Use of polypeptide in the preparation of drugs or cosmetics for promoting collagen secretion
A collagen and cosmetic technology, applied in the field of biomedicine, can solve problems such as inconvenience of use
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Embodiment 1
[0023] Example 1 Effect of Polypeptide on Human Normal Fibroblast Proliferation
[0024] Human skin fibroblasts were cultured in MEM medium (containing 5% fetal bovine serum) in 5% CO 2 , a humidity of 95%, and a constant temperature of 37°C in a carbon dioxide incubator. Take a 96-well plate and seed cells at 1×103 per well. After the cells were attached, 10 μL of 1, 10, and 100 μg / mL JYDB-9 polypeptide solutions were added to each group, and the same volume of PBS buffer was added as a blank control group. After adding the peptide solution, culture the cells for 72 hours, take out the 96-well plate from the incubator, add 10 μL of WST-8 solution to each well, and put it back into the CO 2 Incubate in an incubator for color reaction. Take it out after 4h and measure the OD value at 450nm. The results are shown in Table 1.
[0025] The effect of table 1 polypeptide on human skin fibroblast proliferation
[0026] group OD Blank group (PBS) 0.280±0.027 ...
Embodiment 2
[0029] Example 2 Effect of JYDB-9 polypeptide on the secretion of hyaluronic acid from normal human fibroblasts
[0030] Take a 96-well plate, press 1×10 per well 3 Seed cells and change the medium every 3 days. After the cells were attached, 10 μL of 100 μg / mL solution was added to each group, and the same volume of PBS buffer was added as a blank control group. After culturing for 7 days, the secretion of hyaluronic acid was measured according to the instructions of the kit. The result is as figure 1 shown. see results figure 1 As shown, the secretion of hyaluronic acid in the polypeptide group reached 155.4±2.3, which was significantly higher than that in the blank group, with a significant difference (P<0.05), indicating that the polypeptide had a significant effect on the secretion of hyaluronic acid from human skin fibroblasts cultured in vitro. influences.
Embodiment 3
[0031] Example 3 Effect of Polypeptide on Collagen Gene Expression
[0032] Take a 96-well plate, press 1×10 per well 3Seed cells and change the medium every 3 days. After the cells were attached, 10 μL of 100 μg / mL solution was added to each group, and the same volume of PBS buffer was added as a blank control group. After culturing for 7 days, total RNA was extracted using Trizol reagent. After adding 20% (v / v) chloroform solution, vortex for 30S, and let stand at room temperature for 10min, and then centrifuge at 4°C (12000r / min, 10min). Pipette the upper aqueous phase (400-500 μL) and transfer it to a new centrifuge tube, and add an equal amount of isopropanol. After mixing gently, let stand at 4°C for 10 minutes, and centrifuge again at 4°C (12000r / min, 10min). Wash the sediment with 75% (v / v) ethanol and 25% (v / v) nuclease-free water mixture, wash twice, centrifuge again under the same conditions, extract total RNA, and add 20 μL nuclease-free water to dissolve the...
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