CN116004456B - Lactobacillus reuteri A21325 for inhibiting helicobacter pylori infection and application thereof - Google Patents
Lactobacillus reuteri A21325 for inhibiting helicobacter pylori infection and application thereof Download PDFInfo
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- CN116004456B CN116004456B CN202211690506.0A CN202211690506A CN116004456B CN 116004456 B CN116004456 B CN 116004456B CN 202211690506 A CN202211690506 A CN 202211690506A CN 116004456 B CN116004456 B CN 116004456B
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Lactobacillus reuteri A21325 for inhibiting helicobacter pylori infection, which strain was deposited at the microorganism strain collection in Guangdong province at 12/13 2022, accession number: building 5, no. 59 of Mitsui 100, guangzhou City, guangdong, with a deposit number of GDMCC NO. 63041. The strain can effectively inhibit helicobacter pylori infection of Lactobacillus reuteri A21325 by coagglutination with helicobacter pylori, and the inactivated strain of the strain can also coagglutinate with helicobacter pylori. The method for preparing the functional food and/or health care product by inhibiting helicobacter pylori infection of lactobacillus reuteri A21325.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus reuteri A21325 and application of a composition containing the lactobacillus reuteri A21325 in inhibiting helicobacter pylori infection and relieving inflammation caused by helicobacter pylori.
Background
Helicobacter pylori is a gram-negative microaerophilic bacterium, and to date, helicobacter pylori infection has been considered the most common bacterial infection worldwide. Helicobacter pylori was classified as a class I carcinogen by the world health organization in 1994. According to the intestinal gastric cancer occurrence pattern proposed by corea, the development path of helicobacter pylori from infection to cancer occurrence is: helicobacter pylori infects normal gastric mucosa-chronic non-atrophic gastritis-intestinal metaplasia-dysplasia-intestinal gastric cancer, inflammatory factors play an important role in the pathogenesis of helicobacter pylori, which directly or indirectly causes up-regulation of inflammatory factor expression, and participates in the lesion evolution of gastritis and gastric cancer through various ways.
Since helicobacter pylori is the main causative agent causing gastritis and gastric cancer, eradication of helicobacter pylori infection by various means is the main direction of treatment. Currently, the most common current therapy for H.pylori is eradication of pathogens by a combination of several antibiotics and proton pump inhibitors, such as antibiotic triple therapy, quadruple therapy, etc. Antibiotic therapy can eradicate helicobacter pylori and simultaneously cause serious side effects such as helicobacter pylori resistance, intestinal flora imbalance, intestinal dysfunction and the like.
Thus, in addition to current antibiotic therapies, the use of probiotics to inhibit and alleviate helicobacter pylori infection is of increasing concern. More and more researches show that specific probiotics can inhibit the growth and colonization of helicobacter pylori, and play an important role in the adjuvant therapy of helicobacter pylori. However, the existing probiotics have higher requirements on the activity of the probiotics when the probiotics play a role in inhibiting or antagonizing helicobacter pylori, and the probiotics must be capable of effectively inhibiting or antagonizing helicobacter pylori under the condition of higher viable bacteria number. In addition, the existing probiotics mainly inhibit or antagonize helicobacter pylori, and have no obvious effect of relieving inflammation caused by helicobacter pylori.
Disclosure of Invention
A first object of the present invention is to provide Lactobacillus reuteri A21325 which is effective in inhibiting helicobacter pylori infection by co-agglutination with helicobacter pylori, and an inactivated strain of the strain is also capable of co-agglutinating with helicobacter pylori.
A second object of the present invention is to provide the use of Lactobacillus reuteri A21325 for inhibiting helicobacter pylori infection, in particular for reducing the expression of the inflammatory factor TNF-alpha while promoting the upregulation of the expression of the anti-inflammatory factor IL-10, in inhibiting inflammation caused by helicobacter pylori infection.
A third object of the present invention is to provide the use of Lactobacillus reuteri A21325 for inhibiting helicobacter pylori infection in the preparation of a product for inhibiting helicobacter pylori infection.
