Probiotic solid composition with helicobacter pylori inhibitory activity and preparation method thereof

Abstract

The invention relates to the technical field of biology, and particularly discloses a probiotic solid composition with helicobacter pylori inhibitory activity and a preparation method thereof, the probiotic solid composition comprises 50-70 parts of probiotic freeze-dried powder, 1-10 parts of tremella polysaccharide and 20-40 parts of fructo-oligosaccharide; the strains have a synergistic effect in the aspects of thallus surface adhesion, target cell site competition and the like, the intestinal microenvironment can be adjusted through short-chain fatty acid, natural antibacterial substances and other metabolites secreted by the strains, the complementary antibacterial effect is achieved, and the multiple mechanisms are synergistic, so that the effect of improving the antibacterial effect is achieved. The probiotic composition can weaken or compete for the growth environment of helicobacter pylori more comprehensively in vivo, so that the colonization probability of helicobacter pylori is reduced, the probiotic composition can improve the intestinal flora imbalance state of a host, inflammatory response is weakened by regulating and controlling the immune function of the intestinal tract, meanwhile, proliferation of pathogenic bacteria is inhibited, and the effect of inhibiting helicobacter pylori is achieved. Therefore, the resistance of the host to helicobacter pylori infection is improved on the whole.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology, and particularly relates to a probiotic solid composition with Helicobacter pylori inhibitory activity and a preparation method thereof. Background Art

[0002] Helicobacter pylori (H. pylori) infection is one of the key factors causing gastrointestinal diseases such as chronic gastritis, peptic ulcer and gastric cancer. At present, the clinical treatment of H. pylori infection mainly relies on antibiotic therapy. However, long-term or improper use of antibiotics is likely to lead to the emergence of drug-resistant strains, reducing the treatment effect, and at the same time may cause side effects such as intestinal flora imbalance and gastrointestinal dysfunction, bringing additional health risks to patients.

[0003] With the continuous in-depth study of the relationship between intestinal microecology and human health, probiotics, as a safe and natural biological regulator, have gradually attracted attention for their potential in inhibiting H. pylori infection. In the prior art, some probiotic products have been tried for adjuvant treatment of H. pylori infection, but these products generally have the following deficiencies:

[0004] Single strain selection or lack of synergistic effect: Many probiotic products contain only a single strain or a few strains. The mechanism of action of these strains in inhibiting H. pylori is relatively single, and it is difficult to form an effective synergistic antibacterial effect. There may be potential synergistic effects among different strains in aspects such as adhesion on the cell surface, competition for target cell sites, and secretion of antibacterial substances, but the prior art has not fully utilized these synergistic effects, resulting in limited overall antibacterial effect.

[0005] Lack of comprehensive regulation of the intestinal microenvironment: The role of probiotics in the intestine is not limited to directly inhibiting H. pylori, but also includes regulating the intestinal microenvironment, enhancing the intestinal barrier function, and regulating intestinal immunity. However, the probiotic products in the prior art often ignore these comprehensive regulatory effects and only focus on the direct inhibitory effect of probiotics on H. pylori, thus limiting their effect in preventing and treating H. pylori infection.

[0006] Low probiotic activity retention rate: The activity of probiotics is the key to their antibacterial effect. However, during the preparation, storage and transportation processes, probiotics are easily inactivated by external conditions such as humidity, temperature and oxygen. The probiotic products in the prior art often lack effective protection measures, resulting in a low probiotic activity retention rate and making it difficult to exert their expected antibacterial and regulatory effects when reaching the intestinal site.

[0007] Lack of natural protective layer: When probiotics pass through adverse digestive environments such as gastric acid and bile, they are easily damaged and inactivated. Probiotic products in the prior art often lack a natural protective layer to protect probiotics from these adverse environments, thereby reducing the survival rate and activity of probiotics in the intestine.

