Application of dihydromyricetin in preparation of skin-care products or drugs for preventing and treating optical skin injuries
A technology of dihydromyricetin and photodamage is applied in the field of dihydromyricetin to prepare skin care products or medicines for preventing and treating skin photodamage, which can solve problems such as no skin damage effect, reduce skin itching, inhibit ROS generation, and improve the The effect of preventive action
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Embodiment 1D
[0031] Effect of embodiment 1DHM on the cell viability of HaCaT cells irradiated by UVA
[0032] 1. Solution preparation
[0033] (1) Preparation of DHM: Dissolve 2.1 mg of dihydromyricetin powder in 65.6 μL DMSO (the final volume of DMSO should not exceed 0.1%), prepare a 100 mmol / L stock solution, and store it in a 200 μL PCR tube at 10 μL / tube. tubes, protected from light and stored in a -20°C refrigerator. Take it out before use, and dilute to the corresponding concentration with DMEM according to the needs of different concentrations in the experiment.
[0034] (2) HaCaT cell culture medium (complete medium): DMEM medium (high glucose serum-free medium) + 10% FBS (South American fetal bovine serum), the ratio is 9:1; double antibodies (penicillin and streptomycin) to a final concentration of 100mg / L; seal with parafilm and store at 4°C for later use.
[0035] 2. Experimental method
[0036] 2.1 Effects of different doses of UVA on the survival of HaCaT cells
[003...
Embodiment 2
[0055] The influence of embodiment 2DHM on the ROS level in the HaCaT cell of UVA irradiation
[0056] 1. Experimental method
[0057] (1) Preparation of DCFH-DA: Dilute DCFH-DA with serum-free culture medium to make the final concentration 10 μmol / L;
[0058] (2) HaCaT cells in the logarithmic phase were divided into 1x10 5 The cell density of cells / ml is inoculated in 35mm cell culture dish, 2ml per dish;
[0059] (3) Discard the old culture medium after 24 hours of culture, and add medium containing different concentrations of DHM and continue to incubate for 6 hours;
[0060] (4) After 6 hours, discard the medium and replace it with PBS, and then irradiate with ultraviolet light for 60 minutes;
[0061] (5) After the end of ultraviolet irradiation, replace PBS with serum-containing medium and continue to incubate for 30 minutes;
[0062] (6) Discard the supernatant after 30 minutes, wash the cells with PBS 3 times, add diluted DCFH-DA, 2ml per dish and continue to in...
Embodiment 3
[0068] Example 3 Effect of DHM on the expression level of inflammatory factor genes in HaCaT cells irradiated by UVA.
[0069] 1. Experimental method
[0070] (1) Collect the cells grown in the logarithmic phase according to 1x10 5 The cell density of cells / ml was inoculated in a 35mm cell culture dish, 2ml per dish, placed at 37°C, 5% CO 2 Culture in the incubator;
[0071] (2) After culturing for 24 hours, the old culture medium was discarded after the cells grew to a monolayer, and different concentrations of dihydromyricetin were added to the experimental group (10 μmol / L, 5 μmol / L, 2.5 μmol / L, 1.25 μmol / L L, 0 μmol / L), the cell control group was added with serum-containing medium and incubated at 37°C for 6 h. After 6 h, the medium was discarded and PBS was added to cover the cells, and then ultraviolet irradiation was performed for 60 min. After the ultraviolet irradiation, the PBS was discarded and removed. The serum-containing medium was added again, and then the ...
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