苦水玫瑰精油对B16细胞中黑色素合成的影响及机制研究

doi:10.3969/j.issn.1001-1803.2022.03.008

Abstract

Abstract:

The effects of Kushui rose essential oil on melanin synthesis in mouse B16 melanoma cells were investigated and its mechanism of action was explored to provide an experimental basis for the development and application of Kushui rose essential oil as a natural active substance. The volatile components of the essential oil were analyzed by gas chromatography-mass spectrometry （GC-MS）. The cell model with high melanin expression induced by α-melanocyte-stimulating hormone （α-MSH） was constructed using in vitro cultured mouse B16 cells. The effects of Kushui rose essential oil on melanocyte viability were determined by CCK8 method; the changes in melanin content and tyrosinase （Tyr） activity in B16 cells were analyzed by enzyme-linked immunosorbent assay （ELISA） and dopa oxidation method, respectively; the expression of cyclic adenosine monophosphate （cAMP）-responsive element-binding protein （CRE-binding protein, CREB）, p38 MAP kinase （p38 MAPK） and extracellular signal-regulated kinase （ERK） in three signaling pathways was measured. The expression of CREB, p38 MAPK, ERK and microphthalmia-associated transcription factor （MITF） in B16 cells was determined by quantitative real-time PCR （qRT-PCR） and western blotting using Kushui rose essential oil at a mass concentration of 2 g/L for 12 h, 24 h and 48 h, respectively. qRT-PCR results showed that the expression of MITF gene was significantly inhibited after 48 h treatment of B16 cells with Kushui rose essential oil. The expression of MITF gene was down-regulated by 76%, and the expression of CREB and p38 MAPK was also significantly inhibited, in which the inhibition rates reached 54% and 41%, respectively, while the expression of ERK1 and ERK2 gradually increased with time; similar results were obtained by western blot, and the trend of the expression levels of CREB and p38 MAPK was consistent with the expression level of MITF, which decreased with time, while the expression level of ERK gradually increased with time. It demonstrates that the Kushui rose essential oil reduces intracellular melanin production through three signaling pathways: cAMP/PKA, p38 MAPK and ERK/MAPK, and thus inhibit the expression of MITF.

Key words: Kushui rose essential oil, melanin content, B16 melanoma cell, signaling pathway

CLC Number:

TQ658

Wu Yani,Lv Xiaofan,Wang Ying,Tang Yin. Effects of Kushui rose （Rosa setate × Rosa rugosa） essential oil on melanin synthesis in B16 cells and its mechanism[J].China Surfactant Detergent & Cosmetics, 2022, 52(3): 278-286.

Figures/Tables 10

Tab. 1

Fig. 1

Tab. 2

GC-MS component identification and quantitative results of Kushui rose essential oil"

序号	化学成分	保留时间/min	相对含量/%
1	α-蒎烯（α-Pinene）	9.908	0.05
2	庚醛（Heptanal）	11.662	0.14
3	罗勒烯（Ocimene）	13.217	0.05
4	乙酸己酯（Hexyl acetate）	13.708	0.05
5	庚醇（2-Heptanol）	14.698	0.78
6	乙酸叶醇酯（Cis-3-hexenyl acetate ）	14.840	0.07
7	甲基庚烯酮（5-Hepten-2-one）	15.422	0.08
8	正己醇（1-Hexanol）	15.563	0.16
9	玫瑰醚（Rose oxide）	15.828	1.13
10	顺-3-己烯-1-醇（3-Hexen-1-ol）	16.415	0.02
11	2-庚酮（2-Heptanone）	16.698	0.03
12	壬醛（Nonanal）	16.832	0.02
13	1-辛烯-3-醇（1-Octen-3-ol）	17.968	0.01
14	庚醇（1-Heptanol）	18.087	0.01
15	橙花醚（Nerolin ）	18.720	0.04
16	薄荷酮（Cyclohexanone ）	19.425	0.01
17	芳樟醇（Linalool）	20.303	2.16
18	金合欢烯（Farnesene）	21.437	0.09
19	甲酸香草酯（Citronellyl formate）	22.150	0.57
20	香橙烯（Aromandendrene）	23.003	1.99
21	乙酸香茅酯（Citronellyl acetate）	23.042	9.63
22	月桂烯（Myrcene）	24.440	0.46
23	乙酸苄酯（Benzyl acetate）	24.682	0.02
24	乙酸香叶酯（Geranyl acetate）	25.083	0.34
25	香茅醇（Citronellol）	25.238	57.61
26	杜松烯（Cedrene）	25.303	0.06
27	姜黄烯（Curcumene）	25.601 2	0.07
28	水杨酸甲酯（Methyl salicylate）	25.857	0.03
29	橙花醇（Nerol）	25.967	3.46
30	乙酸苯乙酯（Phenylethyl acetate）	26.518	0.07
31	大马酮（Damascenone）	26.677	0.06
32	香叶醇（Geraniol）	26.888	11.68
33	苯甲醇（Benzyl alcohol）	27.682	0.18
34	人参炔醇（Falcarinol）	27.870	0.03
35	苯乙醇（Phenethyl alcohol）	28.400	3.91
36	正庚酸（Heptanoic acid）	29.013	0.09
37	甲基丁香酚（Methyleugenol）	30.293	1.08
38	2-十二烷酮（2-Dodecanone）	30.403	0.67
39	红没药醇（Bisabolol）	33.301 2	0.41
40	十一酸乙酯（Ethyl undecanoate ）	33.620	0.07
41	金合欢烯（cis-á-Farnesene）	33.785	0.03
42	金合欢醇（Dodecatrienol）	34.295	0.43
43	紫罗酮（Ionone）	34.303	0.08
44	香叶酸（Geranic acid）	34.567	0.43
45	对伞花烃（p-Cymene）	34.588	0.01
46	金合欢醇（trans-Farnesol）	34.740	1.93
47	橙花叔醇（Nerolidol ）	38.583	0.01
48	苯甲酸苄酯（Benzyl Benzoate）	38.907	0.27

