Probiotic agent for resisting helicobacter pylori infection and application thereof

Abstract

The invention relates to a probiotic agent capable of resisting helicobacter pylori infection and application of the probiotic agent. The bacterial strains in the probiotic agent capable of resisting helicobacter pylori infection are composed of a Bifidobacterium animalis subsp. Lactis (Bifidobacterium animalis subsp. Lactis). Lactis BLa80 bacterial strain and a Lactobacillus acidophilus LA18 bacterial strain. The invention further relates to a preparation method of the probiotic agent capable of resisting helicobacter pylori infection and application of the probiotic agent capable of resisting helicobacter pylori infection. The two strains can cooperate with each other and promote each other, and have a synergistic effect in the aspect of resisting helicobacter pylori infection, specifically, the level of an Hp-IgG antibody and the level of endotoxin of a mouse infected with helicobacter pylori are remarkably reduced; the levels of serum inflammatory factors such as IL-17A, IL-10, IL-23, IFN-beta and IFN-gamma of mice infected with helicobacter pylori are regulated, and inflammatory response caused by helicobacter pylori infection is inhibited; the activity of pepsin, the activity of cellulase and the activity of alpha-amylase are obviously improved, so that the digestive ability of the stomach is enhanced.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and relates to a probiotic agent for resisting Helicobacter pylori infection and application thereof. Background Art

[0002] Helicobacter pylori (HP) is a type of aerobic bacteria that is spiral or arc-shaped under a microscope and is closely related to the occurrence of chronic gastritis, peptic ulcers and gastric cancer. It has high nutritional requirements during its growth and reproduction, and can resist gastric acid. After entering the human body, it can survive and reproduce in the gastric mucosal epithelial cells. In the process, it produces a large number of invasive factors and toxins, which promote the occurrence of peptic ulcers and even gastric cancer. The pathogenic mechanism of Helicobacter pylori for gastritis is mainly that after colonization in the stomach, it can produce a large number of virulence factors. These virulence factors can cause the gastric mucosal epithelial cells to degenerate, thereby destroying the normal gastric mucosal barrier, causing gastric acid to erode the gastric mucosa, and then causing gastritis; while the pathogenic mechanism of Helicobacter pylori for peptic ulcer is that while destroying the gastric mucosal barrier, Helicobacter pylori may also stimulate the secretion of gastric acid, aggravating the damage of the gastric mucosa and duodenal mucosa, and then leading to the occurrence of gastric ulcers or duodenal ulcers.

[0003] Probiotics have shown great potential in preventing and treating Helicobacter pylori infection. As living microorganisms, probiotics can bring health benefits to the host in a variety of ways when ingested in sufficient amounts. The mechanism of action of probiotics against Helicobacter pylori mainly includes direct antibacterial effect, competitive inhibition of Helicobacter pylori colonization, and improvement of host anti-inflammatory and barrier function. By producing antibacterial substances, competing with Helicobacter pylori for nutrients and adhesion sites, and regulating the host immune response, probiotics can effectively inhibit the growth and colonization of Helicobacter pylori, thereby reducing or preventing Helicobacter pylori infection.

[0004] The application of probiotics in the prevention and treatment of gastric diseases caused by Helicobacter pylori is still in its early stages, and there are still few strategies. Therefore, the development of more probiotics against Helicobacter pylori can provide new ideas and methods for the treatment of gastric diseases caused by Helicobacter pylori. Summary of the invention

[0005] In view of the deficiencies in the prior art, the object of the present invention is to provide a probiotic agent for resisting Helicobacter pylori infection and its application, and in particular to a probiotic agent for resisting Helicobacter pylori infection and its application in the preparation of a medicament for resisting Helicobacter pylori infection.

[0006] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides a probiotic for resisting Helicobacter pylori infection, wherein the strains in the probiotic for resisting Helicobacter pylori infection are composed of Bifidobacterium animalis subsp. Lactis BLa80 strain with a preservation number of CGMCC No.15410 and Lactobacillus acidophilus LA18 strain with a preservation number of CCTCCNO:M 2022572.

