Whitening Effect of Storage Protein 2 from Silkworm Hemolymph
Author(s)
Z-Hun Kim,
Jung Wook Hwang,
Joon Ho Lee,
Hojin Kim,
Dae-Seog Lim,
Sangjin Kang,
Heui Sam Lee,
Yong-Soo Choi*
Affiliation(s)
Department of Applied Bioscience, CHA University, Seongnam, Korea. Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA, Suwon, Korea.
Department of Applied Bioscience, CHA University, Seongnam, Korea. Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA, Suwon, Korea.
ABSTRACT
This study investigated the effects of silkworm hemolymph-derived storage protein 2 (SP2) on the whitening process in mouse B16F1 melanoma cells. After the cells were treated with various concentrations of SP2 (0.1 - 1.0 mg/mL), cytotoxicity, melanin contents, and differences in mRNA and protein expression associated with melanogenesis were observed. No cytotoxicity was observed when cells were treated with SP2, even with increased SP2 concentrations of up to 2.0 mg/mL. When treated with various SP2 concentrations in the cells, the protein and mRNA expression of tyrosinase were dose-dependently decreased, respectively, and inhibition of tyrosinase was further increased by 50.0% with increasing SP2 concentration of 1.0 mg/mL. Expression mRNAs coding tyrosinase related protein-1 and protein-2 (TRP-1 and TRP-2) was also significantly decreased in a dose dependent manner. When measuring the melanin content in melanoma cells, SP2 at 1 mg/mL inhibited melanin synthesis by 73.5% compared with non-treated cells. The inhibitory effect was 2.8-fold higher than that obtained using arbutin as a positive control. This study demonstrates that SP2, as a whitening material, is capable of suppressing melanin synthesis through the downregulation of proteins and genes in the melanin biosynthesis pathway.
Cite this paper
Kim, Z. , Hwang, J. , Lee, J. , Kim, H. , Lim, D. , Kang, S. , Lee, H. and Choi, Y. (2014) Whitening Effect of Storage Protein 2 from Silkworm Hemolymph. Advances in Bioscience and Biotechnology, 5, 758-767. doi: 10.4236/abb.2014.59089.
Kim, Z. , Hwang, J. , Lee, J. , Kim, H. , Lim, D. , Kang, S. , Lee, H. and Choi, Y. (2014) Whitening Effect of Storage Protein 2 from Silkworm Hemolymph. Advances in Bioscience and Biotechnology, 5, 758-767. doi: 10.4236/abb.2014.59089.
References
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http://dx.doi.org/10.1016/j.phymed.2013.04.015 [4] Burnett, C.L., Bergfeld, W.F., Belsito, D.V., Hill, R.A., Klaassen, C.D., Liebler, D.C., Marks, J.G., Shank, R.C., Slaga, T.J. and Snyder, P.W. (2010) Final Report of the Safety Assessment of Kojic Acid as Used in Cosmetics. International Journal of Toxicology, 29, 244S-273S.
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http://dx.doi.org/10.1016/S0141-0229(03)00149-2 [17] Izumi, S., Fujie, J., Yamada, S. and Tomino, S. (1981) Molecular Properties and Biosynthesis of Major Plasma Proteins in Bombyx mori. Biochimica et Biophysica Acta-Protein Structure, 670, 222-229.
http://dx.doi.org/10.1016/0005-2795(81)90013-1 [18] Pietrzyk, A.J., Bujacz, A., Mueller-Dieckmann, J., Lochynska, M., Jaskolski, M. and Bujacz, G. (2013) Two Crystal Structures of Bombyx mori Lipoprotein 3-Structural Characterization of a New 30-kDa Lipoprotein Family Member. PLoS ONE, 8, Article ID: e61303.
http://dx.doi.org/10.1371/journal.pone.0061303 [19] Kim, E.J. and Park, T.H. (2004) Increase of Host Cell Longevity by the Expression of 30K Protein Originating from Silkworm Hemolymph in an Insect Cell-Baculovirus System. Enzyme and Microbial Technology, 35, 581-586.
http://dx.doi.org/10.1016/j.enzmictec.2004.08.024 [20] Rhee, W.J., Kim, E.J. and Park, T.H. (2002) Silkworm Hemolymph as a Potent Inhibitor of Apoptosis in Sf9 Cells. Biochemical and Biophysical Research Communications, 295, 779-783.
