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Lee, Kim, Jeon, Lee, Hong, and Jin: Inhibition of Melanogenesis by Abietatriene from Vitex Trifolia Leaf Oil
Original Article

Natural Product Sciences 2016;22(4):252-258.

Published online: 20 January 2016

DOI: https://doi.org/10.20307/nps.2016.22.4.252

Inhibition of Melanogenesis by Abietatriene from Vitex Trifolia Leaf Oil

Hong Gu Lee†, Tae Yoon Kim†, Jung Hoon Jeon1, Hwa Sang Lee1, Yoon Ki Hong2, Mu Hyun Jin1, ∗

1Research & Development Center, LG Household & Healthcare Ltd.; 175, Gajeong-ro, Youseong-gu, Daejeon 305–343, South Korea.

2Herbal Experiment Station, Jeonbuk A.R.E.S. Medicinal Resources Research Institute.; 108, Haengjeonggongan-gil, Unbong-eup, Namwon-si, Jeollabuk-do, Korea.

∗Author for correspondence Mu Hyun Jin, Research & Development Center, LG Household & Healthcare Ltd., 175 Gajeong-ro, Youseong-gu, Daejeon 305-343, South Korea Tel: +82-42-860-8725; E-mail: mhjin@lgcare.com

† These authors contributed equally to this work.

Received 30 March 2016       Revised 14 June 2016       Accepted 18 June 2016

Copyright © 2016 The Korean Society of Pharmacognosy

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Vitex trifolia L. has been used traditionally to treat various illnesses, such as inflammation, headache, migraine, and gastrointestinal infections. We analyzed and evaluated the composition of V. trifolia leaf oil. Based on the results, we isolated abietatriene from V. trifolia leaf oil and investigated the effect of V. trifolia leaf oil and its active compound abietatriene on melanogenesis in B16F10 melanoma cells. They significantly decreased melanin contents and melanogenic factors, such as tyrosinase, TRP-1, TRP-2, and MITF dose-dependently in both protein and mRNA levels. Protein and mRNA expressions were determined by Western blot analysis and quantitative real time RT-PCR. Findings indicate that V. trifolia leaf oil and abietatriene reduce melanogenesis by regulating the expression of melanogenic factors. These results suggest that V. trifolia leaf oil and abietatriene could comprise a useful therapeutic agent for treating hyperpigmentation and used as effective skin-whitening agents.

Keywords: Vitex trifolia, Abietatriene, Melanogenesis, B16F10 melanoma

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Fig. 1.
Structure of Compound 1.
nps-22-252f1.tif
Fig. 2.
The cell viability of V. trifolia leaf oil and abietatriene. B16F10 melanoma cells were treated with V. trifolia leaf oil and abietatriene (1 – 100 μg/mL) for 48 h. Cell viability was measured by CCK-8 assay. Data are expressed as mean ± SD of three independent experiments. ∗p < 0.05, ∗∗p < 0.01 compared with DMSO control (−).
nps-22-252f2.tif
Fig. 3.
Effect of V. trifolia leaf oil fractions 1 – 8 on melanin content. B16F10 cells were treated with V. trifolia leaf oil fractions 1 – 8, V. trifolia leaf oil and abietatriene or arbutin for 48 h. (A) Melanin content of V. trifolia leaf oil fractions 1 – 8 (10, 20 μg/mL). (B) Melanin content of V. trifolia leaf oil fractions 1 – 8, V. trifolia leaf oil and abietatriene 20 μg/mL or arbutin 100 μg/mL. Melanin content was determined by spectrophotometry. Data are expressed as % of DMSO control (−).
nps-22-252f3.tif
Fig. 4.
Effect of V. trifolia leaf oil and abietatriene on melanin content. B16F10 cells were treated with V. trifolia leaf oil and abietatriene (1 – 20 μg/mL) or arbutin 100 μg/mL for 48 h. Melanin content was determined by spectrophotometry. Data are expressed as % of control (DMSO) and expressed as mean ± SD of three independent experiments. ∗p < 0.05, ∗∗p < 0.01 compared with DMSO control (−).
nps-22-252f4.tif
Fig. 5.
Effect of V. trifolia leaf oil and abietatriene on the protein expression of melanogenic factors. B16F10 cells were treated with V. trifolia leaf oil and abietatriene (1, 10, and 20 μg/mL) and incubated for 48 h. Cell lysates were evaluated by Western blot with antibodies against tyrosinase, TRP-1, TRP-2, and MITF. Equal protein loading was confirmed by staining with antibodies against â-actin. (A) The protein expression of melanogenic proteins. (B) Relative protein expression of each melanogenic proteins. Band intensity was quantified using ImageJ software. The results are expressed as % of control and each column represents the mean ± SD of three independent experiments. ∗p < 0.05, ∗∗p < 0.01 compared with DMSO control (−).
nps-22-252f5.tif
Fig. 6.
Effect of V. trifolia leaf oil and abietatriene on the mRNA expression of melanogenic factors. B16F10 cells were treated with V. trifolia leaf oil and abietatriene (1, 10 and 20 μg/mL) and incubated for 24 h. Cell lysates were evaluated by qPCR with Taqman probe against tyrosinase, TRP-1, TRP-2, and MITF. Equal mRNA loading was confirmed by GAPDH. The results are expressed as % of control, the mean ± SD of three independent experiments. ∗p < 0.05, ∗∗p < 0.01 compared with DMSO control (−).
nps-22-252f6.tif
Table 1.
Main composition of the essential oil isolated from leaf of V. trifolia
RT (min) Compounds Peak area (%)
13.038 α-pinene 11.38
15.506 β-pinene  2.84
15.815 sabinene 10.25
18.256 eucaluptol  8.60
28.433 camphene 12.69
41.495 Manoyl oxide 16.11
50.807 abietatriene  9.03
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