Lactobacillus reuteri (Limosilactobacillus reuteri) A21325 for inhibiting helicobacter pylori infection, which strain was deposited at the Cantonese province microorganism strain collection at 12/13 2022 at the deposit address: building 5, no. 59 of Mitsui 100, guangzhou City, guangdong, with a deposit number of GDMCC NO. 63041.
The lactobacillus reuteri A21325 for inhibiting helicobacter pylori infection is characterized by comprising the following colony characteristics: on MRS solid culture medium, colony is round, milky white, raised, smooth, and opaque. After gram staining, the color development is observed by a microscope, and the strain is rod-shaped and gram positive.
The Lactobacillus reuteri A21325 for inhibiting helicobacter pylori infection is obtained by screening fresh fecal samples from healthy centenarian aged people in Shangxi Shanglin county.
The screening of the Lactobacillus reuteri A21325 for inhibiting helicobacter pylori infection comprises the following steps:
3.1 weighing 1.0g of fresh fecal sample, adding the fresh fecal sample into 9.0mL of sterile physiological saline, and shaking and uniformly mixing to prepare bacterial suspension. The bacterial suspension is subjected to gradient dilution by using sterile PBS buffer solution, and the dilution solution with proper dilution is coated on MRS chromogenic medium. Anaerobic culturing at 37deg.C for 2-4d, picking out strain with yellow color-changing ring, streaking on MRS solid culture medium, purifying, and preserving the purified strain on slant.
3.2 the test in 3.1The strain is picked up and inoculated with MRS broth culture medium for anaerobic culture at 37 ℃ for 16-24h. Centrifuging to collect thallus of strain to be tested, washing with PBS buffer solution for 2 times, re-suspending with PBS buffer solution and adjusting thallus concentration to 1×10 8 CFU/ml to obtain the bacterial suspension to be tested.
3.3 helicobacter pylori of the indicator bacteria was inoculated on Columbia blood agar plates and cultured in a three-gas incubator (5% oxygen, 10% carbon dioxide, 85% nitrogen) at 37℃for 72-96 hours. Helicobacter pylori cells were collected, washed 2 times with PBS buffer, resuspended with PBS buffer and the cell concentration was adjusted to 1X 10 8 CFU/ml gave a helicobacter pylori suspension.
3.4 mixing the bacteria to be detected with helicobacter pylori suspension in equal amount, shaking for 3-5min, and culturing at 37 ℃ for 2h. The upper liquid was aspirated to determine the OD600 absorbance and the coagglomeration rate was calculated. Selecting the strain with high coagglomeration rate for standby.
3.5 Single colonies of the strain obtained in 3.4 were inoculated into MRS broth, and anaerobically cultured at 37℃for 16-24 hours. Centrifuging to collect thallus of strain to be tested, washing with PBS buffer solution for 2 times, re-suspending with PBS buffer solution and adjusting thallus concentration to 1×10 8 CFU/ml, placing the bacterial liquid in a water bath at 75 ℃ and heating for 30 minutes to obtain the inactivated bacterial suspension to be tested.
3.6 Co-agglutination rates of the inactivated strains were examined by the method of 3.4, and strains with high co-agglutination rates were selected.
3.7 performing an intervention test of helicobacter pylori AGS cell adhesion by using the strain selected in 3.6, and screening the strain capable of reducing the helicobacter pylori adhesion rate.
3.8 the strain screened in 3.7 is used for carrying out the detection of AGS cell IL-8, and the strain which can effectively relieve the inflammation caused by helicobacter pylori is screened.
3.9 preparing freeze-dried bacterial powder from the bacterial strain screened in 3.8, feeding the freeze-dried bacterial powder and helicobacter pylori bacterial suspension to a C57 mouse, sampling and detecting indexes such as urease, TNF-alpha, IL-10 and the like in gastric tissue homogenate of the mouse after 8 weeks, and screening out bacterial strains capable of reducing the urease, the TNF-alpha and improving the IL-10.
