Probiotic composition and its application in anti-Helicobacter pylori

Abstract

The present invention discloses a probiotic composition and its application in anti-Helicobacter pylori. In the present invention, Weissella coagulans powder, Lactobacillus rhamnosus powder, Lactobacillus gasseri powder, nano-molybdenum selenide and / or 3-indole-lactic acid are prepared into probiotic microcapsules by an endogenous emulsification method. Compared with the prior art, the endogenous emulsification method adopted in the present invention can effectively encapsulate probiotics inside the microcapsules, protecting the probiotics from the influence of the external environment; it can control the particle size and morphology of the microcapsules, making them have better stability and release characteristics. Moreover, the probiotic composition of the present invention has a significant inhibitory effect on urease.

Description

Technical Field

[0001] The present invention relates to the technical field of probiotics, and particularly relates to a probiotic composition and its application in anti-Helicobacter pylori. Background Art

[0002] Helicobacter pylori is a Gram-negative bacterium mainly living in the human stomach and duodenum. At present, antibiotics play a key role in the treatment of Helicobacter pylori. The anti-Helicobacter pylori treatment regimen usually uses a combination of a proton pump inhibitor, bismuth agent and two antibiotics, and common combinations include amoxicillin + clarithromycin, etc. Antibiotics mainly exert bactericidal effects by interfering with processes such as cell wall synthesis and protein synthesis of Helicobacter pylori. However, there are also disadvantages in the treatment of Helicobacter pylori with antibiotics. One is the problem of drug resistance. With extensive use, the drug resistance rate of Helicobacter pylori to some antibiotics gradually increases, and the emergence of drug-resistant strains increases the risk of treatment failure. The generation of drug resistance is mainly due to gene mutations, unreasonable use and gene transfer, etc. The second is adverse reactions, which may cause gastrointestinal discomfort, allergic reactions, abnormal liver function, etc., affecting the treatment compliance of patients and even leading to treatment interruption. The third is dysbacteriosis. Long-term or large-dose use of antibiotics will disrupt the normal flora balance in the human intestine, cause symptoms such as diarrhea, increase the risk of infection with other pathogenic bacteria, and may also affect the human immune system and metabolic function.

[0003] Probiotics may play an adjuvant therapeutic role through mechanisms such as regulating the intestinal flora, enhancing the intestinal mucosal barrier function, and inhibiting the growth and adhesion of Helicobacter pylori. After the treatment of Helicobacter pylori infection, it is easy to relapse. Probiotics can help maintain the balance of the intestinal flora and enhance the body's immunity, thereby reducing the recurrence rate of Helicobacter pylori infection.

[0004] CN109662321A discloses a composite probiotic microcapsule and its preparation method. The core material thereof contains composite probiotics (Bifidobacterium, Lactobacillus plantarum, Lactobacillus casei), and the wall material includes konjac glucomannan and sodium alginate. This composite wall material can effectively improve the protection effect on probiotics. CN115006432A discloses a composite probiotic microcapsule and its preparation method. This invention uses the endogenous emulsification method to prepare probiotic microcapsules with a composite probiotic containing Bifidobacterium, Lactobacillus plantarum, and Lactobacillus casei as the core material and konjac glucomannan and sodium alginate as the wall material. This method is simple to operate, low in cost, high in embedding rate and cost-effective, and the composite wall material can effectively improve the protection effect on probiotics. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide a method for preparing probiotic microcapsules by endogenous emulsification method using probiotics (Weizmannia coagulans powder, Lactobacillus rhamnosus powder, Lactobacillus gasseri powder), which can protect probiotics from the influence of the external environment and has a significant inhibitory effect on urease.

