[논문]The effects of green tea (Camellia sinensis) flower extract on melanin synthesis in B16-F10 melanoma cells

Dissanayake, Chanuri-Yashara; Moon, Hae-Hee; Yang, Kyeong-Mi; Lee, Younjae; Han, Chang-Hoon

doi:10.14405/kjvr.2018.58.2.65

The effects of green tea (Camellia sinensis) flower extract on melanin synthesis in B16-F10 melanoma cells 원문보기

Korean journal of veterinary research = 대한수의학회지, v.58 no.2, 2018년, pp.65 - 72

Dissanayake, Chanuri-Yashara (College of Veterinary Medicine, Jeju National University) , Moon, Hae-Hee (College of Veterinary Medicine, Jeju National University) , Yang, Kyeong-Mi (Dahrum International Co., Ltd., Jeju National University Medium & Small Business Center) , Lee, Younjae (College of Veterinary Medicine, Jeju National University) , Han, Chang-Hoon (College of Veterinary Medicine, Jeju National University)

Abstract ▼ AI-Helper

The present study observed the effects of a green tea (Camellia sinensis) flower extract (GTFE) on melanin synthesis in B16-F10 melanoma cells. GTFE exhibited antioxidant activity on 2,2-diphenyl-1-picrylhydrazyl and inhibited mushroom tyrosinase activity in a dose-dependent manner. Furthermore, GTFE significantly diminished ${\alpha}-melanocyte$ stimulating hormone (${\alpha}-MSH$) stimulated cellular melanin content and tyrosinase activity throughout the concentration range evaluated. Based on RNA sequencing analysis, differential gene expression patterns observed in ${\alpha}-MSH$ stimulated B16-F10 melanoma cells were normalized by the addition of GTFE. In particular, the expression levels of melanoregulin and tyrosinase genes which are key regulating genes in melanin synthesis were up-regulated by 3.5 and 3 fold respectively by ${\alpha}-MSH$, and were normalized to control levels by the addition of GTFE. The results suggest that GTFE inhibits melanin synthesis in ${\alpha}-MSH$ stimulated B16-F10 melanoma cells by normalizing expression of genes that are essential for melanin synthesis. Overall, the results suggest that GTFE could be applied in the development of a whitening agent for the treatment of dermal hyperpigmentation.

주제어

AI 본문요약
AI-Helper

* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.

제안 방법

All experiments were done in triplicates or more, and the results were expressed as mean ± standard error for each sample concentration.
DPPH assay was performed to evaluate the antioxidant activity of GTFE. GTFE displayed DPPH free radical scavenging activity in a dose dependent manner.
Following 24 h of cell incubation, DMEM medium was removed and cells were treated with 0, 25, 50, 100, 200, 400 and 1,000 µg/mL of GTFE for 24 h.
Total RNA was separated from melanoma cells after 48 h cultivation and cells were dissolved in 500 µL of Trizol reagent (Invitrogen, USA) according to manufacturer’s instructions, and stored at −80℃ until use. RNA quality was assessed by Agilent 2100 Bioanalyzer using the RNA 6000 Nano Chip (Agilent Technologies, The Netherlands), and RNA quantification was carried out using ND-2000 Spectrophotometer (Thermo Fisher Scientific, USA). SENSE 3' mRNA-Seq Library Prep Kit (Lexogen, Austria) was used to perform the construction of a library for control and test RNAs according to the manufacturer’s instructions.

대상 데이터

Dried green tea flowers were purchased from Research Institute of Plant Resource (Korea) and was ground to a powder. GTF 100 g powder was soaked in 1 L of 80% ethanol (ethanol/water = 80/20, v/v) and was incubated in 60℃ water bath for 1 h to facilitate extraction.

데이터처리

The statistical differences among groups were analyzed with one way analysis (ANOVA) followed by Turkey’s test.

이론/모형

Cellular toxicity of GTFE on B16-F10 melanoma cells was measured using MTT assay. GTFE showed no cellular toxicity on B16-F10 melanoma cell at the concentration up to 200 µg/mL (Fig.
0 [12]. The read count data were processed based on Quantile normalization method using the Genowiz ver. 4.0.5.6 (Ocimum Biosolutions, India). Cytoscape (ver.

성능/효과

Analysis of RNA sequencing data showed that 8 genes related to melanogenesis including Atp6v1g2, Try, Slc1a4, Mreg, Cited1, Hspa5, Slc3a2 and Pdia4 genes were up-regulated by 44.9, 3.5, 3.0, 3, 2.8, 2.6, 2.2 and 2.0 folds compared to the control group by addition of α-MSH.
Based on RNA sequencing results, among 23,282 of total genes in B16-F10 cells, 2,420 genes were upregulated (higher than 2.0) and 3,236 genes were down-regulated (lower than 0.5) in α-MSH treated group compared to control group (Table 1).
In conclusion, this study explored that GTFE inhibits the cellular tyrosinase activity and melanin synthesis in α-MSH stimulated B16-F10 cells by regulating genes related to melanogenesis.
In particular, the expression levels of Mreg and Tyr genes which are key regulating genes in melanin synthesis were up-regulated by 3.5 and 3 fold compared to control group by α-MSH and were normalized to control level by the addition of GTFE (Fig. 4A).
Therefore, the results indicate that components of GTFE normalized the expression of genes related to melanogenesis which were disturbed by α-MSH.