http://dx.doi.org/10.1016/S0006-291X(02)00746-5 [21] Rhee, W.J., Lee, E.H., Park, J.H., Lee, J.E. and Park, T.H. (2007) Inhibition of HeLa Cell Apoptosis by Storage-Protein 2. Biotechnology Progress, 23, 1441-1446.
http://dx.doi.org/10.1021/bp0702065 [22] Jung, J.H., Hwang, J.W., Kim, H., Cha, H.M., Kim, D.I. and Choi, Y.S. (2013) Whitening and Anti-Aging Activities of Soluble Silkworm Gland Hydrolysate. Korean Society for Biotechnology and Bioengineering Journal, 28, 196-201. [23] Costin, G.E. and Hearing, V.J. (2007) Human Skin Pigmentation: Melanocytes Modulate Skin Color in Response to Stress. The Journal of the Federation of American Societies for Experimental Biology, 21, 976-994.
http://dx.doi.org/10.1096/fj.06-6649rev [24] Hwang, J.W., Lee, H.S., Kim, H., Kim, K.O. and Choi, Y.S. (2012) Manufacture and Characterization of Silkworm Gland Hydrolysate. Journal of Sericin and Entomology Science, 50, 76-81. [25] Choi, M.H., Cha, H.M., Kim, S.M., Choi, Y.S. and Kim, D.I. (2013) Effects of Silkworm Gland Hydrolysate on Albumin-Erythropoietin Production in Transgenic Chinese Hamster Ovary Cells. Korean Society for Biotechnology and Bioengineering Journal, 28, 86-91. [26] Cheon, Y., Hwang, J.W., Lee, H.S., Yun, S., Choi, Y.S. and Kang, S. (2013) Anti-Oxidant and Anti-Aging Activities of Sericinjam Gland Hydrolysate Extract in Human Dermal Fibroblasts. Journal of the Society of Cosmetic Scientists of Korea, 39, 9-17. [27] Yu, W., Wang, M., Zhang, H., Quan, Y. and Zhang, Y. (2013) Expression and Functional Analysis of Storage Protein 2 in the Silkworm, Bombyx mori. International Journal of Genomics, 2013, Article ID: 145450. [28] Chang, T.S. and Chen, C.T. (2012) Inhibitory Effect of Homochlorcyclizine on Melanogenesis in α-Melanocyte Stimulating Hormone-Stimulated Mouse B16 Melanoma Cells. Archives of Pharmacal Research, 35, 119-127.
http://dx.doi.org/10.1007/s12272-012-0113-z [29] Lee, S.A., Son, Y.O., Kook, S.H., Choi, K.C. and Lee, J.C. (2011) Ascorbic Acid Increases the Activity and Synthesis of Tyrosinase in B16F10 Cells through Activation of p38 Mitogen-Activated Protein Kinase. Archives of Dermatological Research, 303, 669-678.
http://dx.doi.org/10.1007/s00403-011-1158-4 [30] Rózanowska, M., Sarna, T., Land, E.J. and Truscott, T.G. (1999) Free Radical Scavenging Properties of Melanin: Interaction of Eu- and Pheo-Melanin Models with Reducing and Oxidising Radicals. Free Radical Biology and Medicine, 26, 518-525.
http://dx.doi.org/10.1016/S0891-5849(98)00234-2 [31] Nappi, A., Vass, E., Frey, F. and Carton, Y. (1995) Superoxide Anion Generation in Drosophila during Melanotic Encapsulation of Parasites. European Journal of Cell Niology, 68, 450-456. [32] Momtaz, S., Mapunya, B., Houghton, P., Edgerly, C., Hussein, A., Naidoo, S. and Lall, N. (2008) Tyrosinase Inhibition by Extracts and Constituents of Sideroxylon inerme L. Stem Bark, Used in South Africa for Skin Lightening. Journal of Ethnopharmacology, 119, 507-512.
http://dx.doi.org/10.1016/j.jep.2008.06.006 [33] Kumano, Y., Sakamoto, T., Egawa, M., Iwai, I., Tanaka, M. and Yamamoto, I. (1998) In Vitro and in Vivo Prolonged Biological Activities of Novel Citamin C Derivative, 2-O-α-D-Glucopyranosyl-L-Ascorbic Acid (AA-2G), in Cosmetic Fields. Journal of Nutritional Science and Vitaminology, 44, 345-359.