[0006] To achieve the above object, the present invention provides a method for preparing a probiotic composition, comprising the following steps:

[0007] S1 Add 3 - 6 g of sodium alginate, 10 - 20 g of calcium carbonate, 3 - 6 g of Weizmannia coagulans powder, 3 - 6 g of Lactobacillus rhamnosus powder, 3 - 6 g of Lactobacillus gasseri powder, and 6 - 12 g of nano - molybdenum selenide to 150 - 300 mL of sterile water, stir and mix for 20 - 50 min to obtain the aqueous phase;

[0008] S2 Add 4 - 8 mL of Tween 80 to 400 - 800 mL of soybean oil, stir and mix for 20 - 40 min to obtain the oil phase;

[0009] S3 Drop the aqueous phase into the oil phase according to a volume ratio of 1:2 - 4. After dropping, stir and mix for 10 - 25 min; add 1 - 3 mL of glacial acetic acid and stir for 20 - 40 min; add 400 - 600 mL of 0.1 - 0.2 mol / L acetate buffer solution with a pH of 5.5, stir at a speed of 20 - 60 rpm for 3 - 6 min; let stand for 1 - 3 h, take the lower - layer precipitate, wash it with absolute ethanol and sterile water respectively; freeze - dry to obtain the probiotic composition.

[0010] A preferred technical solution is that the method for preparing the probiotic composition comprises the following steps:

[0011] S1 Add 3 - 6 g of sodium alginate, 10 - 20 g of calcium carbonate, 3 - 6 g of Weizmannia coagulans powder, 3 - 6 g of Lactobacillus rhamnosus powder, 3 - 6 g of Lactobacillus gasseri powder, and 6 - 12 g of 3 - indole - lactic acid to 150 - 300 mL of sterile water, stir and mix for 20 - 50 min to obtain the aqueous phase;

[0012] S2 Add 4 - 8 mL of Tween 80 to 400 - 800 mL of soybean oil, stir and mix for 20 - 40 min to obtain the oil phase;

[0013] S3 Drop the aqueous phase into the oil phase according to a volume ratio of 1:2 - 4. After dropping, stir and mix for 10 - 25 min; add 1 - 3 mL of glacial acetic acid and stir for 20 - 40 min; add 400 - 600 mL of 0.1 - 0.2 mol / L acetate buffer solution with a pH of 5.5, stir at a speed of 20 - 60 rpm for 3 - 6 min; let stand for 1 - 3 h, take the lower - layer precipitate, wash it with absolute ethanol and sterile water respectively; freeze - dry to obtain the probiotic composition.

[0014] The most preferred technical solution, the preparation method of the probiotic composition, comprises the following steps:

[0015] S1 Add 3 - 6 g of sodium alginate, 10 - 20 g of calcium carbonate, 3 - 6 g of Weizmannia coagulans powder, 3 - 6 g of Lactobacillus rhamnosus powder, 3 - 6 g of Lactobacillus gasseri powder, 3 - 6 g of nano - molybdenum selenide, and 3 - 6 g of 3 - indole - lactic acid to 150 - 300 mL of sterile water, stir and mix for 20 - 50 min to obtain the aqueous phase;

[0016] S2 Add 4 - 8 mL of Tween 80 to 400 - 800 mL of soybean oil, stir and mix for 20 - 40 min to obtain the oil phase;

[0017] S3 Drop the aqueous phase into the oil phase at a volume ratio of 1:2 - 4. After dropping, stir and mix for 10 - 25 min; add 1 - 3 mL of glacial acetic acid and then stir for 20 - 40 min; add 400 - 600 mL of 0.1 - 0.2 mol / L acetate buffer solution with pH 5.5, stir at a speed of 20 - 60 rpm for 3 - 6 min; let it stand for 1 - 3 h, take the lower - layer precipitate, wash it with anhydrous ethanol and sterile water respectively; freeze - dry to obtain the probiotic composition.

[0018] Preferably, the Weizmannia coagulans powder is selected as Weizmannia coagulans BC99; the Lactobacillus rhamnosus powder is selected as Lactobacillus rhamnosus LRa05; the Lactobacillus gasseri powder is selected as Lactobacillus gasseri LG08.

