A method for preparing probiotic bacteria and its use for eradicating helicobacter pylori in the oral cavity

Abstract

The application provides a preparation method of probiotics and application of the probiotics in removing Helicobacter pylori in the oral cavity. More specifically, a polypeptide HP-si-125 with good inhibitory activity against Helicobacter pylori is obtained through a bacteriostatic experiment screening. The polypeptide and the probiotics can be used alone or in combination and have good effects on treating Helicobacter pylori in a mouse model. The polypeptide and the probiotics can be prepared into mouthwash or drugs for treating Helicobacter pylori in the oral cavity and have good application values.

Description

Technical Field

[0001] This application relates to the field of biology, and more specifically to a method for preparing probiotics and their use in eradicating Helicobacter pylori in the oral cavity. Background Technology

[0002] Helicobacter pylori (H. pylori) is a unipolar, multi-flagellated, spiral-shaped Gram-negative bacterium, measuring (0.3-1.0) μm × (2.0-5.0) μm. On the surface of gastric mucosal epithelial cells, H. pylori typically presents as a spiral or arc shape, but under unfavorable conditions, it can become rod-shaped or spherical. H. pylori is a microaerophilic bacterium, requiring 5%–8% oxygen in its environment. Humans are the primary host of H. pylori, and it is currently the only known microorganism that can be isolated and cultured from the human stomach. Understanding its transmission process is essential for the prevention and control of H. pylori.

[0003] Studies have shown that oral Helicobacter pylori plays a crucial role in the transmission of Helicobacter pylori. Epidemiological surveys indicate that populations living in crowded conditions have higher rates of Helicobacter pylori infection, suggesting that Helicobacter pylori is likely transmitted directly from one individual to another. The oral-oral and fecal-oral routes are considered the most probable transmission routes for Helicobacter pylori. In addition to direct detection of Helicobacter pylori, the detection of Helicobacter pylori antigens and antibodies in saliva may also provide a basis for the diagnosis of gastric diseases. Studies have shown that the presence or absence of oral Helicobacter pylori antigens in different types of gastric diseases is associated with the degree of gastritis activity and some precancerous lesions of the gastric mucosa. The detection rate of oral Helicobacter pylori antigens is significantly higher in patients with chronic active gastritis or those with moderate to severe intestinal metaplasia or dysplasia. Therefore, effectively eradicating Helicobacter pylori in the oral cavity is an essential step in the complete treatment of Helicobacter pylori infection.

[0004] The most common treatment for Helicobacter pylori is triple therapy, which consists of a proton pump inhibitor and two antibiotics. This is currently the most frequently used and effective treatment for eradicating Helicobacter pylori, but the recurrence rate is relatively high. This may be because Helicobacter pylori still exists in the oral cavity after treatment and is swallowed into the stomach with saliva. Studies have found that in patients with upper gastrointestinal diseases, although Helicobacter pylori may have been eradicated in the stomach after anti-Helicobacter pylori treatment, it may still exist in the oral cavity. This may be related to the fact that oral Helicobacter pylori exists in plaque with a unique biofilm structure, making it difficult for drugs to act on it.

