CN115119940A - Application of Lactobacillus acidophilus LA85 and Bifidobacterium lactis BLA80 in inhibiting helicobacter pylori - Google Patents
Application of Lactobacillus acidophilus LA85 and Bifidobacterium lactis BLA80 in inhibiting helicobacter pylori Download PDFInfo
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- CN115119940A CN115119940A CN202111614935.5A CN202111614935A CN115119940A CN 115119940 A CN115119940 A CN 115119940A CN 202111614935 A CN202111614935 A CN 202111614935A CN 115119940 A CN115119940 A CN 115119940A
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- helicobacter pylori
- lactobacillus acidophilus
- bifidobacterium lactis
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Abstract
The invention belongs to the technical field of microorganisms, and particularly relates to application of lactobacillus acidophilus LA85 and bifidobacterium lactis BLA80 in inhibiting helicobacter pylori. The two strains of Lactobacillus acidophilus LA85 and Bifidobacterium lactis BLA80 can inhibit the growth of helicobacter pylori, and the diameters of the inhibition zones of the helicobacter pylori are respectively 25mm and 16 mm. And also has the ability to reduce the urease activity and the ability to reduce the adhesion of helicobacter pylori to AGS cells, respectively. Through animal model experiments, the mixed probiotics consisting of lactobacillus acidophilus LA85 and bifidobacterium lactis BLA80 can inhibit the level of inflammatory factors after helicobacter pylori infection and relieve inflammation caused by helicobacter pylori infection. The mixed probiotics consisting of lactobacillus acidophilus LA85 and bifidobacterium lactis BLA80 has stronger inflammation relieving effect than that of any single bacterium.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to application of lactobacillus acidophilus LA85 and bifidobacterium lactis BLA80 in inhibiting helicobacter pylori.
Background
Helicobacter pylori is a gram-negative bacterium that spirals or arcs and has very stringent growth requirements. The first successful isolation from human gastric mucosal biopsies in 1983 is the only microorganism species currently known to survive in the human stomach. In 2017, 10, 27 days, a carcinogen list published by the international cancer research institution of the world health organization is preliminarily collated for reference, and helicobacter pylori (infection) is in a class of carcinogen list. A large number of researches show that helicobacter pylori is a main pathogenic bacterium causing diseases such as chronic gastritis, peptic ulcer, gastric cancer and gastric mucosa-associated lymphoid tissue lymphoma. About more than 50% of the population worldwide become infected with H.pylori. Currently, the treatment of helicobacter pylori in clinic mainly adopts the combination of antibiotics, and is easy to repeat. In addition, improper use of antibiotics can cause adverse reactions, such as increased drug resistance in humans, and gastrointestinal flora disorders. The application of probiotics is particularly important for effectively and greenly inhibiting helicobacter pylori to relieve various diseases caused by the infection of the helicobacter pylori.
Probiotics are living microorganisms that, when ingested in sufficient quantities, produce beneficial effects on human health. The probiotics has various effects of promoting nutrient solution substance absorption, inhibiting the growth of intestinal pathogenic bacteria, improving intestinal micro-ecology, regulating organism immunity, reducing side effects caused by drug therapy and the like, and plays a vital role in human health. Lactic acid bacteria are the most widely used, longest-lasting and most diverse probiotic species, of which lactobacilli and bifidobacteria are more widely used. A great deal of research at home and abroad shows that probiotics have common consensus on the aspect of treating colon diseases and have important positions on the aspects of treating digestive tract (including stomach), gastric cancer, functional dyspepsia and the like.
Disclosure of Invention
The invention aims to provide application of lactobacillus acidophilus LA85 and bifidobacterium lactis BLA80 in inhibiting helicobacter pylori.
Lactobacillus acidophilus LA85, which is classified and named Lactobacillus acidophilus, is deposited in China general microbiological culture Collection center (CGMCC) at No. 1 of the China Committee for culture Collection of microorganisms at No. 01 of 3 of Beijing area of West Lu of the sunward area of Beijing, and the deposition number is CGMCC No. 21802.
Lactobacillus acidophilus LA85 belongs to human lactic acid bacteria, has high capability of resisting gastrointestinal tract stress, has obvious inhibiting effect on conditional pathogenic bacteria Escherichia coli, salmonella and staphylococcus aureus, is sensitive to common antibiotic drugs, has no drug resistance, has high growth rate, short growth cycle and high viable count, and can effectively regulate the immune response of organisms.
