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Bifidobacterium longum NX-4 and application thereof in preparing drugs for treating and/or preventing allergic diseases

A technology for allergic diseases and Bifidobacterium longum, applied in the field of microbiology, can solve problems such as dependence on import of probiotic strains, lack of scientific research evidence for the function of probiotics, and affecting the promotion of probiotics and their products.

Pending Publication Date: 2021-01-01
FOSHAN LANGSEE BIOTECHNOLOGY CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The probiotic strains used by domestic production enterprises have long relied on imports, and foreign strains may not be suitable for the physiological conditions of the gastrointestinal tract of Chinese residents
In addition, the function of probiotics lacks strong scientific research evidence, which seriously affects the promotion of probiotics and their products

Method used

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  • Bifidobacterium longum NX-4 and application thereof in preparing drugs for treating and/or preventing allergic diseases
  • Bifidobacterium longum NX-4 and application thereof in preparing drugs for treating and/or preventing allergic diseases
  • Bifidobacterium longum NX-4 and application thereof in preparing drugs for treating and/or preventing allergic diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Isolation, Identification and Preservation of Bifidobacterium longum NX-4

[0024] (1) Separation: Inoculate infant feces into TPY solid medium, MRS solid medium, BDS solid medium, and BS solid medium respectively after gradient dilution, culture anaerobically at 37°C for 48 hours, and pick the Single colonies were streaked to obtain pure colonies. The pure colony on the plate was inoculated in BS liquid medium, cultured anaerobically at 37°C for 12-16h, added 20% glycerol, and stored in a -80°C refrigerator.

[0025] (2) Morphological identification of bacterial strains: Gram-stained strains were observed under a microscope, Gram-positive bacteria were purple, and Gram-negative bacteria were red.

[0026] (3) Molecular biological identification of the strain: Genomic DNA was extracted from the obtained strain, and the full-length 16S rDNA fragment was amplified by PCR using 16S rDNA universal primers 27F and 1492R, followed by sequencing to identify the spec...

Embodiment 2

[0032] Preparation of embodiment 2 Bifidobacterium longum NX-4 fermentation supernatant (extracellular secretion), bacterium suspension (thalline)

[0033] Bifidobacterium longum NX-4 was activated and cultured and inoculated in BS liquid medium. After culturing at 37°C for 15 hours, the concentration of fermentation bacteria was adjusted to 2×10 8 CFU / mL, centrifuged at 4°C, 8000r / min for 10min to obtain the culture supernatant and cell pellet, the supernatant was filtered through a 0.22μm filter membrane to obtain the fermentation supernatant (extracellular secretion); the cell pellet was filtered through PBS twice After the first wash, the cells were resuspended with PBS, and the cell concentration was adjusted to 2×10 8 CFU / mL to obtain bacterial suspension (cell); fermentation supernatant (extracellular secretion) and bacterial suspension (bacterial body) were heated at 121°C for 15 minutes to make heat-inactivated fermentation supernatant (extracellular secretion ), bac...

Embodiment 3

[0034] Example 3 Effect of Bifidobacterium longum NX-4 on Compound 48 / 80 stimulating zebrafish to secrete Tryptase

[0035] Sodium cromoglycate and N-benzoyl-DL-arginine p-nitrobenzamide hydrochloride (BAPNA) were purchased from Beijing Huawei Raycus Chemical Co., Ltd., and Compound 48 / 80 was purchased from Sigma, USA.

[0036] The healthy wild-type AB line zebrafish that had grown to 5dpf (days post fertilization) was selected and placed in a 96-well cell culture plate, 10 cells / well, the normal group was added with PBS, the model group was added with PBS, and the positive control group was added with sodium cromolyn solution ( 100 μg / mL), the bacterial suspension group was added with non-inactivated or inactivated bacterial suspension, the fermentation supernatant group was added with non-inactivated or inactivated fermentation supernatant, and the solvent zero adjustment group (without adding zebrafish) was added with PBS, 100 μL per well, after incubation at 28°C for 24 ho...

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Abstract

The invention discloses bifidobacterium longum NX-4 and application thereof in preparation of drugs for treating and / or preventing allergic diseases, and belongs to the technical field of microorganisms. The invention discloses a bifidobacterium longum NX-4 strain, and the preservation number is CGMCC No. 20112. Bifidobacterium longum NX4 inactivated and non-inactivated fermentation supernatant and bacterial suspension can obviously inhibit Compoud 48 / 80 from stimulating zebra fish to secrete Tryptase in vivo, and have the potential of being applied to relieving or preventing anaphylactic reaction in vivo. The bifidobacterium longum NX-4 disclosed by the invention has a huge potential application prospect in the aspect of relieving and / or preventing allergic reaction.

Description

technical field [0001] The present invention relates to the technical field of microbes, and more specifically relates to bifidobacterium longum NX-4 and its application in the preparation of medicines for treating and / or preventing allergic diseases. Background technique [0002] Worldwide, allergic diseases have become an important public health problem, affecting more than 25% of the world's population, both in developed and developing countries. In recent years, the incidence of allergic diseases in the world has been increasing year by year, seriously affecting people's quality of life. Mast cells are the main effector cells of allergic reactions. After being stimulated by allergens, they will degranulate and release histamine, tryptase (Tryptase) and other mediators to cause symptoms and signs of allergic reactions. At present, the treatment of allergic diseases mainly relies on drug therapy: such as mast cell stabilizers, antihistamines, etc. However, the treatment ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/745A61P37/08C12R1/01
CPCA61K35/745A61P37/08C12R2001/01C12N1/205
Inventor 张召郑康帝马燕陈涛
Owner FOSHAN LANGSEE BIOTECHNOLOGY CO LTD
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