Preparation method of probiotics and application of probiotics in removing helicobacter pylori in oral cavity

Abstract

The invention provides a preparation method of probiotics and application of the probiotics to removal of helicobacter pylori in an oral cavity. More specifically, one polypeptide HP-si-125 with relatively good inhibitory activity on helicobacter pylori is obtained through antibacterial experiment screening. After the polypeptide and the probiotics are used independently or jointly, a good helicobacter pylori treatment effect can be achieved on a mouse model. The polypeptide and the probiotics can be prepared into mouthwash or medicines for treating helicobacter pylori in the oral cavity, and have good application value.

Description

Technical Field

[0001] The present application relates to the biological field, and more specifically to a method for preparing probiotics and use of the probiotics in removing Helicobacter pylori in the oral cavity. Background Art

[0002] Helicobacter pylori (H.pylori) is a unipolar, multi-flagellated, spirally curved Gram-negative bacterium with a size of (0.3-1.0) μm × (2.0-5.0) μm. Helicobacter pylori often appears in a typical spiral or arc shape on the surface of gastric mucosal epithelial cells, and can become rod-shaped or spherical under unfavorable conditions. Helicobacter pylori is a microaerophilic bacterium and requires 5% to 8% environmental oxygen. Humans are the main host of Helicobacter pylori, and Helicobacter pylori is also the only known microorganism that can be isolated and cultured from the human stomach. To prevent and control Helicobacter pylori, it is essential to understand its transmission process.

[0003] Studies have shown that oral Helicobacter pylori plays an important role in the transmission of Helicobacter pylori. Epidemiological surveys have shown that people living in crowded conditions have a higher rate of Helicobacter pylori infection, indicating that Helicobacter pylori is likely to be transmitted directly from one individual to another. The oral-oral route and the fecal-oral route are considered to be the most likely routes of Helicobacter pylori transmission. In addition to direct detection of Helicobacter pylori, detection of Helicobacter pylori antigen and antibody in saliva may also provide a basis for the diagnosis of gastric disease. Studies have shown that oral Helicobacter pylori antigen detection was performed on different types of gastric diseases, and it was found that the presence or absence of oral Helicobacter pylori was related to the degree of gastritis activity and some gastric mucosal precancerous lesions. The detection rate of oral Helicobacter pylori antigen is significantly increased in patients with chronic active gastritis or with moderate to severe intestinal metaplasia or atypical hyperplasia. Therefore, effective removal of Helicobacter pylori in the mouth is also a necessary step for the complete treatment of Helicobacter pylori infection.

[0004] The most common treatment for Helicobacter pylori is triple therapy, which is a proton pump inhibitor plus two antibiotics. It is currently the most commonly used and most effective treatment for eradicating Helicobacter pylori, but the probability of disease recurrence after treatment is high. This may be caused by the fact that Helicobacter pylori still exists in the mouth after treatment and is swallowed into the stomach with saliva. Studies have found that after anti-Helicobacter pylori treatment for patients with upper gastrointestinal diseases, although Helicobacter pylori has been eradicated in the stomach, it still exists in the mouth. This may be related to the fact that oral Helicobacter pylori exists in plaques with a unique biofilm structure that is difficult to react to drugs.

[0005] Since oral Helicobacter pylori is a potential source of recurrence or reinfection of gastric Helicobacter pylori infection, some scholars have proposed the use of drugs combined with basic periodontal treatment to reduce the recurrence rate of gastric Helicobacter pylori infection. The eradication rate of gastric Helicobacter pylori in patients who have undergone basic periodontal treatment and used mouthwash is significantly higher than that in patients who have not undergone basic periodontal treatment and have not used mouthwash. This suggests that the eradication rate of Helicobacter pylori by drug treatment is related to the patient's periodontal condition and oral hygiene. The results showed that Hp infection was indeed present in the gingival fluid and periodontal pocket exudate of patients with periodontitis, and the positive rate of Hp infection was positively correlated with the degree and activity of inflammation, indicating that Hp infection was directly related to periodontitis; the medical history of 144 Hp-positive periodontitis patients was questioned, and the results showed that periodontitis with Hp infection was closely related to Hp infection in the stomach and duodenum. Most studies have shown that the oral cavity is a long-term gathering place for Hp, and has a certain correlation with Hp in the stomach. The two are homologous, and oral Hp may be an important reservoir for Hp infection in the stomach. Oral Hp may play a role in transmission, especially its persistence in dental plaque. Through the study of the correlation between Hp and the periodontitis group and the control group, it was confirmed that Hp and periodontitis have a significant correlation. The positive rate of Hp infection is positively correlated with the degree and activity of inflammation. Ignoring the presence of Hp in multiple places in the oral cavity and digestive tract is the cause of repeated attacks of digestive tract diseases and periodontitis. It can be seen that while eradicating Hp infection in the stomach, the possibility of Hp infection in the oral cavity should also be considered. It is necessary to strengthen oral hygiene education, actively treat oral diseases, inhibit the formation of dental plaque, and thus greatly reduce the high 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 at present, which is beneficial to the eradication of Helicobacter pylori infection in the stomach. Summary of the invention

