[논문]B16F10 피부 흑색종세포에서 갯사상자 추출물의 멜라닌 합성 저해 효과

조현진; 카라데니즈 파티; 오정환; 서영완; 공창숙

doi:10.5352/jls.2022.32.5.331

초록
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멜라닌은 자외선과 같은 외부자극이 가해지면 피부 기저층에 존재하는 멜라닌 세포에서 피부를 보호하기 위한 방어기전으로써 생성이 된다. 하지만 과도한 자외선 노출로 멜라닌이 필요이상으로 생성이 되면 기미, 주근깨, 검버섯과 같은 색소침착 및 색소성 피부장애를 유발할 수 있다. 최근에는 부작용이 적은 식물 추출물을 대상으로 미백소재를 찾기 위한 연구들이 활기를 띠고 있다. 이에 본 연구에서는 국내 서식 염생식물인 갯사상자 추출물을 이용하여 B16F10 흑색종 세포에서 피부 색소 멜라닌 생성 억제에 미치는 효과를 확인하였다. 갯사상자 추출물 처리시 tyrosine 및 L-DOPA 산화를 농도 의존적으로 저해하였으며 세포 내의 멜라닌 생성을 담당하는 tyrosinase, tyrosinase-related protein-1, -2 발현을 억제하였다. 이는 갯사상자 추출물이 α-MSH에 의한 세포신호 전달 경로인 GSKβ/β-catenin 및 PKA/CREB 조절에 의한 것으로 밝혀졌다. 따라서 갯사상자 추출물은 GSKβ/β-catenin 및 PKA/CREB 기전을 통해 멜라닌 합성을 억제하여 미백 효능 지닌 천연물 유래 기능성 화장품 소재로서 활용 가능할 것으로 사료된다.

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Melanin is a pigment produced by melanocytes to protect the skin from external stimuli, mainly ultraviolet (UV) rays. However, abnormal and excessive production of melanin causes hyperpigmentation disorders, such as freckles, age spots, and discoloration. Natural cosmeceuticals are a new trend for t...

Melanin is a pigment produced by melanocytes to protect the skin from external stimuli, mainly ultraviolet (UV) rays. However, abnormal and excessive production of melanin causes hyperpigmentation disorders, such as freckles, age spots, and discoloration. Natural cosmeceuticals are a new trend for treating or preventing hyperpigmentation due to fewer side effects and biocompatibility. In this context, the current study focused on Cnidium japonicum, a halophyte with several uses in folk medicine, to evaluate its potential as a skin-whitening agent. The effect of C. japonicum extract (CJE) on melanin production was analyzed in melanogenesis-stimulated B16F10 melanoma cells. The results showed that CJE successfully inhibited the oxidation of tyrosine and L-DOPA by tyrosinase and subsequently decreased the production of the key enzymes responsible for melanin production: tyrosinase, tyrosinase-related protein-1, and protein-2. This effect was confirmed by decreased intracellular and extracellular melanin levels in B16F10 melanoma cells after CJE treatment. Further experiments to elucidate the action mechanism revealed that CJE treatment suppressed melanin production by inhibiting the activation of glycogen synthase kinase 3 β (GSKβ)/β-catenin and protein kinase A (PKA)/cAMP-response element binding protein (CREB) pathways, which are the upstream activators of melanogenesis. In conclusion, the present study suggests that C. japonicum is a potential natural source of bioactive substances for the development of novel cosmeceuticals that can act against hyperpigmentation.

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표/그림 (5)

그림 Fig. 1. Effect of CJE on the in vitro tyrosinase activity (A) and L-DOPA oxidation (B). Kojic acid, a tyrosinase inhibitor,was used a positive control. a-fMeans with different letters are significantly different (p<0.05) by Duncan’s multiplesrange test.
그림 Fig. 2. Effect of CJE on the viability (A), intracellular tyrosinase content (B), and extracellular (C) and intracellular (D) melanin content of α-MSH-stimulated B16F10 murine melanoma cells. Cells were treated with or without CJE at given concentrations for 72 hr. Tyrosinase content was measured as a relative percentage of the activity of the intracellular tyrosinase obtained from untreated control group. ^a-fMeans with different letters are significantly different (p<0.05) by Duncan's multiples range test.
그림 Fig. 3. Effect of CJE on the mRNA (A) and protein (B) expression levels of melanogenesis-related factors in α-MSH-stimulated B16F10 murine melanoma cells. Cells were treated with or without CJE at given concentrations for 72 hr. Bands were densiometrically quantified, normalized against internal loading control β-actin and given as relative percentage of untreated control group. ^a-fMeans with different letters are significantly different (p<0.05) by Duncan's multiples range test.
그림 Fig. 4. Effect of CJE on the activation of GSK3β (A) and PKA (B) signaling cascades given as protein expression levels in α-MSH-stimulated B16F10 murine melanoma cells. Cells were treated with or without CJE at given concentrations for 72 hr. Bands were densiometrically quantified, normalized against internal loading control β-actin and given as relative percentage of untreated control group. ^a-fMeans with different letters are significantly different (p<0.05) by Duncan's multiples range test.
그림 Fig. 5. Suggested intracellular anti-melanogenic action mechanism of C. japonicum extract (CJE).