[0019] Weizmannia coagulans BC99 has the characteristics of being resistant to high temperature and high pressure and can maintain its activity in a high - temperature environment. It can compete with Helicobacter pylori for living space and nutrients in the gastrointestinal tract, thereby reducing the colonization of Helicobacter pylori. For example, on the surface of the intestinal mucosa, Weizmannia coagulans occupies the binding sites, making it difficult for Helicobacter pylori to attach and reducing its chance of infection. On the other hand, Weizmannia coagulans BC99 may secrete some metabolites with antibacterial activity, which play an inhibitory role in the growth of Helicobacter pylori. These metabolites may include organic acids, bacteriocins, etc. They can damage the cell - membrane structure of Helicobacter pylori or interfere with its metabolic process, inhibiting its growth and reproduction.

[0020] Lactobacillus rhamnosus LRa05 can stimulate the intestinal immune system, promote the secretion of immune factors, enhance the barrier function of the intestinal mucosa, and thus reduce the invasion of Helicobacter pylori. On the other hand, Lactobacillus rhamnosus LRa05 helps maintain the dominant position of beneficial bacteria in the intestine and improve the intestinal microecological environment by regulating the activity of immune cells. A healthy intestinal flora can inhibit the growth of Helicobacter pylori and prevent its overgrowth. Moreover, it can also compete with Helicobacter pylori for adhesion sites on the intestinal mucosa, prevent the attachment of Helicobacter pylori, and thus reduce the risk of infection.

[0021] Lactobacillus gasseri LG08 can produce some antibacterial peptides or other antibacterial substances to directly inhibit the growth of Helicobacter pylori. These antibacterial substances have high specificity and activity, can specifically attack Helicobacter pylori, and have less impact on normal human cells and beneficial bacteria.

[0022] The endogenous emulsification method is a method for preparing microcapsules. Its basic principle is to utilize the two-phase or multi-phase existing in the system to form microcapsules through processes such as phase separation or coagulation. In the process of preparing probiotic microcapsules, the endogenous emulsification method usually disperses probiotics in one of the aqueous phase or oil phase, and then mixes it with the other phase to form an emulsion system by stirring or other means. In the emulsion system, the probiotics are encapsulated in tiny droplets, and these droplets can be further processed, such as solidification, drying, etc., to form microcapsules. The endogenous emulsification method can effectively encapsulate probiotics inside the microcapsules, protect the probiotics from the influence of the external environment, such as gastric acid, bile, etc. It can control the particle size and morphology of the microcapsules, making them have better stability and release characteristics.

[0023] Studies have shown that 3-indole-lactic acid can increase the richness and diversity of the intestinal flora, reduce the relative abundance of potential pathogenic bacteria, and promote the growth of beneficial bacteria such as Lactobacillus. The inventor found that 3-indole-lactic acid can also effectively inhibit the urease activity of Helicobacter pylori, thereby hindering the colonization of bacteria in the gastric epithelium or mucus layer.

[0024] Preferably, the preparation method of the nano-molybdenum selenide is as follows:

[0025] Disperse 1-3 g of sodium molybdate in 400-600 mL of an ethanol aqueous solution and stir for 10-20 min; add 40-60 mL of a hydrazine aqueous solution containing 1-2 g of selenium powder, stir for 20-40 min, then heat it in a sealed manner at 180-220 °C for 8-12 h, and naturally cool to room temperature; centrifuge at 15000-21000 rpm for 5-15 min, remove the supernatant and take the precipitate, and grind it after drying to obtain nano-molybdenum selenide. The volume ratio of ethanol to water in the ethanol aqueous solution is 1:1. The concentration of the hydrazine aqueous solution is 60 wt%.

[0026] Mo in nano-molybdenum selenide 4+ is oxidized to form MoO4 2- , which can interfere with the normal metabolic process of Helicobacter pylori and inhibit its growth and reproduction. Selenium in nano-molybdenum selenide can promote the growth of beneficial bacteria while inhibiting the growth of Helicobacter pylori. Importantly, compared with antibiotics, nano-molybdenum selenide can maintain the balance of the intestinal flora and have less adverse effects on the community structure of the intestinal flora.

[0027] Preferably, the freeze-drying parameters are: the pre-freezing temperature is set at -20 °C, and after the sample temperature drops to -20 °C, it is maintained for 1.5 h. The sublimation temperature is set at 12 °C, the desorption temperature is 34 °C, the vacuum degree is 25 Pa, and the drying time is 24 h.