[0007] The present invention overcomes the defects of the prior art and provides a probiotic composition for removing Helicobacter pylori in the oral cavity.

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

[0009] Specifically, the preparation of the bacterial suspension of Lactobacillus acidophilus LA05 with a preservation number of CGMCC No. 23546 is as follows: Lactobacillus acidophilus LA05 is inoculated in a modified MRS liquid culture medium, cultured at 38° C. for activation, and activated twice continuously to obtain an activated liquid; the activated liquid is inoculated in an MRS liquid culture medium at an inoculum amount of 2% (v / v), and cultured at 38° C. The bacterial liquid concentration is 10 8The Lactobacillus plantarum deposit number is GDMCC No: 62256. Preparation of bacterial suspension: inoculate Lactobacillus plantarum LP1Z from the glycerol tube onto a modified MRS agar plate, culture it anaerobically at 37°C for 48h, pick a single colony of Lactobacillus plantarum LP1Z and inoculate it into a modified MRS medium, culture it anaerobically at 37°C, and the bacterial liquid concentration is 10 8 The fermentation is stopped when the CFU is reached. The fermentation liquid of Lactobacillus acidophilus LA05 and the fermentation liquid of Lactobacillus plantarum LP1Z are mixed in 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 drug for treating anti-Helicobacter pylori.

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

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

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

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

[0016] Specifically, the medicament or kit of the present invention contains a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. The use of such media and agents for pharmaceutically active substances is well known in the art. Except for any conventional media or agents that are incompatible with the active ingredient, the present invention also relates to its use in the therapeutic composition disclosed herein. Supplementary active ingredients can also be incorporated into the pharmaceutical composition.

[0017] On the other hand, the present invention also relates to the use of one or more disclosed polypeptides in the preparation of anti-Helicobacter pylori drugs. In some embodiments, the polypeptide is administered alone or in combination with any other desired therapeutic agent using a suitable delivery method. The disclosed therapeutically useful agents can be administered by, for example, one of the following routes: oral, for example, in the form of sugar coating, coated tablets, pills, semisolid substances, soft or hard capsules, solutions, emulsions or suspensions; parenteral, for example, in the form of injectable solutions; rectal suppositories; by inhalation, for example, in the form of powder preparations or sprays; and / or transdermal or intranasal. For the preparation of such tablets, pills, semisolid substances, coated tablets, lozenges and hard gelatin capsules, the therapeutic product can be mixed with pharmacologically inert, inorganic or organic pharmaceutical carrier substances, for example, lactose, sucrose, glucose, gelatin, malt, silica gel, starch or its derivatives, talc, stearic acid or its salts, skimmed milk powder, etc. For the preparation of soft capsules, pharmaceutical carrier substances such as vegetable oils, petroleum, animal oils or synthetic oils, waxes, fats and polyols can be used. In order to prepare liquid solutions and syrups, pharmaceutical carrier materials such as water, alcohol, saline solution, dextrose solution, polyols, glycerol, vegetable oils, petroleum and animal or synthetic oils can be used. For suppositories, pharmaceutical carrier materials such as vegetable oils, petroleum, animal or synthetic oils, waxes, fats and polyols can be used. For aerosol preparations, compressed gases suitable for this purpose, such as oxygen, nitrogen and carbon dioxide, can be used. Pharmaceutically acceptable agents can also include additives for preservation and stabilization, emulsifiers, sweeteners, flavor rings, salts for changing osmotic pressure, buffers, encapsulation additives and / or antioxidants. For those skilled in the art, many configurations and variations will be apparent after considering the disclosure of the present invention.

