CN113215045B

CN113215045B - Lactobacillus gasseri LGV03 and application thereof

Info

Publication number: CN113215045B
Application number: CN202110523517.9A
Authority: CN
Inventors: 邱峰
Original assignee: Seventh Affiliated Hospital Of Southern Medical University Third People's Hospital Of Nanhai District Foshan City
Current assignee: Seventh Affiliated Hospital Of Southern Medical University Third People's Hospital Of Nanhai District Foshan City
Priority date: 2021-05-13
Filing date: 2021-05-13
Publication date: 2023-03-28
Anticipated expiration: 2041-05-13
Also published as: CN113215045A

Abstract

The invention discloses a Lactobacillus gasseri LGV03 and application thereof, belonging to the technical field of microorganisms. The preservation number of the Lactobacillus gasseri LGV03 is CGMCC No.22010. Fermentation supernatant and bacterial suspension of the Lactobacillus gasseri LGV03 can obviously inhibit Compound 48/80 from stimulating the zebra fish to secrete Tryptase in vivo, can inhibit degranulation of mast cells in vivo, and has the potential of being applied to preventing anaphylactic reaction in vivo. The lactobacillus gasseri LGV03 disclosed by the invention has a huge potential application prospect in the aspect of preventing anaphylactic reaction.

Description

Lactobacillus gasseri LGV03 and application thereof

Technical Field

The invention relates to the technical field of microorganisms, in particular to a lactobacillus gasseri LGV03 and application thereof.

Background

Allergic diseases are becoming a global public health problem, and various allergic diseases including asthma, allergic rhinitis and conjunctivitis, drug allergy, food allergy, insect bite allergy, skin eczema (atopic dermatitis), urticaria or severe systemic anaphylaxis (anaphylactic shock) affect people's health and daily life to various degrees, and some serious cases even endanger life, both in developed countries and developing countries. Therefore, allergic diseases are classified as one of three major diseases which are mainly prevented and treated in the 21 st century. According to epidemiological investigations worldwide, about 22% of the population suffers from different kinds of allergic diseases.

Allergy is a hypersensitivity reaction to harmless environmental antigens such as pollen and food. Typical anaphylaxis is when allergen induces Th2 cell to secrete IL-4, IL-5, IL-10 and IL-13, which can effectively activate B cell to produce IgE antibody, igE is combined with mast cell, and the mast cell degranulation is caused to release serine protease, histamine, cytokine and other mediators, and the anaphylaxis against specific antigen is initiated. Studies have shown that mast cells, a key effector cell in allergic reactions, secrete not only inflammatory mediators but also considerable amounts of various cytokines upon antigen activation of specific signal transduction pathways, and play a critical role in the development and progression of allergic diseases. Tryptase (Tryptase) is an important protease in mast cells, and it is similar to trypsin. Tryptase (Tryptase) is a specific marker for identifying human mast cells because storage and expression in mast cells are highly selective and cannot be detected in other cell types. In addition, tryptase (Tryptase) can be released extracellularly by degranulation of mast cells, and therefore, extracellular Tryptase (Tryptase) can serve as a marker for activation and degranulation of mast cells.

At present, the means for treating allergic diseases is limited, and only some antiallergic medicines are taken at the time of the disease, so that the occurrence of allergic reaction cannot be completely avoided. With the increasingly intensive scientific research on intestinal microorganisms, the relationship between intestinal microorganisms and health is receiving more and more attention from people, especially the relationship between intestinal microorganisms and allergy. It has been shown that food allergy is associated with the microbial composition of the intestine, and that certain bacteria in the intestine protect humans from food allergy. Thus, intervention in the gut microflora may provide a new strategy for preventing allergic sensitization.

