Nelumbo nucifera Leaf Extracts Inhibit Melanogenesis in B16 Melanoma Cells and Guinea Pigs through Downregulation of CREB/MITF Activation

Po-Ju Lai^{1, 2}, Erl-Shyh Kao³, Sin-Rong Chen¹, Yu-Ting Huang¹, Chau-Jong Wang^{1, 4,} and Hui-Pei Huang^{1, 5,}

¹Institute of Biochemistry, Microbiology and Immunology, Chung-Shan Medical University, Taichung, Taiwan

²Department of Dermatology, Chung-Shan Medical University Hospital, Taichung, Taiwan

³Department of Beauty Science, National Taichung University of Science and Technology, Taichung, Taiwan

⁴Department of Medical Research, Chung Shan Medical University Hospital, Taichung, Taiwan;Department of Health Policy and Management, Chung Shan Medical University, Taichung, Taiwan

⁵Clinical Laboratory, Chung Shan Medical University Hospital, Taichung, Taiwan;Department of Biochemistry, School of Medicine, Chung Shan Medical University, Taichung, Taiwan

Cite this paper:
Po-Ju Lai, Erl-Shyh Kao, Sin-Rong Chen, Yu-Ting Huang, Chau-Jong Wang and Hui-Pei Huang. Nelumbo nucifera Leaf Extracts Inhibit Melanogenesis in B16 Melanoma Cells and Guinea Pigs through Downregulation of CREB/MITF Activation. Journal of Food and Nutrition Research. 2020; 8(9):459-465. doi: 10.12691/jfnr-8-9-2

Abstract

Nelumbo nucifera leaves extracts (NLE) has been suggested to provide antioxidant, anti-obesity and anticancer effects. However, the research on the anti-melanogenetic effect of NLE was little. Here, we reported that NLE and Gallic acid (GA, major ingredients of NLE) exhibit the hypopigmentary effect in B16F1 cells. NLE and GA showed potent inhibitory effects on tyrosinase, microphthalmia-associated transcription factor (MITF), and tyrosinase-related protein-1 (TRP-1) protein production. The phosphorylation of intracellular protein kinase A (PKA) and cAMP response element-binding protein (CREB) were also decreased, revealing potent anti-melanogenic effects of NLE and GA. However, NLE showed the better effect than GA on reducing melanin formation, implying the importance of the synergism of polyphenolic compounds of NLE. Furthermore, NLE inhibited skin melanogenesis and epidermal hyperplasia of guinea pigs caused by irradiation through attenuating ERK and CREB activation. NLE reduced skin melanogenesis and epidermal hyperplasia induced by UVB in guinea pigs through downregulation ERK and CREB pathways, and the following decreasing of MITF, tyrosinase and TRP-1 expression. These results demonstrated the potential which NLE as an ingredient of hypopigmentary cosmetics.

Keywords:
CREB gallic acid melanogenesis nelumbo nucifera leaves extracts (NLE) tyrosinase

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References:

