A ciliated Lactobacillus rhamnosus with mast cell activity regulation and its use

A mast cell, rhamnose technology, applied in the field of microorganisms

Active Publication Date: 2020-09-08
江苏新申奥生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no reports of strains that can regulate mast cell degranulation and histamine release at home and abroad.
On the other hand, Lactobacillus rhamnosus exhibits large strain variability in immunomodulatory effects

Method used

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  • A ciliated Lactobacillus rhamnosus with mast cell activity regulation and its use
  • A ciliated Lactobacillus rhamnosus with mast cell activity regulation and its use
  • A ciliated Lactobacillus rhamnosus with mast cell activity regulation and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 strain enrichment and cultivation

[0044] Traditional fermented dairy products are made by herdsmen in Tibet, Xinjiang and Inner Mongolia. Weigh 25g of the sample and add 225 mL of normal saline, homogenate, and dilute 10 times with normal saline; take 5 mL of the appropriate dilution factor and add 1 mg of SpaA antibody Coupled magnetic beads were incubated at 37°C for 75 min, placed on a magnetic separation rack, magnetically separated for 2 min, and the supernatant was discarded; washed twice with 500 μL PBS, resuspended by adding 200 μL PBS, and the obtained magnetic bead suspension Directly coat the MRS agar plate, incubate anaerobically at 37°C for 24 hours, and select a plate with evenly distributed colonies for use.

[0045] The test results showed that among the 17 traditional fermented dairy products, only one sample collected from the Tibetan pastoral area was enriched by immunomagnetic beads and coated with MRS solid medium. After 24 hours of cult...

Embodiment 2

[0046] Example 2 Colony isolation of ciliated phenotype Lactobacillus rhamnosus

[0047] The ciliary phenotype Lactobacillus rhamnosus was isolated by colony immunoblotting, and the steps were as follows: select a plate with uniform colony distribution, cut out a nitrocellulose membrane with the same diameter as the petri dish, soak in deionized water for 10 min, and dry at 37°C 10 min. Place the NC membrane on the surface of the medium, let it stand at room temperature for 5 minutes, remove the NC membrane, and dry it at 37°C for 30 minutes; place the transfer membrane in 5% skim milk, and block it at room temperature for 1 hour; wash with PBST buffer 3 times , incubated in anti-SpaA antibody diluent for 1 h; washed 3 times in PBST, incubated in HRP-labeled goat anti-mouse IgG liquid for 1 h; washed 3 times in PBST, and immersed the transfer membrane in freshly prepared DAB chromogenic solution , and develop color in the dark for 10 min. When obvious spots appear on the memb...

Embodiment 3

[0049] Example 3 Physiological and biochemical identification of positive bacterial strains

[0050] Physiological and biochemical identification of the positive strains, the steps are as follows: inoculate the obtained positive colonies into the MRS medium by streaking, 37°C, anaerobic culture for 24 hours, pick a single colony smear on the plate, fix it, and add crystal violet staining solution dropwise , stain for 1 min, and wash with water; add iodine solution dropwise, act for 1 min, and wash with water; add 95% ethanol dropwise for decolorization for 15-20 s, and wash with water; counterstain with safranin for 1 min, and wash with water; microscopically examine and record the cell morphology of the strain. At the same time, single colonies were picked for catalase test, indole, hydrogen sulfide and ammonia, nitrate reduction, arginine hydrolysis, and sugar fermentation tests.

[0051] The results showed that the strains from which the positive colonies originated were Gr...

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Abstract

The present invention relates to a strain of ciliated Lactobacillus rhamnosus with a function of regulating mast cell activity and its application. The Lactobacillus rhamnosus LRYZU021 ( Lactobacillus rhamnosus LRYZU021), was deposited in China Center for Type Culture Collection (CCTCC for short) on October 30, 2017, deposit number: CCTCC NO: M 2017640. The Lactobacillus rhamnosus LRYZU021 of the present invention has a ciliated phenotype, has unique 16S ribosomal DNA sequence characteristics, and has good acid resistance and bile salt resistance, and outstanding intestinal epithelial cell adhesion ability; at the same time, the strain is compatible with Exposure to sensitized mast cells has a significant inhibitory effect on antigen-induced degranulation and histamine release, which is beneficial to alleviate the vasodilation and blood pressure drop caused by mast cell activation in the effect phase of allergic reactions. The microecological preparation prepared by using the strain has broad application prospects in preventing food allergy and alleviating allergic symptoms.

Description

technical field [0001] The invention belongs to the technical field of microbes, and relates to a ciliated Lactobacillus rhamnosus strain capable of regulating the activity of mast cells and its application in microecological preparations for allergy regulation. Background technique [0002] In recent years, with the increasingly complex variety of allergenic foods, food allergies and related diseases have increased rapidly around the world. At present, the actual prevalence of food allergies in the general population has reached 5%-10% in infants and children, and about 1% in adults. -2%, has become a disease that has a huge impact on society and economy. Mast cells play a central role in the pathogenesis of allergic diseases. Allergen-specific IgE antibodies produced in food allergies bind to mast cells, causing mast cells to degranulate and release mediators such as histamine, thereby initiating an allergic response to specific antigens . Lactobacillus rhamnosus ( Lact...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/747A61P37/08C12R1/225
CPCA61K2035/115A61P37/08C12N1/205C12R2001/225
Inventor 杨振泉高璐薛宇饶胜其胡钦李熠李华祥郑香峰
Owner 江苏新申奥生物科技有限公司
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