Probiotics are defined as living microorganisms that improve the balance of the intestinal flora and may benefit various aspects of the physiological response of the host, including immune function. In addition, the prevention and treatment effects of probiotics on allergic diseases such as atopic dermatitis, allergic rhinitis, food allergy, etc. have also been studied in a large amount. There have been several studies showing that oral administration of bifidobacteria or lactobacilli can reduce food allergies. Lactobacillus can attenuate mast cell activation and release inflammatory mediators associated with allergic reactions. The bifidobacterium BGN4 can inhibit the generation of serum IgE and IgG1 in a food allergy mouse model, and in addition, the lactobacillus casei Shirota can inhibit the generation of antigen-specific IgE by stimulating the secretion of IL-12, so that the lactobacillus casei Shirota shows good probiotic efficacy for preventing or treating allergic diseases. The current international probiotic patent application focuses on the traditional research and development strong countries in the United states, the Japan and the Russia, and China lacks functional strains with independent intellectual property rights. Probiotic strains used by domestic production enterprises are imported for a long time, and foreign strains are not necessarily suitable for the gastrointestinal physiological conditions of residents in China. In addition, the function of the probiotics lacks strong scientific research evidence, and the popularization of the probiotics and the products thereof is seriously influenced. Based on the method, aiming at the deep excavation of the functions of the strain resources, the novel probiotic strain which has independent intellectual property rights, has specific functional properties and is suitable for the physiological characteristics of Chinese people is screened out, and the method is particularly important for improving the core competitiveness of probiotic production enterprises in China and promoting the development of probiotic products in China.

Therefore, the problem to be solved by the technical personnel in the field is to provide the lactobacillus gasseri LGV03 and the application thereof.

Disclosure of Invention

In view of the above, the invention provides a lactobacillus gasseri LGV03 and application thereof in preparing a medicament for preventing allergic diseases.

In order to achieve the purpose, the invention adopts the following technical scheme:

the lactobacillus gasseri (Lactobacillus gasseri) LGV03 has the preservation number of CGMCC No.22010, is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms (CGMCC for short), is collected at China academy of sciences institute of China No.3, beijing, naja, north Cheng Yang district, xilu No. 1, has the preservation date of 2021 year, 03 month and 15 days, and is classified and named as Lactobacillus gasseri.

Further, the Lactobacillus gasseri LGV03 is applied to preparation of a medicament for preventing allergic diseases.

Further, the Lactobacillus gasseri LGV03 is applied to preparation of medicines for inhibiting mast cell degranulation.

Further, the lactobacillus gasseri LGV03 is a bacterial suspension or a fermentation supernatant.

The Lactobacillus gasseri LGV03 disclosed by the invention has the effect of inhibiting Compound 48/80 from stimulating Tryptase secretion of zebra fish in vivo, has the potential of inhibiting mast cell degranulation, and shows a good probiotic effect of preventing anaphylactic reaction.

The bacterial strain capable of remarkably inhibiting Compound 48/80 from stimulating the zebra fish to secrete Tryptase in vivo comprises fermentation supernatant (extracellular secretion) and bacterial suspension (thallus) of the bacterial strain.

According to the technical scheme, compared with the prior art, the invention discloses and provides the Lactobacillus gasseri LGV03 and the application thereof in preparing the medicament for preventing the allergic diseases, the Lactobacillus gasseri LGV03 is obtained by separating and screening the secretion of the genital tract of a healthy female, can obviously inhibit Compoud48/80 from stimulating the secretion of Tryptase by zebra fish in vivo, can inhibit the degranulation of mast cells in vivo, has the potential of being applied to the in vivo prevention of the allergic reaction, and provides theoretical reference and guidance basis for developing the probiotic preparation for preventing the allergic reaction by utilizing the Lactobacillus gasseri LGV 03.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the embodiments or the prior art descriptions will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.

