Application of fermented lactobacillus mucus TY-H27 and its lysate in inhibition of inflammation caused by helicobacter pylori infection

Abstract

The application belongs to the technical field of microorganisms, and discloses a kind of fermented lactobacillus mucus TY-H27 and its lysate in the application of inhibiting inflammation caused by helicobacter pylori infection, and the preservation number of the fermented lactobacillus mucus TY-H27 is CGMCC No.37936.The lysate of the fermented lactobacillus mucus TY-H27 of the application can effectively inhibit the inflammatory response caused by helicobacter pylori infection.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, and in particular to the application of a fermenting Lactobacillus mucinus TY-H27 and its lysate in inhibiting inflammation caused by Helicobacter pylori infection. Background Technology

[0002] Helicobacter pylori (Hp) is a Gram-negative pathogenic bacterium that colonizes the gastric mucosa and is closely associated with chronic gastritis, peptic ulcers, gastric mucosa-associated lymphoid tissue lymphoma, and gastric cancer. Hp infection can trigger the release of pro-inflammatory cytokines such as IL-6 and IL-8 from gastric epithelial cells, leading to a persistent inflammatory response, causing gastric mucosal erosion, glandular atrophy, and inflammatory cell infiltration, which is the core pathological link in gastric mucosal damage. Currently, the mainstream clinical treatment for Hp infection is triple or quadruple therapy with antibiotics combined with proton pump inhibitors, which can eliminate Hp and alleviate inflammation to a certain extent. However, current treatments are highly dependent on antibiotics, and with the continuous increase in Hp resistance rates, eradication rates are declining year by year. At the same time, broad-spectrum antibiotics can easily disrupt intestinal flora homeostasis, causing adverse reactions such as diarrhea, abdominal pain, and indigestion, and are not suitable for immunocompromised individuals, those with abnormal liver or kidney function, or those who are intolerant to antibiotics.

[0003] To overcome the shortcomings of traditional drugs, probiotics and metabiotics are gradually becoming important directions for adjunctive intervention against *Helicobacter pylori* (Hp). Some lactobacilli can exert protective effects by inhibiting Hp adhesion, neutralizing virulence factors, and downregulating pro-inflammatory signaling pathways. Among them, metabiotics are beneficial components of non-living microorganisms, including bacterial lysates, extracellular polysaccharides, and peptidoglycans. They have advantages such as well-defined structure, high stability, no risk of live bacterial colonization, and wide applicability, and have become a new type of functional raw material to replace live probiotics.

[0004] However, existing studies on postbiotic anti-Hp still have the following obvious limitations: (1) Most studies focus on soluble components such as fermentation supernatant and extracellular secretions, and there is insufficient exploration of the anti-inflammatory activity of insoluble precipitate components after bacterial lysis; (2) The effects of postbiotics from different strains are significantly different, and there is a lack of highly efficient, stable and precisely formulated lysate preparations for Hp inflammation; (3) There are generally problems such as weak anti-inflammatory effect, insignificant improvement of gastric mucosal damage in vivo, and inability to match the effects of clinical drugs, which make it difficult to meet the needs of industrialization and clinical auxiliary intervention. Summary of the Invention

[0005] This invention relates to a strain of bacteria isolated from homemade milk produced by herders in Deqin County, Diqing Tibetan Autonomous Prefecture, Yunnan Province. After identification, it was determined to belong to *Lactobacillus fermentum* (…). Lactobacillus fermentum The lysate of Lactobacillus mucinus TY-H27 fermented using the present invention can effectively inhibit the inflammatory response induced by Helicobacter pylori infection.

[0006] To achieve the above objectives, the present invention can adopt the following technical solutions: This invention provides a fermenting Lactobacillus fermentum TY-H27, with accession number CGMCC No. 37936.

[0007] In another aspect, the present invention provides a lysate prepared from the above-mentioned fermented Lactobacillus mucinus TY-H27.

[0008] Preferably, the lysate is obtained by ultrasonically disrupting fermented Lactobacillus mucinus TY-H27 and then collecting the precipitate by centrifugation.

[0009] Preferably, the ultrasonic disruption process uses an ultrasonic cell disruptor in an ice bath, with the following parameters: power 200-400W, time 10-20min, and ultrasonic on for 2s / off for 3s.

[0010] In another aspect, the present invention provides a formulation comprising the above-described composition of the present invention, wherein the formulation is a food, a pharmaceutical or a health product.

