CN113413406A - A metazoan compound, its preparation method and application in resisting helicobacter pylori and gastrointestinal diseases - Google Patents

A metazoan compound, its preparation method and application in resisting helicobacter pylori and gastrointestinal diseases Download PDF

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Publication number
CN113413406A
CN113413406A CN202110680811.0A CN202110680811A CN113413406A CN 113413406 A CN113413406 A CN 113413406A CN 202110680811 A CN202110680811 A CN 202110680811A CN 113413406 A CN113413406 A CN 113413406A
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lactobacillus
metazoan
compound
extract
helicobacter pylori
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Chinese (zh)
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王昭月
李靖
孙雨薇
付修远
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Nanjing Aurora Biotechnology Co ltd
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Nanjing Aurora Biotechnology Co ltd
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Abstract

The invention provides a metazoan compound, which comprises bean powder, inactivated probiotics and metabolites thereof; the compound 1 can realize the slow release effect of inactivated probiotics and metabolites thereof; 2. can effectively remove helicobacter pylori in the organism and relieve stomach discomfort caused by the helicobacter pylori, such as stomachache, heartburn, gastric acid, gastric ulcer and the like; 3. not only the action time is long, but also the action area is large; 4. solves the difficult problems of the storage and the transportation of the live bacteria. Meanwhile, the invention also provides a preparation method and application of the compound.

Description

A metazoan compound, its preparation method and application in resisting helicobacter pylori and gastrointestinal diseases
Technical Field
The invention relates to a metazoan compound, in particular to a compound capable of resisting helicobacter pylori and assisting in treatment of stomach-related diseases.
Background
Helicobacter pylori (Hp) is a spiral or S-shaped, microaerophilic gram-negative bacterium belonging to the order Campylobacter, the family helicobacter, the genus helicobacter. Its infection is specific and often localized on the surface of the gastric mucosa. Because the end part of the flagella is provided with a plurality of unipole flagella with sheaths, the flagella can quickly escape from the acid environment in the gastric cavity and further pass through the thick mucous layer to reach and be planted on the surface of the gastric mucosa. Helicobacter pylori infection is a cause of upper gastrointestinal diseases such as chronic gastritis, peptic ulcer, gastric adenocarcinoma, and gastric mucosa-associated lymphoid tissue lymphoma. According to epidemiological investigation, more than half of the population in the world has Hp infection, the infection rate in developing countries and minority memory migrators is higher, and even the infection probability of children is greatly improved. Therefore, effective control of Hp infection is one of the major routes for the control of gastrointestinal diseases.
At present, the bismuth agent tetratherapy (proton pump inhibitor, bismuth agent and 2 antibiotics) is used as the first choice treatment scheme for eradicating Hp in China medically. It is well known that the balance of microbial flora in the body plays an important role in human health. The use of antibiotics in the treatment of eradicating Hp by quadruple therapy not only can cause microbial imbalance in the human body, but also can change the number, the types and the composition of flora in the intestines and the stomach to different degrees. Antibiotics also cause malaise reactions in humans, such as nausea, vomiting, abdominal pain, diarrhea, dry mouth, bitter taste in the mouth, loss of appetite, etc. Moreover, antibiotics also induce the change of Hp from bacilli to coccobacillus, resulting in an increase in drug resistance.
Patent CN112353822A discloses an application of lactococcus lactis particles and probiotics in preparation of a helicobacter pylori infection resisting medicine, bifidobacterium longum freeze-dried powder, bifidobacterium lactis freeze-dried powder, bifidobacterium adolescentis freeze-dried powder, streptococcus thermophilus freeze-dried powder, lactobacillus acidophilus freeze-dried powder, lactobacillus delbrueckii or subspecies bulgaricus freeze-dried powder are adopted, the field planting of Hp can be obviously reduced, the expression of gastric antibacterial protein is increased, inflammation is relieved, particularly in the aspect of increasing the adaptive immunity of the stomach, the lactobacillus lactis particles and the probiotics have a good synergistic effect when being used together, and diseases such as gastritis caused by helicobacter pylori infection can be effectively treated. International patent WO2007/073709 discloses a lactobacillus strain selected to bind free helicobacter pylori and form aggregates in an in vitro simulated gastric fluid environment, thereby preventing the helicobacter pylori from reaching and infecting the epithelial cells of the stomach, and the aggregates exit in a natural manner through the gastrointestinal tract. Although probiotics are adopted as the main component for resisting helicobacter pylori, the flora balance in the gastrointestinal tract is not damaged, and the generation of drug-resistant strains is prevented. However, generally, probiotics suffer from the following disadvantages: the storage period of the probiotics is short, and the requirements on the storage environment are strict; secondly, most probiotics are affected by mechanical operation, acidic environment and bacteriostatic components in the digestive tract when entering the digestive system, so that the number of live bacteria when the probiotics reach the gastrointestinal tract is greatly reduced, and a good probiotic effect cannot be achieved.