Bifidobacterium lactis BLA80 with accession number CGMCC No.22547, its classification name is: bifidobacterium animalis subsp. lactis, preservation time: 2021, 05 month, 17 days; the preservation unit: china general microbiological culture Collection center; and (4) storage address: xilu No. 1 Hospital No. 3, Beijing, Chaoyang, North.
The bifidobacterium lactis BLA80 has strong bile salt hydrolase activity and excellent cholesterol-lowering property, can obviously reduce the content of serum cholesterol, and can be used for preparing medicaments and foods for lowering cholesterol, such as health-care foods or solid beverages and the like. The bifidobacterium lactis BLA80 also has high capability of resisting gastrointestinal tract stress, strong capability of inhibiting common pathogenic bacteria of a human body and capability of effectively regulating the intestinal flora structure and immune response of an organism.
Through further research, the lactobacillus acidophilus LA85 and the bifidobacterium lactis BLA80 can inhibit the growth of helicobacter pylori and reduce the urease activity of the helicobacter pylori.
The Lactobacillus acidophilus LA85 and the Bifidobacterium lactis BLA80 can remarkably inhibit the growth of helicobacter pylori, the inhibition diameters of inhibition zones are respectively 25mm and 16mm, and the lactobacillus acidophilus LA85 and the Bifidobacterium lactis BLA80 have the capability of resisting helicobacter pylori infection.
The Lactobacillus acidophilus LA85 and the Bifidobacterium lactis BLA80 can remarkably reduce the urease activity of helicobacter pylori, and the inhibition rates of the urease activity of the helicobacter pylori are 78.63% and 67.49% respectively.
Thus, the present invention provides firstly the use of Lactobacillus acidophilus LA85 or Bifidobacterium lactis BLA80 for the preparation of a product for inhibiting the growth of helicobacter pylori. The products include pharmaceuticals and food products. The dosage form of the medicine is powder, granules, capsules, tablets, pills or oral liquid.
The Lactobacillus acidophilus LA85 and Bifidobacterium lactis BLA80 are capable of reducing the adhesion of helicobacter pylori to AGS cells. Further reducing the infection degree of the helicobacter pylori, reducing the colonization amount of the helicobacter pylori in a helicobacter pylori infected patient, and preventing and treating gastric cancer or gastritis caused by the helicobacter pylori infection.
Thus, secondly, the present invention provides the use of lactobacillus acidophilus LA85 or bifidobacterium lactis BLa80 in the manufacture of a product for the prevention or treatment of gastritis or gastric cancer. The products include pharmaceuticals and food products. The dosage form of the medicine is powder, granules, capsules, tablets, pills or oral liquid.
The lactobacillus acidophilus LA80 has stronger capability of inhibiting the growth of helicobacter pylori and reducing the activity of urease; the Bifidobacterium lactis BLA80 has stronger capability of reducing the adhesion of helicobacter pylori. In addition, the two strains have the effects of intestinal colonization and intestinal mucosa protection and repair, and the mixed use of the two probiotics can improve the helicobacter pylori infection resistance of a subject, increase the probiotic treatment and reduce the colonization amount of helicobacter pylori in the stomach or the caused inflammation. The invention provides a mixed probiotic composition consisting of two bacterial strains, which is characterized in that the mass ratio of the mixed probiotic composition consisting of the two bacterial strains is 1-5: 1, and particularly, the mass ratio of the composition consisting of the two bacterial strains is preferably 2: 1.
The mixed probiotic composition consisting of the lactobacillus acidophilus LA85 and the bifidobacterium lactis BLA80 can inhibit the level of inflammatory factors after helicobacter pylori infection and relieve inflammation caused by the helicobacter pylori infection in animal model experiments. The mixed probiotic consisting of lactobacillus acidophilus LA85 and bifidobacterium lactis BLA80 has stronger inhibition effect than that of any single bacterium.
The invention also provides application of the mixed probiotics consisting of the lactobacillus acidophilus LA85 and the bifidobacterium lactis BLA80 in preparing products for preventing or treating helicobacter pylori infection. The product comprises a food or a pharmaceutical product.
The food includes but is not limited to probiotic solid beverages, probiotic beverages, tabletted candies, snack foods, dairy products, soy products, fruit and vegetable products, and functional health products.
The food product is particularly comprised in a health food product for alleviating or treating gastric ulcers, duodenal ulcers or gastrointestinal discomfort.