Tab. 2

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Fig. 6

Fig. 7

Fig. 8

References 20

[1]	Gililbro J M, Olsson M J. The melanogenesis and mechanisms of skin-lightening agentsexisting and new approaches[J]. International Journal of Cosmetic Science, 2011, 33(3) : 210. doi: 10.1111/ics.2011.33.issue-3
[2]	Hwang Y S, Kim Y J, Kim M O, et al. Cannabidiol upregulates melanogenesis through CB1 dependent pathway by activating p38 MAPK and p42/44 MAPK[J]. Chem-Biol Int, 2017, 273:107-114. doi: 10.1016/j.cbi.2017.06.005
[3]	Huang H C, Chou Y C, Wub C Y, et al. [8]-Gingerol inhibits melanogenesis in murine melanoma cellsthrough down- regulation of the MAPK and PKA signalpathways[J]. Biochem Biophys Res Comm, 2013, 438(2) : 375-381. doi: 10.1016/j.bbrc.2013.07.079
[4]	Lubbe A, Verpoorte R. Cultivation of medicinal and aromatic plants for specialty industrial materials[J]. Industrial Crops and Products, 2011, 34:785-801. doi: 10.1016/j.indcrop.2011.01.019
[5]	Chen Y M, Su W C, Li C, et al. Anti-melanogenesis of novel kojic acid derivatives in B16F10 cells and zebrafish[J]. International Journal of Biological Macromolecules, 2019, 123:723-731. doi: 10.1016/j.ijbiomac.2018.11.031
[6]	Ahn J H, Jin S H, Kang H Y. LPS induces melanogenesis through p38 MAPK activation in human melanocytes[J]. Archives of Dermatological Research, 2008, 300(6) : 325-329. doi: 10.1007/s00403-008-0863-0
[7]	Ren G X, Xue P, Sun X Y, et al. Determination of the volatile and polyphenol constituents and the antimicrobial, antioxidant, and tyrosinase inhibitory activities of the bioactive compounds from the by-product of Rosa rugosa Thunb.var.plena Regal tea[J]. BMC Complem Altern M, 2018, 18(1) : 307. doi: 10.1186/s12906-018-2374-7
[8]	Huang Chaoqing, Wang Baolin. GC-MS analysis of the chemical composition of the essential oil of Rose of Jingmiao Feng Mountain[J]. Journal of Beijing Agricultural College, 2011, 26(1) : 46-50.
[9]	Song Jia, Meng Qinghua, Pan Xianhua. Correlation study of chemical composition and fragrance characteristics of rose essential oil[J]. Flavors & Fragrances Cosmetics, 2018 (1) : 12-19.
[10]	Ma Lin, Luo Jiaquan, Li Dan, et al. Real-time cell analysis system and Trypan blue staining in the analysis of toxicity of bisphenol A on leydig cells in mice[J]. Practical Medicine and Clinical, 2018, 21(9) : 978-980.
[11]	Wang Peng, Li Suxia, Han Fusen, et al. Inhibitory effect of grape seed extract on melanin synjournal in B16 cells[J]. Journal of the Japanese Chemical Industry, 2013, 43(4) : 280-284.
[12]	Yuan Bing, Li Ranran, Han Mingyong. Experimental study of malignant melanoma regulating lung microenvironment and promoting lung metastasis[J]. Journal of Shandong University (Health Sciences), 2016, 17:13-17.
[13]	Maack A, Pegard A. Populus nigra (Salicaceae) absolute rich in phenolic acids, phenylpropano ds and flavonoids as a new potent tyrosinase inhibitor[J]. Fitoterapia, 2016, 111:95-101. doi: 10.1016/j.fitote.2016.04.001 pmid: 27091790
[14]	Sarangarajan R, Apte S P. The polymerization of melanin: a poorly understood phenomenon with egregious biological implications[J]. Melanoma Res., 2006, 16(1) : 3. pmid: 16432450
[15]	Zhou Xuesen, Jiang Yumei, Bi Yang, et al. Extraction and GC/MS analysis of essential oil and its components from Bitter Water Rose[J]. Food Industry Science and Technology, 2009, 30(11) : 226-229.
[16]	Mei-Lin Tsai, Cai-Di Lin, Keh Ai Khoo. Composition and bioactivity of essential oil from Citrus grandis (L.) Osbeck ‘Mato Peiyu’ Leaf[J]. Molecules, 2017, 22(12) : 21-54. doi: 10.3390/molecules22010021
[17]	W, Chen. Geraniol: A review of a commercially important fragrance material[J]. South African Journal of Botany, 2010 (76) : 643-651.
[18]	Asaba K, Iwasaki Y, Asai M, et al. High glucose activates pituitary proopiomelanocortin gene expression: possible role of free radical-sensitive transcription factors[J]. Diabetes/metabolism Research & Reviews, 2010, 23(4) : 317-323.
[19]	Li G J, Tang F, Zhang Q, et al. Protective effects of polymethoxyflavone-rich cold-pressed orange peel oil against ultraviolet B-induced photoaging on mouse skin. Journal of functional foods[J]. Journal of Functional Foods, 2020, 67:103834. doi: 10.1016/j.jff.2020.103834
[20]	Newton R A, Roberts D W, Leonard J H, et al. Human melanocytes expressing MC1R variant alleles show impaired activation of multiple signaling pathways[J]. Peptides, 2007, 28(12) : 2387-2396. doi: 10.1016/j.peptides.2007.10.003