FIG. 1 is a phylogenetic tree of the 16SrDNA sequences of Lactobacillus gasseri LGV03 and related species according to the invention;

FIG. 2 is a diagram showing the morphology of colonies formed by Lactobacillus gasseri LGV03 on MRS solid medium;

FIG. 3 is a drawing showing the colony morphology of Lactobacillus gasseri LGV03 on a blood agar plate according to the present invention;

wherein, 1, negative control bacterium: CICC10417 listeria inorage; 2, positive control bacteria: cic 10473 staphylococcus aureus; 3, sample: lactobacillus gasseri LGV03;

FIG. 4 is a drawing showing the microscopic morphology observation of the Lactobacillus gasseri LGV03 after gram-staining;

FIG. 5 is a drawing showing the drug sensitivity test of Lactobacillus gasseri LGV03 of the present invention;

wherein, A: penicillin; b: ampicillin; c: meropenem; d: vancomycin; e: erythromycin; f: clindamycin; g: linezolid;

FIG. 6 is a graph showing the effect of fermentation supernatant, bacterial suspension of Lactobacillus gasseri LGV03 of the present invention on the expression level of Compoud48/80 stimulating Tryptase secretion from zebrafish;

FIG. 7 is a graph showing the inhibition rate of fermented supernatant and bacterial suspension of Lactobacillus gasseri LGV03 on the secretion of Tryptase stimulated by Compoud48/80 zebra fish.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1 isolation and identification of Lactobacillus gasseri LGV03

(1) Separation: the genital secretion of healthy women is diluted in a gradient way, and then is respectively inoculated in a TPY solid culture medium, an MRS solid culture medium, a BDS solid culture medium and a BS solid culture medium, anaerobic culture is carried out for 48 hours at 37 ℃, and a single colony on a flat plate is picked and streaked to obtain a pure colony. Inoculating pure bacterial colonies on the plate into an MRS liquid culture medium, carrying out anaerobic culture at 37 ℃ for 12-16 h, adding 20% glycerol, and storing in a refrigerator at-80 ℃.

(2) Molecular biological identification of the strains: extracting genome DNA of the obtained strain, amplifying a 16SrDNA full-length fragment by using 16SrDNA universal primers 27F and 1492R through a PCR technology, and then sequencing to identify the strain species. Wherein the sequences of the universal primers 27F and 1492R are as follows:

27F：5’-AGAGTTTGATCCTGGCTCAG-3’；SEQ ID NO.1；

1492R：5’-GGTTACCTTGTTACGACTT-3’；SEQ ID NO.2。

physiological and biochemical identification of the strain: the colonies were picked to prepare a bacterial suspension and identified with a VITEKANC identification card. The specific physiological and biochemical results are shown in Table 1.

TABLE 1LGV03 physiological and biochemical reaction results

Description of the symbols: "+", positive; "+ ^w ", weak positive; "-", negative.

Phylogenetic analysis: the MEGA software and the adjacent position connection method are adopted to display the 'Lactobacillus gasseri LGV 03' and a 16SrDNA sequence phylogenetic tree of related species, the similarity is repeatedly calculated for 1000 times, the nodes of the phylogenetic tree in the figure only display Bootstrap values larger than 50% of values, and the superscript 'T' represents a model strain.

The experimental results are as follows: the strains selected from the secretions of the genital tract of healthy women in Guangzhou, guangdong province were analyzed by systematic evolution using MEGA software on the basis of 16SrDNA homology for 11 typical strains, and the results are shown in FIG. 1.LGV03 and Lactobacillus gasseri model strain ATCC33323 ^T (CP 000413) constitutes a branch alone, reflecting that the relationship between the two is recent. Thus, strain LGV03 was identified as Lactobacillus gasseri by biochemical and physiological identification, 16SrDNA identification and phylogenetic tree showing that its 16SrDNA sequence is shown in SEQ ID NO. 3.