[0018] Examples of suitable pharmaceutically acceptable preservatives include, but are not limited to, benzalkonium chloride, benzethonium chloride and cetylpyridinium chloride, benzyl bromide, benzyl alcohol, disodium EDTA, phenylmercuric nitrate, phenylmercuric acetate, ethylmercuric sodium thiosalicylate, thimerosal, acetate and phenylmercuric borate, polymyxin B sulfate, chlorhexidine, methyl and propyl parabens, phenylethyl alcohol, quaternary ammonium chloride, sodium benzoate, sodium propionate, stable chlorine oxygen complex, sorbic acid or mixtures thereof. Preferred pharmaceutically acceptable preservatives include disodium EDTA (disodium ethylenediaminetetraacetic acid) and benzalkonium chloride or mixtures thereof. In one embodiment, the pharmaceutically acceptable preservative is present in an amount of about 0.01% to about 2.0% (weight / volume), preferably 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, borates, citrates, phosphates, acetates, saline, tris-HCl (tris (hydroxymethyl) aminomethane hydrochloride), HEPES (N-2-hydroxyethyl piperazine-N'-2-ethanesulfonic acid (N-2-hydroxyethyl piperazine-N1-2-ethane sulfonic acid)), sodium phosphate, sodium borate, saline, citrates, carbonates, phosphates and / or mixtures thereof to obtain the desired osmotic capacity (osmolarity). In one embodiment, the pharmaceutically acceptable buffer is present in an amount of about 0.01% to about 2.0% (weight / volume), preferably about 0.05% to about 1% (weight / volume).

[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 isotonicity regulators 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 isotonicity regulator is present in an amount of about 0.1% to about 5.0% (weight / volume), preferably about 1% to about 3% (weight / volume). BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 Effect of HP-si-125 peptide on urease activity of Helicobacter pylori

[0024] Beneficial Effects

[0025] The present invention provides a method for preparing probiotics and the use thereof in removing Helicobacter pylori in the oral cavity. More specifically, a polypeptide HP-si-125 having good inhibitory activity against Helicobacter pylori is obtained through computer simulation pre-screening and antibacterial experimental screening. The polypeptide and probiotics can have a good effect of treating Helicobacter pylori in a mouse model after being used alone or in combination. The polypeptide and probiotics can be prepared into a mouthwash or a medicine for treating Helicobacter pylori in the oral cavity, and have good application value. DETAILED DESCRIPTION

[0026] Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve. It is particularly important to point out that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The methods and applications of the present invention have been described by preferred embodiments, and relevant personnel can obviously change or appropriately change and combine the methods and applications described herein without departing from the content, spirit and scope of the present invention to achieve and apply the technology of the present invention. The methods, equipment, and materials in the following implementation cases, if not specifically stated, are all conventional methods, equipment, and materials in the art, and can be purchased from the market.

[0027] Example 1 Simulation design and screening of anti-Helicobacter pylori polypeptides

[0028] Design of antimicrobial peptide library: The length of antimicrobial peptides was set within the range of 13-16 amino acids, and the common dominant amino acids of antimicrobial peptides were selected as the basis to design the peptide library. Based on the anti-Helicobacter pylori resistance of pathogenic bacteria, computer combined with simulation screening was performed, and 14 peptides were initially screened for artificial synthesis.

[0029] The identification method of the antimicrobial peptide anti-Helicobacter pylori effect is as follows:

[0030] H. pylori (Helicobacter pylori ATCC43504, purchased from Shanghai Beino Biotechnology Co., Ltd.) frozen at -80°C was revived on Columbia agar plates containing 7% sheep blood and 0.4% H. pylori selective additives (Oxoid). After two generations of activation, the cells were cultured in Columbia blood plates for 48 h, scraped off, centrifuged at 4000 r / min for 10 min, resuspended in BHI, and the density was adjusted to 1×10 9 CFU / mL, and then dilute in series according to experimental needs.