[0011] Preferably, the dosage form of the preparation is tablets, pills, capsules, powders, gels, granules, or liquids.

[0012] In another aspect, the present invention provides the application of the fermented Lactobacillus mucinus TY-H27 lysate of the present invention in the preparation of an agent that inhibits inflammation caused by Helicobacter pylori infection.

[0013] Preferably, the above applications include one or more of the following combinations: (a) Application of fermented Lactobacillus mucinus TY-H27 lysate in the preparation of drugs that inhibit IL-8 secretion from gastric epithelial cells induced by Helicobacter pylori; (b) Application of fermented Lactobacillus mucinus TY-H27 lysate in the preparation of drugs that reduce serum IL-6 and IL-8 levels induced by Helicobacter pylori infection; (c) Application of fermented Lactobacillus mucinus TY-H27 lysate in the preparation of drugs to improve gastric mucosal erosion or glandular atrophy caused by Helicobacter pylori infection; (d) Application of fermented Lactobacillus mucinus TY-H27 lysate in the preparation of drugs that improve neutrophil infiltration caused by Helicobacter pylori infection.

[0014] The preservation information for *Lactobacillus fermentans* TY-H27 provided by this invention is as follows: Preservation institution: China General Microbiological Culture Collection Center (CGMCC); Address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing; Deposit date: March 16, 2026; Accession number: CGMCC No. 37936; Classification and nomenclature: Limosilactobacillus fermentum.

[0015] The beneficial effects of this invention include at least the following: 1. The fermented Lactobacillus mucinus TY-H27 lysate provided by this invention has clear and specific anti-Helicobacter pylori infection inflammatory activity, with clear components and definite effects; 2. The fermented Lactobacillus mucinus TY-H27 lysate provided by the present invention can significantly inhibit the secretion of IL-8 by gastric epithelial cells induced by Helicobacter pylori, reduce the levels of IL-6 and IL-8 in vivo, and its anti-inflammatory effect is not significantly different from that of the clinical standard triple therapy, but it is better in reducing neutrophil infiltration. 3. The fermented Lactobacillus mucinus TY-H27 lysate provided by this invention can significantly improve pathological damage such as gastric mucosal erosion or glandular atrophy caused by Helicobacter pylori, and has a clear protective effect on the gastric mucosa, and has the potential to become a clinical adjuvant intervention product. Attached Figure Description

[0016] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the present invention will be further described below in conjunction with the accompanying drawings and embodiments. The drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort: Figure 1 This is the Gram staining result of TY-H27 in Experiment 1; Figure 2 This is a graph showing the effect of different components of TY-H27 on IL-8 secretion in Hp-infected GES-1 cells in Experiment 2. Figure 3 This is a graph showing the comparison of serum IL-6 and IL-8 levels in mice from different groups in Experiment 3. Figure 4 These are HE-stained pathological images and histological scoring results of the gastric mucosa of mice in each group of Experiment 3; Figure 5 These are the weight change curves of mice in each group during Experiment 4. Detailed Implementation

[0017] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, a clear and complete description will be provided below in conjunction with the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the protection scope of the present invention.

[0018] In this embodiment of the invention, the fermenting Lactobacillus mucinus TY-H27 was deposited at the China General Microbiological Culture Collection Center (CGMCC No. 37936). Helicobacter pylori ATCC 43504 was purchased from the American Center for Type Culture Collection. GES-1 human gastric epithelial cells were purchased from Jiangsu Enzyme Immunoassay Co., Ltd. All other chemical reagents used were purchased from reputable chemical reagent suppliers and were of analytical purity.

[0019] This invention provides a fermenting Lactobacillus fermentum TY-H27, which is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 37936 and deposit date of March 16, 2026.

[0020] This invention also provides a lysate prepared from the above-mentioned fermented Lactobacillus mucinus TY-H27.

[0021] It should be noted that the lysate of fermented Lactobacillus mucinus TY-H27 can effectively inhibit the inflammatory response caused by Helicobacter pylori infection in the body, and can be prepared into a composition that can be used for food, medicine or health products.

[0022] This invention also provides a formulation comprising the lysate of *Lactobacillus mucinus* TY-H27 of the present invention, and the formulation is a food, pharmaceutical or health product.