The dosage form of the medicine is powder, granules, capsules, tablets, pills or oral liquid.
The invention has the beneficial effects that:
1. the lactobacillus acidophilus LA85 and the bifidobacterium lactis BLA80 have high capability of resisting gastrointestinal tract stress, have strong capability of inhibiting common pathogenic bacteria of a human body, can effectively regulate the intestinal flora structure and the immune response of the organism and promote the health of the human body.
2. The two strains of Lactobacillus acidophilus LA85 and Bifidobacterium lactis BLA80 can inhibit the growth of helicobacter pylori, and the diameters of the inhibition zones of the helicobacter pylori are respectively 25mm and 16 mm. And also has the ability to reduce the urease activity and the ability to reduce the adhesion of helicobacter pylori to AGS cells, respectively.
3. The mixed probiotics consisting of lactobacillus acidophilus LA85 and bifidobacterium lactis BLA80 are tested by animal model experiments
4. Small-scale population tests of helicobacter pylori infection prove that the mixed probiotic composition consisting of lactobacillus acidophilus LA85 and bifidobacterium lactis BLA80 can obviously relieve gastrointestinal symptoms of patients infected with helicobacter pylori and obviously improve the level of inflammatory factors in bodies of patients with helicobacter pylori-related chronic gastritis.
Drawings
FIG. 1 the zone of inhibition diameter of probiotic strains against H.pylori infection.
FIG. 2 probiotic strains reduce the uricase activity of H.pylori.
FIG. 3 the improvement of gastrointestinal symptoms of patients before and after the experiment was evaluated.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.
The helicobacter pylori (ATCC 43504) used in this example was derived from the cantonese culture collection. The general culture medium used in this example is commercially available, and in this example Columbia agar is purchased from OXOID, BHI solid medium and liquid medium are purchased from Qingdao Haibo Biotech, Inc., and sterile defibrinated sheep blood is purchased from Shanghai leaf Biotech, Inc.
The media formulations used in the following examples were:
modified MRS medium: 20g of glucose, 10g of tryptone, 10g of beef extract, 5g of yeast extract and K 2 HPO 4 2g, 2g sodium citrate, MgSO4 & 7H 2 O 0.2g、MnSO4·4H 2 O0.2 g, 1000mL distilled water. Sterilizing at 121 deg.C for 20 min.
Columbia blood plate: BHI solid medium and columbia agar were mixed as 1: 3, dissolving in 1L of water, sterilizing at 121 ℃ for 20min, adding 5% (V/V) of sterile defibered sheep blood when cooling to 50-55 ℃, uniformly mixing, and pouring.
Seed culture medium: 20g/L glucose, 10g/L beef extract powder, 5g/L yeast powder, 10g/L peptone, 5g/L sodium acetate, 2g/L sodium citrate, 3g/L dipotassium phosphate, 0.5g/L magnesium, 0.1g/L manganese sulfate, 1g/L Tween 80 and 1 g/LL-cysteine hydrochloride, and sterilizing at 115 ℃ for 20 min.
Fermentation medium: 50g/L lactose, 15g/L yeast powder, 20g/L peptone, 5g/L beef liver extract powder, 5g/L sodium acetate, 3g/L sodium citrate, 3g/L dipotassium hydrogen phosphate, 0.5g/L magnesium sulfate, 0.1g/L manganese sulfate, 1g/L Tween 80, 1 g/LL-cysteine hydrochloride, and sterilizing at 115 ℃ for 20 min.
Example 1 the Effect of Lactobacillus acidophilus LA85 and Bifidobacterium lactis BLA80 on inhibiting the growth of helicobacter pylori
Starting from a glycerol tube, 100ul of helicobacter pylori is coated on a Columbia blood plate, and cultured in a three-gas culture box (85% N2, 10% CO2 and 5% O2) at 37 ℃ for 3-5 days to obtain a single colony; selecting a single colony, inoculating the single colony in a BHI liquid culture medium containing 10% (v/v) fetal calf serum, and culturing in a three-gas incubator (85% N2, 10% CO2 and 5% O2) at 37 ℃ for 72h to obtain a seed solution; inoculating the seed solution into BHI liquid culture medium at an inoculum size of 2% (v/v), culturing in 37 deg.C three-gas incubator (85% N2, 10% CO2, 5% O2) for 72h to obtain helicobacter pylori liquid, adjusting the bacterial suspension to 10% 9 CFU/mL. Inoculating probiotic strains into modified MRS liquid culture medium according to the inoculation amount of 2% (v/v), culturing at 37 deg.C for 18-24h, activating for 2 generations, and respectively preparing bacterial suspensions of Lactobacillus acidophilus LA85 and Bifidobacterium lactis BLA 80.