참고문헌 (37)

Aizawa M, Fukuda M. Small GTPase Rab2B and its specific binding protein Golgi-associated Rab2B interactorlike 4 (GARI-L4) regulate Golgi morphology. J Biol Chem 2015, 290, 22250-22261.

상세보기
Ando H, Kondoh H, Ichihashi M, Hearing VJ. Approaches to identify inhibitors of melanin biosynthesis via the quality control of tyrosinase. J Invest Dermatol 2007, 127, 751-761.

상세보기
Briganti S, Camera E, Picardo M. Chemical and instrumental approaches to treat hyperpigmentation. Pigment Cell Res 2003, 16, 101-110.

상세보기
Campos PM, da Silva Horinouchi CD, da Silveira Prudente A, Cechinel-Filho V, de Almeida Cabrini D, Otuki MF. Effect of a Garcinia gardneriana (Planchon and Triana) Zappi hydroalcoholic extract on melanogenesis in B16F10 melanoma cells. J Ethnopharmacol 2013, 148, 199-204.

상세보기
Chang TS. An updated review of tyrosinase inhibitors. Int J Mol Sci 2009, 10, 2440-2475.

상세보기
Chatterjee A, Saluja M, Agarwal G, Alam M. Green tea: a boon for periodontal and general health. J Indian Soc Periodontol 2012, 16, 161-167.

상세보기
Chi A, Valencia JC, Hu ZZ, Watabe H, Yamaguchi H, Mangini NJ, Huang H, Canfield VA, Cheng KC, Yang F, Abe R, Yamagishi S, Shabanowitz J, Hearing VJ, Wu C, Appella E, Hunt DF. Proteomic and bioinformatic characterization of the biogenesis and function of melanosomes. J Proteome Res 2006, 5, 3135-3144.

상세보기
Choi B, Heo JH, Kwon HJ, Lee ES, Sohn S. Tocotrienols enhance melanosome degradation through endosome docking/fusion proteins in B16F10 melanoma cells. Food Funct 2013, 4, 1481-1488.

상세보기
Chung KW, Park YJ, Choi YJ, Park MH, Ha YM, Uehara Y, Yoon JH, Chun P, Moon HR, Chung HY. Evaluation of in vitro and in vivo anti-melanogenic activity of a newly synthesized strong tyrosinase inhibitor (E)-3-(2,4 dihydroxybenzylidene)pyrrolidine-2,5-dione (3-DBP). Biochim Biophys Acta 2012, 1820, 962-969.

상세보기
Damek-Poprawa M, Diemer T, Lopes VS, Lillo C, Harper DC, Marks MS, Wu Y, Sparrow JR, Rachel RA, Williams DS, Boesze-Battaglia K. Melanoregulin (MREG) modulates lysosome function in pigment epithelial cells. J Biol Chem 2009, 284, 10877-10889.

상세보기
Funasaka Y, Chakraborty AK, Komoto M, Ohashi A, Ichihashi M. The depigmenting effect of a-tocopheryl ferulate on human melanoma cells. Br J Dermatol 1999, 141, 20-29.

상세보기
Gentleman RC, Carey VJ, Bates DM, Bolstad B, Dettling M, Dudoit S, Ellis B, Gautier L, Ge Y, Gentry J, Hornik K, Hothorn T, Huber W, Iacus S, Irizarry R, Leisch F, Li C, Maechler M, Rossini AJ, Sawitzki G, Smith C, Smyth G, Tierney L, Yang JY, Zhang J. Bioconductor: open software development for computational biology and bioinformatics. Genome Biol 2004, 5, R80.

상세보기
Guyonneau L, Murisier F, Rossier A, Moulin A, Beermann F. Melanocytes and pigmentation are affected in dopachrome tautomerase knockout mice. Mol Cell Biol 2004, 24, 3396-3403.

상세보기
Huang HC, Lien HM, Ke HJ, Chang LL, Chen CC, Chang TM. Antioxidative characteristics of Anisomeles indica extract and inhibitory effect of ovatodiolide on melanogenesis. Int J Mol Sci 2012, 13, 6220-6235.

상세보기
Joshi PG, Nair N, Begum G, Joshi NB, Sinkar VP, Vora S. Melanocyte-keratinocyte interaction induces calcium signalling and melanin transfer to keratinocytes. Pigment Cell Res 2007, 20, 380-384.

상세보기
Kim DS, Park SH, Kwon SB, Li K, Youn SW, Park KC. (-)-Epigallocatechin-3-gallate and hinokitiol reduce melanin synthesisvia decreased MITF production. Arch Pharm Res 2004, 27, 334-339.