Therefore, in the treatment of helicobacter pylori, a substitute which does not generate side effects like antibiotics and can solve the problems of low utilization rate and difficult storage of probiotics is urgently needed.
Disclosure of Invention
In order to solve the defects in the prior art, the invention provides a probiotic compound which can not only resist Hp infection of a human body and gastrointestinal related diseases, but also well solve the problems of viable bacteria storage, transportation and the like.
In order to solve the technical problems, the invention provides an anabiotic compound which comprises bean powder, inactivated probiotics and metabolites thereof.
It should be noted that: "probiotic" refers to a kind of active microorganism beneficial to the host, and is the general term for active beneficial microorganism which is planted in the intestinal tract and reproductive system of human body, can produce health efficacy, improve the micro-ecological balance in the host body and play a beneficial role; the term "inactivated probiotics" refers to the killed or living probiotics which are treated by heating, ultraviolet irradiation, or chemical treatment, so that the bacteria retain the original structure and characteristics, but do not have the ability to grow and reproduce. The fermented product of beans, inactivated probiotics and metabolite thereof are collectively called 'metazoan'. The probiotic metabolite is a metabolite taking bean powder as a substrate.
Further, the probiotic is lactobacillus; preferably, the lactobacillus is selected from one or more of lactobacillus rhamnosus, lactobacillus acidophilus, lactobacillus crispatus, lactobacillus fermentum, lactobacillus plantarum, lactobacillus casei, lactobacillus paracasei, lactobacillus jensenii, lactobacillus gasseri, lactobacillus reuteri, lactobacillus salivarius, lactobacillus bifidus, lactobacillus delbrueckii, lactobacillus caucasicus, lactobacillus mallotiformis, lactobacillus pentosus, lactobacillus bulgaricus, lactobacillus helveticus and lactobacillus gasseri; preferably, the lactobacillus is one or the combination of more than two of lactobacillus rhamnosus, lactobacillus acidophilus, lactobacillus plantarum, lactobacillus fermentum, lactobacillus delbrueckii, lactobacillus casei, lactobacillus reuteri, lactobacillus salivarius, lactobacillus paracasei, lactobacillus mallotiformis, lactobacillus helveticus and lactobacillus gasseri;
more preferably, the Lactobacillus rhamnosus may be selected from Lactobacillus rhamnosus GG (ATCC 53103), Lactobacillus rhamnosus Rosell-11, Lactobacillus rhamnosus HN001, Lactobacillus rhamnosus LR06, Lactobacillus rhamnosus F-1, Lactobacillus acidophilus may be selected from Lactobacillus acidophilus Rosell-52, Lactobacillus acidophilus Rosell-418, Lactobacillus acidophilus NCFM, Lactobacillus acidophilus La27, Lactobacillus acidophilus LA28, Lactobacillus acidophilus LA 201 85, Lactobacillus acidophilus Lac361, Lactobacillus acidophilus HA 35137, Lactobacillus acidophilus 36137, Lactobacillus plantarum HA-80, Lactobacillus plantarum HA-493 3, Lactobacillus plantarum LP-3668, Lactobacillus plantarum HA-80, Lactobacillus plantarum HA-493K 3, Lactobacillus plantarum HA-4938, Lactobacillus plantarum LP-3, Lactobacillus plantarum HA-493L 3, Lactobacillus plantarum LP-3, Lactobacillus plantarum HA-103, Lactobacillus plantarum LP-11, Lactobacillus plantarum HA-103, Lactobacillus plantarum LP-3, Lactobacillus plantarum HA-103, Lactobacillus plantarum LP-103, Lactobacillus plantarum HA-3, Lactobacillus plantarum HA-103, Lactobacillus plantarum HA-11, Lactobacillus plantarum LP-103, Lactobacillus plantarum HA-L-F-L-, Lactobacillus fermentum DR9, Lactobacillus delbrueckii may be selected from Lactobacillus delbrueckii spp.lactis Rosell-187, Lactobacillus delbrueckii Subsp.delbrueckii LDD01, Lactobacillus casei may be selected from Lactobacillus casei Zhang, Lactobacillus casei HA-108, Lactobacillus casei LC-G11, Lactobacillus reuteri may be selected from Lactobacillus reuteri DSM17938, Lactobacillus reuteri HA-188, Lactobacillus reuteri DSM17648, Lactobacillus reuteri NCIMB 30242, Lactobacillus reuteri LR 100, Lactobacillus salivarius may be selected from Lactobacillus salviae salacia HA-118, Lactobacillus salviae AP-32, Lactobacillus paracasei may be selected from Lactobacillus casei HA-5714, Lactobacillus casei may be selected from Lactobacillus casei strain Lactobacillus casei HA-572, Lactobacillus casei may be selected from Lactobacillus casei strain CT-like Lactobacillus strain HA-5714, Lactobacillus strain CT, Lactobacillus casei strain HA-G, Lactobacillus strain HA-G, Lactobacillus strain G, Lactobacillus strain.