BHI solid medium and colombia agar were mixed as 1: 3, when the solid culture medium is cooled to 50-55 ℃, adding 5% (V/V) of sterile defibrinated sheep blood, uniformly mixing, and uniformly mixing with the prepared helicobacter pylori bacterial suspension according to a certain proportion to ensure that the number of the helicobacter pylori viable bacteria in the system is 10 7 Of the order of CFU/mL. Pouring into a plate with Oxford cup, cooling and solidifying, taking out the Oxford cup, and pouring 200 μ per holeL probiotic fermentation liquor, namely, lightly covering a plate, and then, culturing in a three-gas culture box (85% N2, 10% CO2 and 5% O2) at 37 ℃ for 48-72h, observing after culturing for a proper time, and measuring the diameter of a bacteriostatic circle by using a vernier caliper. Modified MRS liquid medium was used as negative control and 3 replicates were tested.
TABLE 1 size of inhibition zone of LA85 and BLA80 against H.pylori
As can be seen from Table 1, the negative control had no zone of inhibition against H.pylori, the zone of inhibition against H.pylori by Lactobacillus acidophilus LA85 was up to 25mm in diameter (shown in FIG. 1), and the zone of inhibition against H.pylori by Bifidobacterium lactis BLA80 was up to 16mm in diameter (shown in FIG. 1). The results show that Lactobacillus acidophilus LA85 and Bifidobacterium lactis BLA80 both have the effects of inhibiting the growth of helicobacter pylori and can prevent the infection of helicobacter pylori.
Example 2 the Effect of Lactobacillus acidophilus LA85 and Bifidobacterium lactis BLA80 on reducing urease activity
After culturing helicobacter pylori according to the culture conditions of the above examples, the cells were washed with PBS, centrifuged, and the helicobacter pylori concentration was adjusted to 10 using BHI 8 CFU/mL. After the probiotics were cultured according to the culture conditions of the above examples, fermentation supernatants of lactobacillus acidophilus LA85 and bifidobacterium lactis BLa80 were prepared, respectively. Adding 40 mu L of helicobacter pylori bacterial suspension and 10 mu L of probiotic supernatant into a 96-well plate, uniformly mixing, and putting into a three-atmosphere culture box for incubation for 24 h; then taking out and adding 150 mu L of pre-prepared urease indicator (0.9% NaCl, 20mmo1/L urea, 14ug/mL phenol red, adjusting pH to 6.8 by HC 1), oscillating and measuring the absorbance value of the enzyme-labeled reagent at the wavelength of 550nm by an enzyme-labeled instrument, wherein the blank group is the urease indicator, and the control group is BHI culture medium.
The probiotic bacteria produce metabolites which inhibit the urease activity of helicobacter pylori to inhibit helicobacter pylori. The urease activity of H.pylori can be expressed by its absorbance at a wavelength of 550 nm. As shown in FIG. 2, the results of the urease activity inhibition assay of the two strains LA85 and BLA80 show that the inhibition rate of LA85 urease is 78.63% and the inhibition rate of BLA80 uricase is 67.49%, which indicates that the two strains can effectively inhibit the urease activity of helicobacter pylori, thereby controlling pathogenic bacteria to destroy the acidic environment of stomach and reducing inflammation and ulcer.
Example 3 Effect of Lactobacillus acidophilus LA85 and Bifidobacterium lactis BLA80 on adhesion to helicobacter pylori
Respectively centrifuging helicobacter pylori (Hp) bacterial liquid, Lactobacillus acidophilus LA85 bacterial liquid and Bifidobacterium lactis BLA80 bacterial liquid, collecting thallus, and adjusting bacterial suspension concentration to 10 with F-12 culture medium 7 CFU/mL. Human gastric adenocarcinoma cells (AGS) were resuspended in F-12 medium containing 5% (v/v) fetal bovine serum and added to 96-well plates (2X 10) 4 One/well), 5% CO at 37 ℃ in culture 2 After the AGS cells are in an adherent state, washing the AGS cells for 3 times by using PBS to remove dead cells; the assay was divided into blank group (AGS cell group), model group (Hp group), experimental group (Hp + LA85 group), experimental group (Hp + BLa80 group), wherein AGS cell group was AGS cells not inoculated with LA85 and BLa80 and not infected with Hp; the Hp group is AGS cells infected with Hp only; hp + LA85 group was AGS cells inoculated with LA85 and infected with Hp; the Hp + BLa80 group was AGS cells inoculated with BLa80 and infected with Hp.