The application of the Lactobacillus reuteri A21325 for inhibiting helicobacter pylori infection in preparing functional food and/or health care products.
The application of the Lactobacillus reuteri A21325 for inhibiting helicobacter pylori infection in preparing fermented dairy products, fermented fruit and vegetable products, beverages or snack fermented foods.
The invention has the beneficial effects that:
1. the Lactobacillus reuteri A21325 for inhibiting helicobacter pylori infection can generate coagglutination with helicobacter pylori, thereby inhibiting the colonization and infection of the helicobacter pylori.
2. The inactivated strain of lactobacillus reuteri a21325 may also co-agglutinate with helicobacter pylori.
3. The lactobacillus reuteri A21325 can effectively relieve cell inflammation caused by helicobacter pylori, and is mainly characterized by reducing the expression of inflammatory factors TNF-alpha and promoting the up-regulation of the expression of anti-inflammatory factors IL-10.
Drawings
FIG. 1 shows the AGS cell adhesion rate.
FIG. 2 shows the results of IL-8 concentration detection.
FIG. 3 shows the results of gastric urease assay.
FIG. 4 shows the result of TNF- α detection in stomach tissue.
FIG. 5 shows the results of IL-10 detection in gastric tissue.
Detailed Description
In order to facilitate an understanding of the invention, reference will now be made to certain embodiments and specific language will be used to describe the same. It should be understood, however, that these specific embodiments are not intended to limit the scope of the invention, and that any alterations and further modifications in the described embodiments, and any further applications of the invention, will occur to those skilled in the art.
Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Unless otherwise indicated, the media and test conditions used in the examples of the present invention are those conventional in the art.
The reagents used in this example are all commercially available unless otherwise specified.
In the following examples, the percentages are mass percentages unless otherwise indicated.
Example 1
The screening, purifying and identifying of the helicobacter pylori infection inhibiting lactobacillus reuteri A21325 comprises the following steps:
1.0g of fresh fecal sample of longevity old people is taken and added with 9ml of sterile physiological saline to be mixed evenly by shaking to prepare bacterial suspension. 5mL of the bacterial suspension is added into 45mL of enrichment medium, and anaerobic culture is carried out for 7d at 37 ℃. Gradient diluting the cultured enriched culture solution to 10 with sterile physiological saline -7 0.1ml dilution of 10 was taken -5 、10 -6 、10 -7 Is coated on MRS chromogenic medium containing bromocresol purple, and is cultured anaerobically at 37 ℃ for 2-4 days. Single colony with early yellow color-changing ring and large color-changing ring is picked, streaked on MRS plate, and repeated three times until the strain is purified.
And (3) selecting single bacterial colonies of the purified strain to be identified, preparing bacterial suspension in sterile water, and performing bacterial colony PCR amplification on the strain 16S rDNA by using the bacterial suspension as a PCR template. The PCR primer is synthesized by Shanghai Biotechnology Co.Ltd by adopting 27F and 1492R universal primers. The reaction was carried out using a 25. Mu.l system: 2 Xsan Taq PCR Mix 12.5. Mu.l, template 1. Mu.l, upstream and downstream primers 1. Mu.l each, make up ddH 2 O to 25. Mu.l. The reaction procedure is: pre-denaturation at 94℃for 5min, denaturation at 94℃for 30s, annealing at 55℃for 30s, extension at 72℃for 1min, 30 cycles were performed in total, and extension at 72℃for 10min. The reaction products were submitted to sequencing by Shanghai Bioengineering Co. Sequencing analysis shows that the strain is lactobacillus reuteri (Limosilactobacillus reuteri) and named lactobacillus reuteri A21325, and the 16S rDNA sequence of the strain is shown as SEQ ID NO.1, and the sequence is submitted to NCBI for Blast alignment. The strain is deposited in the Guangdong province microorganism strain collection center with the deposit number of GDMCC NO. 63041.