[0028] Advantages of the present invention:

[0029] Compared with the prior art, the present invention adopts the endogenous emulsification method to effectively encapsulate probiotics inside the microcapsules, protecting the probiotics from the influence of the external environment. It can control the particle size and morphology of the microcapsules, making them have good stability and release characteristics. The probiotic composition of the present invention has a significant inhibitory effect on urease. Detailed implementation mode

[0030] Use the parameters and sources of specific chemical substances.

[0031] Weizmannia coagulans powder, in the embodiment, it is Weizmannia coagulans BC99, with a specification of 300 billion CFU / g, sourced from Microkang Probiotics (Suzhou) Co., Ltd.

[0032] Lactobacillus rhamnosus powder, in the embodiment, it is Lactobacillus rhamnosus LRa05, with a specification of 600 billion CFU / g, sourced from Microkang Probiotics (Suzhou) Co., Ltd.

[0033] Lactobacillus gasseri powder, in the embodiment, it is Lactobacillus gasseri LG08, with a specification of 500 billion CFU / g, sourced from Microkang Probiotics (Suzhou) Co., Ltd.

[0034] Example 1

[0035] A preparation method of a probiotic composition is as follows:

[0036] 4 g of sodium alginate, 16 g of calcium carbonate, 5 g of Weizmannia coagulans powder, 5 g of Lactobacillus rhamnosus powder, 5 g of Lactobacillus gasseri powder, and 10 g of nano-molybdenum selenide are added to 200 mL of sterile water and stirred and mixed for 30 min to obtain the aqueous phase;

[0037] 6 mL of Tween 80 is added to 600 mL of soybean oil and stirred and mixed for 30 min to obtain the oil phase;

[0038] The aqueous phase was added dropwise to the oil phase at a volume ratio of 1:3. After the addition was completed, the mixture was stirred at 200 rpm for 15 min; after adding 2 mL of glacial acetic acid, the mixture was stirred at 200 rpm for 30 min; 600 mL of 0.1 mol / L acetate buffer solution with a pH of 5.5 was added, and the mixture was stirred at 50 rpm for 5 min; left standing for 2 h, the lower layer precipitate was taken, washed 3 times with absolute ethanol, with 300 mL of absolute ethanol used each time, and then washed 3 times with sterile water, with 300 mL of sterile water used each time; freeze-dried to obtain the probiotic composition. The freeze-drying parameters were: the pre-freezing temperature was set at -20 °C, and after the sample temperature dropped to -20 °C, it was maintained for 1.5 h, the sublimation temperature was set at 12 °C, the desorption temperature was 34 °C, the vacuum degree was 25 Pa, and dried for 24 h.

[0039] The preparation method of the molybdenum diselenide nanosheets is as follows:

[0040] 2.1 g of sodium molybdate was dispersed in 500 mL of an ethanol aqueous solution (volume ratio 1:1), and stirred for 15 min; 50 mL of a hydrazine aqueous solution containing 1.6 g of selenium powder (containing 60 wt% hydrazine) was added, stirred for 30 min and then transferred to a stainless steel autoclave with a polytetrafluoroethylene lining, heated at 200 °C for 10 h, and naturally cooled to room temperature; centrifuged at 20000 rpm for 10 min, the supernatant was removed and the precipitate was taken, and after drying, it was ground to obtain molybdenum diselenide nanosheets.

[0041] Example 2

[0042] A preparation method of a probiotic composition is as follows:

[0043] 4 g of sodium alginate, 16 g of calcium carbonate, 5 g of Weizmannia coagulans powder, 5 g of Lactobacillus rhamnosus powder, 5 g of Lactobacillus gasseri powder, and 10 g of 3-indole-lactic acid were added to 200 mL of sterile water and stirred and mixed for 30 min to obtain the aqueous phase;

[0044] 6 mL of Tween 80 was added to 600 mL of soybean oil and stirred and mixed for 30 min to obtain the oil phase;