Hp group was prepared by adding helicobacter pylori resuspension to washed AGS cells at 37 ℃ with 5% CO 2 After 2h of culture in the incubator, washing with PBS for 3 times to remove unadsorbed helicobacter pylori to obtain helicobacter pylori-infected AGS cells; the Hp + LA85 group is prepared by adding 200 μ l of LA85 bacterial suspension into helicobacter pylori-infected AGS cells respectively, culturing at 37 deg.C and 5% CO 2 Culturing for 2h in the incubator to obtain AGS cells inoculated with LA85 and infected with Hp; the Hp + BLA80 group was prepared by adding 200. mu.l of BLA80 bacterial suspension to the AGS cells infected with helicobacter pylori, respectively, and culturing at 37 ℃ with 5% CO 2 Culturing for 2h in the incubator to obtain AGS cells inoculated with BLA80 and infected with Hp; hp + LA85 group and Hp + BLA80 groupAfter the cells were washed 5 times with PBS solution, 200. mu.L of urease reagent (9g/LNaCl, 14. mu.g/mL phenol red, 20mM urea, pH6.8) was added thereto, and the cells were incubated at 37 ℃ with 5% CO 2 Culturing for 2h in the incubator to obtain a culture solution; the absorbance values at a wavelength of 550nm of the different sets of culture solutions were determined by a microplate reader.
TABLE 2 adhesion rates of helicobacter pylori to AGS cells of different groups
As can be seen from Table 2, the adhesion rate of H.pylori to AGS cells was significantly reduced after LA85 treatment, from 100% to 69.38% of Hp in the model group; after treatment with BLa80, the adhesion rate of h.pylori to AGS cells decreased significantly, from 100% to 62.13% of Hp in the model group. The results show that the lactobacillus acidophilus LA85 and the bifidobacterium lactis BLA80 can effectively reduce the adhesion of helicobacter pylori to AGS cells, reduce the colonization or inflammation occurrence of the helicobacter pylori, effectively prevent and treat the diffusion and proliferation of gastric cancer cells caused by the adhesion of the helicobacter pylori, and possibly achieve the effect and the potential of preventing and treating the gastric cancer.
Example 4 preparation of powders of Lactobacillus acidophilus LA85 and Bifidobacterium lactis BLA80
(1) Preparation of lactobacillus acidophilus LA85 powder
Inoculating Lactobacillus acidophilus strain preserved in Glycine max (L.) Gaertn tube into 10mL seed culture medium, and standing at 37 deg.C for 6-8 hr to obtain first-class seed; inoculating the first-stage seeds into 200mL of seed culture medium by an inoculation amount of 5%, and performing static culture at 37 ℃ for 5-6h to obtain second-stage seeds; inoculating the second-stage seeds obtained by culturing with an inoculum size of 2% (v/v) into a fermentation tank (10L/15L), starting fermentation under the conditions that the fermentation temperature is 35 ℃, the stirring speed is 100rpm, the initial pH is adjusted to 6.5, maintaining the pH at 5.8 and the tank pressure at 0.03MPa in the fermentation process by using 23% (m/v) sodium carbonate, culturing for 10h, obtaining a fermentation liquid, centrifuging at 4 ℃ and 8000rpm for 10min, removing a supernatant, and collecting bacterial sludge. And uniformly mixing the bacterial sludge and the freeze-drying protective agent according to the mass ratio of 1:1-1.2, and carrying out vacuum freeze drying to obtain the freeze-dried powder of the lactobacillus acidophilus. The lyoprotectant comprises the following components in percentage by mass: 15% of trehalose, 5% of skimmed milk powder, 2% of sucrose, 2% of glycerol and 0.5% of sorbitol.