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제안 방법

Following the confirmation of the effect of CJE on tyrosinase activity and melanin production, the expressional changes in melanogenesis role players were evaluated in order to elucidate the action mechanism of CJE. The mRNA and protein expressions of tyrosinase enzyme as well as MITF, TRP-1 and TRP-2 were analyzed for this purpose.

대상 데이터

japonicum Miq. plants used in this study was collected from the coast of Donggeom-ri, Gilsang-myeon, Ganghwagun (Gyeonggi-do, Korea) and was kindly provided by Korea Maritime University (Yeongdo, Busan, Korea).

데이터처리

Statistical differences were defined after one way analysis of variance coupled with Duncan’s multiple range post-hoc test.

성능/효과

Therefore, tyrosinase inhibitory effect of CJE was further analyzed via amount of melanin content in α-MSH-stimulated B16F10 melanoma cells.
Treatment with 1 μM Kojic acid as positive control resulted in 51.3% extracellular and 25.3% intracellular melanin content decrease, both of which was under the level of decrease observed by the CJE treatment
Similar trend was observed for extracellular melanin content as the treatment with 5, 10, and 50 μg/ml CJE decreased the secreted melanin amount to 55.4%, 48.8%, and 25.0% of the untreated control group (Fig
3% intracellular melanin content decrease, both of which was under the level of decrease observed by the CJE treatment. These results confirmed the previous tyrosinase inhibitory effect of CJE as the inhibition of tyrosinase was translated into halted melanin production in melanoma cells.
However, treatment with CJE showed a dose-dependent suppression on both mRNA and protein expression of these factors. These results showed that CJE treatment not only inhibits the tyrosinase activity, but it also suppresses the melanogenesis pathway by regulating the expression of related factors in both mRNA and protein levels.
Treatment with 50 μ g/ml CJE was able to decrease the both phosphorylation of GSK3β and the total β-catenin levels. These results indicated that the effect of CJE on melanogenesis might stemmed from its suppressive properties on PKA-dependent activation of MITF.
In conclusion, current study confirmed the anti-melano- genic potential of CJE. It was shown that CJE was able to inhibit the tyrosinase activity on both tyrosine and L-DOPA.
Overall, the results indicated that CJE treatment decreased the amount of active tyrosinase in melanoma cells which was significantly increased by α-MSH stimulation.
The effect of CJE on L-DOPA oxidation was analyzed with a similar method to tyrosine oxidation. Results again showed that CJE treatment decreased the L-DOPA oxidation in a dose-dependent manner (Fig. 1B).

참고문헌 (20)

Bahraman, A. G., Jamshidzadeh, A., Keshavarzi, M., Arabnezhad, M. R., Mohammadi, H. and Mohammadi-Bardbori, A. 2021. α-Melanocyte-stimulating hormone triggers melanogenesis via activation of the aryl hydro- carbon receptor pathway in B16F10 mouse melanoma cells. Int. J. Toxicol. 40, 153-160.
Bellei, B., Flori, E., Izzo, E., Maresca, V. and Picardo, M. 2008. GSK3β inhibition promotes melanogenesis in mouse B16 melanoma cells and normal human melanocytes. Cell. Signal. 20, 1750-1761.
Bellei, B., Pitisci, A., Catricala, C., Larue, L. and Picardo, M. 2011. Wnt/β-catenin signaling is stimulated by α-melanocyte-stimulating hormone in melanoma and melanocyte cells: implication in cell differentiation. Pigment Cell Melanoma Res. 24, 309-325.
Brenner, M. and Hearing, V. J. 2008. The protective role of melanin against UV damage in human skin. Photochem. Photobiol. 84, 539-549.

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Hwang, Y. S., Oh, S. W., Park, S. H., Lee, J., Yoo, J. A., Kwon, K., Park, S. J., Kim, J., Yu, E., Cho, J. Y. and Lee, J. 2019. Melanogenic effects of maclurin are mediated through the activation of cAMP/PKA/CREB and p38 MAPK/CREB signaling pathways. Oxid. Med. Cell. Longev. 2019, 9827519.