Furthermore, the beans can be selected from one or more of soybeans, green beans and black beans. Semen glycines contains protein, fat, carbohydrate, carotene, vitamin B1, vitamin B2, nicotinic acid, etc., and also contains bean oligosaccharide, isoflavone, saponin, nucleic acid, etc., which are valuable health factors for human health. The green bean is rich in unsaturated fatty acid and bean phospholipid, and has effects of maintaining blood vessel elasticity, nourishing brain and preventing fatty liver formation. Researches show that the green beans are rich in various antioxidant components and can eliminate inflammation. The green beans can provide two flavonoid antioxidants of catechin and epicatechin for human bodies. The two substances can effectively remove free radicals in vivo, prevent diseases caused by free radicals, delay aging speed of body, and have antiinflammatory and broad-spectrum antibacterial effects. Green beans also contain two carotenoids: alpha-carotene and beta-carotene. In 2010, the U.S. centers for disease control discovered, based on a follow-up study of up to 14 years, that the higher the alpha-carotene content in the blood, the longer the life of the individual. Beta-carotene is also an antioxidant, has a detoxifying effect, and can reduce the risk of heart disease and cancer. The black bean mainly contains unsaturated fatty acid, has the absorption rate of 95 percent, and has the function of reducing cholesterol in blood besides meeting the requirement of a human body on fat; the black bean contains rich vitamins, yolk, melanin and other substances, wherein the content of B vitamins and vitamin E is high, the black bean has the nutrition and health care effects, and the black bean also contains rich trace elements, so the black bean is essential for keeping the body function complete, delaying the body aging, reducing the blood viscosity and meeting the requirements of the brain on the trace substances.
The bean metazoan provided by the invention can be compounded with other functional components, such as broccoli extract, Korean vegetable extract, liquorice extract, vitamin U, pumpkin seed extract, pepper extract, clove tea extract, propolis extract, one or more of garlic extract, turmeric extract and emblic leafflower fruit extract; the adding mode can be added after the preparation of the bean metazoan compound is finished, or can be prepared after being mixed with the bean powder.
The invention also provides an auxiliary treatment medicine, food, nutriment, dietary supplement or health care product for resisting the helicobacter pylori, which comprises the bean metazoan compound and auxiliary materials. The dosage form can be hard capsules; tablets, such as chewable tablets and troches; solid beverages such as powder, granules, mixed powders; candies, such as hard candies and soft candies; soft capsules; jelly; an energy bar; chocolate or biscuits. In the course of the preparation, customary auxiliaries such as carrier substances, splitting agents, binders, coating agents, swelling agents, lubricants or slip agents, odorants, sweeteners and solubilizers can be added.
The invention also provides application of the metazoan compound in preparing medicines, foods, dietary supplements, nutriments or health-care products for treating or preventing helicobacter pylori infection and gastrointestinal diseases.
The invention also provides a preparation method of the post-biotic compound, which comprises the steps of pretreating bean powder, fully mixing and stirring the bean powder, inactivated or non-inactivated probiotics and aggregates, carrying out high-temperature spray drying treatment, and packaging a finished product; the inactivated probiotics also includes metabolites thereof; as a specific embodiment, the method for pretreating bean flour may be: soaking beans, grinding, and heating; or grinding beans and fermenting.
Or
Fermenting the bean powder and probiotics, performing high-temperature spray drying treatment on the fermentation liquor in the logarithmic phase or the stationary phase of the probiotics, and packaging to obtain the finished product. As some examples of the present invention, the bean powder is fermented using probiotics,
a) bean pretreatment: pulverizing beans, sterilizing at 0.1MPa and 121 deg.C for 20min, cooling, and refrigerating at 4 deg.C;
b) activating probiotic strains: the strain activation process comprises the following steps: taking probiotic freeze-dried powder (8.0 × 10) refrigerated in 4 deg.C environment8total CFU/g) to be transplanted into 40ml of sterile culture solution, and carrying out shake cultivation at 37 ℃ for 10 hours at a shake speed of 120 rpm/min;
c) beans are pretreated and then inoculated with activated probiotics for fermentation to obtain fermentation liquor; the specific fermentation process is as follows: mixing bean flour, glucose and alkaline protease in ultrapure water to form a fermentation substrate, adding activated bacterial liquid into the fermentation substrate, uniformly stirring, and culturing at 37 ℃, wherein the content of each component in percentage by weight is 14 percent of the bean flour, 0.5 percent of the glucose, 0.5 percent of the alkaline protease and 0.5 percent of the bacterial liquid;
d) after 8 hours of fermentation, the suspension was washed three times with saline solution, then prepared with distilled water, heated at 70 ℃ for 10 minutes and lyophilized.