http://dx.doi.org/10.3177/jnsv.44.345 [34] Akihisa, T., Kawashima, K., Orido, M., Akazawa, H., Matsumoto, M., Yamamoto, A., Ogihara, E., Fukatsu, M., Tokuda, H. and Fuji, J. (2013) Antioxidative and Melanogenesis-Inhibitory Activities of Caffeoylquinic Acids and Other Compounds from Moxa. Chemistry & Biodiversity, 10, 313-327.
http://dx.doi.org/10.1002/cbdv.201200357 [35] Saino, N., Romano, M., Rubolini, D., Ambrosini, R., Caprioli, M., Milzani, A., Costanzo, A., Colombo, G., Canova, L. and Wakamatsu, K. (2013) Viability Is Associated with Melanin-Based Coloration in the Barn Swallow (Hirundo rustica). PLoS ONE, 8, Article ID: e60426.
http://dx.doi.org/10.1371/journal.pone.0060426 [36] Kondo, T. and Hearing, V.J. (2011) Update on the Ascorbic Acid Increases the Activity and Synthesis of Tyrosinase in B16F10 Cells through Activation of p38 Mitogen-Activated Protein Kinase Regulation of Mammalian Melanocyte Function and Skin Pigmentation. Expert Review of Dermatology, 6, 97-108.
http://dx.doi.org/10.1586/edm.10.70
[1] (2014) Global Market for Cosmetic Chemicals to Reach $21.2 bn by 2018: Global Industry Analysts. Focus on Surfactants, 3, 5-6.
http://dx.doi.org/10.1016/S1351-4210(14)70070-8 [2] Curto, E.V., Kwong, C., Hermersdörfer, H., Glatt, H., Santis, C., Virador, V., Hearing Jr., V.J. and Dooley, T.P. (1999) Inhibitors of Mammalian Melanocyte Tyrosinase: In Vitro Comparisons of Alkyl Esters of Gentisic Acid with Other Putative Inhibitors. Biochemical Pharmacology, 57, 663-672.
http://dx.doi.org/10.1016/S0006-2952(98)00340-2 [3] 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, 1082-1087.
http://dx.doi.org/10.1016/j.phymed.2013.04.015 [4] Burnett, C.L., Bergfeld, W.F., Belsito, D.V., Hill, R.A., Klaassen, C.D., Liebler, D.C., Marks, J.G., Shank, R.C., Slaga, T.J. and Snyder, P.W. (2010) Final Report of the Safety Assessment of Kojic Acid as Used in Cosmetics. International Journal of Toxicology, 29, 244S-273S.
http://dx.doi.org/10.1177/1091581810374651 [5] Vermaak, I., Kamatou, G.P.P., Komane-Mofokeng, B., Viljoen, A.M. and Beckett, K. (2011) African Seed Oils of Commercial Importance—Cosmetic Applications. South African Journal of Botany, 77, 920-933.
http://dx.doi.org/10.1016/j.sajb.2011.07.003 [6] Uchida, R., Ishikawa, S. and Tomoda, H. (2014) Inhibition of Tyrosinase Activity and Melanine Pigmentation by 2-Hydroxytyrosol. Acta Pharmaceutica Sinica B, 4, 141-145.
http://dx.doi.org/10.1016/j.apsb.2013.12.008 [7] Manosroi, A., Boonpisuttinant, K., Winitchai, S., Manosroi, W. and Manosroi, J. (2010) Free Radical Scavenging and Tyrosinase Inhibition Activity of Oils and Sericin Extracted from Thai Native Silkworms (Bombyx mori). Pharmaceutical Biology, 48, 855-860.
http://dx.doi.org/10.3109/13880200903300212 [8] Rhee, W.J., Park, J.H., Hahn, J.S. and Park, T.H. (2013) Anti-Apoptotic Mechanism of Silkworm Hemolymph in HeLa Cell Apoptosis. Process Biochemistry, 48, 1375-1380.
