
Lee KT , Jung TW , Lee HJ , Kim SG , Shin YS , Whang WK 2011 The antidiabetic effect of ginsenoside Rb2viaactivation of AMPK. Arch. Pharm. Res. 34 1201 - 1208    DOI : 10.1007/s12272-011-0719-6
Sato K , Mochizuki M , Saiki I , Yoo YC , Samukawa K , Azuma I 1994 Inhibition of tumor angiogenesis and metastasis by a saponin ofPanax ginseng, ginsenoside-Rb2. Biol. Pharm. Bull. 17 635 - 639    DOI : 10.1248/bpb.17.635
Shao ZH , Xie JT , Vanden Hoek TL , Mehendale S , Aung H , Li CQ 2004 Antioxidant effects of American ginseng berry extract in cardiomyocytes exposed to acute oxidant stress. Biochim. Biophys. Acta 1670 165 - 171    DOI : 10.1016/j.bbagen.2003.12.001
Strahle U , Scholz S , Geisler R , Greiner P , Hollert H , Rastegar S 2012 Zebrafish embryos as an alternative to animal experiments - a commentary on the definition of the onset of protected life stages in animal welfare regulations. Reprod. Toxicol. 33 128 - 132    DOI : 10.1016/j.reprotox.2011.06.121
Wang L , Lu AP , Yu ZL , Wong RN , Bian ZX , Kwok HH 2014 The melanogenesis-inhibitory effect and the percutaneous formulation of ginsenoside Rb1. AAPS PharmSciTech 15 1252 - 1262    DOI : 10.1208/s12249-014-0138-3
Wang W , Zhao Y , Rayburn ER , Hill DL , Wang H , Zhang R 2007 In vitroanti-cancer activity and structure-activity relationships of natural products isolated from fruits ofPanax ginseng. Cancer Chemother. Pharmacol. 59 589 - 601    DOI : 10.1007/s00280-006-0300-z
Wu M , Hemesath TJ , Takemoto CM , Horstmann MA , Wells AG , Price ER 2000 c-Kit triggers dual phosphorylations, which couple activation and degradation of the essential melanocyte factor Mi. Genes Dev. 14 301 - 312
Xie JT , Aung HH , Wu JA , Attel AS , Yuan CS 2002 Effects of American ginseng berry extract on blood glucose levels in ob/ob mice. Am. J. Chin. Med. 30 187 - 194    DOI : 10.1142/S0192415X02000442
Xie JT , Shao ZH , Vanden Hoek TL , Chang WT , Li J , Mehendale S 2006 Antioxidant effects of ginsenoside Re in cardiomyocytes. Eur. J. Pharmacol. 532 201 - 207    DOI : 10.1016/j.ejphar.2006.01.001
Xie JT , Wang CZ , Ni M , Wu JA , Mehendale SR , Aung HH 2007 American ginseng berry juice intake reduces blood glucose and body weight in ob/ob mice. J. Food Sci. 72 S590 - S594    DOI : 10.1111/j.1750-3841.2007.00481.x
Xie JT , Wang CZ , Zhang B , Mehendale SR , Li XL , Sun S 2009 In vitroandin vivoanticancer effects of American ginseng berry: exploring representative compounds. Biol. Pharm. Bull. 32 1552 - 1558    DOI : 10.1248/bpb.32.1552
Yasumoto K , Yokoyama K , Takahashi K , Tomita Y , Shibahara S 1997 Functional analysis of microphthalmia-associated transcription factor in pigment cell-specific transcription of the human tyrosinase family genes. J. Biol. Chem. 272 503 - 509    DOI : 10.1074/jbc.272.1.503
Yoo YC , Lee J , Park SR , Nam KY , Cho YH , Choi JE 2013 Protective effect of ginsenoside-Rb2 from Korean red ginseng on the lethal infection of haemagglutinating virus of Japan in mice. J. Ginseng Res. 37 80 - 86    DOI : 10.5142/jgr.2013.37.80
Yoon HS , Lee SR , Ko HC , Choi SY , Park JG , Kim JK , Kim SJ 2007 Involvement of extracellular signal-regulated kinase in nobiletin-induced melanogenesis in murine B16/F10 melanoma cells. Biosci. Biotechnol. Biochem. 71 1781 - 1784    DOI : 10.1271/bbb.70088
Yun JW , Seo JA , Jeong YS , Bae IH , Jang WH , Lee J 2011 TRPV1 antagonist can suppress the atopic dermatitislike symptoms by accelerating skin barrier recovery. J. Dermatol. Sci. 62 8 - 15
Zhao X , Wang J , Li J , Fu L , Gao J , Du X 2009 Highly selective biotransformation of ginsenoside Rb1 to Rd by the phytopathogenic fungusCladosporium fulvum(syn.Fulvia fulva). J. Ind. Microbiol. Biotechnol. 36 721 - 726    DOI : 10.1007/s10295-009-0542-y