[0005] Since oral Helicobacter pylori is a potential source of recurrence or reinfection with gastric Helicobacter pylori, some scholars have proposed using medication combined with basic periodontal therapy to reduce the recurrence rate of gastric Helicobacter pylori infection. Patients who underwent basic periodontal therapy and used mouthwash had significantly higher Helicobacter pylori eradication rates than those who did not undergo basic periodontal therapy or use mouthwash. This suggests that the eradication rate of Helicobacter pylori with medication is related to the patient's periodontal condition and oral hygiene. Research results show that *H. pylori* infection is indeed present in the gingival fluid and periodontal pocket exudate of patients with periodontitis, and the positive rate of *H. pylori* infection is positively correlated with the degree and activity of inflammation, indicating a direct relationship between *H. pylori* infection and periodontitis. Further questioning of the medical history of 144 *H. pylori*-positive periodontitis patients revealed a close relationship between *H. pylori* infection in periodontitis and *H. pylori* infection in the stomach and duodenum. Most studies indicate that the oral cavity is a long-term accumulation site for *H. pylori* and has a certain correlation with *H. pylori* in the stomach; the two have homology, and oral *H. pylori* may be an important reservoir for *H. pylori* infection in the stomach. Oral *Helicobacter pylori* (Hp) may play a role in transmission, especially given its persistent presence in dental plaque. Correlation studies between Hp and periodontitis groups and controls confirmed a significant correlation between Hp and periodontitis. The positive rate of Hp infection was positively correlated with the degree and activity of inflammation. Neglecting the multiple locations of Hp residing in the oral cavity and digestive tract is a cause of recurrent digestive diseases and periodontitis. Therefore, while eradicating Hp infection in the stomach, the possibility of oral Hp infection should also be considered. It is necessary to strengthen oral hygiene education, actively treat oral diseases, and inhibit plaque formation, thereby significantly reducing the incidence and reinfection rate of Hp.

[0006] Therefore, developing drugs that can be used to eliminate Helicobacter pylori in the oral cavity is an important research direction, which is beneficial for eradicating Helicobacter pylori infection in the stomach. Summary of the Invention

[0007] This invention overcomes the shortcomings of the prior art and provides a probiotic composition for eliminating Helicobacter pylori in the oral cavity.

[0008] Specifically, the probiotic composition is prepared by Lactobacillus acidophilus CGMCC No. 23546 and Lactobacillus plantarum GDMCC No. 62256 at the same concentration volume ratio of 2:1.

[0009] Specifically, the preparation of the *Lactobacillus acidophilus* LA05 suspension (CGMCC No. 23546) is as follows: *Lactobacillus acidophilus* LA05 is inoculated into a modified MRS liquid medium and activated by culturing at 38°C. This activation is repeated twice to obtain an activated solution. The activated solution is then inoculated into MRS liquid medium at a rate of 2% (v / v) and cultured at 38°C to achieve a bacterial concentration of 10... 8CFU indicates the cessation of fermentation. The preparation of the *Lactobacillus plantarum* suspension with preservation number GDMCC No: 62256: *Lactobacillus plantarum* LP1Z was inoculated from glycerol tubes onto modified MRS Richter agar plates and anaerobically cultured at 37°C for 48 h. Single colonies of *Lactobacillus plantarum* LP1Z were then picked and inoculated into modified MRS medium and anaerobically cultured at 37°C to a bacterial concentration of 10. 8 CFU stands for CFU, which means fermentation has stopped. The fermentation broth of *Lactobacillus acidophilus* LA05 and *Lactobacillus plantarum* LP1Z was mixed at an optimized volume ratio of 2:1 and then freeze-dried to obtain a pharmaceutical composition containing probiotics.

[0010] More specifically, the present invention also provides an anti-Helicobacter pylori polypeptide HP-si-125.

[0011] More specifically, the present invention also provides the use of the anti-Helicobacter pylori polypeptide HP-si-125 in the preparation of a medicament for treating Helicobacter pylori.

[0012] Furthermore, the present invention also provides the use of the anti-Helicobacter pylori polypeptide HP-si-125 and the probiotic composition in the preparation of a medicine box for treating anti-Helicobacter pylori; wherein the probiotic composition is prepared by Lactobacillus acidophilus CGMCC No. 23546 and Lactobacillus plantarum GDMCC No: 62256 at the same bacterial concentration conditions at a volume ratio of 2:1.

[0013] Furthermore, the present invention also provides a mouthwash for treating oral Helicobacter pylori.