Example 2
Co-agglutination experiment of live Lactobacillus and helicobacter pylori
Preparing a bacterial suspension: washing activated bacteria to be tested and helicobacter pylori thallus with sterile PBS buffer solution for 2 times, and adjusting bacterial suspension concentration to 1×10 after re-suspending with PBS buffer solution 8 CFU/ml。
Mixing the bacterial suspension to be detected and helicobacter pylori suspension in equal amount, shaking for 3-5min, standing at 37deg.C for culture, sampling the supernatant for 0 hr and 2 hr for OD 600 And detecting the absorbance value. And calculating the interaction aggregation rate of the bacteria to be detected on helicobacter pylori according to a formula.
A a :0h of to-be-detected bacterial suspension OD 600 A value;
A b :0h helicobacter pylori suspension OD 600 A value;
A 2 : mixed bacterial suspension OD at each detection time point 600 Values.
The results of the test are shown in Table 1, and the rate of the A21325 strain to helicobacter pylori by the interaction and aggregation reached 76% at 2 hours. This suggests that strain A21325 has a strong binding capacity to helicobacter pylori, and that both form aggregates which help to exclude helicobacter pylori from the body.
TABLE 1 capability of the live Lactobacillus to mutually agglutinate helicobacter pylori
| Strain numbering | Co-aggregation Rate (%) |
| DSM17648 | 62 |
| A21325 | 71.89 |
Example 3
Co-agglutination experiments of Lactobacillus inactivated Strain and helicobacter pylori
Preparing a bacterial suspension: washing activated bacteria to be tested and helicobacter pylori thallus with sterile PBS buffer solution for 2 times, and adjusting bacterial suspension concentration to 1×10 after re-suspending with PBS buffer solution 8 CFU/ml, placing the bacterial liquid in a water bath at 75 ℃ and heating for 30 minutes to obtain the inactivated bacterial suspension to be tested. .
Mixing the bacterial suspension to be detected and helicobacter pylori suspension in equal amount, shaking for 3-5min, standing at 37deg.C for culture, sampling the supernatant for 0 hr and 2 hr for OD 600 And detecting the absorbance value. And calculating the interaction aggregation rate of the bacteria to be detected on helicobacter pylori according to a formula.
A a :0h of to-be-detected bacterial suspension OD 600 A value;
A b :0h helicobacter pylori suspension OD 600 A value;
A 2 : mixed bacterial suspension OD for 2h 600 Values.
The results are shown in Table 2, and the rate of the cross-talk aggregation of the inactivated strain A21325 against helicobacter pylori reaches 65.26% at 2 hours. This shows that the inactivated strain of strain A21325 still has a strong binding capacity with helicobacter pylori, which expands the application range and dosage form selection of strain A21325.
TABLE 2 capability of inactivated Strain to mutually-aggregated helicobacter pylori
| Strain numbering | Co-aggregation Rate (%) |
| A21325 | 41.9 |
Example 4
Experiment of helicobacter pylori cell adhesion inhibition by live Lactobacillus
AGS cell preparation: AGS cells were seeded in sterile 24 well plates at a cell seeding rate of 2X 10 5 Cells/well were cultured overnight with F12K+10% foetal calf serum as medium.
Helicobacter pylori suspension preparation: washing cultured fresh helicobacter pylori with PBS buffer solution for 2 times, re-suspending with F12K+10% foetal calf serum culture medium, and adjusting bacterial concentration to 2×10 8 CFU/ml。
Preparing a live bacteria suspension of a bacteria to be detected: washing cultured thallus to be tested with PBS buffer solution for 2 times, re-suspending with F12K+10% foetal calf serum culture medium, and adjusting bacterial liquid concentration to 2×10 8 CFU/ml。
Preparing an inactivated bacterial suspension of a bacteria to be tested: washing cultured thallus to be tested with PBS buffer solution for 2 times, inactivating in 75 deg.C water bath for 30 min, re-suspending the inactivated thallus with F12K+10% fetal bovine serum culture medium, and adjusting the concentration of the bacterial solution to 2×10 8 CFU/ml。
AGS cells cultured overnight in 24-well plates were washed 3 times with PBS buffer to remove non-adherent cells, then 0.25ml of helicobacter pylori suspension and 0.25ml of the test bacterial suspension were added, and co-cultured in a three-gas incubator at 37℃for 2 hours. After the incubation, the cells were washed 3 times with PBS buffer, and then urease reagent was added thereto to measure absorbance at 550 nm. The method is characterized in that AGS cells which are not treated by helicobacter pylori and to-be-detected bacteria are used as blank groups, AGS cells which are treated by helicobacter pylori and not treated by to-be-detected bacteria are used as model groups, AGS cells which are treated by helicobacter pylori and to-be-detected bacteria are used as experimental groups, and the adhesion rate of the helicobacter pylori is characterized by the activity of urease.