[0008] In view of the deficiencies of the prior art, the present invention provides a probiotic solid composition with Helicobacter pylori inhibitory activity and a preparation method thereof. The composition scientifically proportions a variety of probiotic strains and utilizes their synergistic effects to enhance the antibacterial effect; at the same time, by adding prebiotics and natural polysaccharides to construct a matrix embedding system, a natural protective layer is provided for the probiotics, and the activity retention rate of the probiotics during preparation, storage and transportation is improved; in addition, the preparation method also pays attention to the uniform release and stability of the probiotics in the intestine, so as to ensure that the probiotics can play their expected antibacterial and regulatory roles when reaching the intestinal site.

[0009] In response to this, the inventor proposes a probiotic solid composition with Helicobacter pylori inhibitory activity and a preparation method thereof to solve the above problems. Summary of the Invention

[0010] The purpose of the present invention is to provide a probiotic solid composition with Helicobacter pylori inhibitory activity and a preparation method thereof to solve the problems raised in the above background technology.

[0011] To achieve the above purpose, the present invention provides the following technical solutions:

[0012] A probiotic solid composition with Helicobacter pylori inhibitory activity, comprising:

[0013] 50-70 parts of probiotic freeze-dried powder, 1-10 parts of tremella polysaccharide, 20-40 parts of fructooligosaccharide;

[0014] Wherein the probiotic freeze-dried powder is composed of Lactobacillus plantarum, Bifidobacterium longum subsp. longum, Lactobacillus rhamnosus, Lactobacillus acidophilus, Weissella confusa, Lactobacillus mucosae, Lactobacillus crispatus, Lactobacillus casei, Lactobacillus paracasei, Bifidobacterium breve, Bifidobacterium animalis subsp. lactis, and Lactobacillus salivarius.

[0015] Preferably, the probiotic freeze-dried powder is composed of the following mass percentages:

[0016] Lactobacillus plantarum is LP01, LP-ONLLY, 6595, 299V, LP45: 20%–35%

[0017] Lactobacillus rhamnosus is GG, 6594, HN001, JYLR-127: 5%–20%

[0018] Lactobacillus acidophilus is LA11-Onlly and NCFM: 1% - 10%

[0019] Bifidobacterium longum subsp. longum is W11 and BB536: 1% - 10%

[0020] Lactobacillus mucosae is DSM17648: 5% - 15%

[0021] Weizmannia coagulans is MTCC 5856 and GBI-30: 5% - 15%

[0022] Lactobacillus paracasei is JLPF-176 and NTU 101: 5% - 15%

[0023] Lactobacillus casei is L.Casei 21: 1% - 10%

[0024] Lactobacillus crispatus is LCR01: 1% - 10%

[0025] Bifidobacterium animalis subsp. lactis is Bi-07 and HN019 and BB-12 and JYBR-190: 5% - 20%

[0026] Bifidobacterium breve is BR03 and M-16V: 1% - 10%

[0027] Lactobacillus salivarius is AP-32: 1% - 10%.

[0028] Preferably, the proportions of LP01, LP-ONLLY, 6595, 299V, and LP45 in the Lactobacillus plantarum are 10%, 11%, 5%, 14%, and 60% respectively;

[0029] The proportions of GG, 6594, HN001, and JYLR-127 in the Lactobacillus rhamnosus are 30%, 10%, 10%, and 50% respectively;

[0030] The proportions of LA11-Onlly and NCFM in the Lactobacillus acidophilus are 70% and 30% respectively;

[0031] The proportions of W11 and BB536 in the Bifidobacterium longum subsp. longum are 88.89% and 11.11% respectively;

[0032] The proportions of MTCC 5856 and GBI-30 in the Weizmannia coagulans are 87.5% and 12.5% respectively;

[0033] The proportions of JLPF-176 and NTU 101 in the Lactobacillus paracasei are 90% and 10% respectively;

[0034] The proportions of Bifidobacterium animalis subsp. lactis Bi-07, HN019, BB-12, and JYBR-190 are 10%, 5%, 5%, and 80% respectively;

[0035] The proportions of Bifidobacterium breve BR03 and M-16V are 10% and 90% respectively.