[0008] The present invention creatively develops a new probiotic compounding method and a new anti-Helicobacter pylori infection strategy, that is, Bifidobacterium animalis subsp. Lactis BLa80 strain and Lactobacillus acidophilus LA18 strain are compounded and used together, and it is found that the two can cooperate with each other, promote each other, and synergize in the effect of anti-Helicobacter pylori infection, which is specifically manifested in: (1) significantly reducing the level of Hp-IgG antibodies and endotoxin levels in mice infected with Helicobacter pylori; (2) regulating the levels of serum inflammatory factors such as IL-17A, IL-10, IL-23, IFN-β, and IFN-γ in mice infected with Helicobacter pylori, and inhibiting the inflammatory response caused by Helicobacter pylori infection; (3) significantly improving the activity of pepsin, cellulase and α-amylase to enhance gastric digestion. When the amount of bacteria is the same, compared with the bacterial intervention method lacking any one of the bacteria, the compounding of the two bacteria significantly improves the effect of anti-Helicobacter pylori infection. At the same time, Bifidobacterium animalis subspecies lactis and Lactobacillus acidophilus are both probiotics, the products are highly safe and not prone to resistance.

[0009] Preferably, the ratio of the number of viable bacteria of the BLa80 strain to the LA18 strain is 1:10-10:1, for example, 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, etc. Other specific point values ​​within the above numerical range can be selected, and they will not be described here one by one.

[0010] Preferably, the content of live bacteria in the probiotic is not less than 1×10 10 CFU / g or 1×10 10 CFU / mL, for example 1×10 10 CFU / g (CFU / mL), 5×10 10 CFU / g (CFU / mL), 1×10 11 CFU / g (CFU / mL), 3×1011 CFU / g (CFU / mL), 5×10 11 CFU / g (CFU / mL), 1×10 12 CFU / g (CFU / mL), 1×10 13 CFU / g (CFU / mL), etc. Other specific point values ​​within this numerical range can be selected and will not be described here one by one.

[0011] Preferably, the dosage form of the probiotics includes solutions, lyophilized powders, capsules, tablets or granules. The dosage form of the probiotics involved in the present invention is not limited, including the most commonly used solutions, lyophilized powders, or further prepared capsules, tablets or granules.

[0012] Preferably, the probiotic is in the form of a solution, which is prepared by the following method:

[0013] The BLa80 strain and the LA18 strain are inoculated into a culture medium respectively for activation and fermentation culture in sequence to obtain a fermentation broth; the fermentation broth is centrifuged respectively and resuspended with a solvent to obtain a BLa80 bacterial suspension and a LA18 bacterial suspension; the BLa80 bacterial suspension and the LA18 bacterial suspension are mixed according to the ratio of the number of live bacteria to obtain the probiotic agent.

[0014] Preferably, the probiotic is in the form of a freeze-dried powder, which is prepared by the following method:

[0015] The BLa80 strain and the LA18 strain are inoculated into a culture medium respectively for activation and fermentation culture in sequence to obtain a fermentation broth; the fermentation broth is centrifuged respectively, mixed with a protective agent and freeze-dried to obtain BLa80 bacterial powder and LA18 bacterial powder; the BLa80 bacterial powder and the LA18 bacterial powder are mixed according to the ratio of the number of live bacteria to obtain the probiotic agent.

[0016] Preferably, the protective agent is selected from any one or a combination of at least two of skim milk, gelatin, dextrin, gum arabic, dextran, sodium alginate, polyvinyl pyrrolidone, sucrose, lactose, trehalose, sorbitol or xylitol.

[0017] In a second aspect, the present invention provides use of the probiotic agent described in the first aspect in the preparation of a drug for resisting Helicobacter pylori infection.

[0018] Preferably, the medicine further contains excipients.

[0019] Preferably, the auxiliary materials include any one or a combination of at least two of fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, colorants, pH regulators, antioxidants, antibacterial agents or buffers.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] The present invention creatively develops a new probiotic compounding method and a new anti-Helicobacter pylori infection strategy, that is, Bifidobacterium animalis subsp. Lactis BLa80 strain and Lactobacillus acidophilus LA18 strain are compounded and used together, and it is found that the two can cooperate with each other, promote each other, and synergize in the effect of anti-Helicobacter pylori infection, which is specifically manifested in: (1) significantly reducing the level of Hp-IgG antibodies and endotoxin levels in mice infected with Helicobacter pylori; (2) regulating the levels of serum inflammatory factors such as IL-17A, IL-10, IL-23, IFN-β, and IFN-γ in mice infected with Helicobacter pylori, and inhibiting the inflammatory response caused by Helicobacter pylori infection; (3) significantly improving the activity of pepsin, cellulase and α-amylase to enhance gastric digestion. When the amount of bacteria is the same, compared with the bacterial intervention method lacking any one of the bacteria, the compounding of the two bacteria significantly improves the effect of anti-Helicobacter pylori infection. At the same time, Bifidobacterium animalis subspecies lactis and Lactobacillus acidophilus are both probiotics, the products are highly safe and not prone to resistance.

