Nat Prod Sci.  2016 Sep;22(3):168-174. 10.20307/nps.2016.22.3.168.

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

• Seo JO
• Do MH
• Lee JH
• Lee TH
• Wahedi HM
• Park YU
• Kim SY

• KMID: 2355141
• DOI: http://doi.org/10.20307/nps.2016.22.3.168

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.

Keyword

Amaranth; Myo-inositol; Melan-a cells; Melanogenesis

MeSH Terms

Amaranthus*
Chromatography, High Pressure Liquid
MART-1 Antigen*
Melanins*
Melanocytes
Monophenol Monooxygenase
Squalene
Water
MART-1 Antigen
Melanins
Monophenol Monooxygenase
Squalene
Water

Figure

• 

  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.

• 

  Fig. 2. HPLC chromatogram for the determination of myo-inositol in Amaranth ushypocondriacus (A.D). A. Chromatogram of myo-inositol B. Chromatogram of AD.

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  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.

• 

  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).

• 

  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.

• 

  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.

• 

  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.

• 

  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.

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