[0014] Specifically, the mouthwash is characterized by comprising the following components in parts by weight: 90-110 parts water, 2-15 parts sorbitol, 1-8 parts propylene glycol, 1-8 parts xylitol, 1-5 parts PEG-400 polyethylene glycol, 0.5-2 parts dimethyl glycyrrhizate, 1-5 parts sodium benzoate, 0.1-1 part flavoring, 0.5-5 parts HP-si-125 polypeptide, and 1-5 parts lyophilized probiotic composition powder; wherein the amino acid sequence of the HP-si-125 polypeptide is as shown in SEQ ID NO: 8; the lyophilized probiotic composition powder is prepared by freeze-drying Lactobacillus acidophilus CGMCC No. 23546 and Lactobacillus plantarum GDMCC No: 62256 at a volume ratio of 2:1 under the same bacterial concentration conditions.

[0015] More specifically, the mouthwash is composed of the following components in parts by weight: 90 parts water, 10 parts sorbitol, 5 parts propylene glycol, 5 parts xylitol, 5 parts PEG-400 polyethylene glycol, 1 part dimethyl glycyrrhizate, 3 parts sodium benzoate, 1 part fragrance, 1 part HP-si-125 peptide, and 3 parts lyophilized probiotic composition powder.

[0016] Specifically, the medicaments or medicaments of the present invention contain pharmaceutically acceptable carriers. The terms "pharmaceutically acceptable carrier" or "pharmaceuticalally acceptable excipient" include any and all solvents, dispersion media, coatings, antimicrobial and antifungal agents, isotonic agents, and absorption delay agents, etc. The use of such media and reagents for pharmaceutically active substances is well known in the art. Apart from any conventional media or reagents being incompatible with the active ingredient, the present invention also relates to their use in the therapeutic compositions disclosed herein. Additional active ingredients may also be incorporated into the pharmaceutical compositions.

[0017] On the other hand, the present invention also relates to the use of one or more of the disclosed polypeptides in the preparation of anti-Helicobacter pylori drugs. In some embodiments, the polypeptides are administered, alone or in combination with any other desired therapeutic agent, using a suitable delivery method. The disclosed therapeutically useful agents can be administered via, for example, one of the following routes: oral, e.g., in the form of sugar-coated, coated tablets, pills, semi-solid substances, soft capsules or hard capsules, solutions, emulsions or suspensions; parenteral, e.g., in the form of injectable solutions; rectal suppositories; by inhalation, e.g., in the form of powder formulations or sprays; and / or transdermal or intranasal. For the preparation of such tablets, pills, semi-solid substances, coated tablets, lozenges and hard gelatin capsules, the therapeutic product can be mixed with a pharmacologically inert, inorganic or organic drug carrier substance, e.g., with lactose, sucrose, glucose, gelatin, malt, silica gel, starch or derivatives thereof, talc, stearic acid or salts thereof, skim milk powder, etc. For the preparation of soft capsules, drug carrier substances such as vegetable oils, petroleum, animal oils or synthetic oils, waxes, fats and polyols can be used. To prepare liquid solutions and syrups, pharmaceutical carrier substances such as water, alcohols, saline solutions, dextrose solutions, polyols, glycerol, vegetable oils, petroleum, and animal or synthetic oils can be used. For suppositories, pharmaceutical carrier substances such as vegetable oils, petroleum, animal or synthetic oils, waxes, fats, and polyols can be used. For aerosol formulations, compressed gases suitable for this purpose, such as oxygen, nitrogen, and carbon dioxide, can be used. Pharmaceutically acceptable agents may also include additives for preservation and stabilization, emulsifiers, sweeteners, flavoring rings, salts for altering osmotic pressure, buffers, encapsulating additives, and / or antioxidants. Many constructions and variations will be apparent to those skilled in the art upon consideration of the disclosure of this invention.