(2) Preparation method of Bifidobacterium lactis BLA80 powder
Inoculating the bifidobacterium lactis strain preserved in the glycerin pipe into 10mL of MRS culture medium added with 0.5% L-cysteine hydrochloride, and standing at 37 ℃ for anaerobic culture for 18h to obtain first-grade seeds; inoculating the primary seed bacterial liquid with the inoculum size of 2% (V/V) into MRS culture medium added with 0.5% of L-cysteine hydrochloride, culturing for 12h under anaerobic condition at 37 ℃ to obtain secondary seeds, inoculating the secondary seeds with the inoculum size of 2% into the culture medium added with 0.5% of L-cysteine hydrochloride, culturing for 12h under anaerobic condition at 37 ℃ to obtain the tertiary seeds. Preparing 10L of fermentation medium, adding into a 15L anaerobic fermentation tank, sterilizing at 121 deg.C for 20min, cooling, and adjusting pH to 7 with NaOH. 200mL of the bifidobacterium lactis tertiary seed solution is inoculated into a fermentation medium and fermented at the temperature of 37 ℃ and the rotating speed of 100 rpm. In the fermentation process, 25% ammonia water is automatically fed to maintain the pH value of the fermentation liquor to be 5.5-6.0, and nitrogen is introduced in the whole fermentation process. And when the fermentation is carried out for 16h, stopping fermenting when the OD600 of the fermentation liquor is not increased any more, centrifuging the fermentation liquor obtained by stopping the fermentation at 4 ℃ and 8000rpm for 10min, and collecting bacterial sludge. Uniformly mixing the centrifugally collected bacterial sludge and a freeze-drying protective agent according to the mass ratio of 1:1, placing the mixture in a freeze-drying machine, pre-freezing for 2h at the temperature of minus 40 ℃, then heating for 4h under the condition of 10Pa to the temperature of minus 10 ℃, freeze-drying for 15h, then heating for 2h under the condition of 20Pa to the temperature of 25 ℃, and freeze-drying for 10h to obtain the freeze-dried bacterial powder of the bifidobacterium lactis. The freeze-drying protective agent comprises the following components in percentage by mass: 6% of trehalose, 8% of maltodextrin, 2% of sucrose, 0.02% of vitamin C, 3% of glycerol and 0.5-0.8% of L-arginine.
Example 5 improving Effect of Lactobacillus acidophilus LA85 and Bifidobacterium lactis BLA80 Mixed probiotic on mice infected with helicobacter pylori (Hp)
The experiment takes 60 SPF BALB/c male mice, the weight is about 20 g. The animal breeding method has room temperature of 23 + -2 deg.C, humidity of 50% + -10%, day and night alternation of 12h/12h, and is suitable for breeding for 3-5 days under free food and waterFollowed by random grouping. The group was divided into 5 groups, each consisting of a blank group, an Hp model group, a lactobacillus acidophilus LA85 intervention Hp group (LA85 group), a bifidobacterium lactis BLa80 intervention Hp group (BLa80 group), and a mixed probiotic (mass ratio 2:1) consisting of lactobacillus acidophilus LA85 and bifidobacterium lactis BLa80 intervention Hp group (BLa80+ LA85 group), with 12 mice per group. During molding, the blank group was gavaged with physiological saline at a rate of 0.5 ml/d/tube. The other groups were gavaged with Hp at a dose of 0.5ml/d for 1 time per day for 2 weeks. After 1 week of Hp gavage, selecting mice with successful modeling, and gavage the blank group and the Hp model group with physiological saline at a rate of 0.5 ml/d; the probiotic group (LA85 group, BLA80 group and LA85+ BLA80 group) used 10 9 And (3) irrigating the stomach with the CFU/mL probiotic bacterial suspension for 10 days continuously, and killing the mice after the lavage is finished.
TABLE 3 levels of inflammatory factors (pg/mL) after H.pylori infection in groups of mice
The production of cell factors is very critical in the immune pathogenesis of helicobacter pylori infection, and the imbalance of the immune mechanism of the organism and the imbalance of the regulation of inflammatory factors IL-6 and IL-1 beta are important pathogenesis of ulcer diseases caused by helicobacter pylori infection. The test results shown in Table 3 indicate that the serum IL-6 and IL-1 beta contents of the Hp model group are higher than those of the blank group, and the serum IL-6 and IL-1 beta contents of the probiotic and Hp group are lower than those of the Hp model group, which indicates that the probiotic can stimulate the organism to generate cytokines, improve the organism immunity and reduce the occurrence of mouse inflammation. The IL-6 and IL-1 beta content in the serum of LA85+ BLA80 group will reach the level equivalent to that of the blank group. The results show that the effect of the LA85 and BLa80 mixed probiotics is better than that of any single bacterium in relieving or treating diseases such as gastric ulcer, chronic digestive tract ulcer and gastritis caused by helicobacter pylori infection.