[0023] It should be noted that food, pharmaceuticals, or health products are all products known in this field. The choice of dosage form is important; for example, they can be prepared as probiotic beverages, dairy beverages (such as yogurt), or probiotic biscuits, probiotic powder, and other food products. They can also be combined with pharmaceutical excipients to create drugs with therapeutic effects. Similarly, they can be prepared as health products with certain health benefits, depending on the desired outcome. In actual preparation, the aforementioned lysate containing *Lactobacillus fermentum* TY-H27 can be added to food, pharmaceuticals, or health products.

[0024] In some specific embodiments, the dosage form of the formulation is tablets, pills, capsules, powders, gels, granules, or liquids. It should be noted that the aforementioned solid dosage forms such as tablets, pills, granules, or capsules can be product forms such as probiotic tablets, probiotic sugar pills, probiotic powders, or probiotic capsules; the aforementioned liquids can be product forms such as probiotic beverages; and the aforementioned gels can be product forms such as probiotic jellies, probiotic milk foam, or set yogurt. The specific form of the formulation is a form known in the art. Different dosage forms have different excipients. For example, the preparation of tablets mainly uses diluents (such as starch, dextrin, sucrose, or sucrose), absorbents (calcium sulfate, dicalcium phosphate, or light magnesium oxide), binders (povidone, syrup, or hydroxypropyl methylcellulose), wetting agents (water, etc.), or disintegrants (dry starch, sodium hydroxymethyl starch, or crospovidone, etc.); the preparation of liquids mainly uses solubilizers, suspending agents, emulsifiers, or colorants. Different types of formulations can be added according to what is known in the art.

[0025] This invention also provides an application of the fermented Lactobacillus mucinus TY-H27 lysate of this invention in the preparation of a formulation that inhibits inflammation caused by Helicobacter pylori infection.

[0026] It should be noted that the *Lactobacillus fermentum* TY-H27 lysate of this invention can be used in the preparation of formulations, which can be food, pharmaceuticals, or health products. It should be understood that *Lactobacillus fermentum* TY-H27 of this invention, as a probiotic, simultaneously possesses the attributes of functional food, therapeutic drug, and health product. When it exerts the functional effect of food, the above-mentioned formulation is a food; when it exerts the therapeutic effect, the above-mentioned formulation is a pharmaceutical; and when it exerts the health product effect, the above-mentioned formulation is a health product.

[0027] It should also be noted that the above-mentioned formulations can be in the form of powder, tablets, liquid, granules or capsules, etc. The excipients added to different dosage forms are different and can be added according to those known in the art.

[0028] In some specific examples, the above applications include one or more combinations of the following applications: (a) Application of fermented Lactobacillus mucinus TY-H27 lysate in the preparation of drugs that inhibit IL-8 secretion from gastric epithelial cells induced by Helicobacter pylori; (b) Application of fermented Lactobacillus mucinus TY-H27 lysate in the preparation of drugs that reduce serum IL-6 and IL-8 levels induced by Helicobacter pylori infection; (c) Application of fermented Lactobacillus mucinus TY-H27 lysate in the preparation of drugs to improve gastric mucosal erosion or glandular atrophy caused by Helicobacter pylori infection; (d) Application of fermented Lactobacillus mucinus TY-H27 lysate in the preparation of drugs that improve neutrophil infiltration caused by Helicobacter pylori infection.

[0029] I. Isolation, source, and lysate preparation of *Lactobacillus myxoides* TY-H27 1. Experimental Materials Lactobacillus fermentum TY-H27 was isolated from homemade milk from a herdsman's home in Deqin County, Diqing Tibetan Autonomous Prefecture, Yunnan Province, and is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 37936.

[0030] Sampling method: Use a sterile spoon to transfer milk into a 15L sterile screw-capped centrifuge tube containing an appropriate amount of sterile calcium carbonate and soluble starch (the mass ratio of calcium carbonate to soluble starch is 1:1). Stir well, tighten the cap, and then place the tube in a refrigerator. Transport the tube back to the laboratory for immediate purification and separation of lactic acid bacteria.

[0031] 2. Isolation and purification of Lactobacillus fermentans TY-H27 Take 1 mL of the retrieved sample and add it to 9 mL of sterile physiological saline to obtain a 10⁻¹ sample dilution. Then, perform serial dilutions of 10-fold to obtain concentrations of 10⁻¹, etc. -5 10 -6 and 10 -7 Diluents. Take appropriate amounts of the three diluents mentioned above and spread them onto MRS solid medium. Incubate upside down at 37°C for 48 hours. After incubation, observe the colony morphology. Select medium-sized, raised, neatly edged, and round single colonies for streak plating to isolate the strain. Repeat the above steps until a purified strain is obtained.