[1]	Brenner, M., and C. Berking, [Principles of skin pigmentation. Biochemistry and regulation of melanogenesis]. Hautarzt, 61(7). 554-560. 2010.
[2]	Videira, I.F., D.F. Moura, and S. Magina, Mechanisms regulating melanogenesis. The journal Brazilian Annals of Dermatology, 88(1). 76-83. 2013.
[3]	Rees, J.L., Genetics of hair and skin color. The Annual Review of Genetics, 37. 67-90. 2003.
[4]	Taylor, S.C., Epidemiology of skin diseases in people of color. Cutis, 71(4). 271-275. 2003.
[5]	Draelos, Z.D., Skin lightening preparations and the hydroquinone controversy. Dermatologic Therapy, 20(5). 308-313. 2007.
[6]	Hwang, J.H. and B.M. Lee, 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. 2007.
[7]	Zhong, S., Wu, Y., Soo-Mi, A., Zhao, J., Wang, K., Yang, S., Jae-Ho, Y. and Zhu, X., Depigmentation of melanocytes by the treatment of extracts from traditional Chinese herbs: a cell culture assay. Biological and Pharmaceutical Bulletin, 29(9). 1947-1951. 2006.
[8]	Chuang, C.Y., Liu, H. C., Wu, L. C., Chen, C. Y., Cnang, J. T., and Hsu, S. L.,, Gallic acid induces apoptosis of lung fibroblasts via a reactive oxygen species-dependent ataxia telangiectasia mutated-p53 activation pathway. Journal of Agricultural and Food Chemistry, 58(5). 2943-2951. 2010.
[9]	Su, T.R., Lin, J. J., Tsai, C. C., Huang, T. K., Yang, Z. Y., Wu, M. O., Zheng, Y. Q., Su, C. C., and Wu, Y. J., Inhibition of melanogenesis by gallic acid: possible involvement of the PI3K/Akt, MEK/ERK and Wnt/beta-catenin signaling pathways in B16F10 cells. International Journal of Molecular Sciences, 14(10). 20443-20458. 2013.
[10]	Chan, C.F., Wu, C. T., Huang, W. Y., Lin, W. S., Wu, H. W., Huang, T. K., Chang, M. Y., and Lin, Y. S., Antioxidation and Melanogenesis Inhibition of Various Dendrobium tosaense Extracts. Molecules, 23(7). 1810. 2018.
[11]	Sun, L., Guo, Y., Zhang, Y., and Zhuang, Y., Antioxidant and Anti-tyrosinase Activities of Phenolic Extracts from Rape Bee Pollen and Inhibitory Melanogenesis by cAMP/MITF/TYR Pathway in B16 Mouse Melanoma Cells. Frontiers in Pharmacology, 8: p. 104. 2017.
[12]	Roh, H.J., Noh, H. J., Na, C. S., Kim, C. S., Kim, K. H., Hong, C. Y., and Lee, K. R., Phenolic Compounds from the Leaves of Stewartia pseudocamellia Maxim. and their Whitening Activities. Biomolecules & Therapeutics, 23(3). 283-289. 2015.
[13]	Lin, M.C., Kao, S. H., Chung, P. J., Chan, K. C., Yang, M. Y., and Wang, C. J., Improvement for high fat diet-induced hepatic injuries and oxidative stress by flavonoid-enriched extract from Nelumbo nucifera leaf. Journal of Agricultural and Food Chemistry, 57(13). 5925-5932. 2009.
[14]	Ho, H.H., Hsu, L. S., Chan, K. C., Chen, H. M., Wu, C. H., and Wang, C. J., Extract from the leaf of nucifera reduced the development of atherosclerosis via inhibition of vascular smooth muscle cell proliferation and migration. Food and Chemical Toxicology, 48(1). 159-168. 2010.
[15]	Wu, C.H., Yang, M. Y., Chan, K. C., Chung, P. J., Ou, T. T., and Wang, C. J., Improvement in high-fat diet-induced obesity and body fat accumulation by a Nelumbo nucifera leaf flavonoid-rich extract in mice. Journal of Agricultural and Food Chemistry, 58(11). 7075-7081. 2010.
[16]	Chang, C.H., Ou, T. T., Yang, M. Y., Huang, C. C., and Wang C. J., Nelumbo nucifera Gaertn leaves extract inhibits the angiogenesis and metastasis of breast cancer cells by downregulation connective tissue growth factor (CTGF) mediated PI3K/AKT/ERK signaling. Journal of Ethnopharmacology, 188. 111-122. 2016.
[17]	Chen, H.W., , Nelumbo nucifera leaves extract attenuate the pathological progression of diabetic nephropathy in high-fat diet-fed and streptozotocin-induced diabetic rats. J Food Drug Anal, 2019. 27(3). 736-748.
