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. 2020 Mar 20;12(3):832.
doi: 10.3390/nu12030832.

Anti-Melanogenic Effect of Ethanolic Extract of Sorghum bicolor on IBMX-Induced Melanogenesis in B16/F10 Melanoma Cells

Affiliations

Anti-Melanogenic Effect of Ethanolic Extract of Sorghum bicolor on IBMX-Induced Melanogenesis in B16/F10 Melanoma Cells

Hye Ju Han et al. Nutrients. .

Abstract

To evaluate possibility as a skin whitening agent of Sorghum bicolor (S. bicolor), its antioxidant activity and anti-melanogenic effect on 3-isobutyl-1-methylxanthine (IBMX)-induced melanogenesis in B16/F10 melanoma cells were investigated. The result of total phenolic contents (TPC) indicated that 60% ethanol extract of S. bicolor (ESB) has the highest contents than other ethanol extracts. Antioxidant activity was evaluated using the 2,2'-azino-bis-(3-ethylbenzothiazolin-6-sulfonic acid) diammonium salt (ABTS)/1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activities and malondialdehyde (MDA) inhibitory effect. These results showed ESB has significant antioxidant activities. Inhibitory effect against tyrosinase was also assessed using L-tyrosine (IC50 value = 89.25 μg/mL) and 3,4-dihydroxy-L-phenylalanine (L-DOPA) as substrates. In addition, ESB treatment effectively inhibited melanin production in IBMX-induced B16/F10 melanoma cells. To confirm the mechanism on anti-melanogenic effect of ESB, we examined melanogenesis-related proteins. ESB downregulated melanogenesis by decreasing expression of microphthalmia-associated transcription factor (MITF), tyrosinase and tyrosinase-related protein (TRP)-1. Finally, 9-hydroxyoctadecadienoic acid (9-HODE), 1,3-O-dicaffeoylglycerol and tricin as the main compounds of ESB were analyzed using the ultra-performance liquid chromatography-ion mobility separation-quadrupole time of flight/tandem mass spectrometry (UPLC-IMS-QTOF/MS2). These findings suggest that ESB may have physiological potential to be used skin whitening material.

Keywords: 3-isobutyl-1-methylxanthine; B16/F10 melanoma cell; Sorghum bicolor; anti-melanogenesis; hydroxyoctadecadienoic acid.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Total phenolic contents of various ethanolic extracts from Sorghum bicolor. Data were expressed as the means ± SD (n = 3). Each small letter represented statistical difference, and was statistically considered at p < 0.05.
Figure 2
Figure 2
2,2’-azino-bis-(3-ethylbenzothiazolin-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity (A), 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activity (B) and malondialdehyde (MDA) inhibitory effect (C) of the 60% ethanolic extract from Sorghum bicolor (ESB). Data were expressed as the means ± SD (n = 3). Each small letter represented statistical difference, and was statistically considered at p < 0.05.
Figure 2
Figure 2
2,2’-azino-bis-(3-ethylbenzothiazolin-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity (A), 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activity (B) and malondialdehyde (MDA) inhibitory effect (C) of the 60% ethanolic extract from Sorghum bicolor (ESB). Data were expressed as the means ± SD (n = 3). Each small letter represented statistical difference, and was statistically considered at p < 0.05.
Figure 3
Figure 3
Inhibitory effect of the 60% ethanolic extract of Sorghum bicolor (ESB) on melanogenesis-mediated enzymes. Tyrosinase inhibitory effect using L-tyrosine (A) and L-DOPA (B) as substrates, and α-glucosidase inhibitory effect (C). Data were expressed as the means ± SD (n = 5). Each small letter represented statistical difference, and was statistically considered at p < 0.05.
Figure 4
Figure 4
Cell viability (A) and melanin contents (B) in B16/F10 melanoma cells of the 60% ethanolic extract of Sorghum bicolor (ESB). Data were expressed as the means ± SD (n = 3). Each small letter represented statistical difference, and was statistically considered at p < 0.05.
Figure 5
Figure 5
Anti-melanogenic effect of the 60% ethanolic extract of Sorghum bicolor (ESB) on 3-isobutyl-1-methylxanthine (IBMX)-induced melanogenesis in B16/F10 melanoma cells. Representative band images of proteins (A), relative expression of microphthalmia-associated transcription factor (MITF) (B), tyrosinase (C) and tyrosinase and tyrosinase-related protein (TRP)-1 (D). Data were expressed as the means ± SD (n = 3). Each small letter represented statistical difference, and was statistically considered at p < 0.05.
Figure 5
Figure 5
Anti-melanogenic effect of the 60% ethanolic extract of Sorghum bicolor (ESB) on 3-isobutyl-1-methylxanthine (IBMX)-induced melanogenesis in B16/F10 melanoma cells. Representative band images of proteins (A), relative expression of microphthalmia-associated transcription factor (MITF) (B), tyrosinase (C) and tyrosinase and tyrosinase-related protein (TRP)-1 (D). Data were expressed as the means ± SD (n = 3). Each small letter represented statistical difference, and was statistically considered at p < 0.05.
Figure 6
Figure 6
UPLC IMS-QTOF/MS spectra in negative ion mode (A) and MS2 spectra and chemical structure of 1,3-O-dicaffeoylglycerol (B), Tricin (C) and 9-hydroxyoctadecadienoic acid (9-HODE) (D) from the 60% ethanolic extract of Sorghum bicolor (ESB).
Figure 6
Figure 6
UPLC IMS-QTOF/MS spectra in negative ion mode (A) and MS2 spectra and chemical structure of 1,3-O-dicaffeoylglycerol (B), Tricin (C) and 9-hydroxyoctadecadienoic acid (9-HODE) (D) from the 60% ethanolic extract of Sorghum bicolor (ESB).

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