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Polypeptide with antioxidant activity and liver protection activity, polypeptide coding gene as well as preparation method and application of polypeptide

An anti-oxidation and liver-protecting technology, which is applied in the fields of polypeptides and genes encoding the polypeptides and its preparation, can solve the problems of obvious side effects of drugs and poor treatment effects, and achieve the effect of reducing damage and wide application prospects

Active Publication Date: 2018-12-28
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Alcoholic fatty liver is the most significant pathological change in the early stage of alcoholic liver disease. Now it is usually treated with nutritional support, blood lipid lowering, unsaturated fatty acid and phospholipid drugs, but the treatment effect is not good and some drugs have obvious side effects, so there is obvious treatment for it Effective medicines and health care products to control the effect have become the focus of research at home and abroad

Method used

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  • Polypeptide with antioxidant activity and liver protection activity, polypeptide coding gene as well as preparation method and application of polypeptide
  • Polypeptide with antioxidant activity and liver protection activity, polypeptide coding gene as well as preparation method and application of polypeptide
  • Polypeptide with antioxidant activity and liver protection activity, polypeptide coding gene as well as preparation method and application of polypeptide

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: Polypeptide solid-phase synthesis

[0024] 1. Resin selection

[0025] (1) Using the standard Fomc scheme, select 0.0125mmol, 2-chlorotrityl chloride resin resin, according to the sequence characteristics of the amino acid sequence YFLP, add 0.3mol of the first Fmoc protected amino acid, and add DCC and 5% (mass fraction) DMAP to the reaction Shake the reaction, and rinse the resin with NMP to remove excess protected amino acids.

[0026] (2) select 0.0125mmol for use, Wang resin, method is the same as above;

[0027] 2. Synthesis process

[0028] Using the standard Fomc scheme, select 2-chlorotril chloride resin resin with a high coupling rate, and extend the peptide chain from the C-terminal to the N-terminal one by one according to the sequence characteristics of the amino acid sequence YFLP. The amount of each amino acid is 0.1mol, and 0.3mol of Fmoc is added for protection For amino acids, HOBT was added to activate and protect the carboxyl groups ...

Embodiment 2

[0031] Embodiment 2: ORAC value determination of synthetic polypeptide

[0032] Oxygen radical absorbance capacity (ORAC) is a method for evaluating antioxidant capacity based on the hydrogen atom transfer mechanism, which is considered to be close to the antioxidant function of organisms. The ORAC reaction was carried out in a 75mM phosphate buffer solution environment at 37°C, and the synthetic polypeptide (YFLP), reduced glutathione (GSH), fluorescein and AAPH free radicals were all dissolved in a 75mM phosphate buffer solution, and the buffer solution The pH value is 7.4, and its final concentration is 20mg / L, 20mg / L, 78nM and 9.95mM respectively. Trolox was used as a standard antioxidant with a final concentration gradient of 0.5-4.2 μM, and GSH was used as a positive control. Mix the synthetic peptide and GSH with fluorescein respectively, and incubate at 37°C for 20 minutes, then add the azo compound AAPH, detect the fluorescence intensity at the excitation wavelength ...

Embodiment 3

[0035] Example 3: Liver-protecting application of synthetic polypeptide acting on ethanol-induced liver cell injury model

[0036] LO2 cells in the logarithmic growth phase were inoculated into 96-well cell plates, with 5000 cells per well, and placed in CO 2 After culturing in the incubator for 24 hours, the experimental group was added with the culture solution with the concentration of synthetic peptides being 0.25, 0.5, 0.75, 1, 1.25 mmol / L respectively, and the blank control group was added with the same amount of culture solution. The incubator continued to cultivate for 24h. Add 20 μL of MTT solution (5 mg / mL) to each well, incubate in an incubator for 4 hours, discard the supernatant, add 150 μL DMSO to each well, and measure the absorbance value OD of each well at a wavelength of 490 nm. Calculate the survival rate of cells in each group after treatment with different ethanol concentrations:

[0037]

[0038] Calculated as Figure 4 shown. Figure 4 It is a compa...

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Abstract

The invention discloses a polypeptide with antioxidant activity and liver protection activity, a polypeptide coding gene as well as a preparation method and an application of the polypeptide. The amino acid sequence of the polypeptide is Tyr-Phe-Leu-Pro. According to ORAC (oxygen radical absorbance capacity) experiment evaluation for antioxidation capacity of the synthetic peptide, ethanol can induce a certain degree of damage to liver cells (LO2), an LO2 cell damage model is used for evaluating the liver protection effect of the synthetic peptide, and results show that the synthetic peptide can reduce the damage effect of ethanol on the LO2 cells and has wide application prospect in preventing and treating ALD (alcoholic liver disease).

Description

technical field [0001] The invention relates to the fields of health products and medicines, in particular to a polypeptide having both anti-oxidation and liver-protecting activities, a gene encoding the polypeptide, a preparation method and application thereof. Background technique [0002] With the improvement of living standards and changes in dietary structure, the incidence of alcoholic liver disease (alcoholic liver disease, ALD) increased year by year. Alcoholic fatty liver (alcoholic fatty liver, AFL) is a kind of alcoholic liver disease, which is caused by alcoholic liver damage caused by long-term drinking of alcohol. disorder, which may develop into alcoholic hepatitis, liver fibrosis, and even cirrhosis. Alcoholic fatty liver is the most significant pathological change in the early stage of alcoholic liver disease. It is often treated with nutritional support, blood lipid lowering, unsaturated fatty acids and phospholipid drugs, but the treatment effect is not g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K5/107C12N15/11A61K38/07A61P1/16A61P39/06A23L33/18
CPCA23V2002/00A61K38/00A23L33/18A61P1/16A61P39/06C07K5/1016A23V2200/30A23V2250/55
Inventor 任娇艳沙婉倩尚帅明
Owner SOUTH CHINA UNIV OF TECH
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