Method for analyzing regulation effect of deer skin protein peptide on macrophage immune cells
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A technology of macrophage regulation, applied in the field of biotechnology research and development analysis, can solve problems that have not yet been developed, and achieve the effect of fast detection speed, high sensitivity and low cost
Pending Publication Date: 2020-03-10
吉林省东北亚生物科技有限公司
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Problems solved by technology
Among them, the advantages of easy absorption and high activity of peptide components have been widely concerned, but the research on the immune regulation of buckskin peptide on macrophage RAW264.7 has not yet been carried out
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Embodiment 1
[0062] Take 40g of raw deerskin, shred it into small pieces about 0.5cm in length, take distilled water 10 times the quality of the raw deerskin, add it to the shredded raw deerskin, and under the condition of 4°C, Flash extraction was carried out 5 times, each time lasting 1 minute, at a speed of 3600 r / min, centrifuged for 10 minutes, and the supernatant was taken. The yield was 0.148%.
Embodiment 2
[0064] Take 40g of raw buckskin, shred it into small pieces about 0.5cm in length, take distilled water 20 times the quality of the raw buckskin, add it to the shredded raw buckskin, and under the condition of 4°C, Flash extraction was performed 5 times, each time lasting 1 minute, at a speed of 3600 r / min, centrifuged for 20 minutes, and the supernatant was taken. The yield was 0.286%.
Embodiment 3
[0066] Take 40g of raw deerskin, shred it into small pieces about 0.5cm in length, take distilled water 25 times the quality of the raw deerskin, add it to the shredded raw deerskin, and under the condition of 4°C, Flash extraction was carried out 8 times, each time lasting for 1 hour, at a speed of 3600 r / min, centrifuged for 30 minutes, and the supernatant was taken. The yield was 0.447%.
[0067] Example of research on the immune regulation of buckskin peptide on macrophage RAW264.7
[0068] Embodiment one, buckskin peptide protein content determination
[0069]Taking the bovine serum albumin mass concentration (X) as the abscissa and the absorbance OD value (Y) as the ordinate, draw a standard curve as figure 1 As shown, the protein content of the buckskin peptide solution calculated by the formula is 0.3660mg / mL respectively.
[0070] Example 2. Proliferation effect of different mass concentrations of DSPs on RAW264.7 macrophages
[0071] Both LPS and DSPs can promote...
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Abstract
The invention discloses a method for analyzing a regulation effect of deer skin protein peptide on macrophage immune cells, and belongs to the technical field of biotechnology research, development and analysis. By compared with proliferation activities of samples with different concentrations, a result shows that the proliferation index of RAW264.7 cells is increased along with the increase of adeer skin protein peptide concentration, and the concentration is 200[mu]g/mL and is closest to an LPS positive control group; in order to verify the influence of deer skin peptide on immunoregulation, the influence of deer skin protein peptide with different mass concentrations on NO secretion amount, LZM activity and release of cytokines such as IL-6 and TNF-[alpha] is researched, and the resultshows that the NO secretion amount is increased along with the increase of the sample concentration; after the deer skin protein peptide acts for 24 hours, the secretion function of RAW264.7 cytokines is obviously increased, and when the concentration is 50 [mu]g/mL, the levels of all the cytokines are close to those of an LPS group, which indicates that the deer skin protein peptide has the effect of enhancing cytokine secretion; in conclusion, the deer skin protein peptide has a potential effect of enhancing the immunocompetence, and can provide reference for subsequent research.
Description
technical field [0001] The invention belongs to the technical field of research and development and analysis of biotechnology, and in particular relates to an analysis method for the regulating effect of buckskin peptide on macrophage immune cells. Background technique [0002] Deerskin is the skin of Cervus nippon Temminck or red deer Cervus elaphusLinnaeus. It has the effects of invigorating qi, astringent essence, and astringing sores. Cure mainly leucorrhea, metrorrhagia, kidney deficiency and spermatorrhea, leaking sores. Deerskin is a traditional precious Chinese medicinal material. The deerskin collagen, which is processed and refined from it as a raw material, has strong pharmacological activity and is a top-grade medicine. In this paper, the research status of buckskin collagen, including its base source, chemical composition, and pharmacological effects, is comprehensively reviewed. At the same time, on the basis of the current research, it is suggested that in-...
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