상세보기
Kimura, Y., Sumiyoshi, M., Taniguchi, M. and Baba, K. 2008. Antitumor actions of a chromone glucoside cnidimoside A isolated from Cnidium japonicum. J. Nat. Med. 62, 308-313.

상세보기
Lambert, M. W., Maddukuri, S., Karanfilian, K. M., Elias, M. L. and Lambert, W. C. 2019. The physiology of melanin deposition in health and disease. Clin. Dermatol. 37, 402-417.
Lee, S. G., Karadeniz, F., Seo, Y. and Kong, C. S. 2017. Anti-melanogenic effects of flavonoid glycosides from Limonium tetragonum (Thunb.) Bullock via inhibition of tyrosinase and tyrosinase-related proteins. Molecules 22, 1480.

상세보기
Nautiyal, A. and Wairkar, S. 2021. Management of hyperpigmentation: Current treatments and emerging therapies. Pigment Cell Melanoma Res. 34, 1000-1014.

상세보기
Nguyen, N. T. and Fisher, D. E. 2019. MITF and UV responses in skin: From pigmentation to addiction. Pigment Cell Melanoma Res. 32, 224-236.

상세보기
Obaid, R. J., Mughal, E. U., Naeem, N., Sadiq, A., Alsantali, R. I., Jassas, R. S., Moussa, Z. and Ahmed, S. A. 2021. Natural and synthetic flavonoid derivatives as new potential tyrosinase inhibitors: a systematic review. RSC Adv. 11, 22159-22198.

상세보기
Park, J. Y., Lee, J. Y., Lee, D. Y., Kim, H. D., Kim, G. S., Lee, S. E., Seo, K. H., Park, J. Y., Lee, J. Y., Lee, D. Y., Kim, H. D., Kim, G. S., Lee, S. E. and Seo, K. H. 2019. Melanogenesis inhibitory effect of steamed Platycodon grandiflorum on B16F10 Cells. Asian J. Beauty Cosmetol. 17, 37-46.
Pillaiyar, T., Manickam, M. and Namasivayam, V. 2017. Skin whitening agents: Medicinal chemistry perspective of tyrosinase inhibitors. J. Enzyme Inhib. Med. Chem. 32, 403.
Sotiropoulou, G., Zingkou, E. and Pampalakis, G. 2021. Redirecting drug repositioning to discover innovative cosmeceuticals. Exp. Dermatol. 30, 628-644.

상세보기
Vachtenheim, J. and Borovansky, J. 2010. 'Transcription physiology' of pigment formation in melanocytes: Central role of MITF. Exp. Dermatol. 19, 617-627.

상세보기
Venkatesan, H., Soundhararajan, R. and Srinivasan, H. 2022. Interaction of various types of bisphenols with enzymes involved in melanin synthesis. Toxicol. Environ. Health Sci. 14, 19-24.
Wang, R. F., Ko, D., Friedman, B. J., Lim, H. W. and Mohammad, T. F. 2022. Disorders of hyperpigmentation. Part I. Pathogenesis and clinical features of common pigmentary disorders. J. Am. Acad. Dermatol. In Press.
Yang, E. J., Yim, E. Y., Song, G., Kim, G. O. and Hyun, C. G. 2009. Inhibition of nitric oxide production in lipopolysaccharide-activated RAW 264.7 macrophages by Jeju plant extracts. Interdiscip. Toxicol. 2, 245.
Zhao, S. J., Jia, H., Xu, X. L., Bu, W. B., Zhang, Q., Chen, X., Ji, J. and Sun, J. F. 2021. Identification of the role of Wnt/β-catenin pathway through integrated analyses and in vivo experiments in vitiligo. Clin. Cosmet. Investig. Dermatol. 14, 1089.
Zolghadri, S., Bahrami, A., Hassan Khan, M. T., Munoz-Munoz, J., Garcia-Molina, F., Garcia-Canovas, F. and Saboury, A. A. 2019. A comprehensive review on tyrosinase inhibitors. J. Enzyme Inhib. Med. Chem. 34, 279-309.

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[국내논문] B16F10 피부 흑색종세포에서 갯사상자 추출물의 멜라닌 합성 저해 효과
Anti-melanogenic Effects of Cnidium japonicum in B16F10 Murine Melanoma Cells 원문보기

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[국내논문] B16F10 피부 흑색종세포에서 갯사상자 추출물의 멜라닌 합성 저해 효과 Anti-melanogenic Effects of Cnidium japonicum in B16F10 Murine Melanoma Cells 원문보기

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[국내논문] B16F10 피부 흑색종세포에서 갯사상자 추출물의 멜라닌 합성 저해 효과
Anti-melanogenic Effects of Cnidium japonicum in B16F10 Murine Melanoma Cells 원문보기

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