[0022] The BLa80 strain involved in the present invention is classified and named as Bifidobacterium animalis subsp. Lactis, and the preservation unit is the General Microbiology Center of China Microbiological Culture Collection Administration. The preservation time is March 5, 2018, the preservation number is CGMCC No. 15410, and the address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0023] The LA18 strain involved in the present invention is classified and named as Lactobacillus acidophilus, the preservation unit is China Center for Type Culture Collection, the preservation time is May 9, 2022, the preservation number is CCTCC NO: M 2022572, and the address is: Wuhan University, Wuhan, China. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is the statistical result diagram of the weight change of mice in each group;

[0025] Figure 2 This is a statistical result diagram of the anti-Hp-IgG antibody content in the serum of each group of mice;

[0026] Figure 3 This is the statistical result diagram of serum endotoxin levels of mice in each group;

[0027] Figure 4 This is the statistical result graph of IL-10 levels in each group of mice;

[0028] Figure 5 This is a statistical result graph of IL-17A levels in each group of mice;

[0029] Figure 6 This is a statistical graph of IL-23 levels in each group of mice;

[0030] Figure 7 This is the statistical result of interferon IFN-β level in each group of mice;

[0031] Figure 8 This is the statistical result of interferon IFN-γ level in each group of mice;

[0032] Fig. 9 This is the statistical result graph of pepsin activity of mice in each group;

[0033] Fig.10 This is the statistical result graph of cellulase levels in each group of mice;

[0034] Fig.11 This is a statistical graph of α-amylase levels in each group of mice. DETAILED DESCRIPTION

[0035] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0036] The BLa80 strain involved in the following embodiment is classified and named as Bifidobacterium animalis subsp. Lactis, and the depositor is the General Microbiological Center of China Microbiological Culture Collection Administration, the deposit time is March 5, 2018, the deposit number is CGMCC No. 15410, and the address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0037] The LA18 strain involved in the following examples is classified and named Lactobacillus acidophilus, and the preservation unit is China Center for Type Culture Collection. The preservation time is May 9, 2022, the preservation number is CCTCC NO: M 2022572, and the address is: Wuhan University, Wuhan, China.

[0038] The SPF-grade C57BL / 6 mice involved in the following examples were purchased from Shanghai Experimental Animal Center; ordinary feed was purchased from Shanghai Slack Company; and skim milk was purchased from Fonterra Group with specifications of fat ≤ 1.0% and protein ≥ 32.9%.

[0039] The culture medium formula involved in the following examples is as follows:

[0040] MRS medium: peptone 10g / L, beef extract 10g / L, glucose 20g / L, sodium acetate 2g / L, yeast powder 5g / L, diammonium hydrogen citrate 2g / L, K2PO4·3H2O 2.6g / L, MgSO4·7H2O 0.1g / L, MnSO4 0.05g / L, Tween 801mL / L, cysteine ​​hydrochloride 0.5g / L.

[0041] The following method for preparing a bacterial suspension involves: taking out the desired strain from a -80°C refrigerator, inoculating it into an MRS liquid culture medium at an inoculation rate of 2% (v / v), culturing it at 37°C for 24 hours for activation, and continuously activating it twice to obtain an activated solution; inoculating the activated solution into an MRS liquid culture medium at an inoculation rate of 2% (v / v), culturing it at 37°C for 24 hours to obtain a bacterial solution; centrifuging the bacterial solution at 4°C and 5000 rpm for 10 minutes, filtering to obtain bacterial bodies, and resuspending the bacterial bodies in a PBS solution to obtain the obtained product.

[0042] The Lactobacillus acidophilus BNCC185344 strain involved in the following examples was commercially available; Helicobacter pylori SS1 (H. pylori SS1) was purchased from Guangdong Provincial Microbiological Culture Collection Center.

