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Use of antimicrobial peptides for maintaining abundance of intestinal Akkermansia bacteria

A technology of antimicrobial peptides and intestinal flora, applied in the field of antimicrobial peptides, to slow down the progress, inhibit the growth of Spirulina mucus, and promote the growth of Akkermansia

Active Publication Date: 2022-05-27
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, targeting the role of RNASE4 in gut microbiota maintenance is still a blank field

Method used

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  • Use of antimicrobial peptides for maintaining abundance of intestinal Akkermansia bacteria
  • Use of antimicrobial peptides for maintaining abundance of intestinal Akkermansia bacteria
  • Use of antimicrobial peptides for maintaining abundance of intestinal Akkermansia bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1. RNASE4 can change the composition of intestinal flora

[0045] 1.1 Experimental method

[0046] In this example, 16S rDNA amplicon high-throughput sequencing was used to compare the differences in bacterial composition in the fecal DNA samples of 6 wild-type mice and 6 Rnase4 knockout mice matched in gender, age, and body weight. .

[0047] Among them, the Rnase4 knockout mouse was constructed by TALEN technology. The specific construction process is as follows: The artificially modified FokI is used to form a dimer, and under the guidance of the TALEN arm with a DNA recognition domain, the activity of endonuclease is exerted. , specifically cut off the target gene DNA, use TALEN technology to insert the "TG" base in the Rnase4 gene on chromosome 14 of the mouse genome, introduce point mutations to cause frameshift, construct the Rnase4 gene knockout mouse, after sequencing, the Rnase4 gene The genome of knockout mice successfully inserted "TG" bases, causi...

Embodiment 2

[0054] Example 2. RNASE4 has a protective effect during the occurrence of enteritis

[0055] 1.1 Experimental method

[0056] Enteritis model was induced by dextran sulfate sodium (DSS). First, the DSS drug powder was weighed and dissolved in sterilized water to a final concentration of 2.5% (ie, 2.5 g / 100 ml). Then, 8-week-old wild-type and Rnase4 knockout mice were taken, and during the modeling process, the drinking water was changed to 2.5% DSS solution. Changes in body weight, fecal dryness, and blood in the stool were recorded on the 1st to 10th day of induction of enteritis.

[0057] Weight Score: 0, no weight loss; 1, 1-5% decrease; 2, 6-10% decrease; 3, 11-20% decrease; 4, more than 20% decrease; stool score: 0, solid stool; 1, Solid stool, easily deformed; 2, unformed stool; 3, liquid stool; blood in stool score: 0, negative for occult blood test; 1, positive for occult blood test; 2, visible blood in stool; 3, severe blood in stool.

[0058] The disease activity...

Embodiment 3

[0061] Example 3. Intestinal flora plays a role in RNASE4-related enteritis

[0062] 1.1 Experimental method

[0063] In this example, the intestinal flora of Rnase4 knockout mice was transplanted into mice treated with mixed antibiotics by fecal bacteria transplantation. On this basis, the DSS-induced enteritis model was used to analyze the intestinal flora Role in RNASE4-associated enteritis.

[0064] The specific process is as follows: firstly prepare a mixed antibiotic solution (containing 1g / L ampicillin, 1g / L neomycin, 1g / L metronidazole, 0.5g / L vancomycin), and then drink the drinking water of 12 wild-type mice. The water was replaced with the above mixed antibiotic solution for a total of 4 weeks. After 4 weeks of treatment, the feces of each mouse were collected and dissolved in sterile PBS (the dissolving ratio was 100 mg feces / 1 mL PBS), followed by 4 serial serial dilutions at a ratio of 1:10, and each 50 μL was applied to LB plates , perform anaerobic and aerob...

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Abstract

The invention discloses the application of an antibacterial peptide in the preparation of a medicine for regulating intestinal flora, and the antibacterial peptide is RNASE4. The antimicrobial peptide significantly increased the proportion of Akkermansia bacteria in the intestinal tract, while the proportion of Spirulina mucilage significantly decreased in the intestinal tract. The invention also discloses the application of the antibacterial peptide in the preparation of medicine for treating inflammatory bowel disease. The antibacterial peptide is RNASE4; the inflammatory bowel disease includes ulcerative colitis and Crohn's disease.

Description

technical field [0001] The present invention relates to an antibacterial peptide for improving intestinal flora, which contains antibacterial peptide RNASE4 as an active ingredient, and the antibacterial peptide RNASE4 can improve the abundance of intestinal flora, in particular to Akkermansia muciniphila in the intestine abundance. Background technique [0002] Intestinal flora is the most complex and largest micro-ecosystem in the human body. The latest research results show that the human gut contains thousands of about 10 14 Although a microorganism has the same number of human cells, the number of genes encoded by it is hundreds of times the number of human genes. In normal organisms, symbiotic bacteria (referring to the intestinal flora that co-exist with the organism) are mainly distributed in the intestinal lumen and mucosal surface, and evolve together with the host to maintain the integrity of the mucosal barrier system and antagonize pathogens and other harmful ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/46A61P1/04A61P1/00A61P29/00
CPCA61K38/465A61P1/04A61P1/00A61P29/00
Inventor 许正平盛静浩孙钧陈木雄
Owner ZHEJIANG UNIV
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