[0032] 3. Morphological and structural observation The purified strain was Gram-stained, and purple cell morphology was observed under a 10×100 magnification microscope. The results are as follows: Figure 1 As shown. (Through) Figure 1 As can be seen, the cells are rod-shaped, indicating that they are Gram-positive bacteria (G+).

[0033] 4. Preparation of lysates from fermented Lactobacillus mucilaginosus TY-H27 After activation, Lactobacillus fermentum TY-H27 was inoculated into MRS liquid medium and cultured at 37°C until the stationary phase. The cells were collected by centrifugation, washed twice with sterile PBS, resuspended, and then disrupted in an ice bath using an ultrasonic cell disruptor (300W power, 15min total time, 2s on / 3s off). After disruption, the cells were centrifuged (4°C, 12000rpm, 20min) and the precipitate was collected to obtain the TY-H27 lysate.

[0034] II. Effects of different components of fermented Lactobacillus mucinus TY-H27 on IL-8 secretion in Hp-infected GES-1 cells This experiment mainly investigated the effects of different components of fermented Lactobacillus mucinus TY-H27 on IL-8 secretion in Hp-infected GES-1 cells. The experimental procedure is as follows: 1. Experimental Materials Cells: GES-1 human gastric epithelial cells; Strains: Helicobacter pylori ATCC 43504, Lactobacillus fermentum TY-H27; Reagents: DMEM medium, fetal bovine serum, IL-8 ELISA kit, PBS buffer; Instruments: CO2 incubator, ELISA reader, high-speed centrifuge, ultrasonic cell disruptor.

[0035] 2. Preparation of main solutions The following are the specific preparation processes for preparing *H. pylori* bacterial culture, TY-H27 supernatant, TY-H27 lysis buffer, TY-H27 inactivated cells, TY-H27 lysate, and triple drug working solution: Hp bacterial culture: ATCC 43504 was inoculated onto Columbia blood agar plates, cultured microaerophilically, and the bacterial cells were collected and the concentration was adjusted to 1×10⁻⁶ with PBS. 9 CFU / mL.

[0036] TY-H27 supernatant: After TY-H27 culture reaches the stationary phase, centrifuge at 8000 rpm for 10 min at 4℃, collect the sterile supernatant, and filter through a filter membrane.

[0037] TY-H27 lysis buffer: Following the method in Experiment 1, TY-H27 cells were resuspended in PBS and then sonicated. The whole cell lysis buffer was directly taken without centrifugation.

[0038] TY-H27 inactivated bacteria: live bacterial suspension was inactivated by water bath at 65℃ for 30 min, and plate verification showed no live bacterial growth.

[0039] TY-H27 lysate: TY-H27 lysate was prepared according to the method in Experiment 1. The precipitate was resuspended in PBS at a concentration equivalent to 1×10⁻⁶. 9 CFU original bacterial cells / mL.

[0040] Triple drug working solution: Omeprazole + Amoxicillin + Clarithromycin mixture (Omeprazole 140mg / kg + Amoxicillin 28.6mg / kg + Clarithromycin 14.3mg / kg, mg / kg represents the number of grams of drug per kg of mouse body weight), concentration refers to the clinical equivalent in vitro concentration.

[0041] 3. Cell Culture GES-1 cells (human gastric mucosal epithelial cell line) were cultured in DMEM medium containing 10% fetal bovine serum and 1% penicillin antibiotics at 37°C in a 5% CO2 incubator. Cells in the logarithmic growth phase were used for experiments.

[0042] 4. Experimental Grouping and Treatment GES-1 cells were divided into 8 groups and cultured under the same environment (each group consisted of 3 parallel experiments, and the results were taken as the average of each group). The groups were named experimental groups 1 to 8, and the treatment process for each group was as follows: Experimental group 1 served as the control group: This group did not add Hp or any TY-H27 components, but only an equal volume of complete culture medium, and was cultured normally. Experimental group 2 was the model group: this group was given Hp bacterial culture, but no drugs or TY-H27 components were added; Experimental group 3 was the drug group: Hp bacterial culture was added first, followed by triple drug working solution, as a positive control; Experimental group 4 was the TY-H27 supernatant group: Hp bacterial culture was added first, followed by TY-H27 sterile supernatant; Experimental group 5 was the TY-H27 lysis buffer group: Hp bacterial culture was added first, followed by TY-H27 whole cell ultrasonic disruption mixture; Experimental group 6 was the TY-H27 inactivated bacterial group: first add Hp bacterial solution, then add inactivated TY-H27 bacterial suspension.