引物名称	引物序列
CREB	Forward 5’-GAGCAGACAACCAGCAGAGT-3’
CREB	Reverse 5’-ACCTGGGCTAATGTGGCAAT-3’
p38 MAPK	Forward 5’-CCGTTTCAGTCCATCATTCA-3’
p38 MAPK	Reverse 5’-AGGTTGCTGGGCTTTAGGT-3’
ERK1	Forward 5’-GTCTCTGCCCTCGAAAACCA-3’
ERK1	Reverse 5’-AGGTAGGAGCAGGACCAGAT-3’
ERK2	Forward 5’-ATCACATCCTGGGTATTCTTGG-3’
ERK2	Reverse 5’-GGGCTCATCACTTGGGTCA-3’
MITF	Forward 5’-CCCAGGTATGAACACGCACT-3’
MITF	Reverse 5’-GGGAAAATACACGCTGTGAGC-3’
GAPDH	Forward 5’-TCAACGGCACAGTCAAGG-3’
GAPDH	Reverse 5’-TGAGCCCTTCCACGATG-3’

Effects of Kushui rose （Rosa setate × Rosa rugosa） essential oil on melanin synthesis in B16 cells and its mechanism

RichHTML

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 10

References 20

Related Articles 5

Metrics

Comments

Recommended 10

[1]	Che Biao,Gao Yan,Xiao Lei,Huang Jinlian. Research progress of related factors in oral ulcers [J]. China Surfactant Detergent & Cosmetics, 2021, 51(9): 881-889.
[2]	Ren Qianqian,Wu Hua,Jin Jianming. Botanical cosmetic ingredient (IV) Research and development of skin whitening ingredients from plant extracts with melanogenesis signaling pathway inhibiting effect [J]. China Surfactant Detergent & Cosmetics, 2021, 51(7): 590-597.
[3]	WANG Ying,TANG Yin,WU Ya-ni. Effects of blend essential oil on melanogenesis in B16 cells of mice [J]. China Surfactant Detergent & Cosmetics, 2021, 51(3): 214-219.
[4]	Luo Tianji,Mei Zikun,Song Jingjiu,Zhu Jun. Study on synthesis and whitening activity of isoferic acid amides [J]. China Surfactant Detergent & Cosmetics, 2021, 51(11): 1102-1108.
[5]	HUANG Hai-chao,ZHENG Gong-ming,WANG Ru-yi,ZHANG Xian-ce,LI Zhong-jun,LIU Gang-yong,FU Zhong. Inhibition of corilagin on the proliferation of B16 melanoma cells and tyrosinase activity [J]. China Surfactant Detergent & Cosmetics, 2016, 46(1): 44-47.