[0018] Examples of suitable pharmaceutically acceptable preservatives include, but are not limited to, benzalkonium chloride, benzyl chloride and cetylpyridine chloride, benzyl bromide, benzyl alcohol, disodium EDTA, phenylmercuric nitrate, phenylmercuric acetate, sodium ethylmercuric thiosalicylate, thimerosal, acetate and phenylmercuric borate, polymyxin B sulfate, chlorhexidine, methylparaben and propylparaben, phenethyl alcohol, quaternary ammonium chloride, sodium benzoate, sodium propionate, stable chloroxychloride complexes, sorbic acid, or mixtures thereof. Preferred pharmaceutically acceptable preservatives include disodium EDTA (disodium ethylenediaminetetraacetate) and benzalkonium chloride, or mixtures thereof. In one embodiment, the pharmaceutically acceptable preservative is present in an amount from about 0.01% to about 2.0% (weight / volume), preferably from about 0.05% to about 1% (weight / volume).

[0019] Examples of suitable pharmaceutically acceptable buffers include, but are not limited to, sodium chloride, glucose, lactose, and phosphate-buffered saline (PBS), or any combination thereof. Other suitable pharmaceutically acceptable buffers include, but are not limited to, disodium succinate hexahydrate, borate, citrate, phosphate, acetate, physiological saline, tris-HCl (tris(hydroxymethyl)aminomethane hydrochloride), HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid), sodium phosphate, sodium borate, physiological saline, citrate, carbonate, phosphate, and / or mixtures thereof, to obtain the desired osmolarity. In one embodiment, the pharmaceutically acceptable buffer is present in an amount of about 0.01% to about 2.0% (w / v), preferably about 0.05% to about 1% (w / v).

[0020] Examples of suitable pharmaceutically acceptable chelating agents include, but are not limited to, ethylenediaminetetraacetic acid (EDTA), disodium EDTA and its derivatives, citric acid and its derivatives, nicotinamide and its derivatives, sodium deoxycholate and its derivatives, or mixtures of these chelating agents. In one embodiment, the pharmaceutically acceptable chelating agent is present in an amount of about 0.01% to about 2.0% (weight / volume), preferably about 0.05% to about 1% (weight / volume).

[0021] In one embodiment, the pharmaceutically acceptable wetting agent or surfactant is present in an amount of about 0.01% to about 5.0% (weight / volume), preferably about 0.05% to about 2.0% (weight / volume), more preferably about 0.1% to about 1.0% (weight / volume), for example about 0.1%, 0.2%, 0.5%, 1.0% (weight / volume).

[0022] Examples of suitable pharmaceutically acceptable isotonic modifiers include, but are not limited to, D-mannitol, glucose, glycerol, sodium chloride, potassium chloride, calcium chloride, and magnesium chloride, or any combination thereof. Various nitrates, citrates, acetates, or mixtures thereof may also be used. In one embodiment, the pharmaceutically acceptable isotonic modifier is present in an amount of about 0.1% to about 5.0% (weight / volume), preferably about 1% to about 3% (weight / volume). Attached Figure Description

[0023] Figure 1 The effect of HP-si-125 peptide on urease activity in Helicobacter pylori (Figure 1)

[0024] Beneficial effects

[0025] This invention provides a method for preparing probiotics and their application in eradicating Helicobacter pylori in the oral cavity. More specifically, a polypeptide HP-si-125 with good inhibitory activity against Helicobacter pylori was obtained through computer simulation pre-screening and antibacterial experiments. The polypeptide, used alone or in combination with the probiotics, showed good therapeutic effects against Helicobacter pylori in mouse models. The polypeptide and probiotics can be formulated into mouthwash or drugs for the treatment of Helicobacter pylori in the oral cavity, demonstrating significant application value. Detailed Implementation

[0026] Those skilled in the art can refer to the content of this document and appropriately improve the process parameters to achieve the desired results. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit, and scope of this invention to implement and apply the technology of this invention. Unless otherwise specified, the methods, equipment, and materials in the following embodiments are all conventional methods, equipment, and materials in the art and are commercially available.