[0036] A method for preparing a probiotic solid composition with Helicobacter pylori inhibitory activity, comprising the following steps:

[0037] S1. Probiotic pretreatment, used to mix various probiotic powders in a preset ratio to obtain a probiotic premix;

[0038] Used to prevent the inactivation of bacterial strains due to humidity or heat treatment and ensure the active effect of the final product;

[0039] S2. Matrix formulation treatment, used to compound and mix prebiotics with natural polysaccharides to form a probiotic carrier system and obtain a premixed matrix;

[0040] Used to improve the adhesion and stability of probiotics and provide the nutritional basis required for their proliferation at the same time;

[0041] S3. Composite embedding treatment, used to slowly mix the probiotic premix into the premixed matrix so that the probiotics form an embedded structure in the matrix system to obtain a composite;

[0042] Utilize the adhesion characteristics of tremella polysaccharide and the synergistic effect of prebiotics to form a natural protective layer and improve the survival rate of bacterial strains in the gastric environment;

[0043] S4. Low-temperature drying treatment, used to perform low-temperature drying and micronization treatment on the composite to obtain a probiotic solid powder;

[0044] Avoid the damage to the bacterial activity caused by traditional high-temperature drying and improve the biological activity retention rate of the product;

[0045] S5. Aging and stabilization treatment, used to perform a constant-temperature static treatment on the probiotic solid powder to enhance the stability of the composite structure and obtain an aged powder;

[0046] Through the static aging process, improve the stability of active probiotics in the powder and the uniformity of later release;

[0047] S6. Subpackaging and forming treatment, used to package the aged powder according to the dose to obtain a probiotic solid composition with Helicobacter pylori inhibitory activity;

[0048] Automatically subpackage the probiotic solid composition according to the dose of 1–3 g per bag to complete the final finished product of the solid probiotic composite product.

[0049] Preferably, the mixing process of the probiotic premix is carried out at a low temperature below 4°C and stored under vacuum drying conditions to ensure the viability of the bacteria.

[0050] Preferably, in step S3, the composite embedding treatment is carried out with slow stirring for more than 15 minutes at a temperature below 25°C, so that the probiotics are evenly distributed in the natural viscous network formed by tremella polysaccharide and fructooligosaccharide and form a primary embedding protective layer.

[0051] Preferably, in step S4, the low-temperature drying treatment step is carried out by fluidized bed drying method at a temperature below 35°C to ensure that the viability of the probiotics is not damaged by heat, and powders with an average particle size less than 200 microns are obtained through micronization treatment.

[0052] Preferably, in step S5, the ripening and stabilizing treatment step is to store it in a closed environment with a relative humidity below 30% and a temperature of 25°C for 12% - 24 hours to improve the product stability and extend the shelf life.

[0053] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0054] (1) In the present invention, not only is there a synergistic effect among the strains in terms of adhesion on the cell surface and competition for target cell sites, but they can also regulate the intestinal microenvironment through the short-chain fatty acids, natural antibacterial substances and other metabolites secreted by themselves, exerting a complementary antibacterial effect. The synergy of these multiple mechanisms can more comprehensively weaken or compete for the growth environment of Helicobacter pylori in vivo, thereby reducing its colonization probability.

[0055] (2) The probiotic combination in the present invention can improve the disordered state of the host's intestinal flora, weaken the inflammatory response by regulating the intestinal immune function, and at the same time inhibit the proliferation of pathogenic bacteria, thereby improving the host's resistance to Helicobacter pylori infection as a whole. This regulation not only helps to prevent gastrointestinal diseases such as chronic gastritis and ulcers caused by Helicobacter pylori, but also can reduce the risk of serious complications such as gastric cancer to a certain extent.

