Application of eupatilin in preparation of product for resisting skin light injury
A technology of eupholzanthin and photodamage, which is applied in the field of biomedicine to improve skin wrinkle formation and cell oxidative damage
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Embodiment 1
[0036] Induction of photoaging damage in HaCaT cells
[0037] HaCaT cells were seeded in 96-well plates, and when they grew to a density of 50%, 0, 3, 5, 7, 10, 20, 30 mJ / cm 2 After continuing to culture for 24 hours, use Beyontian CCK8 kit to detect the cell survival rate under different ultraviolet radiation doses.
[0038] figure 1 It was shown that with the increase of UVB radiation dose, the survival rate of cells decreased gradually. The dose of UVB radiation should be above 80% of the cell survival rate, so choose the dose of UVB as 5mJ / cm 2 for subsequent experiments.
Embodiment 2
[0040] Toxicity of eupholzanthin to HaCaT cells
[0041] HaCaT cells were seeded in a 96-well plate, and when they grew to a density of 50%, they were treated with 0, 12.5, 25, 50, and 100 μM concentrations of eupholzanthin (using DMSO as a solvent), and after continuing to culture for 24 hours, they were treated with Phi Yuntian CCK8 kit is used to detect the cell survival rate under different doses.
[0042] according to figure 2 As a result, based on the cell viability of 90% as the standard, the maximum non-toxic concentration within the experimental range was 25 μM. At the same time, combined with the literature reports on the toxicity of eupholzanthin, subsequent experiments were carried out at a concentration of eupholzanthin of 30 μM.
Embodiment 3
[0044] Improvement effect on UVB-induced oxidative damage of HaCaT cells
[0045] Reactive oxygen species (ROS), lipid peroxidation product malondialdehyde (MDA), antioxidant enzyme superoxide dismutase (SOD) were used as indicators for research.
[0046] 1. Reactive oxygen species (ROS)
[0047]HaCaT cells were seeded in a 96-well plate, and when they grew to a density of 50%, they were treated with UVB, UVB+ with a concentration of 100 μM eupholzanthin, and UVB+0.1% DMSO, while the cells in the blank control group were not treated with any treatment (marked as UVB0); after each group continued to culture for 24 hours, the Beyontian ROS detection kit was used to detect the improvement effect of eupholzanthin on UVB-induced ROS accumulation.
[0048] The intracellular ROS level of HaCaT cells was characterized by the fluorescence intensity of DCFH-DA probe. image 3 It was shown that after HaCaT cells were irradiated with UVB, the level of intracellular ROS was significantly...
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