[0043] Experimental group 7 was the TY-H27 live bacteria group: first add Hp bacterial solution, then add TY-H27 live bacteria suspension; Experimental group 8 was the TY-H27 lysate group: Hp bacterial culture was added first, followed by the TY-H27 precipitate lysate collected by ultrasonic disruption and centrifugation.

[0044] In experimental groups 2 to 8, the multiplicity of infection (MOI) of adding Hp bacterial solution was 100.

[0045] 5. Cultivation and Detection After adding samples to each group, the cells were cultured for another 12 hours. The cell supernatant was collected, and the IL-8 concentration was calculated strictly according to the instructions of the ELISA kit (Hangzhou Lianke Biotechnology Co., Ltd., EK108-96). Data are expressed as mean ± SD. One-way ANOVA was used, and P < 0.05 was considered statistically significant.

[0046] 6. Experimental Results Test results as follows Figure 2 As shown in Figure A (which represents the metabolite components of each group, and Figure B (which represents the bacterial cell structure components of each group), the discrepancy between the control group and the model group in Figures A and B is due to errors caused by batch-by-batch measurements, which is a normal experimental result.

[0047] according to Figure 2 The overall results showed that, compared with the control group, the IL-8 level in the model group was significantly increased (P<0.05), indicating successful modeling. While the IL-8 levels in the TY-H27 supernatant group and the TY-H27 lysis buffer group were lower than those in the model group, they were still significantly higher than those in the drug group. Figure 2 (See Figure A in the figure); the IL-8 level in the TY-H27 inactivated bacterial group was not significantly different from that in the model group; while the IL-8 level in the TY-H27 live bacterial group and the TY-H27 lysate group was significantly decreased (P<0.05), and there was no significant difference between the TY-H27 lysate group and the TY-H27 live bacterial group and the drug group (P<0.05). Figure 2 This indicates that the TY-H27 lysate can significantly inhibit the Hp-induced inflammatory response in GES-1 cells.

[0048] III. Effects of TY-H27 lysate on serum inflammatory factors and gastric mucosal pathology in *H. pylori*-infected mice 1. Experimental Materials Experimental animals: SPF-grade male C57BL / 6J mice, 4 weeks old, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., housed in an SPF-grade animal facility, used for experiments after 1 week of acclimatization feeding, 10 mice per group, 4 groups in total.

[0049] Strains: Helicobacter pylori ATCC 43504, Lactobacillus fermentum TY-H27.

[0050] Reagents: PBS buffer, omeprazole, amoxicillin, clarithromycin, IL-6 ELISA kit (Hangzhou Lianke Biotechnology Co., Ltd., EK106-96), IL-8 ELISA kit (Hangzhou Lianke Biotechnology Co., Ltd., EK108-96), 4% paraformaldehyde solution, HE staining kit.

[0051] Instruments: electronic balance, high-speed centrifuge, pathological slicer, optical microscope, enzyme-linked immunosorbent assay reader, gavage needle.

[0052] 2. Preparation of main solutions Hp bacterial suspension: ATCC 43504 was inoculated onto Columbia blood agar plates and cultured microaerophilically for 48 h. The bacterial cells were collected, resuspended in sterile PBS, and the concentration was adjusted to 1×10⁻⁶. 9 CFU / mL, prepare and use immediately.

[0053] Triple drug working solution: Based on clinically equivalent dose conversion, omeprazole 140mg / kg, amoxicillin 28.6mg / kg, and clarithromycin 14.3mg / kg (mg / kg indicates the number of grams of drug per kg of mouse body weight). Dissolve and mix with PBS, adjust the volume, and ensure that the gavage volume for each mouse is 0.3mL.

[0054] TY-H27 lysate suspension: Take TY-H27 lysate, resuspend in sterile PBS, and adjust the concentration to the equivalent of 1×10⁻⁶. 9 CFU original bacterial cells / 0.3mL, prepare fresh before use.

