Composition for improving skin
A composition and skin aging technology, applied in the direction of skin care preparations, skin diseases, drug combinations, etc., can solve the problems of weak moisturizing effect of wrinkles, skin irritation and redness, and fragility of component changes, so as to prevent skin wrinkles, improve or Aging, skin wrinkle improvement effect
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Embodiment 1
[0097] Example 1: Confirmation of the effect of promoting the synthesis of collagen type I in human fibroblasts
[0098] When fisetin was added to the culture medium of human fibroblasts, the effect of promoting collagen synthesis was confirmed at the cellular level. The fisetin used was purchased from Sigma Aldrich (USA). The synthetic collagen was quantified using a PICP EIA kit (Procollagen Type I C-Peptide Enzyme Immuno Assay KIT).
[0099] In the experiment, the final concentration of fisetin was adjusted to 10 μg / mL, 20 μg / mL. After adding fisetin to the culture medium (DMEM medium) of human fibroblasts and culturing for 48 hours, take the culture solution, pass through the PICP EIA kit, and use a spectrophotometer to measure the concentration of fisetin in each concentration at 450nm. The degree of collagen synthesis.
[0100] In order to compare the effect, the degree of collagen synthesis was measured by the same method for the fibroblast culture medium to which no...
Embodiment 2
[0104] Example 2: Confirmation of the effect of promoting the synthesis of sulfated glycosaminoglycans in human fibroblasts
[0105] When fisetin was added to the culture medium of human fibroblasts, the effect of promoting the synthesis of sulfated glycosaminoglycans was confirmed at the cellular level. The determination of sulfated glycosaminoglycans was quantified using a sulfated aminoglycan assay kit (Blyscan glycosaminoglycan assay kit, Biocolor).
[0106] In the experiment, the final concentration of fisetin was adjusted to 1 μg / mL, 10 μg / mL, 20 μg / mL. After adding fisetin to the culture medium (DMEM medium) of human fibroblasts and culturing for 24 hours, the culture solution was taken, passed through the aminoglycan sulfate detection kit, and using a spectrophotometer, measured the respective concentrations at 656 nm. The total amount of sulfated glycosaminoglycans in the concentration.
[0107] The increase rate of sulfated glycosaminoglycan production was calculat...
Embodiment 3
[0111] Example 3: Increase of fisetin-based hyaluronan synthase (HSS) gene expression in human keratinocytes
[0112] Human keratinocytes were cultured in DMEM medium containing 10% bovine serum (fetal bovin serum) at 1×10 6 Cells / mL were cultured, respectively injected into 6-well plates, with an injection volume of 3000 μL per well, and cultured for 18 hours. Then, it was washed twice with DMEM medium not containing bovine serum, and treated with fisetin (1 ppm or 10 ppm) for 24 hours. The cells were recovered, and all RNA was extracted by RNeasy mini kit (Qiagen), and then quantified, and 1 μg of RNA was used RNA PCR Kit (Applied Biosystems) was used for reverse transcription. The reverse transcription reaction is carried out by PCR instrument (Biorad) was performed.
[0113] Then, using reverse transcription reaction solution 2 μ L and HAS-2, HAS-3, GAPDH (3-phosphate glyceraldehyde dehydrogenase, Glyceraldehyde-3-phosphate dehydrogenase) The probes (Invitrogen, US...
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