[0043] Example

[0044] This example explores the improvement effect of probiotics on the mouse model of Helicobacter pylori infection:

[0045] (1) Preparation of Helicobacter pylori culture medium: H. pylori SS1 frozen at -80°C was revived on Columbia agar plates containing 5% sterile defibrinated sheep blood. After two generations of activation, the cells were cultured in Columbia blood plates for 48 h and then washed with BHI liquid medium to obtain H. pylori SS1 culture medium. The concentration was adjusted to 1×10 10 CFU / mL, for future use.

[0046] (2) Experimental animals: healthy female SPF grade C57BL / 6 mice, 8 weeks old (90 mice), these mice were housed in a controlled environment, with room temperature maintained at 23-27°C, humidity at 45-55%, and a 12 h light / dark cycle, they were allowed to eat and drink freely. The bedding was changed 1-2 times a week.

[0047] (3) Animal grouping: After one week of adaptive feeding, the mice were randomly divided into nine groups, with 10 mice in each group: blank group, model group, BLa80 group (intervention with BLa80 bacterial solution), LA18 group (intervention with LA18 bacterial solution), BLa80+LA18 group 1 (combined intervention with BLa80 bacterial solution and LA18 bacterial solution, with a live bacteria count ratio of 1:1), BLa80+LA18 group 2 (combined intervention with BLa80 bacterial solution and LA18 bacterial solution, with a live bacteria count ratio of 10:1), BLa80+LA18 group 3 (combined intervention with BLa80 bacterial solution and LA18 bacterial solution, with a live bacteria count ratio of 1:10), BNCC185344 group (intervention with commercially available Lactobacillus acidophilus), and BLa80+BNCC185344 group (combined intervention with BLa80 bacterial solution and commercially available Lactobacillus acidophilus bacterial solution, with a live bacteria count ratio of 1:1).

[0048] (4) Animal modeling and intervention methods:

[0049] After one week of adaptive feeding, mice were fasted for 12 h and deprived of water for 4 h. They were first gavaged with 0.25 mL of 0.2 mol / L sodium bicarbonate solution, and then gavaged with 0.3 mL of SS1 bacterial solution (1×10 10 CFU / mL), and gavage was performed every other day. The above operation was performed 5 times in total.

[0050] The experiment lasted for 6 weeks.

[0051] Blank group: 0.2 mL 0.9% NaCl was gavaged daily from week 1 to week 6, and blank BHI liquid culture medium was gavaged from week 2 according to the above modeling method.

[0052] Model group: 0.2 mL 0.9% NaCl was gavaged daily from week 1 to week 6. In the second week, the infection model was established in the same manner as above, and the feeding conditions remained unchanged.

[0053] Each probiotic intervention group: Daily intragastric administration of 0.2 mL of probiotic solution or compound probiotic solution (total bacterial concentration of 1×10 10 CFU / mL), and the infection model was established in the second week according to the above method, and the breeding conditions remained unchanged.

[0054] (5)Indicator analysis:

[0055] (5.1) During the experiment, the body weight of mice was measured weekly and the diet and drinking water records were recorded. The dynamic body weight data of mice in each group are as follows: Figure 1 As shown. Figure 1 It can be seen that the weight of mice in each group began to increase after the gavage. After the experiment, the weight of mice in each group increased to about 22g, indicating that Helicobacter pylori infection had no significant effect on the weight of mice, and the intervention of probiotics had no significant effect.

[0056] (5.2) Effect of probiotic intervention on serum anti-Hp-IgG antibody levels in infected mice:

[0057] After the last oral gavage, fast for 12 hours, anesthetize with intraperitoneal injection of 4% chloral hydrate solution, collect blood from the eyeball, centrifuge at 4℃3500r / min for 20 minutes, and collect serum in EP tube. Anti-Hp-IgG antibody detection was performed by ELISA. 100μL of the specimen to be tested diluted 1:50 was added to the Hp antigen-coated microwells, reacted at 37℃ for 30 minutes, and washed 3 times. 100μL of rabbit anti-mouse IgG antibody labeled with horseradish peroxidase (HRP) was added to each well, reacted at 37℃ for 30 minutes, and washed 3 times. 100μL of TMB substrate (tetramethylbenzidine) was added to each well, reacted at room temperature in the dark for 15 minutes, and 50μL of H2SO4 stop solution was added. The absorbance of each well was measured by an enzyme reader at λ=450nm, and the serum Hp-IgG antibody content of each group of mice was as follows. Figure 2 shown.