[0056] (3) In the present invention, the matrix embedding system constructed by tremella polysaccharide and fructooligosaccharide can encapsulate the probiotics in the natural polymer network. This protective layer plays a barrier role during the preparation and storage processes, effectively preventing the damage of moisture, oxygen and external adverse conditions to the viability of the bacteria. This process enables the probiotics to remain viable when passing through adverse digestive environments such as gastric acid and bile, so as to exert their expected antibacterial and regulatory effects when reaching the intestinal site. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] Figure 1 It is a flowchart of the preparation method of the probiotic solid composition with Helicobacter pylori inhibitory activity of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0058] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0059] Example 1:

[0060] Please refer to Figure 1 as shown in the application of the multi-strain solid composition combined with static aging process in a preclinical model and the evaluation of its antibacterial effect:

[0061] 1. Formulation composition and process flow:

[0062] 1.1 Formulation composition (by mass parts):

[0063] The proportions of LP01, LP-ONLLY, 6595, 299V, and LP45 in the Lactobacillus plantarum are 10%, 11%, 5%, 14%, and 60% respectively;

[0064] The proportions of GG, 6594, HN001, and JYLR-127 in the Lactobacillus rhamnosus are 30%, 10%, 10%, and 50% respectively;

[0065] The proportions of LA11-Onlly and NCFM in the Lactobacillus acidophilus are 70% and 30% respectively;

[0066] The proportions of W11 and BB536 in the Bifidobacterium longum subsp. longum are 88.89% and 11.11% respectively;

[0067] The proportions of MTCC 5856 and GBI-30 in the Weissella cibaria are 87.5% and 12.5% respectively;

[0068] The proportions of JLPF-176 and NTU 101 in the Lactobacillus paracasei are 90% and 10% respectively;

[0069] The proportions of Bi-07, HN019, BB-12, and JYBR-190 in the Bifidobacterium animalis subsp. lactis are 10%, 5%, 5%, and 80% respectively;

[0070] The proportions of BR03 and M-16V in the Bifidobacterium breve are 10% and 90% respectively:

[0071] In this example, the strain proportions are further adjusted. As shown in Table 1 below, the material composition is as follows:

[0072] Table 1

[0073]

[0074]

[0075] 1.2 Preparation process steps:

[0076] Step 1: Pre-treatment of bacterial powder. Under low temperature (<4°C) conditions, freeze-dried powders of 12 probiotic strains are mixed according to the above ratio to obtain a premixed probiotic mixture. The mixing time is 8 minutes, and a stainless-steel drying mixer is used throughout.

[0077] Step 2: Matrix premixing and dissolution. Tremella polysaccharide and fructooligosaccharide are mixed at room temperature, and a small amount of purified water (not exceeding 5% of the total matrix) is added to assist dissolution. After stirring for 5 minutes (at a rotation speed of 300 rpm), a premixed matrix with good fluidity is obtained.

[0078] Step 3: Embedding and compounding. The premixed bacterial powder is slowly added to the premixed matrix in batches, and a stirrer is used for low-speed stirring (at a rotation speed of 200 rpm) for more than 15 minutes to form a compound embedding product. This process further ensures that the bacterial cells are evenly distributed and fully wrapped by the matrix.

[0079] Step 4: Low-temperature drying and micronization. The compound is fed into a fluidized bed low-temperature drying system (temperature controlled at 32°C, duration 25 minutes) and micronized to obtain a preliminarily dried powder. Parameters: drying air temperature 32°C, wind speed 1.0 m / s, moisture content controlled at ≤5%.

[0080] Step 5: Static ripening. The powder is sealed and stored in an environment with constant temperature (25°C) and low humidity (relative humidity ≤30%) for 24 hours to allow the bacterial cells and the matrix to fully interact and form a stable solid composition.

[0081] Step 6: Finished product packaging and viability detection. Each bag is automatically filled with 1.5 g, and finally, the total viable count in the product is maintained at 1×10 9 CFU / g or more.

[0082] Example 2:

[0083] Five-strain probiotic solid composition and its in vitro antibacterial effect test:

[0084] 1. Formulation composition and preparation:

[0085] 1.1 Formulation composition (by mass parts):

[0086] This example uses the probiotic strains and excipients shown in Table 2 below, and all raw materials are of pharmaceutical grade or food grade:

[0087] Table 2

[0088]

[0089]

[0090] The ratio of probiotics to excipients is 70:30 (ratio 7:3) to ensure the coordination of the bacterial activity and the nutrient matrix.

