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Housefly polypeptide capable of promoting tissue repair as well as preparation method and application thereof

A tissue repair and housefly technology, applied in the field of biomedicine, can solve problems such as unclear functional components, and achieve the effects of improving skin beauty, good stability, and promoting proliferation and migration.

Active Publication Date: 2020-06-12
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The applicant's previous research found that the housefly extract has a good effect on promoting the proliferation and migration of human immortalized epidermal cells (HaCat), but its functional components are still unclear

Method used

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  • Housefly polypeptide capable of promoting tissue repair as well as preparation method and application thereof
  • Housefly polypeptide capable of promoting tissue repair as well as preparation method and application thereof
  • Housefly polypeptide capable of promoting tissue repair as well as preparation method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0030] The preparation method of embodiment 1 housefly repair peptide, it comprises the following steps:

[0031] (1) Preparation of Musca domestica extract: extract the whole housefly that is pulverized or not pulverized with 70% ethanol, and concentrate the extract;

[0032] (2) Extraction: disperse the housefly extract concentrate with 70% methanol solution, add about 1 / 10 volume of chloroform, fully shake, after standing, discard the chloroform layer, and partly centrifuge, concentrate and dry the methanol solution to obtain home Methanol fraction of fly extract;

[0033] (3) Strong oxygen ion exchange resin (SCX) separation: the methanol part of the housefly extract obtained in step (2) was redissolved with 10mM ammonium formate solution (containing 25% acetonitrile, pH 2.7~3), and the SCX small Column, to ensure that the sample is fully adsorbed, wash the SCX small column with 10mM ammonium formate solution (containing 25% acetonitrile, pH 2.7-3) for 7 to 8 times, then ...

Embodiment 2

[0035] Example 2 The sequence analysis of the housefly repair peptide, which comprises the following steps:

[0036] Take the housefly repair peptide prepared in Example 1, and use nano-LC-ESI-MS / MS to analyze the amino acid sequence of the polypeptide. The mass spectrometry conditions are: ESI source, scanning mode: positive ion mode, and the first-level full scan range of the mass spectrometer is m / z 300~2000, the separation width is 3Da; the tandem mass spectrometry adopts the data-dependent secondary mass spectrometry scanning mode of the primary mass spectrometry, and sequentially selects the five ions with the highest ion intensity in the primary mass spectrometry for collision-induced dissociation (CID) secondary Tandem mass spectrometry. The amino acid sequence of the housefly repair peptide determined according to the obtained MS / MS spectrum is: Leu-Ala-Asp-Glu-Glu-Ser-Arg-Glu-Arg-Ala-Thr-Leu-Leu-Gly, such as figure 1 shown.

Embodiment 3

[0037] Example 3 Synthesis of Housefly Repair Peptide

[0038] According to the amino acid sequence obtained in Example 2, the dipeptidyl peptidase-4 inhibitory peptide Leu-Ala-Asp-Glu-Glu-Ser-Arg-Glu-Arg-Ala-Thr-Leu- Leu-Gly, the purity of the synthesized polypeptide is 98% by HPLC analysis, and the molecular weight determined by mass spectrometry is 1558.79Da, which is consistent with the retention time and molecular weight of the purified polypeptide, and the fragments of the secondary mass spectrum are consistent with the purified polypeptide fragments.

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Abstract

The invention discloses a housefly polypeptide capable of promoting tissue repair as well as a preparation method and application of the housefly polypeptide. The amino acid sequence of the polypeptide is Leu-Ala-Asp-Glu-Glu-Ser-Arg-Glu-Arg-Ala-Thr-Leu-Leu-Gly. According to the invention, a modern biochemical technical means is adopted; the polypeptide for promoting tissue repair in houseflies istracked, screened and evaluated; the preparation method comprises the steps of ethanol extraction, extraction, ion exchange resin, reversed-phase high performance liquid chromatography and other purification methods. Experimental results show that the polypeptide can promote proliferation and migration of human immortalized epidermal cells, is low in toxic and side effects, can be used for preparing medicines or skin care products for whitening, removing wrinkles, removing color spots and the like, can obviously improve the skin beauty, and has a good application prospect.

Description

technical field [0001] The invention relates to an active polypeptide, in particular to an active polypeptide isolated from housefly extract and capable of promoting tissue repair and a preparation method thereof, belonging to the technical field of biomedicine. Background technique [0002] Skin wound healing refers to the healing process after the skin tissue is severed or damaged due to external force, including the repair and regeneration of various tissues, such as cell proliferation, differentiation, migration and other processes. Regeneration of the epidermis and other tissues usually occurs within 24 hours after trauma. The basal cells at the wound edge begin to proliferate and migrate to the center of the wound under the clot to form a single layer of epithelium covering the surface of the granulation tissue. Therefore, promoting tissue cell proliferation, migration, and repair is very important for wound healing. [0003] Peptide drugs have unique advantages in pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K1/14C07K1/18C07K1/20A61K38/10A61K8/64A61Q19/00A61P17/00
CPCC07K7/08A61K8/64A61Q19/00A61P17/00A61K38/00
Inventor 刘睿段金廒
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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