[0058] Figure 2 The results showed that compared with the model group, the levels of anti-Hp-IgG antibodies in the serum of mice decreased to varying degrees after intervention with probiotics in each group. The results of BLa80 group, LA18 group, BNCC185344 group and BLa80+BNCC185344 group were higher than the anti-Hp-IgG antibody levels in the uninfected control group of mice, but the results of BLa80 and LA18 combined groups 1 to 3 were significantly lower than the anti-Hp-IgG antibody levels in the infected control group of mice, indicating that its effect was the best, and that BLa80 and LA18 strains synergized in the above-mentioned effects.

[0059] (5.3) Effects of probiotic intervention on serum endotoxin levels in infected mice:

[0060] After the last oral administration, the mice were fasted for 12 hours, anesthetized by intraperitoneal injection of 4% chloral hydrate solution, blood was collected from the eyeball, centrifuged at 4℃3500r / min for 20 minutes, and serum was collected in EP tubes. The serum of mice was collected and tested according to the method of ET detection kit. The endotoxin content of the serum of mice in each group was as follows Figure 3 shown.

[0061] Endotoxin (ET) is the lipopolysaccharide (LPS) component of the outer layer of the cell wall of Gram-negative bacteria. Bacterial decomposition releases LPS, which can stimulate monocytes and macrophages to produce inflammatory mediators and cytokines such as interleukin-1, tumor necrosis factor, and interferon, thereby destroying the integrity of the intestinal epithelium and leading to intestinal epithelial cell inflammation, apoptosis, and systemic inflammatory response. Figure 3The results showed that compared with the model group, the endotoxin levels of each probiotic group decreased to varying degrees. The results of the LA18 group, BNCC185344 group, BLa80+BNCC185344 group and BLa80 group were higher than the endotoxin levels of the uninfected control group mice, but the results of the BLa80+LA18 groups 1 to 3 were significantly lower than the endotoxin levels of the uninfected control group mice and were close to the endotoxin levels of the uninfected control group mice. This shows that the probiotics involved in the present invention can reduce the endotoxins produced by Helicobacter pylori infection and the harm it causes to the body.

[0062] (5.4) Effects of probiotic intervention on serum interleukin levels in infected mice:

[0063] After the last oral administration, the mice were fasted for 12 hours, anesthetized by intraperitoneal injection of 4% chloral hydrate solution, blood was collected from the eyeball, centrifuged at 4℃3500r / min for 20 minutes, and serum was collected in EP tubes. The serum of mice was collected and tested according to the IL-10, IL-17A and IL-23 detection kit methods. The serum interleukin content of mice in each group was as follows Figure 4-Figure 6 shown.

[0064] As can be seen from the figure, compared with the model group, the IL-10 level, IL-17A level and IL-23 level in the serum of mice in each probiotic intervention group were improved, and the improvement effect of the probiotic groups of BLa80+LA18 groups 1 to 3 was more significant.

[0065] (5.5) Effect of probiotic intervention on serum interferon levels in infected mice:

[0066] After the last oral administration, the mice were fasted for 12 hours, anesthetized by intraperitoneal injection of 4% chloral hydrate solution, blood was collected from the eyeball, centrifuged at 4℃3500r / min for 20 minutes, and serum was collected in EP tubes. The serum of mice was collected and tested according to the methods of interferon IFN-β and IFN-γ detection kits. The endotoxin content of the serum of mice in each group was as follows: Figure 7-Figure 8 shown.

[0067] Depend on Figure 7 and Figure 8 The results showed that compared with the model group, the levels of IFN-β and IFN-γ in the serum of mice in each probiotic intervention group were significantly increased, and the increasing effect of BLa80+LA18 groups 1 to 3 was better.

[0068] (5.6) Effects of probiotic intervention on digestive enzyme activities in gastric tissue of infected mice:

[0069] Pepsin is a digestive fluid secreted by the chief cells of the gastric mucosa, which breaks down proteins in food into small peptides. Symptoms such as chronic gastritis, chronic gastric dilatation, and chronic duodenitis will cause a decrease in pepsin secretion. Cellulase is a type of enzyme that can degrade cellulose and hydrolyze protein, breaking down protein in food into small molecules, which is beneficial for digestion and absorption. If cellulase is lacking, symptoms such as indigestion, bloating, and constipation may occur. Cellulase can promote the balance of gastrointestinal flora in animals, optimize the structure of intestinal flora, and protect the health of the digestive tract. Amylase is mainly produced in the pancreas and oral salivary glands, breaking down carbohydrates into monosaccharides that can be absorbed by the human body and generating energy for the human body. In the stomach, amylase will be neutralized by gastric acid, and some of the decomposed starch will continue to enter the small intestine.