[0091] 1.2 Preparation process steps:

[0092] Step 1: Pretreatment of probiotics. Weigh and mix the freeze-dried powders of each strain in proportion in a dry environment below 4°C to obtain a premixed probiotic mixture. The mixing time is 5 minutes, and a low-speed rotating mixer is used.

[0093] Step 2: Premixing treatment of the matrix. Mix tremella polysaccharide and fructooligosaccharide in proportion at room temperature (20°C ± 2°C) and stir at 400 rpm for 10 minutes to make them evenly combined, obtaining a premixed matrix.

[0094] Step 3: Composite embedding. Slowly add the premixed probiotic mixture to the premixed matrix and slowly stir at a low speed (200 rpm) below 25°C for 15 minutes to fully embed the probiotics in the matrix, forming a composite. Utilize the adhesion of tremella polysaccharide and the nutritional effect of fructooligosaccharide to initially embed the bacteria and improve their tolerance to the acidic environment.

[0095] Step 4: Low-temperature drying and granulation. Feed the composite into a fluidized bed drying device for low-temperature drying (temperature controlled below 35°C, air humidity below 10%) for 20 minutes, and at the same time, perform micronization treatment to obtain a solid powder.

[0096] Step 5: Ripening and stabilization. Store the powder in a sealed environment with a relative humidity below 30% and a temperature of 25°C for 12 hours to allow the components in the composite to fully combine, obtaining a stable solid composition.

[0097] Step 6: Automatic packaging and final inspection. Perform quantitative packaging according to the single dose (1 g / bag) using an automatic packaging device. Detect the total viable count in the final product to ensure it is not less than 1×10 9 CFU / g.

[0098] 2. Hp antibacterial effect test:

[0099] 2.1 Experimental scheme design:

[0100] Adopt the in vitro antibacterial experiment method to compare the inhibitory effects of the solid composition in this example, the blank control (PBS), and the conventional probiotic freeze-dried powder (unembedded mixed bacteria) on Hp.

[0101] 2.2 Experimental conditions

[0102] Cultivation of bacterial strains:

[0103] Helicobacter pylori was collected from a 24-hour fresh slant culture, eluted with sterile PBS, and diluted to 5.0×10 5 CFU / mL to 4.5×10 6 CFU / mL. Control conditions: The pH was controlled at 6.8, and the temperature was maintained at 37°C ± 1°C.

[0104] Reaction steps:

[0105] ① First, add 5.0 mL of sample solution (A: suspension obtained by dissolving the prepared solid composition in water or diluting according to the instructions; B: lyophilized suspension of conventional probiotics; C: PBS blank control) into a sterile test tube.

[0106] ② Pretreat the test tube in a water bath at 20°C ± 1°C for 5 min.

[0107] ③ Quickly add 0.1 mL of Hp bacterial suspension (at the above concentration), and mix thoroughly immediately and start timing.

[0108] ④ After the specified reaction time (30 minutes), take 1.0 mL of the mixed solution from each test tube, make 10-fold serial dilutions, and then inoculate on a solid medium, and count the colonies by the plate method.

[0109] ⑤ All tests were carried out after culturing at 36°C ± 1°C for 48 hours for colony counting.

[0110] ⑥ Each group was repeated 3 times, record the average number of colonies, and calculate the antibacterial rate.

[0111] 2.3 Explanation of data and results of the comparative example:

[0112] The following Table 2 shows the colony counts obtained from the experiment and the calculated antibacterial rates, where:

[0113] (Antibacterial rate = [(number of colonies in the control - number of colonies in the treatment group) / number of colonies in the control] × 100%)

[0114] Table 2

[0115] Experimental group Average number of colonies (CFU / mL) Standard deviation Bacteriostatic rate (%) A (this example group) <![CDATA[2.0×10 4 > <![CDATA[±1.0×10 3 > 80.0 B (conventional probiotic group) <![CDATA[6.5×10 4 > <![CDATA[±2.0×10 3 > 30.0 C (PBS control group) <![CDATA[1.0×10 5 > <![CDATA[±3.0×10 3 > ——

[0116] Data acquisition method:

[0117] The plate counting method was used, and a digital counter was used to record the number of colonies. Each test used an electronic temperature controller and a digital water bath to ensure accurate control of temperature and time; data processing was performed using Excel statistical analysis, and the mean and standard deviation were the results of independent repeated experiments (n = 3).