[0031] After H.pylori was activated for two generations, 10 8 CFU H.pylori was spread on fresh Columbia medium without antibiotics, immediately placed in an Oxford cup, 100 μL of the polypeptide to be tested (100 μg / mL SEQ ID NO: 1-14) was added, fixed for 1 hour under microaerobic conditions, placed in a three-gas incubator and cultured for 72 hours, and then the size of the inhibition zone was measured. The experiment was repeated 3 times. The results are shown in Table 1.

[0032] Table 1 Inhibitory effect of each peptide on Helicobacter pylori

[0033]

[0034]

[0035] - indicates no inhibitory effect.

[0036] As can be seen from Table 1, the blank culture medium control as a negative control has no ability to inhibit the growth of H. pylori, and no inhibition zone appears. HP-si-125 peptide has the best inhibitory effect on Helicobacter pylori, so this peptide was selected for subsequent experiments.

[0037] Example 2: Verification of the effect of HP-si-125 polypeptide

[0038] Fresh Helicobacter pylori ATCC43504 was washed twice with PBS and the concentration of H. pylori was adjusted to 2×10 7CFU / mL. In a 96-well plate, take 50μL H.pylori and 10μL HP-si-125 polypeptide of different concentrations (10μg / mL, 100μg / mL, 200μg / mL, three concentrations of low, medium and high), mix well, and place in a three-gas incubator for co-culture for 48h. Then take out and add 150μL urease test solution, shake and use a 550nm microplate reader to measure the absorbance value. The blank is the result of the urease reagent solution assay; the negative control is BHI instead of the polypeptide assay; the positive control is 100μg / mL metronidazole tablets. Urease indicator formula: 0.9% NaCl, 20mmol / L urea, 14μg / mL phenol red. Use HCl to adjust the pH to 6.8 (the color change point of phenol red) and measure its density with a spectrophotometer at OD550nm. The results are as follows. Figure 1 shown.

[0039] from Figure 1 It can be seen that 24 hours after Helicobacter pylori was treated without adding peptides or drugs, the activity of urease was significantly increased compared with the blank control group (P<0.01), and after adding peptides and positive control treatment, the activity of urease was significantly reduced compared with the negative control group (P<0.05), especially the high concentration peptide group, the activity of urease was reduced by 2.7 times, the effect was significant. The reason why Helicobacter pylori can survive in the stomach with very low acidity is because Helicobacter pylori can produce urease. Inhibiting the urease activity of Helicobacter pylori will make Helicobacter pylori lose the favorable environment for survival, thereby inhibiting the colonization and growth of Helicobacter pylori in the stomach. Figure 1 The results showed that the polypeptide of the present invention has significant activity in inhibiting Helicobacter pylori.

[0040] Example 3 Peptide Safety Testing

[0041] The hemolytic effect of antimicrobial peptides and probiotics was detected using sheep red blood cells; 10 μl of HP-si-125 peptide solution with a concentration of 1 mg / ml was dripped on the surface of the blood plate, and distilled water was used as a control. It was placed at 37°C for more than 24 hours, and then the presence of hemolysis circle was observed. The results showed that no hemolysis circle appeared in all the tested antimicrobial peptides. Preliminary results show that the antimicrobial peptides do not have hemolytic toxicity.

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

[0043] Improved MRS liquid medium: glucose 20g / L, peptone 10g / L, beef extract 10g / L, yeast extract 5g / L, whey powder 10g / L, tomato paste 10g / L, K2HP042g / L, diammonium hydrogen citrate 2g / L, sodium acetate 5g / L, Tween801g / L, MgS04·7H200.5g / L, MnS04·4H200.05g / L, L-cysteine ​​hydrochloride 0.5g / L. Use 1mol / LNaOH to adjust the pH to 7.0, and sterilize at 121℃ for 20min.

[0044] Preparation of Lactobacillus acidophilus LA05 (preservation number is CGMCC No. 23546, see CN202210413078.0) bacterial suspension: Lactobacillus acidophilus LA05 is inoculated into a modified MRS liquid culture medium, cultured at 38° C. for 18 h for activation, and activated twice in succession to obtain an activated solution; the activated solution is inoculated into an MRS liquid culture medium at an inoculum amount of 2% (v / v), cultured at 38° C., and fermentation is stopped when the bacterial solution concentration is 108 CFU.