[0045] The aqueous phase was added dropwise to the oil phase at a volume ratio of 1:3. After the addition was completed, the mixture was stirred at a speed of 200 rpm for 15 min; 2 mL of glacial acetic acid was added and the mixture was stirred at a speed of 200 rpm for 30 min; 600 mL of 0.1 mol / L acetate buffer solution with a pH of 5.5 was added, and the mixture was stirred at a speed of 50 rpm for 5 min; it was allowed to stand for 2 h, the lower layer precipitate was taken, washed 3 times with anhydrous ethanol, with 300 mL of anhydrous ethanol used each time, and then washed 3 times with sterile water, with 300 mL of sterile water used each time; freeze-dried to obtain the probiotic composition. The freeze-drying parameters were: the pre-freezing temperature was set at -20°C, and after the sample temperature dropped to -20°C, it was maintained for 1.5 h. The sublimation temperature was set at 12°C, the desorption temperature was 34°C, the vacuum degree was 25 Pa, and the drying time was 24 h.

[0046] Example 3

[0047] A preparation method of a probiotic composition is as follows:

[0048] 4 g of sodium alginate, 16 g of calcium carbonate, 5 g of Weizmannia coagulans powder, 5 g of Lactobacillus rhamnosus powder, 5 g of Lactobacillus gasseri powder, 5 g of nano-molybdenum selenide, and 5 g of 3-indole-lactic acid were added to 200 mL of sterile water and stirred and mixed for 30 min to obtain the aqueous phase;

[0049] 6 mL of Tween 80 was added to 600 mL of soybean oil and stirred and mixed for 30 min to obtain the oil phase;

[0050] The aqueous phase was added dropwise to the oil phase at a volume ratio of 1:3. After the addition was completed, the mixture was stirred at a speed of 200 rpm for 15 min; 2 mL of glacial acetic acid was added and the mixture was stirred at a speed of 200 rpm for 30 min; 600 mL of 0.1 mol / L acetate buffer solution with a pH of 5.5 was added, and the mixture was stirred at a speed of 50 rpm for 5 min; it was allowed to stand for 2 h, the lower layer precipitate was taken, washed 3 times with anhydrous ethanol, with 300 mL of anhydrous ethanol used each time, and then washed 3 times with sterile water, with 300 mL of sterile water used each time; freeze-dried to obtain the probiotic composition. The freeze-drying parameters were: the pre-freezing temperature was set at -20°C, and after the sample temperature dropped to -20°C, it was maintained for 1.5 h. The sublimation temperature was set at 12°C, the desorption temperature was 34°C, the vacuum degree was 25 Pa, and the drying time was 24 h.

[0051] The preparation method of the nano-molybdenum selenide is as follows:

[0052] Disperse 2.1 g of sodium molybdate in 500 mL of an ethanol-water solution (volume ratio 1:1), and stir for 15 min; add 50 mL of a hydrazine aqueous solution (containing 60 wt% hydrazine) containing 1.6 g of selenium powder, stir for 30 min, then transfer to a stainless steel autoclave with a polytetrafluoroethylene liner, heat at 200 °C for 10 h, and naturally cool to room temperature; centrifuge at 20000 rpm for 10 min, discard the supernatant and take the precipitate, dry and grind to obtain molybdenum selenide nanoparticles.

[0053] Comparative Example 1

[0054] A preparation method of a probiotic composition is as follows:

[0055] Add 4 g of sodium alginate, 16 g of calcium carbonate, 5 g of Weizmannia coagulans powder, 5 g of Lactobacillus rhamnosus powder, and 5 g of Lactobacillus gasseri powder to 200 mL of sterile water, stir and mix for 30 min to obtain the aqueous phase;

[0056] Add 6 mL of Tween 80 to 600 mL of soybean oil, stir and mix for 30 min to obtain the oil phase;

[0057] Drop the aqueous phase into the oil phase according to a volume ratio of 1:3, after dropping, stir and mix for 15 min; add 2 mL of glacial acetic acid and stir for 30 min; add 600 mL of 0.1 mol / L acetate buffer solution with pH 5.5, stir at a speed of 50 rpm for 5 min; let stand for 2 h, take the lower layer precipitate, wash it 3 times with absolute ethanol, with 300 mL of absolute ethanol used each time, and then wash it 3 times with sterile water, with 300 mL of sterile water used each time; freeze-dry to obtain the probiotic composition. The freeze-drying parameters are: set the pre-freezing temperature to -20 °C, keep it for 1.5 h after the sample temperature drops to -20 °C, set the sublimation temperature to 12 °C, the desorption temperature to 34 °C, the vacuum degree to 25 Pa, and dry for 24 h.

