Peptide for reducing hair loss and promoting hair growth, and cosmetic composition and pharmaceutical composition comprising same
A cosmetic composition and hair growth technology, applied in cosmetics, cosmetic preparations, hair care, etc., can solve problems such as poor hair loss, erectile dysfunction, etc., to promote keratinocyte proliferation, regulate hair follicle growth, improve The effect of cell regeneration
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Embodiment 1
[0086] Example 1: Confirmation of the activation effect of the peptide on the Wnt / β-catenin signal transduction pathway
[0087] In order to analyze the activation of the Wnt / β-catenin signaling pathway in cells treated with the peptides of SEQ ID NO: 5 and SEQ ID NO: 10 shown in Table 1 below, β-catenin, glycogen synthesis Western blot of enzyme kinase 3β and p-glycogen synthase kinase 3β protein expression.
[0088] Human follicular dermal papilla cells (HFDPC) were mixed with 1×10 5 or 5×10 4 The number of cells was uniformly inoculated on a 6-well plate for culture, in Dulbecco's Modified Eagle Medium (DMEM, Dulbecco's Modified Eagle Media, Gibco BRL), at a temperature of 37°C, 5% CO 2 Conditions were incubated for 24 hours in the incubator. After that, multiple peptides of SEQ ID NO: 5 and SEQ ID NO: 10 were dissolved in water to a concentration of 10 mM to prepare a concentrated solution, which was diluted to a concentration of 200 uM, 100 uM, and 20 uM in a culture ...
Embodiment 2
[0092] Example 2: Finding the optimal concentration for activation of Wnt / β-catenin signal transduction pathway
[0093] The optimal concentration for activating the Wnt / β-catenin signal transduction pathway was found by treating the peptides of SEQ ID NO: 5 and SEQ ID NO: 10 according to the concentration of the present invention. The specific experimental method is as follows: in the case of SEQ ID NO:5, the concentration is diluted to 0.1 μM, 0.5 μM, 1 μM, 5 μM, 10 μM, 20 μM, 40 μM, 60 μM, 80 μM, 100 μM, in addition, the same as in Example 1 same in Figure 3a The result is shown. In the case of SEQ ID NO: 10, the concentration was diluted to 0.1 μM, 0.5 μM, 1 μM, 5 μM, 10 μM, 20 μM, 40 μM, 50 μM, except that it was the same as in Example 1, in Figure 3b The result is shown.
[0094] exist Figure 3a and Figure 3bIt can be seen that the peptides of SEQ ID NO: 5 and SEQ ID NO: 10 of the present invention were confirmed to increase the expression of active β-catenin pr...
Embodiment 3
[0095] Example 3: Confirmation of DKK-1 Activity Inhibition
[0096] The SEQ ID NO:5 and SEQ ID NO:10 peptides of the invention were processed to analyze whether the Wnt / β-catenin signaling pathway inhibited by DKK-1 was reactivated.
[0097] Specifically, human dermal papilla cells were divided into 5×10 4 The number of cells was uniformly seeded on a 12-well plate for culture, in Dulbecco's modified Eagle's medium, at a temperature of 37 ° C, 5% CO 2 Conditions were incubated for 24 hours in the incubator. After culturing, each well was treated with recombinant protein DKK-1 to inhibit the Wnt / β-catenin signal transduction pathway. At the same time, after additional treatment with the SEQ ID NO:5 and SEQ ID NO:10 peptides of the present invention at an appropriate concentration, Cultivate for a specified time.
[0098] After the culture was completed, the medium was removed, the cells were disrupted with sodium dodecyl sulfate sample buffer, and the proteins were separate...
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