http://dx.doi.org/10.1016/j.procbio.2013.06.018 [9] Mondal, M., Trivedy, K. and Kumar, S.N. (2007) The Silk Proteins, Sericin and Fibroin in Silkworm, Bombyx mori Linn.,—A Review. Caspian Journal of Evironmental Siences, 5, 63-76. [10] Chon, J.W., Kweon, H., Jo, Y.Y., Park, M.K., Son, Y.H. and Lee, H.S. (2012) A Study on the Development of Functional Cosmetics Using Silk-Grand Power of Silkworm. Journal of the Society of Cosmetic Scientists of Korea, 38, 163-169. [11] Haddad, A.L., Matos, L.F., Brunstein, F., Ferreira, L.M., Silva, A. and Costa, D. (2003) A Clinical, Prospective, Randomized, Double-Blind Trial Comparing Skin Whitening Complex with Hydroquinone vs. Placebo in the Treatment of Melasma. International Journal of Dermatology, 42, 153-156. http://dx.doi.org/10.1046/j.1365-4362.2003.01621.x [12] Murata, K., Takahashi, K., Nakamura, H., Itoh, K. and Matsuda, H. (2014) Search for Skin-Whitening Agent from Prunus Plants and the Molecular Targets in Melanogenesis Pathway of Active Compounds. Natural Product Communications, 9, 185-188. [13] Winder, A., Kobayashi, T., Tsukamoto, K., Urabe, K., Aroca, P., Kameyama, K. and Hearing, V. (1993) The Tyrosinase Gene Family—Interactions of Melanogenic Proteins to Regulate Melanogenesis. Cellular & Molecular Biology Research, 40, 613-626. [14] Jackson, I.J., Chambers, D., Tsukamoto, K., Copeland, N., Gilbert, D., Jenkins, N. and Hearing, V. (1992) A Second Tyrosinase-Related Protein, TRP-2, Maps to and Is Mutated at the Mouse Slaty Locus. The EMBO Journal, 11, 527-535. [15] Yen, F.L., Wang, M.C., Liang, C.J., Ko, H.H. and Lee, C.W. (2012) Melanogenesis Inhibitor(s) from Phyla Nodiflora Extract. Evidence-Based Complementary and Alternative Medicine, 2012, Article ID: 867494. [16] Park, H.J., Kim, E.J., Koo, T.Y. and Park, T.H. (2003) Purification of Recombinant 30K Protein Produced in Escherichia coli and Its Anti-Apoptotic Effect in Mammalian and Insect Cell Systems. Enzyme and Microbial Technology, 33, 466-471.
http://dx.doi.org/10.1016/S0141-0229(03)00149-2 [17] Izumi, S., Fujie, J., Yamada, S. and Tomino, S. (1981) Molecular Properties and Biosynthesis of Major Plasma Proteins in Bombyx mori. Biochimica et Biophysica Acta-Protein Structure, 670, 222-229.
http://dx.doi.org/10.1016/0005-2795(81)90013-1 [18] Pietrzyk, A.J., Bujacz, A., Mueller-Dieckmann, J., Lochynska, M., Jaskolski, M. and Bujacz, G. (2013) Two Crystal Structures of Bombyx mori Lipoprotein 3-Structural Characterization of a New 30-kDa Lipoprotein Family Member. PLoS ONE, 8, Article ID: e61303.
http://dx.doi.org/10.1371/journal.pone.0061303 [19] Kim, E.J. and Park, T.H. (2004) Increase of Host Cell Longevity by the Expression of 30K Protein Originating from Silkworm Hemolymph in an Insect Cell-Baculovirus System. Enzyme and Microbial Technology, 35, 581-586.
http://dx.doi.org/10.1016/j.enzmictec.2004.08.024 [20] Rhee, W.J., Kim, E.J. and Park, T.H. (2002) Silkworm Hemolymph as a Potent Inhibitor of Apoptosis in Sf9 Cells. Biochemical and Biophysical Research Communications, 295, 779-783.
http://dx.doi.org/10.1016/S0006-291X(02)00746-5 [21] Rhee, W.J., Lee, E.H., Park, J.H., Lee, J.E. and Park, T.H. (2007) Inhibition of HeLa Cell Apoptosis by Storage-Protein 2. Biotechnology Progress, 23, 1441-1446.
http://dx.doi.org/10.1021/bp0702065 [22] Jung, J.H., Hwang, J.W., Kim, H., Cha, H.M., Kim, D.I. and Choi, Y.S. (2013) Whitening and Anti-Aging Activities of Soluble Silkworm Gland Hydrolysate. Korean Society for Biotechnology and Bioengineering Journal, 28, 196-201. [23] Costin, G.E. and Hearing, V.J. (2007) Human Skin Pigmentation: Melanocytes Modulate Skin Color in Response to Stress. The Journal of the Federation of American Societies for Experimental Biology, 21, 976-994.