[0027] Example 1: Simulation Design and Screening of Anti-Helicobacter pylori Peptides

[0028] Design of the antimicrobial peptide library: A peptide library was designed by setting the length of antimicrobial peptides within the range of 13-16 amino acids and selecting common dominant amino acids for antimicrobial peptides as the basis. Computer-aided simulation screening was conducted based on pathogenic bacteria's anti-Helicobacter pylori activity, and 14 peptides were initially screened for artificial synthesis.

[0029] The method for identifying the efficacy of antimicrobial peptides against Helicobacter pylori is as follows:

[0030] H. pylori (ATCC43504, purchased from Shanghai Beinuo Biotechnology Co., Ltd.) frozen at -80℃ was revived on Columbia agar plates containing 7% sheep blood and 0.4% H. pylori selective additive (Oxoid). After two generations of activation, the cells were cultured in Columbia blood plates for 48 hours. The cells were scraped off, centrifuged at 4000 rpm for 10 min, resuspended in BHI, and the density was adjusted to 1×10⁻⁶. 9 CFU / mL, then serially diluted as needed for the experiment.

[0031] After two generations of H. pylori activation, 10 8 CFU H. pylori was plated onto fresh, antibiotic-free Columbia agar medium and immediately placed in an Oxford cup. 100 μL of the test peptide (100 μg / mL SEQ ID NO: 1-14) was added, and the mixture was fixed under microaerophilic conditions for 1 hour. After incubation in a tri-gas incubator for 72 hours, the size of the inhibition zone was measured. The experiment was repeated three times. The results are shown in Table 1.

[0032] Table 1. Inhibitory effects of various peptides on Helicobacter pylori

[0033]

[0034]

[0035] - indicates no inhibitory effect.

[0036] As shown in Table 1, the blank culture medium control, serving as a negative control, did not inhibit the growth of *H. pylori*, and no inhibition zone appeared. The HP-si-125 peptide exhibited the best inhibitory effect against *H. pylori*; therefore, this peptide was selected for subsequent experiments.

[0037] Example 2: Efficacy verification of HP-si-125 peptide

[0038] Fresh Helicobacter pylori ATCC43504 was washed twice with PBS, and the H. pylori concentration was adjusted to 2 × 10⁻⁶ using BHI. 7CFU / mL. In a 96-well plate, 50 μL of H. pylori and 10 μL of HP-si-125 peptide at different concentrations (10 μg / mL, 100 μg / mL, and 200 μg / mL, low, medium, and high concentrations) were mixed and incubated in a tri-gas incubator for 48 h. Then, 150 μL of urease test solution was added, and after shaking, the absorbance was measured using a microplate reader at 550 nm. The blank is the result of the urease reagent solution test; the negative control is BHI instead of peptide assay; the positive control is 100 μg / mL metronidazole tablets. The urease indicator formula is: 0.9% NaCl, 20 mmol / L urea, and 14 μg / mL phenol red. The pH was adjusted to 6.8 (the color change point of phenol red) with HCl, and the density was measured at OD550 nm using a spectrophotometer. The results are as follows. Figure 1 As shown.

[0039] from Figure 1 It can be seen that after 24 hours without the addition of peptides or drugs, the urease activity of Helicobacter pylori was significantly increased compared to the blank control group (P<0.01). However, after treatment with peptides and the positive control, the urease activity was significantly decreased compared to the negative control group (P<0.05), especially in the high-concentration peptide group, where the urease activity decreased by 2.7 times, demonstrating a significant effect. Helicobacter pylori can survive in the low-acidity stomach because it produces urease. Inhibiting the urease activity of Helicobacter pylori deprives it of its favorable environment for survival, thereby inhibiting its colonization and growth in the stomach. Figure 1 The results showed that the polypeptide of the present invention has significant inhibitory activity against Helicobacter pylori.