Example 6 Experimental validation of the human combination of LA85 and BLa80 for the treatment of H.pylori infection
42 positive H.pylori-infected subjects were enrolled in the trial, who had not taken antibiotics or probiotic products within 1 month prior to inclusion in the study, had no history of smoking, no history of drinking, no history of gastrointestinal surgery, no historical severe disease or psychiatric illness. The 42 patients were randomized into 2 groups, of which, placebo group was 20 and probiotic group (LA85+ BLa 80) 22. The placebo group takes placebo 2 times in the morning and evening, and the probiotic group takes probiotic powder (mixed probiotic consisting of lactobacillus acidophilus LA85 and bifidobacterium lactis BLA80 in a mass ratio of 2:1) 2 times in the morning and evening, and the whole experiment lasts for 1 month.
1. Evaluation of gastrointestinal symptom amelioration before and after experiment
The Gastrointestinal Symptom Rating Scale (GSRS) is filled in two groups of patients before and after the experiment begins, the average value of each group is calculated to represent the gastrointestinal health condition of each group, the improvement condition of the gastrointestinal symptoms of the patients before and after the experiment is evaluated, and the measurement result is shown in figure 2.
As can be seen from FIG. 3, the GSRS score of the placebo group, helicobacter pylori positive infected subjects did not change much before and after the experiment, while the GSRS score of the probiotic group (LA85+ BLA80 group), helicobacter pylori positive infected subjects before the start of the experiment, was significantly reduced. The experimental results show that the probiotic group (LA85+ BLA80 group) can remarkably relieve gastrointestinal symptoms of patients infected by helicobacter pylori.
2. Evaluation of the eradication of helicobacter pylori-positive patients before and after the experiment
TABLE 4 elimination before and after the experiment of H.pylori-positive patients
| Group of | Number of people | Positive for | Negative of | Negative rate (%) |
| |
20 | 17 | 3 | 15.00 |
| Probiotic group | 22 | 8 | 16 | 72.72 * |
Note: indicates significant difference compared to placebo group (p < 0.05).
The result shows that the negative rate of the placebo group is 15.00% after taking the probiotic powder for 1 month; the negative rate of the probiotic group is as high as 72.72%, and the result shows that the mixed probiotic consisting of lactobacillus acidophilus LA85 and bifidobacterium lactis BLA80 can obviously reduce the helicobacter pylori infection degree of a helicobacter pylori infected patient.
3. Levels of inflammatory factors (pg/mL) in H.pylori positive patients before and after the experiment
The levels of IL-6 and IL-8 in the body serum of helicobacter pylori positive infected persons in a placebo group and a probiotic group are measured by an ELISA kit before and after an experiment.
TABLE 5 levels of inflammatory factors (pg/mL) in H.pylori positive patients before and after the experiment
Note: indicates that each group of the same index has significant difference compared with the group before intervention (p <0.05)
As can be seen from the results in Table 5, the IL-6 level and the IL-8 level in the probiotic group (LA85+ BLA80 group) were reduced to 9.26pg/mL and 7.46pg/mL, respectively, after the intervention, and the IL-6 level and the IL-8 level secreted by the body were significantly reduced. Indicating that the probiotic bacteria can regulate the immune response caused by helicobacter pylori infection after being dried. The mixed probiotics consisting of lactobacillus acidophilus LA85 and bifidobacterium lactis BLA80 can obviously improve the level of inflammatory factors in a patient with helicobacter pylori related chronic gastritis.
4. Serum PG levels in H.pylori positive subjects before and after the experiment
Before and after the experiment, the PG level in the body serum of the helicobacter pylori positive infected person in the placebo group and the probiotic group is determined by referring to the specification of an ELISA kit.
TABLE 6 serum PG levels in H.pylori positive subjects before and after the experiment
Note: indicates that each group of the same index has significant difference compared with the group before intervention (p <0.05)
The serum PG can reflect the strength of the secretory function of the gastric mucosa and further reflect the state of the gastric mucosa. As can be seen from Table 6, there was no significant change in the PGI level, a decrease in the PGII level, and an increase in the PGI/PGII ratio in the placebo group after the intervention, but no significant difference in the change in the PGI/PGII ratio before and after the intervention. The dry prognosis probiotic group has high PGI level, obviously reduced PGII level and obviously increased PGI/PGII ratio which reaches 10.67. The experimental results show that the probiotic group (LA85+ BLA80 group) has the function of regulating the gastric mucosal immunity.
