Composition for inhibiting melanin generation, whitening essence, and preparation method and application thereof
A technology for inhibiting melanin and composition, which is applied in the field of daily chemicals, can solve the problems of single action of arbutin, skin irritation, and easy discoloration of kojic acid, etc., and achieve the effect of inhibiting the formation of melanin, activating the function, and reducing the formation of melanin
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Embodiment 1-6
[0076] Phellodendron Phellodendri extract was diluted with phosphate buffer to obtain 6 groups of test solutions with different concentrations (final concentrations), which were filtered with a 0.22 μm filter head. When performing the cell viability test, the mass concentration of Phellodendron Phellodendri extract is shown in Table 1 below.
[0077] Cell Viability Test
[0078] CCK-8 reagent contains WST-8: chemical name 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfone Acid phenyl)-2H-tetrazole monosodium salt, which is reduced to highly water-soluble Yellow formazan product, the amount of formazan produced is directly proportional to the number of viable cells. The light absorption value was measured at a wavelength of 450nm by an enzyme-linked immunosorbent assay instrument, which can indirectly reflect the number of living cells. This method has been widely used in the activity detection of some biologically active factors, large-scale antitumor drug scr...
Embodiment 7-12
[0086] Phellodendron Phellodendri extract was diluted with phosphate buffer to obtain 6 groups of test solutions with different concentrations (final concentrations), which were filtered with a 0.22 μm filter head. When performing the cell viability test, the mass concentration of Phellodendron Phellodendri extract is shown in Table 2 below.
[0087] Melanocyte norepinephrine receptor inhibition test
[0088] When norepinephrine activates melanocyte norepinephrine receptors, it generates cAMP via adenylyl cyclase (AC) activation. cAMP in turn activates PKA, which activates the gene expression of MITF through the phosphorylation of CREB. Finally, by interacting with TYR, TRP-1 and TRP-2 promoter regions, MITF gene is transcribed to produce melanin-related proteins. Therefore, detecting the concentration of intracellular cAMP can reflect the degree of activation of norepinephrine receptors.
[0089] Dilute melanoma B16 cells to 2.5*10 4 cells / ml, transfer the cell suspensi...
Embodiment 13-17
[0098] Phellodendron Phellodendri extract and Glycyrrhizae extract were diluted with phosphate buffer to obtain 5 test solutions with different concentrations (final concentrations). When testing, the mass concentrations of Phellodendron Phellodendri Extract and Licorice Root Extract are shown in Table 3 below.
[0099] B16 cell melanin change rate test
[0100] Dilute melanoma B16 cells to 2.5*10 4 cells / ml, move the cell suspension to a 9cm culture dish, and add 100 μl of the cell suspension to each well of a flat-bottomed 96-well plate with a multi-channel pipette. The culture plate was placed in a 5% carbon dioxide incubator, cultured in a humid environment at 37°C for 24 hours, and Phellodendron Phellodendri extract and Glycyrrhizae extract (according to the mass concentration in Table 3) were added. After acting for 4 hours, add 10*10 -9 mol / L norepinephrine and 1*10 -6 mol / L α-MSH, and after 48 hours, the culture medium was aspirated, digested with 0.25% trypsin, ...
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