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Anti-inflammatory sericin peptide and its application

A sericin and protein peptide technology, applied in the field of biomedicine, can solve the problems of reporting the function of sericin inflammatory factors and achieve the effect of inhibiting inflammatory factors

Active Publication Date: 2021-10-22
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, neither macromolecular sericin nor hydrolyzed sericin has reported the function of sericin against LPS-induced inflammatory factors

Method used

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  • Anti-inflammatory sericin peptide and its application
  • Anti-inflammatory sericin peptide and its application
  • Anti-inflammatory sericin peptide and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1, the extraction of sericin

[0017] A method for extracting sericin, comprising the steps of:

[0018] (1) clean the silkworm cocoons with water;

[0019] (2) Cut into cocoon pieces with a size of 2*2cm, and heat them in an oven at 120°C for 2.5 hours;

[0020] (3) Add Milli-Q water according to the bath ratio of 1:30, and treat at 120°C for 1h and 2h respectively;

[0021] (4) filter cocoon sheet residue, collect protein extract;

[0022] (5) Freeze-dry the protein extract to make a powder, and store it in a -20°C refrigerator.

[0023] The sericin protein obtained by high temperature treatment for 1 h is coded as HS1, and the sericin protein obtained by high temperature treatment for 2 h is coded as HS2.

Embodiment 2

[0024] Embodiment 2, sericin protein anti-inflammatory effect

[0025] 1. Cell culture method:

[0026] Cells were cultured at 37°C, 5% CO 2 In a constant temperature incubator, NIH3T3 and HaCaT cell lines were cultured with DMEM medium containing 10% fetal bovine serum, and RAW264.7 cell lines were cultured with 1640s medium containing 10% fetal bovine serum. All cell lines were changed medium every day and subcultured every other day.

[0027] Three kinds of cells were treated with HS1 and HS2 prepared in Example 1, and three commercially available sericin proteins (numbered S1, S2, S3) were used as controls, and then inflammatory factors were detected by real-time quantitative PCR and ELISA.

[0028] 2. ELISA detection

[0029] (1) After the cells are processed, absorb the cell culture medium, centrifuge at 13,500 rpm for 10 minutes, and take the supernatant; use it now or store it at -20°C for later use.

[0030] (2) Take a microwell plate, add 100 μL of cell culture s...

Embodiment 3

[0073] Embodiment 3, sericin protein hydrolysis and anti-inflammatory effect

[0074] In order to understand the function of the polypeptide after hydrolysis of sericin protein, LC-MS / MS detection is performed after hydrolysis of sericin protein. The detection method of LC-MS / MS is as follows:

[0075] Using the FASP enzymatic hydrolysis method, the process is carried out according to enzymatic hydrolysis-desalting-mass spectrometry-protein identification, and the specific steps are as follows:

[0076] Enzymolysis:

[0077] (1) Take the protein solution, add DTT with a final concentration of 100mM, heat at 95-100°C for 5min, and cool to room temperature.

[0078] (2) Add 200 μL of UA buffer and mix well. Transfer to a 30kD ultrafiltration centrifuge tube, centrifuge at 14000g for 15min, and discard the filtrate.

[0079] (3) Repeat step (2).

[0080] (4) Add 100 μL of 100 mM IAA buffer solution, shake at 600 rpm for 1 min.

[0081] (5) Place at room temperature, protect ...

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Abstract

The invention discloses an anti-inflammatory sericin peptide and its application. The amino acid sequence of the sericin peptide is shown in SEQ ID NO.1. The modified protein peptide can be obtained by pretreating cocoon slices and then extracting them with water. Sericin is obtained by hydrolysis; it can also be artificially synthesized; the obtained sericin has the effect of inhibiting inflammatory factors, so it can be used to prepare anti-inflammatory preparations.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the anti-inflammatory sericin peptide and the application of the anti-inflammatory sericin peptide. Background technique [0002] Sericin was originally a waste material from the silk reeling industry, but its application has gradually attracted attention in recent years. First of all, the safety of sericin has been approved by a series of studies, and it has the premise of application in tissue engineering materials and daily skin care. Secondly, its antioxidant, whitening and moisturizing effects are widely used. Sericin is a functional ingredient in the catalog of cosmetic raw materials, but the molecular weight of sericin-1 protein is about 400k. As a macromolecular protein, sericin generally needs to be hydrolyzed during application. However, neither macromolecule sericin nor hydrolyzed sericin has reported the function of sericin against LPS-induced inflammatory factors. And ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/435A61K38/17A61P29/00
CPCA61K38/00A61P29/00C07K14/43586
Inventor 夏庆友孙悦婷赵萍
Owner SOUTHWEST UNIV
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