[0058] Test Example 1

[0059] Helicobacter pylori antibacterial detection

[0060] (1) Bacterial solution preparation: Cultivate Helicobacter pylori to the logarithmic phase, take 5 mL of the bacterial solution, centrifuge at 5000 g for 5 min, resuspend the bacterial cells with TSB medium, and adjust the bacterial solution concentration to 1×10 6 CFU / mL;

[0061] (2) Take a 96-well plate, add 100 μL of the bacterial solution from the above step (1) to each well, 95 μL of TSB medium, and add 5 μL of the test solution respectively. The blank control is to add 5 μL of dimethyl sulfoxide. Each treatment is set with 3 replicates. Place the 96-well plate in a shaker at 37 °C and 150 rpm for 48 h; the preparation method of the test solution is as follows: Resuspend 1 g of the probiotic composition in the example and the control example with 100 mL of sterile water respectively.

[0062] (3) Use the OD 600 value to reflect the bacterial solution concentration, and the inhibition rate (%) is calculated according to the following formula: (OD 600 control bacterial solution - OD 600 treated bacterial solution) / OD 600 control bacterial solution × 100%.

[0063] Table 1

[0064] Bacteriostatic rate (%) Example 1 39.8 Example 2 43.9 Example 3 49.2 Control Example 1 28.7

[0065] As can be seen from the above table, the probiotic compositions in Examples 1-2 have significantly better inhibitory effects on Helicobacter pylori than those in Control Example 1. Obviously, adding 3-indole-lactic acid or molybdenum selenide nanoparticles during the preparation of probiotic microcapsules by the endogenous emulsification method can effectively improve the inhibitory effect of probiotic microcapsules on Helicobacter pylori. The possible reason is that: 3-indole-lactic acid has certain lipophilicity and may penetrate into the periplasmic space between the inner and outer membranes through the membrane surface proteins of Helicobacter pylori, resulting in damage to the integrity of the outer membrane and leakage of intracellular substances, thus achieving the inhibition of Helicobacter pylori. Mo 4+ in molybdenum selenide nanoparticles is oxidized to form MoO4 2- , which can interfere with the normal metabolic process of Helicobacter pylori and inhibit its growth and reproduction.

[0066] Test Example 2

[0067] Determination of the inhibitory activity of Helicobacter pylori urease

[0068] Turn on the ultraviolet light in the ultra-clean workbench for more than 30 min before the experiment starts, and then turn on the blower for 5 min before starting the experimental operation. Adjust the concentration of the activated Helicobacter pylori bacterial solution to 10 8 CFU / mL. Take 2 mL of the bacterial solution and add it to a screw-cap tube, add the test solution respectively, vortex for 5 min, and culture it microaerophilically (85% N2, 10% CO2, 5% O2) at 37 °C for 6 h. Then take 100 μL of each group of solutions into a 96-well plate, and add an equal amount of urea-phenol red reagent to initiate a biochemical reaction. After reacting for 30 min, put it into an enzyme-linked immunosorbent assay (ELISA) reader and measure its absorbance at 550 nm. Each experimental group is carried out with 3 parallel experiments.

[0069] Urease inhibition rate (%) = (Absorbance of control group - Absorbance of experimental group) / Absorbance of control group × 100%

[0070] Table 2

[0071] Urease inhibition rate (%) Example 1 45.9 Example 2 60.1 Example 3 69.3 Control Example 1 34.8

[0072] In Helicobacter pylori, urease is one of its important virulence factors. Helicobacter pylori can produce a large amount of urease, which can decompose urea in the stomach to produce ammonia, thereby neutralizing gastric acid and creating a suitable microenvironment for the survival of bacteria in the stomach. In addition, urease is also involved in the adhesion and colonization process of Helicobacter pylori.