TGCCTAATACATGCAAGTCGAGCGAGCTTGCCTAGATGAATTTGGTGCTTGCACCAGATGAAACTAGATACAAGCGAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCAAGAGACTGGGATAACACCTGGAAACAGATGCTAATACCGGATAACAACACTAGACGCATGTCTAGAGTTTAAAAGATGGTTCTGCTATCACTCTTGGATGGACCTGCGGTGCATTAGCTAGTTGGTAAGGTAACGGCTTACCAAGGCAATGATGCATAGCCGAGTTGAGAGACTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAGCTCTGTTGGTAGTGAAGAAAGATAGAGGTAGTAACTGGCCTTTATTTGACGGTAATTACTTAGAAAGTCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGTGCAGGCGGTTCAATAAGTCTGATGTGAAAGCCTTCGGCTCAACCGGAGAATTGCATCAGAAACTGTTGAACTTGAGTGCAGAAGAGGAGAGTGGAACTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTCTCTGGTCTGCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAGTGCTAAGTGTTGGGAGGTTTCCGCCTCTCAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCAGTGCAAACCTAAGAGATTAGGTGTTCCCTTCGGGGACGCTGAGACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCATTAGTTGCCATCATTAAGTTGGGCACTCTAATGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACGGTACAACGAGAAGCGAACCTGCGAAGGCAAGCGGATCTCTGAAAGCCGTTCTCAGTTCGGACTGTAGGCTGCAACTCGCCTACACGAAGCTGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTCTGTAACACCCAAAGCCGGTGGGATAACCTTTATAGGAG；SEQ ID NO.3。

(3) Culture characteristics of the Strain

An LGV03 colony is picked by an inoculating loop, is subjected to zonal streaking and is inoculated on an MRS agar plate, and is cultured for 48 hours under an aerobic condition at 37 ℃; in addition, LGV03 colonies are picked and inoculated on a blood agar plate, and are cultured for 48h under anaerobic conditions at 37 ℃ by taking CICC10417 Listeria innococcus (Listeria innocus) as a negative control bacterium and CICC10473 Staphylococcus aureus (Staphylococcus aureus) as a positive control bacterium; the growth of colonies in MRS plates and the presence or absence of hemolytic ring formation on blood agar plates were observed.

The experimental results are as follows: inoculating a single LGV03 colony to an MRS solid culture medium, and carrying out anaerobic culture at 37 ℃ for 48h, wherein the colony is white, round, wet in surface, opaque and neat in edge (figure 2); the negative control bacterium cic 10417 Listeria inorage (Listeria innocus) showed no hemolytic ring on blood agar plates, while the positive control bacterium cic 10473 Staphylococcus aureus (Staphylococcus aureus) showed hemolytic ring on blood agar plates, and lactobacillus gasseri LGV03 showed no hemolytic ring on blood agar plates (fig. 3).

(4) And (3) identifying the strain morphology: when the screened LGV03 strain is observed under a microscope after gram staining, gram-positive bacteria are purple, and gram-negative bacteria are red.

The experimental results are as follows: LGV03 cells were rod-shaped, 0.4-0.5. Mu. M.times.0.9-6.3. Mu.m, aligned singly or in pairs, and gram-positive (FIG. 4).

(5) Drug sensitivity test: and taking the LGV03 strain cultured above, and determining the sensitivity of the LGV03 strain to common antibacterial drugs by adopting an E-test method.

The experimental results are as follows: as can be seen from table 2 and fig. 5, LGV03 is sensitive to penicillin, ampicillin, meropenem, vancomycin, erythromycin, linezolid, and not to clindamycin.

TABLE 2LGV03 drug sensitivity test results

Antibacterial agent	MIC value (μ g/mL)	Drug sensitivity
			Penicillin (PEN)	0.048	Sensitivity of
Ampicillin (AM)	0.19	Sensitivity to
			Meropenem (MP)	0.38	Sensitivity of
Vancomycin (VA)	0.75	Sensitivity of
			Erythromycin (EM)	0.064	Sensitivity of
Clindamycin (CM)	6	Drug resistance
			Linezolid (LZ)	0.75	Sensitivity to

Example 2 preparation of fermentation supernatant (extracellular secretion) and bacterial suspension (thallus) of Lactobacillus gasseri LGV03

Activating and culturing Lactobacillus gasseri LGV03, inoculating into MRS liquid culture medium, culturing at 37 deg.C for 15 hr, and regulating fermentation bacteria concentration to 2 × 10 ⁸ Centrifuging at 4 deg.C and 8000r/min for 10min to obtain culture supernatant and thallus precipitate, and filtering with 0.22 μm filter membrane to obtain fermentation supernatant (extracellular secretion); after the cell pellet was washed twice with PBS, the cell pellet was resuspended in PBS to adjust the cell concentration to 2X 10 ⁸ CFU/mL gave a suspension (thallus).