[0045] Preparation of Lactobacillus plantarum LP1Z (deposit number is GDMCC No: 62256, see CN202210263672.6) bacterial suspension: inoculate Lactobacillus plantarum LP1Z from a glycerol tube onto a modified MRS agar plate, culture anaerobically at 37° C. for 48 h, pick a single colony of Lactobacillus plantarum LP1Z and inoculate it into a modified MRS medium, culture it anaerobically at 37° C., and stop fermentation when the bacterial solution concentration is 108 CFU.

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

[0047] Example 5 Animal Experiments of Probiotic Compositions and Peptides

[0048] The experimental animals were (SPF) C57BL / 6 mice, aged 6-8 weeks, weighing 18-20 g. Each mouse was given 0.5 ml (containing 5×10 8 CFU) of Helicobacter pylori ATCC43504, once every other day, for a total of 3 times. Two weeks after modeling, the modeling animals were randomly sampled, and the mice were killed. The stomachs were taken for rapid urease test, smear Gram staining, pathological sections and bacterial culture tests to confirm that the mice were successfully infected.

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

[0050] Model group: 0.4 ml normal saline (NS) was administered intragastrically to each rat.

[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 50 mg / kg;

[0053] Probiotic group: 150 mg / kg of freeze-dried powder of the probiotic composition prepared in Example 4;

[0054] Peptide combined with probiotics group: HP-si-125 polypeptide 50 mg / kg; 2 hours after administration, 150 mg / kg of the probiotic composition lyophilized powder prepared in Example 4 was administered;

[0055] Each group of animals was given the drug once a day by gavage for 14 days. All mice were killed 4 weeks after the last administration. The stomach was dissected and cut into 4 parts along the greater curvature of the stomach. The rapid urease test, Gram staining, pathological sections and bacterial culture were performed to detect Hp. Those with negative results in all four detection methods were defined as Hp negative, and those with positive results in one or more detection methods were defined as Hp infection. The experimental results are shown in Table 2.

[0056] Table 2 In vivo anti-Helicobacter pylori activity results of each group

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

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

[0059] As can be seen from 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 the present invention were 80% and 70%, respectively, which were significantly different from the model group. In particular, the polypeptide combined with the probiotic group had a significant synergistic therapeutic effect, and the clearance rate reached 100%. This shows that the combined use has a better effect.

[0060] Example 6 Preparation and efficacy verification of mouthwash for removing oral Helicobacter pylori

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

[0062] 20 patients with Helicobacter pylori infection, 10 of whom were found to have Helicobacter pylori in their oral cavity, used the mouthwash of the present invention three times a day for 30 consecutive days. The oral mucosa of the patient was scraped with a sterile spatula, and 1 ml of saliva was collected. After sufficient mixing, the presence of H. pylori in the oral cavity was detected. The results showed that no Helicobacter pylori was detected in the oral cavity, showing a good therapeutic effect.

[0063] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as 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, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. However, these modifications or replacements do not make the essence of the corresponding technical solutions 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 according to claim 1 in the preparation of a drug for inhibiting anti-Helicobacter pylori activity.

3. Use of the anti-Helicobacter pylori polypeptide HP-si-125 and the probiotic composition as claimed in claim 1 in the preparation of a drug kit for treating anti-Helicobacter pylori; the probiotic composition is prepared by Lactobacillus acidophilus CGMCC No. 23546 and Lactobacillus plantarum GDMCC No: 62256 in a volume ratio of 2:1 under the same bacterial concentration conditions.

4. The use according to claim 3, characterized in that The medicine kit 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 of water, 2-15 parts of sorbitol, 1-8 parts of propylene glycol, 1-8 parts of xylitol, 1-5 parts of PEG-400 polyethylene glycol, 0.5-2 parts of dimethyl glycyrrhizinate, 1-5 parts of sodium benzoate, 0.1-1 parts of flavor, 0.5-5 parts of HP-si-125 polypeptide; 1-5 parts of probiotic composition freeze-dried powder; wherein the amino acid sequence of the HP-si-125 polypeptide is shown in SEQ ID NO: 8; the probiotic composition freeze-dried powder is prepared by configuring Lactobacillus acidophilus CGMCC No. 23546 and Lactobacillus plantarum GDMCC No: 62256 in a volume ratio of 2:1 under the same bacterial concentration conditions and then freeze-drying.

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