Application of chick embryo bioactive peptide in preparing product for protecting cell activity
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A chicken embryo active peptide and cell technology, which is applied in the application field of the product to achieve the effects of improving fine lines, active protection and removing red blood.
Active Publication Date: 2016-03-09
耿威
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Problems solved by technology
However, there are still few studies on peptides in chicken embryos at present, and it should be further developed and utilized in the research of cosmetics and health products for fire-fighting and anti-allergy.
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Embodiment 1
[0054] (1) Hatching chicken embryos: Take SPF grade fertilized eggs and put them into the incubator for incubation. The incubation temperature is controlled at 37.5°C, the relative humidity is 60%, and the eggs are automatically turned every 2 hours. After hatching to 16 days, take out the fertilized eggs , put it in a freezer at -20°C, stop the incubation and save it for later use.
[0055] (2) Homogenization and cell crushing: surface disinfection of the hatched eggs, removal of egg shells, taking out the chicken embryos, homogenizing the chicken embryos with a tissue homogenizer at 4°C, pouring them into a sterilized sealed box, Put it in a freezer at -30°C for 72 hours, and after thawing, perform cell ultrasonic disruption in an environment of 4°C, add an equal amount of water for injection, and place it in a freezer at 4°C for use.
[0056] (3) Enzymatic hydrolysis and centrifugation: Take the refrigerated solution after cell crushing, heat it in a water bath to 50°C, adj...
Embodiment 2
[0067] Cell viability was determined by MTT colorimetry. The detection principle is that succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-purple crystalline formazan (Formazan) and deposit in the cells, while dead cells do not have this function. Dimethyl sulfoxide (DMSO) can dissolve formazan in cells, and its light absorption value is measured at a wavelength of 550nm by an enzyme-linked immunosorbent detector, which can indirectly reflect the number of living cells. Within a certain range of cell numbers, the amount of MTT crystal formation is proportional to the number of living cells.
[0068] MTT assay for cell viability:
[0069] Human fibroblasts (HDF cells) (the sixth generation) were treated with 1×10 4 Individuals / well were inoculated in 96-well plates;
[0070] The experiment was divided into experimental group and blank control group;
[0071] After 24 hours, the experimental group was given different...
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Abstract
The invention relates to the technical field of biology, and in particular relates to chick embryo bioactive peptide, as well as a preparation method and application thereof. The chick embryo bioactive peptide provided by the invention comprises the chick embryo bioactive peptide and free amino acid, wherein the molecular weight of the chick embryo bioactive peptide is less than 10kDa. The chick embryo bioactive peptide provided by the invention can protect the activity of human fibroblast, and in comparison with a matched group without adding the chick embryo bioactive peptide provided by the invention, the activity of the human fibroblast of an experimental group adding the chick embryo bioactive peptide is protected. Compared with a blank group without ultraviolet radiation, the activity of the human fibroblast of the experimental group is basically equal to, even slight higher than, that of the blank group. The preparation method of the chick embryo bioactive peptide, provided by the invention is simple to operate, the cytomembrane is crushed to the greatest extent through homogenate, freeze thawing and ultrasonication, so that bioactive peptide in a cell is well dissolved out, and then through the biotechnologies of centrifugation, ultrafiltration and the like, the extracting efficiency is improved.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to the application of chicken embryo active peptides in the preparation of products for protecting cell viability. Background technique [0002] As the largest organ of the human body, the skin covers the whole body, is the protective layer of the body, and plays an extremely important role in the human body. The facial skin can not only resist the invasion of external bacteria into the body, but also a measure of a person's health and beauty. Common facial skin problems include: fine lines, redness, scars, dull complexion, etc. The main causes of these skin problems are: the facial skin has been exposed to the air for many years, the irradiation of ultraviolet rays, and the natural harmful substances of dirt, dust and bacteria in the air stimulate the skin surface. In addition to self-secreted oil, sweat, dead cells, etc., these factors will affect the normal function of the skin, ca...
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