Further, the probiotic is lactobacillus; preferably, the lactobacillus is selected from one or more of lactobacillus rhamnosus, lactobacillus acidophilus, lactobacillus crispatus, lactobacillus fermentum, lactobacillus plantarum, lactobacillus casei, lactobacillus paracasei, lactobacillus jensenii, lactobacillus gasseri, lactobacillus reuteri, lactobacillus salivarius, lactobacillus bifidus, lactobacillus delbrueckii, lactobacillus caucasicus, lactobacillus mallotiformis, lactobacillus pentosus, lactobacillus bulgaricus, lactobacillus helveticus and lactobacillus gasseri; preferably, the lactobacillus is one or the combination of more than two of lactobacillus rhamnosus, lactobacillus acidophilus, lactobacillus plantarum, lactobacillus fermentum, lactobacillus delbrueckii, lactobacillus casei, lactobacillus reuteri, lactobacillus salivarius, lactobacillus paracasei, lactobacillus mallotiformis, lactobacillus helveticus and lactobacillus gasseri;
more preferably, the Lactobacillus rhamnosus may be selected from Lactobacillus rhamnosus GG (ATCC 53103), Lactobacillus rhamnosus Rosell-11, Lactobacillus rhamnosus HN001, Lactobacillus rhamnosus LR06, Lactobacillus rhamnosus F-1, Lactobacillus acidophilus may be selected from Lactobacillus acidophilus Rosell-52, Lactobacillus acidophilus Rosell-418, Lactobacillus acidophilus NCFM, Lactobacillus acidophilus La27, Lactobacillus acidophilus LA28, Lactobacillus acidophilus LA 201 85, Lactobacillus acidophilus Lac361, Lactobacillus acidophilus HA 35137, Lactobacillus acidophilus 36137, Lactobacillus plantarum HA-80, Lactobacillus plantarum HA-493 3, Lactobacillus plantarum LP-3668, Lactobacillus plantarum HA-80, Lactobacillus plantarum HA-493K 3, Lactobacillus plantarum HA-4938, Lactobacillus plantarum LP-3, Lactobacillus plantarum HA-493L 3, Lactobacillus plantarum LP-3, Lactobacillus plantarum HA-103, Lactobacillus plantarum LP-11, Lactobacillus plantarum HA-103, Lactobacillus plantarum LP-3, Lactobacillus plantarum HA-103, Lactobacillus plantarum LP-103, Lactobacillus plantarum HA-3, Lactobacillus plantarum HA-103, Lactobacillus plantarum HA-11, Lactobacillus plantarum LP-103, Lactobacillus plantarum HA-L-F-L-, Lactobacillus fermentum DR9, Lactobacillus delbrueckii may be selected from Lactobacillus delbrueckii spp.lactis Rosell-187, Lactobacillus delbrueckii Subsp.delbrueckii LDD01, Lactobacillus casei may be selected from Lactobacillus casei Zhang, Lactobacillus casei HA-108, Lactobacillus casei LC-G11, Lactobacillus reuteri may be selected from Lactobacillus reuteri DSM17938, Lactobacillus reuteri HA-188, Lactobacillus reuteri DSM17648, Lactobacillus reuteri NCIMB 30242, Lactobacillus reuteri LR 100, Lactobacillus salivarius may be selected from Lactobacillus salviae salacia HA-118, Lactobacillus salviae AP-32, Lactobacillus paracasei may be selected from Lactobacillus casei HA-5714, Lactobacillus casei may be selected from Lactobacillus casei strain Lactobacillus casei HA-572, Lactobacillus casei may be selected from Lactobacillus casei strain CT-like Lactobacillus strain HA-5714, Lactobacillus strain CT, Lactobacillus casei strain HA-G, Lactobacillus strain HA-G, Lactobacillus strain G, Lactobacillus strain.
Furthermore, the beans are selected from one or more of soybeans, green beans and black beans.
The invention achieves the following beneficial technical effects:
first, in the compound, the bean powder is aggregate, many micropores are distributed in the bean powder, and the inactivated probiotics and its metabolites can be attached on the micropores or the outer wall. After entering the organism, more inactivated probiotics and metabolites thereof are exposed due to the digestion effect of protease in stomach, thereby realizing the slow release effect of the inactivated probiotics and the metabolites thereof;
secondly, the compound can effectively resist the infection of helicobacter pylori in human body and various gastrointestinal discomfort caused by the helicobacter pylori, such as stomach heartburn, gastric acid, stomachache and the like. The reason for this effect is probably that 1. after the compound enters the stomach, the inactivated probiotics and metabolites such as polysaccharide, conjugated protein and other adhesins attached to the surface of the compound gather the helicobacter pylori on the compound; 2. products of bean fermentation such as various polypeptides, long-chain fatty acids, organic acids and the like, inactivated probiotics and metabolites thereof such as various enzymes and the like can destroy the colonization environment of helicobacter pylori to a certain extent and weaken the colonization of the helicobacter pylori on gastric epithelial cells; this is advantageous to some extent for the aggregation of the colonized H.pylori. The dietary fiber in the compound can promote the gastrointestinal peristalsis, thereby helping the efflux of the agglomerated helicobacter pylori. 3. After the helicobacter pylori is discharged out of the body, the damaged gastric mucosa obtains a good treatment or repair environment. Unsaturated fatty acid in bean fermentation product, and hydrophobic substances such as protein and polysaccharide on the surface of inactivated probiotic bacteria adhere to injured gastric mucosa to prevent reinfection. Thereby rapidly and effectively relieving various stomach discomforts caused by the helicobacter pylori, such as stomachache, heartburn, gastric acid, gastric ulcer and the like.