Example 3 Effect of Lactobacillus gasseri LGV03 on the stimulation of Tryptase secretion by Compoud48/80 Zebra fish

Cromolyn sodium and Na-benzoyl-DL-arginine-p-nitroamide hydrochloride (BAPNA) were purchased from Shanghai leaf Biotech, inc., compoud48/80 was purchased from Sigma-Aldrich.

Healthy wild-type AB line zebrafish developed to 5dpf (days post fertilization) were picked and placed 10 strips/well in 96-well cell culture plates. The experiment set comprises a normal group, a model group, an intervention group (a positive control group, a bacterial suspension group and a fermentation supernatant group) and a solvent zero-adjustment group, wherein each group is provided with 6 multiple wells. Adding PBS into a normal group, adding PBS into a model group, adding a sodium cromoglycate solution (100 mu g/mL) into a positive control group, adding a bacterial suspension into a bacterial suspension group, adding a fermentation supernatant into a fermentation supernatant group, adding PBS into a solvent zeroing group (without adding zebra fish), and adding 100 mu L of PBS into each hole; after incubation for 24h at 28 ℃ in a biochemical incubator, 150 mu LPBS is added to a normal group, compoud48/80 (8 mu g/mL) is added to a model group, a bacterial suspension group, a fermentation supernatant group, a positive control group and a solvent zero-adjustment group respectively, 150 mu L of the solution is added to each well, after incubation for 2h at 28 ℃, 150 mu L of the solution is taken from each well and is placed in a centrifuge tube of 2mL respectively, 150 mu LBAPNA solution (20 mg/mL) is added to each centrifuge tube, after incubation for 48h at 37 ℃ in the biochemical incubator, 200 mu L of the solution is taken from each centrifuge tube and is placed in a cell culture plate of 96 wells, and the absorbance (OD value) is measured at 405nm by an enzyme labeling instrument. The expression level and inhibition rate of Tryptase are as follows:

the data were statistically processed using SPSS19.0 software, and the experimental data were expressed as x + -SEM data and analyzed by one-way anova. Compared to the normal group: ^### p<0.005; compared to the model group: * P is<0.05，**P<0.01，***P<0.005。

The results of the expression level and inhibition rate of Tryptase are shown in FIGS. 6 and 7, and the results show that the Tryptase secretion amount (172.96 +/-8.02%) of the model group is obviously increased compared with that of the normal group (44.64 +/-4.44%), which indicates that the Compoud48/80 stimulation is effective.

The Tryptase secretion amount of the sodium cromoglycate group is 55.18 +/-3.03%, the Tryptase inhibition rate is 68.10 +/-1.75%, and the difference is obvious compared with that of a model group (the Tryptase secretion amount is 172.96 +/-8.02%, and the Tryptase inhibition rate is 0) (P is less than 0.005). Therefore, the cromolyn sodium can inhibit the degranulation of mast cells in vivo, has an anti-sensitization effect, and is consistent with clinical results. The secretion amounts of Tryptose of the fermentation supernatant and the bacterial suspension group of the Lactobacillus gasseri LGV03 are respectively 79.78 +/-3.89% and 122.00 +/-6.52%, and the inhibition rates of the Tryptose are respectively 53.88 +/-2.25% and 29.47 +/-3.77%, so that the differences (P < 0.005) between the secretion amounts of the Tryptose and the inhibition rates of the Tryptose are obvious compared with those of a model group (the secretion amounts of the Tryptose are 172.96 +/-8.02% and the inhibition rates of the Tryptose are 0). Therefore, the results show that the fermentation supernatant and the bacterial suspension of the lactobacillus gasseri LGV03 can inhibit the degranulation of mast cells in vivo and show good effect of preventing anaphylactic reaction.