[0070] Mice were killed by cervical vertebrae fracture, and the stomach tissues were obtained. After grinding with liquid nitrogen, they were homogenized with Tris-HCl buffer (pH 7.4) and enzyme cutting agent. The homogenate was centrifuged at 1800g and 4°C for 10 min, and the supernatant was collected and analyzed for pepsin activity, cellulase activity, and α-amylase activity. The results are as follows: Figure 9-11 shown.

[0071] As can be seen from the figure, compared with the blank group, the pepsin activity, cellulase activity and α-amylase activity of the mice in the model group were significantly reduced, indicating that Helicobacter pylori infection has an inhibitory effect on the digestive enzyme activity of mice; after the intervention of each probiotic group, the pepsin activity, cellulase activity and α-amylase activity of the mice were improved to varying degrees, and the effects of BLa80+LA18 groups 1 to 3 were better, indicating that the probiotic agent involved in the present invention can reverse the decreased pepsin activity, cellulase activity and α-amylase activity caused by Helicobacter pylori infection, and the BLa80 strain and LA18 strain have a synergistic promoting effect on the above effects.

[0072] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0073] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0074] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A probiotic for resisting Helicobacter pylori infection, characterized in that: The strains in the probiotic for preventing Helicobacter pylori infection are composed of Bifidobacterium animalis subsp. Lactis BLa80 strain with a preservation number of CGMCC No.15410 and Lactobacillus acidophilus LA18 strain with a preservation number of CCTCC NO: M2022572.

2. The probiotic agent for resisting Helicobacter pylori infection according to claim 1, characterized in that: The ratio of the number of viable bacteria of the BLa80 strain to that of the LA18 strain is 1:10-10:

1.

3. The probiotic agent for resisting Helicobacter pylori infection according to claim 1, characterized in that: The content of live bacteria in the probiotics is not less than 1×10 10 CFU / g or 1×10 10 CFU / mL.

4. The probiotic agent for resisting Helicobacter pylori infection according to claim 1, characterized in that: The dosage form of the probiotics includes solution, lyophilized powder, capsule, tablet or granule.

5. The probiotic agent for resisting Helicobacter pylori infection according to claim 4, characterized in that: The dosage form of the probiotic is a solution, which is prepared by the following method: The BLa80 strain and the LA18 strain are inoculated into a culture medium respectively for activation and fermentation culture in sequence to obtain a fermentation broth; the fermentation broth is centrifuged respectively and resuspended with a solvent to obtain a BLa80 bacterial suspension and a LA18 bacterial suspension; the BLa80 bacterial suspension and the LA18 bacterial suspension are mixed according to the ratio of the number of live bacteria to obtain the probiotic agent.

6. The probiotic agent for resisting Helicobacter pylori infection according to claim 4, characterized in that: The dosage form of the probiotic is a freeze-dried powder, which is prepared by the following method: The BLa80 strain and the LA18 strain are inoculated into a culture medium respectively for activation and fermentation culture in sequence to obtain a fermentation broth; the fermentation broth is centrifuged respectively, mixed with a protective agent and freeze-dried to obtain BLa80 bacterial powder and LA18 bacterial powder; the BLa80 bacterial powder and the LA18 bacterial powder are mixed according to the ratio of the number of live bacteria to obtain the probiotic agent.

7. The probiotic agent for resisting Helicobacter pylori infection according to claim 6, characterized in that: The protective agent is selected from any one of skim milk, gelatin, dextrin, gum arabic, dextran, sodium alginate, polyvinyl pyrrolidone, sucrose, lactose, trehalose, sorbitol or xylitol, or a combination of at least two thereof.

8. Use of the probiotic according to any one of claims 1 to 7 in the preparation of a drug for resisting Helicobacter pylori infection.

9. The use according to claim 8, characterized in that: The medicine also contains auxiliary materials.

10. The use according to claim 9, characterized in that: The auxiliary materials include any one of fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, colorants, pH regulators, antioxidants, antibacterial agents or buffers, or a combination of at least two of them.

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