[0118] As can be seen from the above, in the present embodiment group (A), due to the adoption of the low-temperature embedding and composite protection process, probiotics can more effectively release natural antibacterial substances under the condition of co-culturing with Hp, thereby significantly reducing the colony count. The antibacterial rate is as high as 80%, showing obvious advantages compared with the conventional probiotic treatment (the antibacterial rate is only 30%), and at the same time, the PBS control has no antibacterial effect. This result fully proves the technical effect of the present solution;

[0119] In the present solution, a variety of probiotic strains that have been verified to have an inhibitory effect on Helicobacter pylori are selected, such as Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus reuteri, etc. There are synergistic effects among the strains not only in terms of adhesion on the cell surface and competition for target cell sites, but also can regulate the intestinal microenvironment through the short-chain fatty acids, natural antibacterial substances and other metabolites secreted by themselves, and play complementary antibacterial effects. The synergy of such multiple mechanisms can more comprehensively weaken or compete with the growth environment of Helicobacter pylori in vivo, thereby reducing its colonization probability.

[0120] 3. Antibacterial effect of the preclinical model:

[0121] The present invention adopts the detection experiment of Helicobacter pylori urease activity in an in vitro simulated gastric mucosa model ( 13 C-urea simulation method)

[0122] to construct a simulated gastric mucosa model infected with Helicobacter pylori, and through 13 the rate change of 13 CO2 released by the C-labeled urease-catalyzed reaction, quantitatively evaluate the effect of the probiotic solid composition on the urease activity and metabolic ability of Helicobacter pylori, so as to indirectly reflect the anti-Hp activity of the probiotics.

[0123] The materials and equipment are shown in Table 3 below

[0124] Table 3

[0125]

[0126]

[0127] Experimental steps

[0128] I. Model construction

[0129] Seed gastric epithelial cells in Matrigel on the bottom of a 6-well plate and culture until a monolayer structure is formed;

[0130] Inoculate the Hp strain onto the cell surface (MOI = 100:1) and culture for 24 hours;

[0131] Wash off the non-adherent bacteria with PBS to prepare a simulated infection model.

[0132] II. The grouping of reagent treatments is shown in Table 4 below

[0133] Table 4

[0134]

[0135] Continue culturing for 2 hours after treatment.

[0136] III. Add 13 C-labeled urea and seal the reaction

[0137] Add 13 C-labeled urea with a final concentration of 10 mmol / L to each well;

[0138] Seal the reaction system and connect it to a gas collection device;

[0139] React at 37 °C for 60 minutes and collect gas samples every 10 minutes;

[0140] Use an infrared CO2 detector to detect the total CO2 released, or measure the change in 13CO2 concentration with a mass spectrometer; calculate the release rate and cumulative amount as an indirect indicator of urease activity.

[0141] IV. Data recording and calculation method

[0142] Unit of CO2 release rate: μmol / min

[0143] Inhibition rate = [(release amount in blank group - release amount in treatment group) / release amount in blank group] × 100%. The fitting result data table (the experiment was repeated 3 times and the results were averaged) is shown in Table 5 below

[0144] Table 5

[0145]

[0146] The calculation results of enzyme activity inhibition rate are shown in Table 6 below

[0147] Table 6

[0148] Group Total release amount in 60 min (μmol) Inhibition rate (relative to group C) Group A (Example 1) 11.0 42.7％ Group B (Example 2) 14.5 24.5％ Group C (PBS control) 19.2 0

[0149] Explanation of experimental results

[0150] Group A (the probiotic composition of Example 1 of the present invention) significantly reduced the Hp urease activity in the in vitro simulated environment, 13 and the CO2 release rate was significantly lower than that of the blank control group;

[0151] Compared with Group B (the probiotic composition of Example 2 of the present invention), the urease inhibitory effect of the combination in Example 1 of the present invention was stronger, as shown by a lower 13 CO2 cumulative release amount;

[0152] It shows that it exerts a comprehensive inhibitory effect through mechanisms such as competitive inhibition, enzyme activity interference, or disruption of the Hp metabolic chain.

