Application of leech peptide HE-D in skin care and whitening
A technology of HE-D and leech peptide, which is applied in the field of skin care products, can solve the problems affecting the whitening effect and appearance of products, and achieve the effect of inhibiting macrophage migration activity, good safety and easy absorption
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Embodiment 1
[0057] Example 1 Detection of tyrosinase inhibitory activity
[0058] (1) Inhibitory effect of different concentrations of leech peptide HE-D solutions on enzyme activity
[0059] Five different concentration gradient solutions (0.0075 mg / ml, 0.015 mg / ml, 0.03mg / ml, 0.06 mg / ml, 0.12mg / ml) were prepared for Hirudopeptide HE-D. Add PBS, tyrosinase solution, and leech peptide HE-D to test tubes A, B, C, and D respectively. The specific addition information is as follows. After placing in a constant temperature water bath at 25°C for 10 min, immediately add 1 mL of 2w / v% L-Dopa solution and mix well. After another 5 min in constant temperature water bath at 25 ℃, the absorbance was measured at 475 nm immediately. Repeat for each group.
[0060] A: 2 mL PBS+1 mL tyrosinase solution;
[0061] B: 3 mL PBS;
[0062] C: 1 mL sample solution + 1 mL PBS + 1 mL tyrosinase solution;
[0063] D: 1 mL sample solution + 2 mL PBS;
[0064] Inhibition rate of tyrosinase activity (%)=[(A-...
Embodiment 2
[0074] Embodiment 2 inhibits the synthesis of melanin
[0075] 1. MTT method to detect the toxicity of hirudopeptide HE-D to B16 melanoma cells
[0076] (1) Cell culture: the cell density is 10 5 / ml, 100 μl of cell suspension per well, and 100 μl of PBS in the edge wells.
[0077] (2) Dosing treatment: Add 25 μg / mL, 50 μg / mL, 100 μg / mL, 200 μg / mL, and 400 μg / mL of leech HE-D respectively, set up 5 duplicate wells, and set up a set of blank controls.
[0078] (3) MTT method: under dark conditions, 10 μl MTT per well. After culturing in the incubator for 24 hours, aspirate the culture solution and add 100 μl DMSO to each well. The OD value was measured at 570nm with a microplate reader, and the cell viability was calculated. The result is as figure 1 As shown, the results show that the hirud peptide HE-D has no toxicity to B16 cells within 24 hours within the test concentration range of 0-400 μg / ml.
[0079] 2. Determination of melanin content in B16 melanoma cells by NaO...
Embodiment 3
[0086] Example 3 Essence
[0087] According to the formula in Table 3, stir to dissolve the materials in the formula evenly, and then bottle it.
[0088]
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