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Seo, Do, Lee, Lee, Wahedi, Park, and Kim: Modulation of Melanin Synthesis by Amaranthus spp. L Seed Extract in Melan-a Cells
Original Article

Natural Product Sciences 2016;22(3):168-174.

Published online: 17 January 2016

DOI: https://doi.org/10.20307/nps.2016.22.3.168

Modulation of Melanin Synthesis by Amaranthus spp. L Seed Extract in Melan-a Cells

Jae Ok Seo1, Moon Ho Do1, Jae Hak Lee2, Taek Hwan Lee3, Hussain Mustatab Wahedi1, Yong Un Park1, Sun Yeou Kim1, 4, 5, ∗

1College of Pharmacy, Gachon University, Yeonsu-gu, Incheon, Republic of Korea

2Korea Plant Resource Institute, Paju-si, Gyeonggi-do, Republic of Korea

3College of Pharmacy, Yonsei University, Yeonsu-gu, Incheon, Republic of Korea

4Gachon Medical Research Institute, Gil Medical Center, Incheon, Republic of Korea

5Gachon Institute of Pharmaceutical Science, Gachon University, Yeonsu-gu, Incheon, Republic of Korea

∗Author for correspondence Sun Yeou Kim, College of Pharmacy, Gachon University, 191 Ham-bakmoero, Yeonsu-gu, Incheon 406-799, Republic of Korea. Tel: +82-32-899-6411; E-mail: sunnykim@gachon.ac.kr

Received 5 November 2015       Revised 23 February 2016       Accepted 29 February 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

Anti-melanogenic effects of amaranth (AT), one of the key source of squalene, were investigated in melanocytes. Amaranth seed powder was extracted with water and melan-a cells were treated with various concentrations of AT. By using HPLC, content of myo-inositol, one of potential active components, was measured in the crude extract of AT.AT reduced the melanin content in melan-a melanocytes and down-regulated melanogenic enzyme activity such as tyrosinase, TRP-1 and TRP-2. By regulating melanogenic enzyme activity, AT may be a potential natural source for whitening agent. Myo-inositol was detected in AT by HPLC and may be one of the active compounds from AT involved in the regulation of anti-melanogenesis. In this study, we demonstrated that AT has anti-melanogenesis properties. This new function of amaranth may be useful in the development of new skin-whitening products and its value as food.

Keywords: Amaranth, Myo-inositol, Melan-a cells, Melanogenesis

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Fig. 1.
Scheme of sample preparation. Simply present the steps of extraction of AT. A: Amaranthus spp. L Total extract, B: Hexane soluble fraction, C: Residue, D: EtoAc soluble fraction.
nps-22-168f1.tif
Fig. 2.
HPLC chromatogram for the determination of myo-inositol in Amaranth ushypocondriacus (A.D). A. Chromatogram of myo-inositol B. Chromatogram of AD.
nps-22-168f2.tif
Fig. 3.
Effects of Amaranth seed extract (AT) on melanin content and cell viability in melan-a cells. Melanin contents (A) and cell viability (B) were measured. CTL indicates normal conditions and Arbu indicate the use of arbutin as a positive control (μM). All sample (μ g/mL) were treated for 72 h. Data are expressed as mean ± S.D. of three experiments. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 compared with control.
nps-22-168f3.tif
Fig. 4.
Effects of AT on tyrosinase activity in melan-a cells. Melan-a cell originated tyrosinase activity was measured. CTL indicates normal conditions and KA indicate the use of kojic acid as a positive control (μM). Each samples were treated with μg/ mL. The results are expressed as mean ± S.D. of three experiments (∗p < 0.05).
nps-22-168f4.tif
Fig. 5.
Effects of Amaranth ushybridus (A.C) and Amaranth ushypocondriacus (A.D) on the expression of melanogenic enzymes in melan-a cells. To confirm the expression of melanogenic enzymes, melan-a cells were treated with 0.1, 1 and 10 μg/mL of A.C and A.D. Arbutin was used as a positive control (μ M). (A) Western blot analysis. Densitometric analysis of Tyrosinase (B), TRP-1 (C), TRP-2 (D) and MITF (F). Each band was normalized to that of α-tubulin.
nps-22-168f5.tif
Fig. 6.
Effects of each fraction of amaranth seed extract (AT) on melanin content and cell viability in melan-a cells Melanin contents (A) and cell viability (B) were measured. CTL indicates normal conditions and Arbu indicate the use of arbutin as a positive control (μM). All sample (μg/mL) were treated for 72 h. Data are expressed as mean ± S.D. of three experiments. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 compared with control.
nps-22-168f6.tif
Supplement 1.
Effects of Amaranth species on melanin content and cell viability in mouse melanoma B16F10 cells. Cell viability (A) and melanin contents (B) were measured. B16F10 cells were pre-stimulated by α-MSH (100 ng/ml). After treatment of α-MSH for 30 min, all sample were treated for 72 h.
nps-22-168fs1.tif
Supplement 2.
Effects of myo-inositol on melanin content and cell viability in Melan-a cells. Cell viability (A) and melanin contents (B) were measured. myo-inositol was treated for 72 h.
nps-22-168fs2.tif
Table 1.
Contents (μ g/mg) of myo-inositol in Amaranth ushypocondriacus (A.D).
  r2 Content
Myo-inositol 0.9955 49.036 ± 1.032
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