[0153] As can be seen from the above, the present invention adopts a stable embedding process, low-temperature drying and ripening treatment to make the product release more uniform active antibacterial substances, and has a significant antibacterial effect on Hp;

[0154] The probiotic combination can improve the dysbiosis state of the host's intestinal flora. By regulating the intestinal immune function, weakening the inflammatory response, and at the same time inhibiting the proliferation of pathogenic bacteria, it can overall improve the host's resistance to Helicobacter pylori infection. This regulation not only helps to prevent gastrointestinal diseases such as chronic gastritis and ulcers caused by Helicobacter pylori, but also can reduce the risk of serious complications such as gastric cancer to a certain extent;

[0155] The matrix embedding system constructed by tremella polysaccharide and fructooligosaccharide can encapsulate probiotics in the natural polymer network. This protective layer plays a barrier role during the preparation and storage processes, effectively preventing the destruction of the bacterial activity by moisture, oxygen and external adverse conditions. In theory, this process can enable probiotics to remain active when passing through adverse digestive environments such as gastric acid and bile, so as to play their expected antibacterial and regulatory roles when reaching the intestinal site;

[0156] The active bacteria released by the probiotic solid composition can competitively occupy the adhesion sites of the target cells of Helicobacter pylori. This competitive effect not only reduces the contact opportunity between the pathogenic bacteria and the gastric mucosa, but also further inhibits the colonization and amplification of Helicobacter pylori by locally releasing natural antibacterial factors after adhesion;

[0157] During the coexistence with Helicobacter pylori, the short-chain fatty acids and other antibacterial factors secreted by probiotics can directly act on the cell wall and membrane structure of the pathogenic bacteria, interfering with its metabolism. This effect reduces the proliferation ability of Helicobacter pylori from the source and is of great significance for maintaining the balance of the gastrointestinal flora.

[0158] In the description of this specification, the description with reference to terms such as "one embodiment", "some embodiments", "example", "specific example" or "some examples" means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in a suitable manner in any one or more embodiments or examples. In addition, without contradiction, those skilled in the art can combine and combine the different embodiments or examples described in this specification and the features of different embodiments or examples.

[0159] In the accompanying drawings of the disclosed embodiments of the present invention, only the structures related to the disclosed embodiments are involved. For other structures, reference may be made to the general design. Without conflict, the same embodiment and different embodiments of the present invention may be combined with each other.

[0160] Although the embodiments of the present invention have been shown and described, those of ordinary skill in the art can understand that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A probiotic solid composition having Helicobacter pylori inhibitory activity, characterized in that, Comprising: 50 - 70 parts of probiotic freeze-dried powder, 1 - 10 parts of tremella polysaccharide, 20 - 40 parts of fructooligosaccharide; Wherein the probiotic freeze-dried powder is composed of Lactobacillus plantarum, Bifidobacterium longum subsp. longum, Lactobacillus rhamnosus, Lactobacillus acidophilus, Weizmannia coagulans, Lactobacillus mucosae, Lactobacillus crispatus, Lactobacillus casei, Lactobacillus paracasei, Bifidobacterium breve, Bifidobacterium animalis subsp. lactis, and Lactobacillus salivarius.