[0073] As can be seen from the above table, the urease inhibition effect of the probiotic composition in Example 2 is significantly better than that in Example 1 and Control Example 1. Obviously, adding 3-indole-lactic acid during the preparation of probiotic microcapsules by the endogenous emulsification method has a significantly better urease inhibition rate than adding molybdenum selenide nanoparticles or not adding. The possible reason is that 3-indole-lactic acid can cause deprotonation of acidic functional groups (such as phosphate groups) on the cell wall and cell membrane of Helicobacter pylori, increasing the negative charge on the bacterial surface. This will affect the stability of the cell membrane and reduce the interaction between the cell membrane and the cell wall, thereby interfering with the normal function of urease. It may also be that 3-indole-lactic acid can interfere with the metabolic pathway of Helicobacter pylori, affecting the synthesis or regulation of urease, resulting in a decrease in urease activity.

Claims

1. Use of a probiotic composition in the preparation of a drug for anti-Helicobacter pylori and inhibiting the urease activity of Helicobacter pylori. The preparation method of the probiotic composition comprises the following steps: S1: Add 3 - 6 g of sodium alginate, 10 - 20 g of calcium carbonate, 3 - 6 g of Weizmannia coagulans powder, 3 - 6 g of Lactobacillus rhamnosus powder, 3 - 6 g of Lactobacillus gasseri powder, 3 - 6 g of nano-molybdenum selenide, and 3 - 6 g of 3-indole-lactic acid to 150 - 300 mL of sterile water, stir and mix for 20 - 50 min to obtain an aqueous phase; the Weizmannia coagulans powder is selected as Weizmannia coagulans BC99; the Lactobacillus rhamnosus powder is selected as Lactobacillus rhamnosus LRa05; the Lactobacillus gasseri powder is selected as Lactobacillus gasseri LG08; S2: Add 4 - 8 mL of Tween 80 to 400 - 800 mL of soybean oil, stir and mix for 20 - 40 min to obtain an oil phase; S3: Drop the aqueous phase into the oil phase according to a volume ratio of 1:2 - 4. After dropping, stir and mix for 10 - 25 min; add 1 - 3 mL of glacial acetic acid and then stir for 20 - 40 min; add 400 - 600 mL of 0.1 - 0.2 mol / L acetate buffer solution with pH 5.5, stir at a speed of 20 - 60 rpm for 3 - 6 min; let it stand for 1 - 3 h, take the lower layer precipitate, wash it with absolute ethanol and sterile water respectively; freeze-dry to obtain the probiotic composition; The preparation method of the nano-molybdenum selenide is as follows: Disperse 1 - 3 g of sodium molybdate in 400 - 600 mL of ethanol aqueous solution, stir for 10 - 20 min; add 40 - 60 mL of hydrazine aqueous solution containing 1 - 2 g of selenium powder, stir for 20 - 40 min, then heat it in a closed state at 180 - 220 °C for 8 - 12 h, and naturally cool to room temperature; centrifuge at 15000 - 21000 rpm for 5 - 15 min, remove the supernatant and take the precipitate, dry it and then grind it to obtain nano-molybdenum selenide.

2. Use of the probiotic composition according to claim 1 in the preparation of a medicament for anti-Helicobacter pylori and inhibiting the urease activity of Helicobacter pylori, characterized in that: The volume ratio of ethanol to water in the ethanol aqueous solution is 1:

1.

3. Use of the probiotic composition according to claim 1 in the preparation of a medicament for anti-Helicobacter pylori and inhibiting the urease activity of Helicobacter pylori, characterized in that: The concentration of the hydrazine aqueous solution is 60 wt%.

4. Use of the probiotic composition according to claim 1 in the preparation of a medicament for anti-Helicobacter pylori and inhibiting the urease activity of Helicobacter pylori, characterized in that, The freeze-drying parameters are: set the pre-freezing temperature to -20 °C, keep it for 1.5 h after the sample temperature drops to -20 °C, set the sublimation temperature to 12 °C, the analysis temperature to 34 °C, the vacuum degree to 25 Pa, and dry for 24 h.