Thirdly, as mentioned above, the aggregate of the complex is partially hydrolyzed into polypeptide by the action of pepsin, so that the probiotic bacteria placed in the micropores of the aggregate are released. Therefore, the compound of the invention not only can effectively prolong the action time, but also can enlarge the action area due to the sustained release effect as the aggregate passes through the stomach, the intestine and other parts. In the intestinal tract, the released metazoan can help the beneficial bacteria in the intestinal tract to grow and reproduce, thereby regulating the microecological balance of the beneficial bacteria in the intestinal tract.
Fourthly, the inactivated probiotics not only have the growth and reproduction capability, but also keep the original structure and characteristics, thereby solving the difficult problems existing in the storage and transportation processes of the live bacteria.
Drawings
FIG. 1 shows the staining pattern of the post-legume biotype complex of the present invention.
Detailed Description
The invention is further described with reference to specific examples. The following examples are only for illustrating the technical solutions of the present invention more clearly, and the protection scope of the present invention is not limited thereby.
The invention is further described with reference to the following figures and examples.
Example 1
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus Rosell-11 thallus and metabolites thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 2
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus LR06 thallus and metabolite thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 3
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus F-1 thallus and metabolites thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 4
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus Rosell-418 thalli and metabolites thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 5
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus LA27 thallus and metabolite thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 6
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus HA-122 thalli and metabolites thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 7
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus LA-G80 thallus and metabolite thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 8
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus HA-119 thallus and metabolites thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 9
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus L-137 thallus and metabolite thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 10
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus HA-108 thalli and metabolites thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 11
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus LC-G11 thallus and metabolites thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 12
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus HA-188 thalli and metabolites thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 13
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus DSM17648 thalli and metabolites thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 15
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus NCIMB 30242 thallus and metabolite thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 16
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus HA-118 thalli and metabolites thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 17
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus HA-196 thalli and metabolites thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 18
The invention provides a metazoan compound, which comprises bean powder, inactivated lactobacillus LPC-G110 thallus and metabolites thereof; wherein, in 1g of the compound, the number of lactobacillus is 100 hundred million.
Example 19
The invention provides a metazoan compound, which comprises bean powder, inactivated probiotics thallus and metabolites thereof; wherein the probiotics comprise Lactobacillus Rosell-11 and Lactobacillus HA-119; in 1g of the complex, the number of the lactobacillus Rosell-11 bacteria is 50 hundred million, and the number of the lactobacillus HA-119 bacteria is 50 hundred million.
Example 20
The invention provides a metazoan compound, which comprises bean powder, inactivated probiotics thallus and metabolites thereof; wherein the probiotic bacteria are Lactobacillus HA-122, Lactobacillus HA-119, Lactobacillus L-137, and Lactobacillus DSM 17648; in 1g of the compound, the number of lactobacillus HA-122 bacteria is 25 hundred million, the number of lactobacillus HA-119 bacteria is 25 hundred million, the number of lactobacillus L-137 bacteria is 25 hundred million, and the number of lactobacillus DSM17648 bacteria is 25 hundred million.
Example 21
The invention provides a metazoan compound, which comprises bean powder, inactivated probiotics thallus and metabolites thereof; wherein the probiotic bacteria are Lactobacillus Rosell-418, Lactobacillus LA27, Lactobacillus L-137, and Lactobacillus HA-188; in 1g of the compound, the number of lactobacillus Rosell-418 bacteria is 25 hundred million, the number of lactobacillus LA27 bacteria is 25 hundred million, the number of lactobacillus HA-188 bacteria is 25 hundred million, and the number of lactobacillus L-137 bacteria is 25 hundred million.
Example 22
The invention provides a metazoan compound, which comprises bean powder, inactivated probiotics thallus and metabolites thereof; wherein the probiotic bacteria are Lactobacillus Rosell-418, Lactobacillus LA27, Lactobacillus L-137, and Lactobacillus HA-188; in 1g of the compound, the number of lactobacillus Rosell-418 bacteria is 25 hundred million, the number of lactobacillus LA27 bacteria is 25 hundred million, the number of lactobacillus HA-188 bacteria is 25 hundred million, and the number of lactobacillus L-137 bacteria is 25 hundred million.
Example 23
The invention provides a metazoan compound, which comprises bean powder, inactivated probiotics thallus and metabolites thereof; wherein the probiotic bacteria comprise Lactobacillus HA-122, Lactobacillus HA-119, Lactobacillus HA-108, Lactobacillus HA-188, Lactobacillus HA-118, and Lactobacillus HA-196; in 1g of the compound, the number of lactobacillus HA-122 bacteria is 20 hundred million, the number of lactobacillus HA-119 bacteria is 20 hundred million, the number of lactobacillus HA-188 bacteria is 20 hundred million, the number of lactobacillus HA-108 bacteria is 20 hundred million, the number of lactobacillus HA-118 bacteria is 20 hundred million, and the number of lactobacillus HA-196 bacteria is 20 hundred million.