2. The probiotic solid composition with Helicobacter pylori inhibitory activity according to claim 1, characterized in that, The probiotic freeze-dried powder is composed by mass percentage as follows: Lactobacillus plantarum includes LP01, LP - ONLLY, 6595, 299V, and LP45: 20% - 35% Lactobacillus rhamnosus includes GG, 6594, HN001, and JYLR - 127: 5% - 20% Lactobacillus acidophilus includes LA11 - Onlly and NCFM: 1% - 10% Bifidobacterium longum subsp. longum includes W11 and BB536: 1% - 10% Lactobacillus mucosae includes DSM17648: 5% - 15% Weizmannia coagulans includes MTCC 5856 and GBI - 30: 5% - 15% Lactobacillus paracasei includes JLPF - 176 and NTU 101: 5% - 15% Lactobacillus casei includes L.Casei 21: 1% - 10% Lactobacillus crispatus includes LCR01: 1% - 10% Bifidobacterium animalis subsp. lactis includes Bi - 07, HN019, BB - 12, and JYBR - 190: 5% - 20% Bifidobacterium breve includes BR03 and M - 16V: 1% - 10% Lactobacillus salivarius includes AP - 32: 1% - 10%.

3. The probiotic solid composition with Helicobacter pylori inhibitory activity according to claim 2, characterized in that, The proportions of LP01, LP - ONLLY, 6595, 299V, and LP45 in the Lactobacillus plantarum are respectively: 10%, 11%, 5%, 14%, and 60%; The proportions of GG, 6594, HN001, and JYLR - 127 in the Lactobacillus rhamnosus are respectively: 30%, 10%, 10%, and 50%; The proportions of LA11 - Onlly and NCFM in the Lactobacillus acidophilus are respectively: 70% and 30%; The proportions of W11 and BB536 in the Bifidobacterium longum subsp. longum are respectively: 88.89% and 11.11%; The proportions of MTCC 5856 and GBI - 30 in the Weizmannia coagulans are respectively: 87.5% and 12.5%; The proportions of JLPF - 176 and NTU 101 in the Lactobacillus paracasei are respectively: 90% and 10%; The proportions of Bi - 07, HN019, BB - 12, and JYBR - 190 in the Bifidobacterium animalis subsp. lactis are respectively 10%, 5%, 5%, and 80%; The proportions of BR03 and M - 16V in the Bifidobacterium breve are respectively: 10% and 90%.

4. The preparation method of the probiotic solid composition with Helicobacter pylori inhibitory activity according to any one of claims 1-3, characterized in that, Including the following steps: S1. Probiotic pretreatment, used to mix various probiotic powders according to a preset ratio to obtain a probiotic premix; S2. Matrix formulation treatment, used to compound and mix prebiotics and natural polysaccharides to form a probiotic carrier system and obtain a premixed matrix; S3. Composite embedding treatment, which is used to slowly mix the probiotic premix into the premix matrix so that the probiotics form an embedded structure in the matrix system to obtain a composite; S4. Low-temperature drying treatment, which is used to perform low-temperature drying and micronization treatment on the composite to obtain probiotic solid powder; S5. Ripening and stabilizing treatment, which is used to perform a constant-temperature static treatment on the probiotic solid powder to enhance the stability of the composite structure and obtain ripened powder; S6. Sub-packaging and forming treatment, which is used to package the ripened powder by dosage to obtain a probiotic solid composition with Helicobacter pylori inhibitory activity.

5. The preparation method of the probiotic solid composition with Helicobacter pylori inhibitory activity according to claim 4, characterized in that, The mixing process of the probiotic premix is carried out at a low temperature below 4°C and stored under vacuum drying conditions.

6. The preparation method of the probiotic solid composition with Helicobacter pylori inhibitory activity according to claim 4, characterized in that, In the composite embedding treatment in step S3, slow stirring is carried out at a temperature below 25°C for more than 15 minutes, so that the probiotics are evenly distributed in the natural viscous network formed by tremella polysaccharide and fructooligosaccharide and form a primary embedding protective layer.

7. The preparation method of the probiotic solid composition with Helicobacter pylori inhibitory activity according to claim 4, characterized in that, In the low-temperature drying treatment step in step S4, it is carried out below 35°C by the fluidized bed drying method, and powder with an average particle size less than 200 microns is obtained through micronization treatment.

8. The preparation method of the probiotic solid composition with Helicobacter pylori inhibitory activity according to claim 4, characterized in that, The ripening and stabilizing treatment step in step S5 is to store it in a closed environment with a relative humidity below 30% and a temperature of 25°C for 12% - 24 hours.

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