The adhesion rate is calculated by subtracting the OD value of the blank group from the OD value of the model group to be 100%, and the adhesion rate of the experimental group is calculated by the following formula:as shown in FIG. 1, compared with the HP model group, the adhesion rate of helicobacter pylori to cells after the treatment of A21325 live bacteria is reduced to 47.03%, the adhesion rate of helicobacter pylori to cells after the treatment of A21325 inactivated strain is reduced to 53.7%, and the adhesion rate of helicobacter pylori to cells can be obviously reduced by both live bacteria or inactivated strain of Lactobacillus reuteri A21325 strain.
Example 5
AGS cell IL-8 secretion assay
AGS cell preparation: AGS cells were seeded in sterile 24 well plates at a cell seeding rate of 1X 10 5 Cells/well were cultured overnight with F12K+10% foetal calf serum as medium.
Helicobacter pylori suspension preparation: washing cultured fresh helicobacter pylori with PBS buffer solution for 2 times, re-suspending with F12K+10% foetal calf serum culture medium, and adjusting bacterial concentration to 1×10 7 CFU/ml。
Preparing a bacterial suspension to be tested: washing cultured thallus to be tested with PBS buffer solution for 2 times, re-suspending with F12K+10% foetal calf serum culture medium, and adjusting bacterial liquid concentration to 1×10 7 CFU/ml。
AGS cells cultured overnight in 24-well plates were washed 3 times with PBS buffer to remove non-adherent cells, then 0.5ml of helicobacter pylori suspension was added, and co-cultured in a three-gas incubator for 2 hours. After the completion of the cultivation, the non-adherent helicobacter pylori was removed by washing 3 times with PBS buffer, 0.5ml of the suspension of the bacteria to be tested was added, and the mixture was co-cultivated in a three-gas incubator for 24 hours. AGS cells not infected with helicobacter pylori are used as a blank group, AGS cells infected with helicobacter pylori but not treated with test bacteria are used as a model group, and AGS cells infected with helicobacter pylori and treated with test bacteria are used as an experimental group. After the completion of the culture, the cell culture broth was collected, centrifuged at 4000rpm/min at 4℃for 10min, and the supernatant was collected. The supernatants were assayed for IL-8 content according to the method provided by ELISA kit (purchased from Shanghai Jiang Lai Biolabs). The results of the IL-8 assay are shown in FIG. 2, and compared with the control group, the concentration of IL-8 in the supernatant of the HP model group AGS cells infected with helicobacter pylori is remarkably increased by 4.5 times that of the control group; after being treated by the A21325 strain, the secretion amount of the AGS cell IL-8 is 2.6 times that of a control group, and is reduced by 42.2 percent compared with an HP model group. The result shows that the lactobacillus reuteri A21325 can obviously inhibit the infection of helicobacter pylori to cells, and can reduce the secretion of a cell pro-inflammatory factor IL-8 and relieve the cell inflammation caused by HP.
Example 6
Tolerance of strains to artificial gastrointestinal fluids
Preparing a bacterial suspension: transferring and activating the strain from the preservation inclined plane for 2 times, inoculating MRS culture medium, culturing for 16-24 hr, and regulating bacterial suspension concentration to 1×10 9 CFU/ml。
Artificial gastric juice resistance experiment: 1ml of bacterial suspension is added into 9ml of artificial gastric juice with pH value of 3.0, and anaerobic culture is carried out for 3 hours at 37 ℃; the culture solutions were subjected to gradient dilution at 0h and 3h, respectively, and then plated on MRS plates for plate counting. The survival rate was calculated according to the formula.
