Application of astragaloside to preparation of drug
A technology of astragaloside IV and medicine, which is applied in the field of application of astragaloside IV, can solve the problems of no astragaloside IV, accelerate skin regeneration and anti-scar, achieve the goal of reasonable and practical method, reduce scar generation, and promote skin regeneration Effect
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Embodiment 1
[0041] Embodiment 1: skin keratinocyte scratch test
[0042] (1) 24-well wound healing assay (wound healing assay kit, Cell Biolabs, INC) was preheated at room temperature for 10 minutes;
[0043] (2) Put the insert into a 24-well plate under sterile conditions;
[0044] (3) Skin keratinocytes (purchased from ATCC, Shanghai Branch) were mixed with 4×10 5 The density of each well was inoculated in a 24-well plate, 0.5ml per well (add the cell suspension from both sides of the insert, 250 μL on each side), and high Sugar culture medium (both purchased from Gibico, Brl, USA) was cultured for 13 hours;
[0045] (4) Carefully remove the inserts in the 24-well plate to avoid the irregularity of the trauma area caused by the migration of the inserts;
[0046] (5) Discard the previous culture solution, wash away the dead cells with PBS (phosphate buffer solution, pH=7.3), add DMEM high-sugar culture solution containing astragaloside IV (astragaloside IV was purchased from the Natio...
Embodiment 2
[0058] Embodiment 2: skin fibroblast scratch test
[0059] The cell scratch test refers to culturing cells on a petri dish or a flat plate, and drawing a line in the central area with a cell scraper after the cells are fused. The cells in this line are removed by mechanical force, and then the cells are continued to be cultured to observe The migration of cells to the cell-free scratch area was used to judge the migration ability of cells. It is another classic model for measuring cell migration in vitro.
[0060] (1) Draw a vertical diameter line in the center of the outer bottom wall of each well of a 24-well cell culture plate.
[0061] (2) Get well-grown skin fibroblasts (the fibroblast cell line was purchased from ATCC, Shanghai branch), and use 10 5 The density of cells / well was inoculated in a marked 24-well plate, and cultured with DMEM high-glucose medium containing 10% inactivated fetal bovine serum.
[0062] (3) After the cells have adhered to the wall and fused,...
Embodiment 3
[0077] Example 3: Test for the ability of skin fibroblasts to inhibit the secretion of TGF-β1
[0078] Before testing the effect of astragaloside IV on the secretion of TGF-β1 from fibroblasts, firstly, MTT assay was used to investigate whether astragaloside IV had a cell growth inhibitory effect on fibroblasts.
[0079] Experimental procedure: the fibroblasts were seeded on a 96-well plate at the same dose, 0.1 mL / well, and the seeding concentration of cells per well was 1×10 4 individual / mL. Incubate in the incubator for 24 hours. After the cells adhere to the wall, add astragaloside IV diluted with RPMI-1640 culture medium to different concentrations, 0, 12.5 μmol / L, 25 μmol / L, 50 μmol / L, 100 μmol / L, 200 μmol / L, a total of 6 concentration gradients, 3 parallel wells were set for each concentration (ie, 3 wells were repeated for each concentration), and incubated for another 24 hours. MTT (tetramethylazozole, Huamei Bioengineering Co., Ltd.) was dissolved in PBS (pH=7.3) ...
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