[18]	Yang, M.Y., Hung, T. W., Wang, C. J., and Tseng, T. H., Inhibitory Effect of Nelumbo nucifera Leaf Extract on 2-Acetylaminofluorene-induced Hepatocarcinogenesis Through Enhancing Antioxidative Potential and Alleviating Inflammation in Rats. Antioxidants (Basel), 8(9). 329. 2019.
[19]	Huang, H.C., Chou, Y. C., Wu, C. Y., and Chang, T. M., [8]-Gingerol inhibits melanogenesis in murine melanoma cells through down-regulation of the MAPK and PKA signal pathways. Biochemical and Biophysical Research Communications, 438(2). 375-381. 2013.
[20]	Chung, S.Y., Liu, H. C., Wu, L. C., Chen, C. Y., Chang, J. T., and Hsu, S. L., Fermented rice bran downregulates MITF expression and leads to inhibition of alpha-MSH-induced melanogenesis in B16F1 melanoma. Bioscience, Biotechnology, and Biochemistry, 73(8). 1704-1710. 2009.
[21]	Ling, Z.Q., B.J. Xie, and E.L. Yang, Isolation, characterization, and determination of antioxidative activity of oligomeric procyanidins from the seedpod of Nelumbo nucifera Gaertn. Journal of Agricultural and Food Chemistry, 53(7). 2441-2445. 2005.
[22]	Liu, W., Yi, D. D., Guo, J. L., Xiang, Z. X., Deng, L. F., and He, L., Nuciferine, extracted from Nelumbo nucifera Gaertn, inhibits tumor-promoting effect of nicotine involving Wnt/beta-catenin signaling in non-small cell lung cancer. Journal of Ethnopharmacology, 165. 83-93. 2015.
[23]	Yang, F., Tuxhorn, J. A., Ressler, S. J., McAlhany, S. J., Dang, T. D., and Rowley, D. R., Stromal expression of connective tissue growth factor promotes angiogenesis and prostate cancer tumorigenesis. Cancer Research, 65(19). 8887-8895. 2005.
[24]	Yu, S., C.E. Lan, and H.S. Yu, Mechanisms of repigmentation induced by photobiomodulation therapy in vitiligo. Experimental Dermatology, 28 Suppl 1. 10-14. 2019.
[25]	Jeong, Y.J., Lee, J. Y., Park, J., and Park, S. N., An inhibitory mechanism of action of a novel syringic-acid derivative on alpha-melanocyte-stimulating hormone (alpha-MSH)-induced melanogenesis. Life Science, 191. 52-58. 2017.
[26]	Kumar, K.J., Vani, M. G., Wang, S. Y., Liao, J. W., Hsu, L. S., Yang, H. L., and Hseu, Y. C., In vitro and in vivo studies disclosed the depigmenting effects of gallic acid: a novel skin lightening agent for hyperpigmentary skin diseases. Biofactors, 39(3). 259-270. 2013.
[27]	Choubey, S., Varughese, L. R., Kumar, V., and Beniwal, V., Medicinal importance of gallic acid and its ester derivatives: a patent review. Pharmaceutical patent analyst, 4(4). 305-315. 2015.
[28]	Wu, W.B., Chiang, H. S., Fang, J. Y., Chen, S. K., Huang, C. C., and Hung, C. F., (+)-Catechin prevents ultraviolet B-induced human keratinocyte death via inhibition of JNK phosphorylation. Life Science, 79(8). 801-7. 2006.
[29]	Choi, M.H., Jo, H. G., Yang, J. H., Ki, S. H., and Shin, H. J., Antioxidative and Anti-Melanogenic Activities of Bamboo Stems (Phyllostachys nigra variety henosis) via PKA/CREB-Mediated MITF Downregulation in B16F10 Melanoma Cells. International Journal of Molecular Sciences, 19(2). 409. 2018.
[30]	Park, S.H., Kim, D. S., Park, S. H., Shin, J. W., Youn, S. W., and Park, K. C., Inhibitory effect of p-coumaric acid by Rhodiola sachalinensis on melanin synthesis in B16F10 cells. Pharmazie, 63(4). 290-295. 2008.
[31]	PK, M., P. S, and V. P., Synergy in herbal medicinal products: Concept to realization. Indian Journal of Pharmaceutical Education and Research, 45. 210-217. 2011.
[32]	Bohn, T., Carriere, F., Day, L., Deglaire, A., Egger, L., Freitas, D., Golding, M., Le Feunteun, S., Macierzanka, A., Menard, O., Miralles, B., Moscovici, A., Portmann, R., Recio, I., Remond, D., Sante-Lhoutelier, V., Wooster, T. J., Lesmes, U., Mackie, A. R., and Dupont, D., Correlation between in vitro and in vivo data on food digestion. What can we predict with static in vitro digestion models? Critical Reviews in Food Science and Nutrition, 58(13). 2239-2261. 2018.