Survival rate of artificial gastric juice= (3 h plate colony count/0 h plate colony count) ×100%
Artificial intestinal juice resistance experiment: 1mL of bacterial suspension is added into 9mL of artificial gastric juice with pH value of 3.0, anaerobic culture is carried out for 3 hours at 37 ℃, 1mL of bacterial suspension is added into 9mL of artificial intestinal juice with pH value of 8.0, and anaerobic culture is carried out for 8 hours at 37 ℃; and adding artificial intestinal juice, taking culture solution at 0h and 8h respectively, performing gradient dilution, coating on an MRS plate, performing plate counting, and calculating the survival rate according to a formula.
Survival rate of artificial intestinal juice= (8 h plate colony count/0 h plate colony count) ×100%
The survival statistics are shown in table 3. Lactobacillus reuteri a21325 can well withstand gastric acid environments and also has good tolerance to intestinal fluids.
TABLE 3 results of test for artificial gastrointestinal fluid tolerance by strain
Example 7
Preparation of freeze-dried bacterial powder and microecological bacterial agent
Lactobacillus reuteri a21325 is inoculated into an MRS solid culture medium for activation, and the activated strain is inoculated into an MRS liquid culture medium to prepare seed liquid. The prepared seed solution is inoculated into MRS liquid culture medium with an inoculum size of 2 percent, and is cultured for 16 hours at 37 ℃ for mass fermentation. After the fermentation is finished and microscopic examination is confirmed that no pollution exists, the bacterial liquid is centrifuged for 10min at 4000-6000r/min at 4 ℃ to collect bacterial cells.
Washing the collected thalli with sterile physiological saline for 2 times, adding conventional freeze-drying protective agents such as skim milk powder and the like into the thalli according to a ratio of 1:1, and uniformly mixing to prepare bacterial suspension. Transferring the bacterial liquid into a freeze-drying container, cooling and freezing in a gradient way, and freeze-drying by using a vacuum freeze dryer. And crushing and grinding the freeze-dried thalli to obtain lactobacillus reuteri A21325 freeze-dried fungus powder.
The lactobacillus reuteri A21325 freeze-dried bacterial powder and common auxiliary materials are mixed and then are pressed into tablets or filled into capsules to prepare micro-ecological bacterial agents such as powder, tablets, capsules and the like.
Example 8
Effect of a21325 strain and other commercially available strains in preventing or alleviating helicobacter pylori infection in mice
In order to verify that lactobacillus reuteri a21325 of the present invention has the effect of preventing or alleviating helicobacter pylori infection in animals, experiments were conducted by selecting 6-8-week-old C57BL/6 mice, and 1 commercially available lactobacillus plantarum having the effect of antagonizing helicobacter pylori (here, lactobacillus plantarum a is used instead of a specific strain name) was purchased as a reference, and the effects of lactobacillus reuteri a21325 strain and commercially available lactobacillus plantarum a strain on preventing or alleviating helicobacter pylori infection in mice were compared.
The blank control group is not filled with gastric helicobacter and helicobacter pylori, the HP model group is only filled with gastric helicobacter pylori bacterial liquid and is not filled with gastric helicobacter, and the experimental group is filled with gastric helicobacter pylori bacterial liquid and is filled with gastric Lactobacillus to be detected after 1h interval. Dissolving the freeze-dried powder of the lactobacillus to be detected by using sterile normal saline, and then, lavaging the mice for 7 days, then, lavaging helicobacter pylori liquid once a day, continuously lavaging for 5 days, and then, lavaging the lactobacillus to be detected for 49 days. After the feeding, the mice are sacrificed by cervical fracture, and the stomach tissues are dissected and taken by a sterilized scalpel and scissors, and the stomach tissues are treated according to the following ratio of 1:5 adding sterile physiological saline, homogenizing, and detecting indexes of the obtained gastric tissue homogenate.