[0040] Example 3: Peptide Safety Testing

[0041] The hemolytic activity of antimicrobial peptides and probiotics was detected using sheep blood erythrocytes. 10 μl of a 1 mg / ml HP-si-125 peptide solution was added to the surface of a blood agar plate, with distilled water as a control. The plate was incubated at 37°C for at least 24 hours, and the presence or absence of hemolytic zones was observed. Results showed that no hemolytic zones were observed for any of the tested antimicrobial peptides. Preliminary results indicate that the antimicrobial peptides do not exhibit hemolytic toxicity.

[0042] Example 4: Preparation of a pharmaceutical composition containing probiotics

[0043] Modified MRS liquid culture medium: glucose 20 g / L, peptone 10 g / L, beef extract 10 g / L, yeast extract 5 g / L, whey powder 10 g / L, tomato paste 10 g / L, K2HPO4 2 g / L, diammonium citrate 2 g / L, sodium acetate 5 g / L, Tween 80 1 g / L, MgSO4·7H2O 0.5 g / L, MnSO4·4H2O 0.05 g / L, L-cysteine ​​hydrochloride 0.5 g / L. Adjust the pH to 7.0 with 1 mol / L NaOH and sterilize at 121℃ for 20 min.

[0044] Preparation of Lactobacillus acidophilus LA05 (preservation number CGMCC No.23546, see CN202210413078.0) bacterial suspension: Lactobacillus acidophilus LA05 was inoculated into a modified MRS liquid medium and cultured at 38℃ for 18h to activate it. This activation was repeated twice to obtain the activated solution. The activated solution was inoculated into MRS liquid medium at an inoculation rate of 2% (v / v) and cultured at 38℃. Fermentation was stopped when the bacterial concentration reached 10⁸ CFU.

[0045] Preparation of Lactiplantibacillus plantarum LP1Z (preservation number GDMCC No: 62256, see CN202210263672.6) bacterial suspension: Lactiplantibacillus plantarum LP1Z was inoculated from glycerol tubes onto modified MRS Rich Agar plates and anaerobically cultured at 37℃ for 48 h. Single colonies of Lactiplantibacillus plantarum LP1Z were picked and inoculated into modified MRS medium and anaerobically cultured at 37℃. Fermentation was stopped when the bacterial concentration reached 10⁸ CFU.

[0046] The fermentation broths of Lactobacillus acidophilus LA05 and Lactobacillus plantarum LP1Z were mixed at an optimized volume ratio of 2:1 and then freeze-dried to obtain a pharmaceutical composition containing probiotics.

[0047] Example 5: Animal experiments with probiotic composition and peptides

[0048] The experimental animals were (SPF) C57BL / 6 mice, 6-8 weeks old and weighing 18-20g. Each mouse was administered 0.5ml (containing 5×10⁻⁶ ml of the solution) by gavage. 8 Helicobacter pylori ATCC43504 bacterial suspension (CFU) was administered every other day for a total of 3 times. Two weeks after modeling, mice were randomly sampled, sacrificed, and their stomachs were harvested for rapid urease test, Gram staining of smears, pathological sections, and bacterial culture to confirm successful infection.

[0049] The prepared Helicobacter pylori-infected male mice were randomly divided into 5 groups, with 10 mice in each group.

[0050] Model group: each animal was given 0.4 ml of normal saline (NS) by gavage.

[0051] Positive group: CBS 6.15mg / kg, tetracycline 50mg / kg, metronidazole 22.5mg / kg; (CBS: colloidal bismuth subcitrate).

[0052] Peptide group: HP-si-125 peptide 50mg / kg;

[0053] Probiotic group: 150 mg / kg of lyophilized probiotic composition prepared in Example 4;

[0054] The peptide combined with probiotic group: HP-si-125 peptide 50mg / kg; after a 2-hour interval following administration, 150mg / kg of the lyophilized probiotic composition prepared in Example 4 was administered.