[0055] Sterile PBS: Autoclaved, cooled and ready for use as a control for oral administration to mice and for dilution of related reagents.

[0056] 3. Adaptive feeding of animals After purchasing the mice, they were kept in separate cages and given standard rodent food and sterile drinking water. They were allowed free access to food and water and were given an acclimatization period of one week. During this period, the mice were observed daily for their mental state, diet, and fecal characteristics. Mice that were lethargic or had abnormal weight (deviating from the average weight by ±10%) were removed to ensure that the experimental animals were in a consistent condition.

[0057] 4. Experimental Grouping and Treatment After the adaptive feeding period, the mice were randomly divided into 4 groups of 10 mice each, named control group, model group, drug group, and lysate group, respectively. The grouping and specific treatments are as follows: Control group: No *H. pylori* suspension was administered via gavage; instead, an equal volume of sterile PBS (0.3 mL / animal) was administered via gavage every other day for a total of 5 times. After modeling, sterile PBS (0.3 mL / animal) was administered via gavage daily for 14 consecutive days.

[0058] Model group: *H. pylori* bacterial suspension was administered by gavage every other day for a total of 5 times to establish an *H. pylori* infection inflammatory model. After model establishment, sterile PBS (0.3 mL / animal) was administered by gavage daily for 14 consecutive days.

[0059] Drug group: Same as the model group, Hp bacterial suspension was administered by gavage once every other day for a total of 5 times. After modeling, triple drug working solution (0.3 mL / animal) was administered by gavage daily for 14 consecutive days, with the administration time fixed at 9-10 am each day.

[0060] Lysate group: Consistent with the model group, *H. pylori* suspension was administered by gavage once every other day for a total of 5 times. After modeling, TY-H27 lysate suspension (equivalent to 1×10⁻⁶) was administered by gavage daily. 9 CFU original bacterial cells / 0.3mL, 0.3mL / animal), administered by gavage for 14 consecutive days, with the administration time consistent with the drug group to ensure uniform experimental conditions.

[0061] In the model group, drug group, and lysate group, the amount of *H. pylori* suspension administered by gavage was 1×10⁻⁶. 9 CFU / mL, 0.3mL / animal.

[0062] It should be noted that: in all groups above, all mice were administered gavage using sterile gavage needles and slowly to avoid aspiration into the trachea, which could lead to death; during the modeling and intervention periods, the mice were observed at fixed times every day and weighed once a week to record weight changes.

[0063] 5. Sample collection and testing Fourteen days after the intervention, samples were collected from all mice, following the steps outlined below: Serum sample collection: Blood was collected from the eyeballs of mice and placed in sterile centrifuge tubes to separate the serum for the detection of IL-6 and IL-8 concentrations.

[0064] Gastric tissue sample collection: Mice were euthanized by cervical dislocation, quickly dissected, and gastric tissue was separated. The gastric mucosa was rinsed with sterile PBS to remove food residue and blood. A portion of the gastric tissue was fixed in paraformaldehyde solution for HE staining and pathological scoring; the other portion of the gastric tissue was preserved for later use.

[0065] Detection method: (1) Detection of serum IL-6 and IL-8: Strictly follow the instructions of the ELISA kit to calculate the concentration of IL-6 and IL-8 in serum.

[0066] (2) Gastric mucosal pathological examination: After fixation, the gastric tissue was dehydrated, embedded in paraffin, sectioned, stained with HE, and observed under an optical microscope. Two pathology professionals scored the tissue in a double-blind manner. The scoring criteria were based on the "Rat Gastric Mucosal Injury Scoring Criteria". The degree of inflammatory infiltration and the degree of mucosal injury were scored (0-4 points, the higher the score, the more severe the inflammation and injury).

[0067] 6. Experimental Results (1) Results of inflammatory factor detection: Based on Figure 3 The results showed that, compared with the control group, the serum IL-6 and IL-8 concentrations of mice in the model group were significantly increased (P<0.05), indicating that Hp infection successfully induced an inflammatory response in mice. Compared with the model group, the serum IL-6 and IL-8 concentrations of mice in the drug group and the lysate group were significantly decreased (P<0.05), and there was no significant difference between the two groups, indicating that the anti-inflammatory effect of TY-H27 lysate was comparable to that of the triple drug.