Urease assay was performed in 96-well plates, gastric homogenates were centrifuged at 4000rpm for 10min at 4℃and 10. Mu.L of supernatant was added to sterile 96-well plates, 190. Mu.L of urease assay solution was added thereto, and after shaking reaction, absorbance at 550nm was measured using an microplate reader. Urease test agent formulation was 0.9% NaCl,20mmol/L urea, 14. Mu.g/mL phenol red, and pH was adjusted to 6.8 with HCl. The relative activities of the urease groups are shown in FIG. 3, taking the activity of the urease of the HP model group as 100%. Urease is secreted by helicobacter pylori, the urease in the stomach tissue of the mice in the model group after being infected by HP is obviously increased compared with a blank control group, and the urease in the stomach tissue of the mice after being perfused with the Lactobacillus reuteri A21325 is reduced to 69.4% of the HP model group, and the effect is better than that of the commercially available Lactobacillus plantarum A (79.2%). This indicates that lactobacillus reuteri a21325 can better inhibit helicobacter pylori infection colonization in animal mice.
The TNF-alpha content of the stomach tissue of the mice was measured using a kit from Shanghai Jiang Lai Bio Inc., the stomach tissue homogenate was centrifuged at 4℃and 4000rpm for 10 minutes, and the supernatant was measured according to the instructions provided by the kit, with the relative concentration of TNF-alpha in the HP model group being 100%, as shown in FIG. 4. The HP model group mice were infected with helicobacter pylori and the inflammatory response of gastric tissue produced an increase in TNF-alpha content over the blank group. After the mice were perfused with lactobacillus reuteri a21325, the TNF- α content of the stomach tissue of the mice was reduced to 54.38% of that of the HP model group, with better effect than that of the commercially available lactobacillus plantarum a (60.1%). The lactobacillus reuteri A21325 can well inhibit helicobacter pylori infection, effectively reduce the rise of inflammatory factor expression caused by helicobacter pylori infection, and alleviate inflammation.
The IL-10 content of the stomach tissue of the mice was measured by using a Jiangsu enzyme immune company kit, the stomach tissue homogenate was centrifuged at 4℃and 4000rpm for 10 minutes, and the supernatant was measured according to the instructions provided by the kit, and the IL-10 concentration of the HP model group was 100%, and the relative concentration of each group of IL-10 was shown in FIG. 5. The HP model group mice showed no significant change in gastric tissue after infection with helicobacter pylori but no significant change in the anti-inflammatory factor IL-10 compared to the placebo group. After the mice were perfused with lactobacillus reuteri a21325, the IL-10 content of the stomach tissue of the mice was increased over that of the HP model group and the blank group, with a 1.53-fold higher content than that of the HP model group, while the commercially available lactobacillus plantarum a was at the same level as that of the blank group and the HP model group. This indicates that the mice stimulated gastric tissue to increase the expression of anti-inflammatory factor IL-10 under the action of Lactobacillus reuteri A21325 after inflammatory reaction of gastric tissue infected by helicobacter pylori, inhibit the progress of inflammatory reaction, and alleviate inflammation.
Example 9
Fermented cow milk preparation containing lactobacillus reuteri a21325
Pasteurizing cow milk or skimmed cow milk by conventional method, cooling to 37deg.C, adding Lactobacillus reuteri A21325 powder, and stirring. Fermenting at 37-40deg.C for 8 hr to obtain fermented milk containing Lactobacillus reuteri A21325.
Example 10
Preparation of solid beverage containing lactobacillus reuteri a21325
Lactobacillus reuteri a21325 is as shown in example 7, a Cheng Luoyi lactobacillus reuteri a21325 strain freeze-dried powder is prepared, inulin 15%, fructo-oligosaccharide 15%, isomaltooligosaccharide 15%, galacto-oligosaccharide 15%, trehalose 10%, stachyose 10%, lactobacillus reuteri a21325 freeze-dried powder 10%, cherry fruit powder 5% and blueberry fruit powder 5% are weighed, mixed uniformly by a mixer, and the mixture is packaged and sealed by a powder packaging machine.