Example 24
The invention provides an auxiliary therapeutic medicine for resisting helicobacter pylori, which comprises bean powder, inactivated probiotics and metabolites thereof, and the medicine is a hard capsule.
Example 25
The invention provides an auxiliary therapeutic medicine for resisting helicobacter pylori, which comprises bean powder, inactivated probiotics and metabolites thereof, and the medicine is a tablet.
Example 26
The invention provides a food for resisting helicobacter pylori, which comprises bean powder, inactivated probiotics and metabolites thereof; the food is candy.
Example 27
The invention provides a food for resisting helicobacter pylori, which comprises bean powder, inactivated probiotics and metabolites thereof; the food is jelly.
Example 28
The invention provides a food for resisting helicobacter pylori, which comprises bean powder, inactivated probiotics and metabolites thereof; the food product is an energy bar.
Example 29
The invention provides a food for resisting helicobacter pylori, which comprises bean powder, inactivated probiotics and metabolites thereof; the food is chocolate.
Example 30
The invention provides a food for resisting helicobacter pylori, which comprises bean powder, inactivated probiotics and metabolites thereof; the food is biscuit.
Example 31
The invention provides a dietary supplement for resisting helicobacter pylori, which comprises bean powder, inactivated probiotics and metabolites thereof, and the dietary supplement is a solid beverage.
Example 32
The invention provides a nutriment for resisting helicobacter pylori, which comprises bean powder, inactivated probiotics and metabolites thereof; the nutritional product is an energy bar.
Example 33
The invention provides a health product for resisting helicobacter pylori, which comprises bean powder, inactivated probiotics and metabolites thereof, wherein the health product is a soft capsule.
Example 34
The invention provides a preparation method of a post-legume prebiotic compound, which comprises the following specific processes:
a) bean pretreatment: pulverizing beans, sterilizing at 0.1MPa and 121 deg.C for 20min, cooling, and refrigerating at 4 deg.C;
b) activation of probiotic (taking lactobacillus reuteri HA-188 as an example) strain: the strain activation process comprises the following steps: taking probiotic freeze-dried powder (8.0 × 10) refrigerated in 4 deg.C environment8total CFU/g) to be transplanted into 40ml of sterile culture solution, and carrying out shake cultivation at 37 ℃ for 10 hours at a shake speed of 120 rpm/min;
c) beans are pretreated and then inoculated with activated probiotics for fermentation to obtain fermentation liquor; the specific fermentation process is as follows: mixing bean flour, glucose and alkaline protease in ultrapure water to form a fermentation substrate, adding activated bacterial liquid into the fermentation substrate, uniformly stirring, and culturing at 37 ℃, wherein the content of each component in percentage by weight is 14 percent of the bean flour, 0.5 percent of the glucose, 0.5 percent of the alkaline protease and 0.5 percent of the bacterial liquid;
d) after 8 hours of fermentation, the suspension was washed three times with saline solution, then prepared with distilled water, heated at 70 ℃ for 10 minutes and lyophilized.
Beans are used as basic food of traditional gourmet food, are rich in protein, carbohydrate, dietary fiber, minerals, vitamins and the like, and are the main sources of isoflavone. The frequent consumption of beans can reduce the risk of cardiovascular and cerebrovascular diseases, diabetes, cancer and digestive tract diseases. However, the administration of beans to humans generally causes problems in that they contain indigestible carbohydrates, trypsin inhibitors, coagulans and other proteins, which are major allergens. Therefore, the application of the legumes is greatly limited. Gastrointestinal diseases such as gastritis, superficial gastritis, gastric ulcer, and various stomach disorders caused by helicobacter pylori are caused by the impaired digestive function of the stomach. The legume products are rarely used in the treatment of gastrointestinal diseases. However, long-term research by the applicant shows that the problem can be solved by fermenting beans with probiotics, and the probiotics can be applied to resisting helicobacter pylori infection and various stomach diseases caused by helicobacter pylori.
2. Gram stain characterization:
1) the instrument comprises the following steps: precision balance ME204E/02, hot plate magnetic stirrer IT-09B15, biomicroscope ML31, alcohol lamp, 100ML beaker, glass rod, slide, cover slip, etc.
2) Reagent: gram stain (fast method), distilled water.
3) Dyeing: weighing 1g of compound by an electronic balance, adding 50ml of water, and stirring for 60min in a constant-temperature water bath at 25 ℃ by adopting a magnetic stirrer at the speed of 640 rpm; the solution was gently dipped with a sterile glass rod and spread on a glass slide to ensure a thin and uniform layer. Standing and drying at room temperature. Then, the glass is heated and fixed by an alcohol lamp, the temperature is not too high, and the condition that the back of the glass slide is contacted with the back of the hand and is not scalded is taken as the standard. Then adding gentian violet liquid for dyeing for 10 seconds, washing with water and drying; adding iodine solution for dyeing for 10 seconds, washing and drying; adding a decoloring solution for decoloring for 10-20 seconds, and washing with water and spin-drying; adding a yellow-sand solution for redyeing for 10 seconds, and washing with water. And (5) drying and performing microscopic examination. (Observation with an oil lens, objective lens 100X, eyepiece lens 10X). The results are shown in FIG. 1.