[0055] Animals in each group were administered the drug via gavage once daily for 14 days. Four weeks after the last administration, all mice were sacrificed. The stomachs were dissected and cut longitudinally into four sections along the greater curvature. These sections were then subjected to rapid urease testing, Gram staining, pathological sectioning, and bacterial culture for *Helicobacter pylori* (Hp) detection. Mice were defined as Hp-negative if all four detection methods yielded negative results, and as Hp-infected if one or more methods yielded positive results. The experimental results are shown in Table 2.

[0056] Table 2 Results of in vivo anti-Helicobacter pylori activity in each group

[0057] Group Number of infections Clearance Model group 10 0％ positive group 2 <![CDATA[80% # ]]> Peptide group 2 <![CDATA[80% # ]]> Probiotic group 4 <![CDATA[70% # ]]> Peptide combined with probiotic group 0 <![CDATA[100% #* ]]>

[0058] #Compared with the model group (P<0.01), *Compared with the positive control group (P<0.05).

[0059] As shown in Table 2, the clearance rate of Helicobacter pylori infection in the positive control group receiving triple therapy was 80%, which was significantly different from the model group. The clearance rates of the polypeptide group and the probiotic group of this invention were 80% and 70%, respectively, which were also significantly different from the model group. In particular, the polypeptide combined with probiotic group showed a significant synergistic therapeutic effect, with a clearance rate of 100%. This indicates that the combination has a better effect.

[0060] Example 6: Preparation and efficacy verification of mouthwash for eliminating Helicobacter pylori in the oral cavity

[0061] The mouthwash is composed of the following components in parts by weight: 90 parts water, 10 parts sorbitol, 5 parts propylene glycol, 5 parts xylitol, 5 parts PEG-400 polyethylene glycol, 1 part dimethyl glycyrrhizate, 3 parts sodium benzoate, 1 part fragrance, 1 part HP-si-125 peptide, and 3 parts lyophilized powder of the probiotic composition prepared in Example 4.

[0062] Twenty patients (10 with Helicobacter pylori infection detected in their oral cavity) used the mouthwash of this invention three times a day for 30 consecutive days. Oral mucosa samples were scraped using a sterile spatula, and 1 ml of saliva was collected, thoroughly mixed, and then tested for the presence of H. pylori in the oral cavity. The results showed that no Helicobacter pylori was detected in the oral cavity, indicating a good treatment effect.

[0063] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

[0064] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. An anti-Helicobacter pylori polypeptide HP-si-125, characterized in that, The amino acid sequence is shown in SEQ ID NO:

8.

2. Use of the anti-Helicobacter pylori polypeptide HP-si-125 as described in claim 1 in the preparation of a medicament for inhibiting Helicobacter pylori activity.

3. The use of the anti-Helicobacter pylori polypeptide HP-si-125 and probiotic composition as described in claim 1 in the preparation of a medicine box for treating Helicobacter pylori; wherein the probiotic composition is prepared by Lactobacillus acidophilus CGMCC No. 23546 and Lactobacillus plantarum GDMCC No. 62256 at the same bacterial concentration conditions at a volume ratio of 2:

1.

4. The use as described in claim 3, characterized in that, The medicine box also contains a pharmaceutically acceptable carrier.

5. A mouthwash for treating oral Helicobacter pylori, characterized in that, The mouthwash is composed of the following components in parts by weight: 90-110 parts water, 2-15 parts sorbitol, 1-8 parts propylene glycol, 1-8 parts xylitol, 1-5 parts PEG-400 polyethylene glycol, 0.5-2 parts dimethyl glycyrrhizate, 1-5 parts sodium benzoate, 0.1-1 part flavoring, 0.5-5 parts HP-si-125 polypeptide, and 1-5 parts lyophilized probiotic composition powder; wherein the amino acid sequence of the HP-si-125 polypeptide is as shown in SEQ ID NO: 8; the lyophilized probiotic composition powder is prepared by freeze-drying Lactobacillus acidophilus CGMCC No. 23546 and Lactobacillus plantarum GDMCC No: 62256 at a volume ratio of 2:1 under the same bacterial concentration conditions.

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