Claims (6)
1. Lactobacillus reuteri (Limosilactobacillus reuteri) a21325 for inhibiting helicobacter pylori infection, characterized in that the strain was deposited at the collection of microorganism strains in the cantonese province at 12/13/2022 at the deposit address: building 5, no. 59 of Mitsui 100, guangzhou City, guangdong, with a deposit number of GDMCC NO. 63041.
2. Lactobacillus reuteri a21325 for inhibiting helicobacter pylori infection according to claim 1, characterized in that the colony is characterized in that: on MRS solid culture medium, colony is round, milky white, raised, smooth, and clean in edge, and is opaque, and after gram staining, the strain is rod-shaped and gram positive.
3. The helicobacter pylori infection-inhibiting lactobacillus reuteri a21325 according to claim 1, wherein the helicobacter pylori infection-inhibiting lactobacillus reuteri a21325 is selected from fresh fecal samples from healthy centella asiatica in Shangxi county.
4. Use of lactobacillus reuteri a21325 for inhibiting helicobacter pylori infection according to claim 1 for the preparation of a functional food and/or a health care product.
5. Use of lactobacillus reuteri a21325 for inhibiting helicobacter pylori infection according to claim 1 for the manufacture of a fermented dairy product, a fermented fruit and vegetable product, a beverage or a snack fermented food.
6. Use of lactobacillus reuteri a21325 for inhibiting helicobacter pylori infection according to claim 1 in any one or more of the following (i) - (v);
preparing a product for inhibiting helicobacter pylori infection;
(II) preparing a product for inhibiting the activity of helicobacter pylori urease;
(III) preparing a product for reducing the adhesion rate of helicobacter pylori to cells;
(IV) preparing a product for inhibiting the secretion amount of IL-8 or TNF-alpha in helicobacter pylori infected cells;
(V) preparation of a product for promoting secretion of IL-10 in helicobacter pylori infected cells.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112430553A (en) * | 2020-11-25 | 2021-03-02 | 山东向日葵生物工程有限公司 | Lactobacillus reuteri SF-L-25 with helicobacter pylori inhibiting function and application thereof |
| CN113337431A (en) * | 2021-06-04 | 2021-09-03 | 青岛诺森生物技术有限责任公司 | Lactobacillus reuteri NSL0501 for inhibiting helicobacter pylori as well as biological agent and application thereof |
| CN114214230A (en) * | 2021-12-09 | 2022-03-22 | 江南大学 | A strain of Lactobacillus northern acid with the ability to co-polymerize Helicobacter pylori and its application |
| CN114480231A (en) * | 2022-04-18 | 2022-05-13 | 南京益瑞兰生物科技有限公司 | Lactobacillus reuteri for resisting helicobacter pylori infection and application thereof |
| CN115252654A (en) * | 2022-07-12 | 2022-11-01 | 国珍健康科技（北京）有限公司 | A kind of composition for inhibiting Helicobacter pylori and its preparation method and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113337431A (en) * | 2021-06-04 | 2021-09-03 | 青岛诺森生物技术有限责任公司 | Lactobacillus reuteri NSL0501 for inhibiting helicobacter pylori as well as biological agent and application thereof |
| CN114214230A (en) * | 2021-12-09 | 2022-03-22 | 江南大学 | A strain of Lactobacillus northern acid with the ability to co-polymerize Helicobacter pylori and its application |
| CN114480231A (en) * | 2022-04-18 | 2022-05-13 | 南京益瑞兰生物科技有限公司 | Lactobacillus reuteri for resisting helicobacter pylori infection and application thereof |
| CN115252654A (en) * | 2022-07-12 | 2022-11-01 | 国珍健康科技（北京）有限公司 | A kind of composition for inhibiting Helicobacter pylori and its preparation method and application |
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