The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Sequence listing

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<120> Lactobacillus gasseri LGV03 and application thereof

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tgcctaatac atgcaagtcg agcgagcttg cctagatgaa tttggtgctt gcaccagatg 60

aaactagata caagcgagcg gcggacgggt gagtaacacg tgggtaacct gcccaagaga 120

ctgggataac acctggaaac agatgctaat accggataac aacactagac gcatgtctag 180

agtttaaaag atggttctgc tatcactctt ggatggacct gcggtgcatt agctagttgg 240

taaggtaacg gcttaccaag gcaatgatgc atagccgagt tgagagactg atcggccaca 300

ttgggactga gacacggccc aaactcctac gggaggcagc agtagggaat cttccacaat 360

ggacgcaagt ctgatggagc aacgccgcgt gagtgaagaa gggtttcggc tcgtaaagct 420

ctgttggtag tgaagaaaga tagaggtagt aactggcctt tatttgacgg taattactta 480

gaaagtcacg gctaactacg tgccagcagc cgcggtaata cgtaggtggc aagcgttgtc 540

cggatttatt gggcgtaaag cgagtgcagg cggttcaata agtctgatgt gaaagccttc 600

ggctcaaccg gagaattgca tcagaaactg ttgaacttga gtgcagaaga ggagagtgga 660

actccatgtg tagcggtgga atgcgtagat atatggaaga acaccagtgg cgaaggcggc 720

tctctggtct gcaactgacg ctgaggctcg aaagcatggg tagcgaacag gattagatac 780

cctggtagtc catgccgtaa acgatgagtg ctaagtgttg ggaggtttcc gcctctcagt 840

gctgcagcta acgcattaag cactccgcct ggggagtacg accgcaaggt tgaaactcaa 900

aggaattgac gggggcccgc acaagcggtg gagcatgtgg tttaattcga agcaacgcga 960

agaaccttac caggtcttga catccagtgc aaacctaaga gattaggtgt tcccttcggg 1020

gacgctgaga caggtggtgc atggctgtcg tcagctcgtg tcgtgagatg ttgggttaag 1080

tcccgcaacg agcgcaaccc ttgtcattag ttgccatcat taagttgggc actctaatga 1140

gactgccggt gacaaaccgg aggaaggtgg ggatgacgtc aagtcatcat gccccttatg 1200

acctgggcta cacacgtgct acaatggacg gtacaacgag aagcgaacct gcgaaggcaa 1260

gcggatctct gaaagccgtt ctcagttcgg actgtaggct gcaactcgcc tacacgaagc 1320

tggaatcgct agtaatcgcg gatcagcacg ccgcggtgaa tacgttcccg ggccttgtac 1380

acaccgcccg tcacaccatg agagtctgta acacccaaag ccggtgggat aacctttata 1440

ggag 1444

Claims

1. The application of Lactobacillus gasseri LGV03 in the preparation of drugs for preventing allergic diseases is characterized in that the preservation number of the Lactobacillus gasseri LGV03 is CGMCC No.22010.

2. Use of lactobacillus gasseri LGV03 for the preparation of a medicament for the prevention of allergic disease according to claim 1, wherein said lactobacillus gasseri LGV03 is a bacterial suspension.

3. Use of lactobacillus gasseri LGV03 according to claim 1 for the preparation of a medicament for inhibiting mast cell degranulation.

4. Use of a lactobacillus gasseri LGV03 for the manufacture of a medicament for inhibiting mast cell degranulation according to claim 3, wherein said lactobacillus gasseri LGV03 is a bacterial suspension.