As can be seen from FIG. 1, the bean powder forms aggregates with different particle sizes after being fermented by Lactobacillus reuteri HA-188. After treatment, the probiotics keep an intact cell structure and are distributed on the surface or inside the aggregate.
Experimental example one test for anti-helicobacter pylori infection in mice
The applicant demonstrated the effect of probiotic fermented legume fermentation products against helicobacter pylori by a mouse model.
1. As fermentation products for testing the anti-helicobacter pylori infection effect: the compound was made into 100ml of liquid, wherein the amount of the compound was 1 g.
2. Establishing a helicobacter pylori infection mouse model: BALB/c mice, 6-8 weeks old, pathogen Free grade (SPF grade), male, 20-25g in weight, purchased from Nanjing university model animal institute; before Hp infection, the stomach is first perfused with mixed antibiotic solution to eliminate interference factors, 0.3 mL/piece, 1 time/day, 3 days in total, and the mixed antibiotic solution includes ampicillin 10mg/mL, gentamicin 1.2mg/mL, and azithromycin 10 mg/mL. One week later gavage of 0.3ml freshly cultured Hp solution (1 x 10)9CUF/ml). After 48 hours, the mice were tested for Hp infection using Hp urease kit to confirm successful modeling of the mice.
3. Animal grouping: the mice successfully modeled were divided into 5 groups on average, and each group was divided into 50 mice, namely a blank control group, a group 1 (bean powder group), a group 2 (compound group of example 23), a group 3 (lactobacillus reuteri DSM17648 bacterial solution), and a positive control group; the mice were administered by gavage 24 hours after successful modeling, once a day, 0.3ml once, with 4 weeks as a course of treatment. Wherein: the blank control group is irrigated with clear water with the same amount, and the positive control group is administered with the combined medicine of the proton pump inhibitor, the amoxicillin and the clarithromycin with the same amount.
4. And (3) counting the growth condition of the mice: the growth state of the mice after administration is evaluated by referring to the 'clinical research index principle of treating spleen deficiency syndrome by traditional Chinese medicines', the evaluation indexes comprise mental state, activity state, hair state and excrement condition, each index accounts for 25% of the weight, and the scoring standard is shown in table 1. Mice were separated into observation cages every 7 days, and the growth of the mice was scored, and the evaluation score was the average of scores of 10 mice, and the results are shown in table 2.
TABLE 1 mouse growth evaluation index distribution chart
Figure BDA0003122434550000191
TABLE 2 mouse growth score scoring table during dosing
Group of Before administration Day 7 Day 14 Day 21 Day 28
Blank control group 1.53 1.49 1.63 1.60 1.65
Group 1 1.60 1.59 1.57 1.62 1.60
Group 2 1.57 1.42 1.04 0.87 0.48
Group 3 1.56 1.48 1.34 1.14 0.81
Positive control group 1.58 1.39 1.06 0.95 0.54
As can be seen from table 2, the growth status of the mice in groups 2, 3 and the positive control group improved significantly during the administration period, and especially after 28 days, the symptoms substantially disappeared. In group 2, compared with the blank control group, the bean metagenome is improved more obviously, and even converted more obviously than the positive control group; the group 1, namely the bean powder group, mainly comprises bean powder, the growth state of the mice is not obviously changed compared with that of a blank control group, and the fact that the beans have no improvement effect on the helicobacter pylori infection of the mice is shown. Therefore, the improved bean metazoan compound can improve the condition that the growth state of mice is not good due to the infection of helicobacter pylori.
5. Statistics of mouse negative turning condition: after one treatment course is finished, the mice are killed by a vertebral dislocation sacrifice method, a mouse stomach tissue is longitudinally cut, and the Hp infection condition of the mice is detected by using a Hp urease kit. The results are shown in Table 3:
TABLE 3 statistics of treatment of mice infected with helicobacter pylori
Figure BDA0003122434550000192
Figure BDA0003122434550000201
As can be seen from table 3, group 1, i.e., the bean powder group, did not turn negative in mice after one treatment period, indicating that the bean powder did not have anti-helicobacter pylori efficacy; group 2, i.e. the group of posterogenous beans, when the second week of treatment, mice begin to turn negative, and the negative-turning rate reaches 92% after the treatment course is over, and is higher than that of the positive control group; group 3, lactobacillus reuteri DSM17648, also had a certain negative conversion rate, but was lower than the positive control group. Lactobacillus helveticus DSM17648 is a Lactobacillus strain screened by ORGANO BALANCE, Germany in 2002, and research proves that the bacterial liquid of the strain can reduce the number of helicobacter pylori in vivo after entering the stomach, and related patent is applied (PCT/DE 2006/001842). The action mechanism is as follows: the surface of the strain is provided with adhesion molecules for identifying and adhering helicobacter pylori, and the adhesion molecules can be physically bound and wound with the helicobacter pylori in the stomach to form insoluble polymers, and then are discharged out of the body through the digestive tract. Therefore, it can only reduce the number of helicobacter pylori, but cannot radically cure it, so its negative conversion rate is low.