원문보기 상세보기
Kim YC, Choi SY, Park EY. Anti-melanogenic effects of black, green, and white tea extracts on immortalized melanocytes. J Vet Sci 2015, 16, 135-143.

상세보기
Langmead B, Salzberg SL. Fast gapped-read alignment with Bowtie 2. Nat Methods 2012, 9, 357-359.

상세보기
Lee JH, Hsieh CF, Liu HW, Chen CY, Wu SC, Chen TW, Hsu CS, Liao YH, Yang CY, Shyu JF, Fischer WB, Lin CH. Lipid raft-associated stomatin enhances cell fusion. FASEB J 2017, 31, 47-59.

상세보기
Lin JY, Fisher DE. Melanocyte biology and skin pigmentation. Nature 2007, 445, 843-850.

상세보기
Lin YS, Wu SS, Lin JK. Determination of tea polyphenols and caffeine in tea flowers (Camellia sinensis) and their hydroxyl radical scavenging and nitric oxide suppressing effects. J Agric Food Chem 2003, 51, 975-980.

상세보기
Lobo V, Patil A, Phatak A, Chandra N. Free radicals, antioxidants and functional foods: Impact on human health. Pharmacogn Rev 2010, 4, 118-126.

상세보기
Mustapha N, Bzeouich IM, Ghedira K, Hennebelle T, Chekir-Ghedira L. Compounds isolated from the aerial part of Crataegus azarolus inhibit growth of B16F10 melanoma cells and exert a potent inhibition of the melanin synthesis. Biomed Pharmacother 2015, 69, 139-144.

상세보기
Nair SS, Chaubal VA, Shioda T, Coser KR, Mojamdar M. Over-expression of MSG1 transcriptional co-activator increases melanin in B16 melanoma cells: a possible role for MSG1 in melanogenesis. Pigment Cell Res 2001, 14, 206-209.

상세보기
No JK, Soung DY, Kim YJ, Shim KH, Jun YS, Rhee SH, Yokozawa T, Chung HY. Inhibition of tyrosinase by green tea components. Life Sci 1999, 65, PL241-246.
Rabeta M, Vithyia M. Effect of different drying methods on the antioxidant properties of Vitex negundo Linn. tea. Int Food Res J 2013, 20, 3171-3176.
Roh JS, Han JY, Kim JH, Hwang JK. Inhibitory effects of active compounds isolated from safflower (Carthamus tinctorius L.) seeds for melanogenesis. Biol Pharm Bull 2004, 27, 1976-1978.

상세보기
Sasaki M, Kizawa K, Igarashi S, Horikoshi T, Uchiwa H, Miyachi Y. Suppression of melanogenesis by induction of endogenous intracellular metallothionein in human melanocytes. Exp Dermatol 2004, 13, 465-471.

상세보기
Serafini M, Ghiselli A, Ferro-Luzzi A. In vivo antioxidant effect of green and black tea in man. Eur J Clin Nutr 1996, 50, 28-32.

상세보기
Shon MS, Kim RH, Kwon OJ, Roh SS, Kim GN. Beneficial role and function of fisetin in skin health via regulation of the $CCN_{2}$ /TGF- ${\beta}$ signaling pathway. Food Sci Biotechnol 2016, 25 (Suppl 1), 133-141.

상세보기
Solano F, Briganti S, Picardo M, Ghanem G. Hypopigmenting agents: an updated review on biological, chemical and clinical aspects. Pigment Cell Res 2006, 19, 550-571.

상세보기
Tang H, Zhou P. Heat shock protein Hspa5 interacts with and protects tyrosinase activity. Braz Arch Biol Technol 2015, 58, 547-552.

상세보기
Tsuboi T, Kondoh H, Hiratsuka J, Mishima Y. Enhanced melanogenesis induced by tyrosinase gene-transfer increases boron-uptake and killing effect of boron neutron capture therapy for amelanotic melanoma. Pigment Cell Res 1998, 11, 275-282.

상세보기
Wang S, Ma Z, Xu X, Wang Z, Sun L, Zhou Y, Lin X, Hong W, Wang T. A role of Rab29 in the integrity of the trans-Golgi network and retrograde trafficking of mannose-6-phosphate receptor. PLoS One 2014, 9, e96242.

상세보기
Wellings SR, Siegel BV. Role of Golgi apparatus in the formation of melanin granules in human malignant melanoma. J Ultrastruct Res 1959, 3, 147-154.

상세보기
Yamakoshi J, Otsuka F, Sano A, Tokutake S, Saito M, Kikuchi M, Kubota Y. Lightening effect on ultravioletinduced pigmentation of guinea pig skin by oral administration of a proanthocyanidin-rich extract from grape seeds. Pigment Cell Res 2003, 16, 629-638.

상세보기
Yang Z, Xu Y, Jie G, He P, Tu Y. Study on the antioxidant activity of tea flowers (Camellia sinensis). Asia Pac J Clin Nutr 2007, 16 (Suppl 1), 148-152.

저자의 다른 논문 :

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증