http://dx.doi.org/10.1096/fj.06-6649rev [24] Hwang, J.W., Lee, H.S., Kim, H., Kim, K.O. and Choi, Y.S. (2012) Manufacture and Characterization of Silkworm Gland Hydrolysate. Journal of Sericin and Entomology Science, 50, 76-81. [25] Choi, M.H., Cha, H.M., Kim, S.M., Choi, Y.S. and Kim, D.I. (2013) Effects of Silkworm Gland Hydrolysate on Albumin-Erythropoietin Production in Transgenic Chinese Hamster Ovary Cells. Korean Society for Biotechnology and Bioengineering Journal, 28, 86-91. [26] Cheon, Y., Hwang, J.W., Lee, H.S., Yun, S., Choi, Y.S. and Kang, S. (2013) Anti-Oxidant and Anti-Aging Activities of Sericinjam Gland Hydrolysate Extract in Human Dermal Fibroblasts. Journal of the Society of Cosmetic Scientists of Korea, 39, 9-17. [27] Yu, W., Wang, M., Zhang, H., Quan, Y. and Zhang, Y. (2013) Expression and Functional Analysis of Storage Protein 2 in the Silkworm, Bombyx mori. International Journal of Genomics, 2013, Article ID: 145450. [28] Chang, T.S. and Chen, C.T. (2012) Inhibitory Effect of Homochlorcyclizine on Melanogenesis in α-Melanocyte Stimulating Hormone-Stimulated Mouse B16 Melanoma Cells. Archives of Pharmacal Research, 35, 119-127.
http://dx.doi.org/10.1007/s12272-012-0113-z [29] Lee, S.A., Son, Y.O., Kook, S.H., Choi, K.C. and Lee, J.C. (2011) Ascorbic Acid Increases the Activity and Synthesis of Tyrosinase in B16F10 Cells through Activation of p38 Mitogen-Activated Protein Kinase. Archives of Dermatological Research, 303, 669-678.
http://dx.doi.org/10.1007/s00403-011-1158-4 [30] Rózanowska, M., Sarna, T., Land, E.J. and Truscott, T.G. (1999) Free Radical Scavenging Properties of Melanin: Interaction of Eu- and Pheo-Melanin Models with Reducing and Oxidising Radicals. Free Radical Biology and Medicine, 26, 518-525.
http://dx.doi.org/10.1016/S0891-5849(98)00234-2 [31] Nappi, A., Vass, E., Frey, F. and Carton, Y. (1995) Superoxide Anion Generation in Drosophila during Melanotic Encapsulation of Parasites. European Journal of Cell Niology, 68, 450-456. [32] Momtaz, S., Mapunya, B., Houghton, P., Edgerly, C., Hussein, A., Naidoo, S. and Lall, N. (2008) Tyrosinase Inhibition by Extracts and Constituents of Sideroxylon inerme L. Stem Bark, Used in South Africa for Skin Lightening. Journal of Ethnopharmacology, 119, 507-512.
http://dx.doi.org/10.1016/j.jep.2008.06.006 [33] Kumano, Y., Sakamoto, T., Egawa, M., Iwai, I., Tanaka, M. and Yamamoto, I. (1998) In Vitro and in Vivo Prolonged Biological Activities of Novel Citamin C Derivative, 2-O-α-D-Glucopyranosyl-L-Ascorbic Acid (AA-2G), in Cosmetic Fields. Journal of Nutritional Science and Vitaminology, 44, 345-359.
http://dx.doi.org/10.3177/jnsv.44.345 [34] Akihisa, T., Kawashima, K., Orido, M., Akazawa, H., Matsumoto, M., Yamamoto, A., Ogihara, E., Fukatsu, M., Tokuda, H. and Fuji, J. (2013) Antioxidative and Melanogenesis-Inhibitory Activities of Caffeoylquinic Acids and Other Compounds from Moxa. Chemistry & Biodiversity, 10, 313-327.
http://dx.doi.org/10.1002/cbdv.201200357 [35] Saino, N., Romano, M., Rubolini, D., Ambrosini, R., Caprioli, M., Milzani, A., Costanzo, A., Colombo, G., Canova, L. and Wakamatsu, K. (2013) Viability Is Associated with Melanin-Based Coloration in the Barn Swallow (Hirundo rustica). PLoS ONE, 8, Article ID: e60426.
http://dx.doi.org/10.1371/journal.pone.0060426 [36] Kondo, T. and Hearing, V.J. (2011) Update on the Ascorbic Acid Increases the Activity and Synthesis of Tyrosinase in B16F10 Cells through Activation of p38 Mitogen-Activated Protein Kinase Regulation of Mammalian Melanocyte Function and Skin Pigmentation. Expert Review of Dermatology, 6, 97-108.
http://dx.doi.org/10.1586/edm.10.70