Composition with anti-aging activity as well as preparation and application thereof
A composition, anti-aging technology, applied in the field of medicine, can solve the problems of burden, pension and heavy medical system
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Embodiment 1
[0044] Example 1: Extraction and Identification of Primary Human Skin Fibroblasts
[0045] Primary human dermal fibroblasts (Human dermal fibroblast, HDF) were extracted from the foreskin tissue of adult healthy men. The tissue was sterilized with 75% ethanol for 1-2min. Then change to a new PBS buffer solution containing penicillin-streptomycin for washing. After cleaning, use tweezers and scissors to remove the subcutaneous connective tissue; cut the skin tissue into a size of 1cm×1cm, put it in a petri dish containing buffered saline solution containing dispase II, and incubate overnight at 4°C; take the tissue out of the refrigerator, Put it into an ultra-clean bench, and use tweezers and scissors to separate the epidermis and dermis. Subsequently, the separated dermal tissue was cut into small pieces of about 3mm×3mm, carefully placed into a culture bottle, the dermis was inoculated into the culture bottle, and then 2 mL of complete medium was added and placed in an inc...
Embodiment 2
[0048] Human skin fibroblasts were subcultured in vitro to construct a spontaneous natural cellular aging model. The specific operation is as follows: spread the extracted human fibroblasts in a 6-well plate (50×10 4 cells / well) were maintained in DMEM medium containing 10% FBS. When the cell density reached 80-90%, the cells were digested and collected with trypsin, and 1 / 4 of the cells were taken out and continued to be cultured in a 6-well plate. After the cells were passaged 40 times in the above manner, the number and proportion of senescent cells were determined by SAβ-Gal staining (pH 6.0). It should be noted that the control group (natural aging) was subcultured with normal medium, and the medium of the other groups contained one of 8-gingerol (μM), quercetin (10 μM) and vitamin E (VE). one or more species.
[0049] The specific steps of SA-β-Gal staining are as follows: After washing the cells twice with PBS solution (about 30 seconds each time), add fixative solut...
Embodiment 3
[0054] In this example, the ginger extract was prepared through the following scheme. The specific steps are as follows. After the ginger was washed several times with water, high-quality ginger was selected and dried in the shade. Mince the ginger into 0.5×0.5cm tissue pieces, add 30 times the amount of 70% ethanol solution, heat reflux extraction 4 times, each extraction for 1.5 hours, combine the extracts, filter under reduced pressure to obtain the filtrate, and concentrate by rotary evaporation Dried to obtain ginger extract.
[0055] The seabuckthorn leaf extract is prepared according to the following scheme. The specific steps are as follows. After washing the seabuckthorn leaves with water for several times, select complete and high-quality seabuckthorn leaves and dry them in the shade. After mechanical pulverization, add 40 times the amount of deionized water, heat reflux extraction 4 times, each time for 1.5 hours, combine the extracts, filter under reduced pressure ...
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