[0068] (2) Pathological examination results of gastric mucosa: 1) According to Figure 4Figure A (a, b, c, and d represent the pathological examination results of the gastric mucosa in the control group, model group, drug group, and lysate group, respectively) shows that the gastric mucosa of mice in the control group was intact, the glands were neatly arranged, and there was no inflammatory cell infiltration. The gastric mucosa of mice in the model group showed extensive erosion and hemorrhage, disordered gland arrangement, atrophy of the intrinsic glands, and a large number of neutrophil infiltrations. The pathological score was significantly increased (P<0.05). The gastric mucosa of mice in the lysate group was basically restored to normal thickness, the glands were neatly arranged, there was no obvious erosion or hemorrhage, and no neutrophil infiltration was observed.

[0069] 2) According to Figure 4 As shown in Figures B and C (where Figures B and C represent the inflammatory infiltration score and the mucosal injury score, respectively), the pathological score of the lysate group was significantly lower than that of the model group (P<0.05), and not much different from that of the triple drug group. Moreover, the lysate group was more effective than the drug group in reducing neutrophil infiltration.

[0070] Based on the above test results, the TY-H27 lysate in this invention can significantly improve the inflammatory response in Hp-infected mice and repair gastric mucosal pathological damage. The effect is comparable to that of triple therapy, and it has a greater advantage in improving inflammatory infiltration.

[0071] IV. Safety Evaluation of TY-H27 Pyrolysis Products Throughout the entire process, the weight changes, diet, activity status, and fecal characteristics of mice in each group were recorded. After the intervention, the mice were dissected to observe the appearance of major organs such as the heart, liver, spleen, lungs, and kidneys, and to determine whether there were any abnormal lesions such as swelling, congestion, or necrosis.

[0072] The results of weight changes in each group of mice are as follows: Figure 5 As shown, according to Figure 5 The results showed that there was no significant difference in body weight change between the TY-H27 lysate group and the control group. During the actual observation process, no mice in any group exhibited diarrhea, lethargy, loss of appetite, or abnormal activity. Post-dissection, the major organs appeared normal, without swelling, congestion, necrosis, or other lesions, indicating that the TY-H27 lysate has good in vivo safety and no obvious toxic side effects.

[0073] The specific embodiments described above further illustrate the purpose, technical solution, and beneficial effects of the present invention. It should be understood that the above description is only a specific implementation method of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the scope of the present invention should be included within the protection scope of the present invention.

Claims

1. A fermenting *Lactobacillus fermentum* TY-H27, characterized in that, Its accession number is CGMCC No.37936.

2. The lysate prepared from fermented Lactobacillus mucinus TY-H27 according to claim 1.

3. The pyrolysis product according to claim 2, characterized in that, The lysate was obtained by ultrasonically disrupting fermented Lactobacillus mucinus TY-H27 followed by centrifugation to collect the precipitate.

4. The pyrolysis product according to claim 3, characterized in that, The ultrasonic disruption process was performed using an ultrasonic cell disruptor in an ice bath. The disruption parameters were: power 200-400W, time 10-20min, and ultrasonic on for 2s / off for 3s.

5. A formulation comprising the lysate of any one of claims 2 to 4, wherein the formulation is a food, pharmaceutical, or health product.

6. The formulation according to claim 5, characterized in that, The dosage form of the preparation is tablets, pills, capsules, powders, gels, granules, or liquids.

7. The use of the fermented Lactobacillus mucinus TY-H27 lysate according to any one of claims 2 to 4 in the preparation of a medicament for inhibiting inflammation caused by Helicobacter pylori infection.

8. The application according to claim 7, characterized in that, Applications include one or more of the following combinations: (a) Application of fermented Lactobacillus mucinus TY-H27 lysate in the preparation of drugs that inhibit IL-8 secretion from gastric epithelial cells induced by Helicobacter pylori; (b) Application of fermented Lactobacillus mucinus TY-H27 lysate in the preparation of drugs that reduce serum IL-6 and IL-8 levels induced by Helicobacter pylori infection; (c) Application of fermented Lactobacillus mucinus TY-H27 lysate in the preparation of drugs to improve gastric mucosal erosion or glandular atrophy caused by Helicobacter pylori infection; (d) Application of fermented Lactobacillus mucinus TY-H27 lysate in the preparation of drugs that improve neutrophil infiltration caused by Helicobacter pylori infection.

9. The application according to claim 7 or 8, characterized in that, The dosage form of the drug is tablet, pill, capsule, powder, gel, granule or liquid.

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