The bean metazoan compound comprises bean polypeptide, long-short chain fatty acid, organic acid and the like, inactivated probiotics and metabolites thereof such as various enzymes and the like, and can destroy the colonization environment of helicobacter pylori to a certain extent and weaken the colonization of the helicobacter pylori on gastric epithelial cells; this is advantageous to some extent for the aggregation of the colonized helicobacter pylori. In addition, the dietary fiber in the compound can promote the gastrointestinal peristalsis, thereby helping the efflux of the agglomerated helicobacter pylori. After the helicobacter pylori is discharged out of the body, the damaged gastric mucosa obtains a good treatment or repair environment. The unsaturated fatty acid in the bean fermentation product, and hydrophobic substances such as protein, phosphatidic acid, and capsule on the surface of inactivated probiotics adhere to the injured gastric mucosa to prevent reinfection. Thereby rapidly and effectively relieving various stomach discomforts caused by the helicobacter pylori, such as stomachache, heartburn, gastric acid, gastric ulcer and the like. Therefore, the bean powder fermentation product realizes the synergistic effect with the inactivated probiotics and the metabolite thereof, and has higher negative conversion rate.
The present invention has been disclosed in terms of the preferred embodiment, but is not intended to be limited to the embodiment, and all technical solutions obtained by substituting or converting equivalents thereof fall within the scope of the present invention.

Claims (10)

1. An metazoan complex, characterized by: comprises bean powder, inactivated probiotics and metabolites thereof.
2. The metazoan complex of claim 1, wherein: the probiotic is lactobacillus; preferably, the lactobacillus is selected from one or more of lactobacillus rhamnosus, lactobacillus acidophilus, lactobacillus crispatus, lactobacillus fermentum, lactobacillus plantarum, lactobacillus casei, lactobacillus paracasei, lactobacillus jensenii, lactobacillus gasseri, lactobacillus reuteri, lactobacillus salivarius, lactobacillus bifidus, lactobacillus delbrueckii, lactobacillus caucasicus, lactobacillus mallotiformis, lactobacillus pentosus, lactobacillus bulgaricus, lactobacillus helveticus and lactobacillus gasseri.
3. The metazoan complex of claim 1, wherein: the bean species is selected from one or more of semen glycines, semen Phaseoli Radiati, and semen Sojae Atricolor.
4. The metazoan complex of claim 1, wherein: the composition further comprises one or more of broccoli extract, Korean vegetable extract, Glycyrrhrizae radix extract, vitamin U, semen Cucurbitae extract, fructus Piperis extract, flos Caryophylli tea extract, propolis extract, Bulbus Allii extract, Curcuma rhizome extract, fructus Phyllanthi extract, and cortex Ulmi Pumilae extract.
5. An adjunctive therapeutic drug, food, nutraceutical, dietary supplement or nutraceutical comprising the metazoan complex of any of claims 1-4.
6. Use of the metazoan complex of any one of claims 1-4 in the manufacture of a medicament, food, dietary supplement, nutraceutical or nutraceutical for the treatment or prevention of helicobacter pylori infection and gastrointestinal disorders.
7. The anti-helicobacter pylori adjunctive therapy medicinal food, nutraceutical, dietary supplement or nutraceutical according to claim 6, wherein: the dosage form is hard capsule; tablets including chewable tablets and troches; solid beverages including powders, granules, mixed powders; candies including hard and soft candies; soft capsules; jelly; an energy bar; chocolate or biscuits.
8. A method for preparing an metazoan complex, comprising: pretreating bean powder, mixing with inactivated or non-inactivated probiotics, drying, and packaging; the inactivated probiotics also includes metabolites thereof;
or
Fermenting the bean powder and probiotics, drying the fermentation liquor in the logarithmic phase or the stationary phase of the probiotics, and packaging to obtain the finished product.
9. The method of claim 8, wherein: the probiotic is lactobacillus; the lactobacillus is selected from one or the combination of more than two of lactobacillus rhamnosus, lactobacillus acidophilus, lactobacillus crispatus, lactobacillus fermentum, lactobacillus plantarum, lactobacillus casei, lactobacillus paracasei, lactobacillus jensenii, lactobacillus gasseri, lactobacillus reuteri, lactobacillus salivarius, lactobacillus bifidus, lactobacillus delbrueckii, lactobacillus gasseri, lactobacillus mallotiformis, lactobacillus pentosus, lactobacillus bulgaricus, lactobacillus helveticus and lactobacillus gasseri.
10. The method of claim 7, wherein: the beans are selected from one or more of semen glycines, semen Phaseoli Radiati, and semen Sojae Atricolor.
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