The above embodiments are provided to facilitate better understanding of the present invention, the embodiments of the present invention are not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof and are included in the scope of the present invention.
Claims (8)
1. Use of lactobacillus acidophilus LA85 for the preparation of a product for inhibiting the growth of helicobacter pylori, the product including a pharmaceutical product and a food product including, but not limited to, a probiotic solid beverage, a probiotic beverage, a tabletted candy, a snack food, a dairy product, a soy product, a fruit and vegetable product, or a functional health product; the lactobacillus acidophilus LA85 is deposited in China general microbiological culture Collection center of China Committee for culture Collection of microorganisms at the time of 2021, 02/01, with the address of No. 3 Xilu-Shih No. 1 of Beijing, Chaoyang, and the deposition number of CGMCC No. 21802.
2. The use according to claim 1, wherein the food product comprises a health food product for alleviating or treating gastric ulcers, duodenal ulcers or gastrointestinal discomfort.
3. Use of bifidobacterium lactis BLa80 in the manufacture of a product for inhibiting the growth of helicobacter pylori, the product including a pharmaceutical product and a food product including, but not limited to, a probiotic solid beverage, a probiotic beverage, a tabletted candy, a snack food, a dairy product, a soy product, a fruit and vegetable product or a functional health product; the bifidobacterium lactis BLA80 has the preservation number of CGMCC No.22547 and the preservation time of: 2021, 05 month, 17 days; the preservation unit: china general microbiological culture Collection center; the preservation address is as follows: xilu No. 1 Hospital No. 3, Beijing, Chaoyang, North.
4. The use according to claim 3, wherein the food product comprises a health food for alleviating or treating gastric ulcers, duodenal ulcers or gastrointestinal discomfort.
5. Use of lactobacillus acidophilus LA85 for the preparation of a product for preventing or treating gastritis or gastric cancer, the product comprising a pharmaceutical product and a food product, the food product comprising but not limited to a probiotic solid beverage, a probiotic beverage, a tabletted candy, a snack food, a dairy product, a soy product, a fruit and vegetable product or a functional health product; the lactobacillus acidophilus LA85 is deposited in China general microbiological culture Collection center of China Committee for culture Collection of microorganisms at the time of 2021, 02/01, with the address of No. 3 Xilu-Shih No. 1 of Beijing, Chaoyang, and the deposition number of CGMCC No. 21802.
6. Use of bifidobacterium lactis BLa80 for the manufacture of a product for the prevention or treatment of gastritis or gastric cancer, said product comprising a pharmaceutical product and a food product, said food product comprising but not limited to a probiotic solid beverage, a probiotic beverage, a tabletted candy, a snack food, a dairy product, a soy product, a fruit and vegetable product or a functional health product; the bifidobacterium lactis BLA80 has the preservation number of CGMCC No.22547 and the preservation time of: 2021, 05 month, 17 days; the preservation unit: china general microbiological culture Collection center; and (4) storage address: xilu No. 1 Hospital No. 3, Beijing, Chaoyang, North.
7. Use of a mixed probiotic of lactobacillus acidophilus LA85 and bifidobacterium lactis BLa80 for the manufacture of a product for the prevention or treatment of helicobacter pylori infection, including a food product or a pharmaceutical product, including but not limited to a probiotic solid beverage, a probiotic beverage, a tabletted confection, a snack food, a dairy product, a soy product, a fruit and vegetable product, or a functional health product; the lactobacillus acidophilus LA85 is preserved in China general microbiological culture Collection center of China general microbiological culture Collection management Committee on the day 01 of 02 months in 2021, the address is No. 3 of Xilu No. 1 of Beijing, Chaoyang, and the preservation number is CGMCC No. 21802; the bifidobacterium lactis BLA80 has the preservation number of CGMCC No.22547 and the preservation time of: 2021, 05 month, 17 days; the preservation unit is as follows: china general microbiological culture Collection center; and (4) storage address: xilu No. 1 Hospital No. 3, Beijing, Chaoyang, North.
8. Use according to claim 7, characterized in that the food product comprises in particular a health food for alleviating or treating gastric ulcers, duodenal ulcers or gastrointestinal discomfort.
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