CN112236154A - Composition and application thereof - Google Patents
Composition and application thereof Download PDFInfo
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- CN112236154A CN112236154A CN201880093354.1A CN201880093354A CN112236154A CN 112236154 A CN112236154 A CN 112236154A CN 201880093354 A CN201880093354 A CN 201880093354A CN 112236154 A CN112236154 A CN 112236154A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
Abstract
A composition and its use are provided, the composition comprising Lactobacillus gasseri and/or its metabolites, Lactobacillus acidophilus and/or its metabolites and Bifidobacterium pseudocatenulatum and/or its metabolites. The composition can be used for treating inflammatory diseases including ulcerative enteritis and related diseases.
Description
The application relates to the field of microbial preparations, in particular to a microorganism or a microorganism-related composition and application thereof.
Inflammatory Bowel Disease (IBD) is a chronic inflammatory bowel disease with unknown etiology, which is easily recurrent and seriously affects the quality of life of patients. Modern medicine considers that factors that cause Inflammatory Bowel Disease (IBD) include genetics, diet, infection, autoimmunity, psychological factors, and environment. Inflammatory bowel diseases include ulcerative enteritis (UC) and Crohn's Disease (CD), both of which are inflammation-related diseases.
Ulcerative Colitis (UC) is an important type of Inflammatory Bowel Disease (IBD), the cause of which is unknown, and the main lesion is in the submucosa of the colonic mucosa, which is a chronic intestinal disease. Based on the current research, the causes of UC are mainly considered to be host genetic susceptibility, intestinal flora and immune reaction of intestinal mucosa, the clinical and pathological manifestations are continuous abdominal pain, diarrhea and mucous bloody stool, and the disease condition is repeated, and the number of people suffering from UC in China shows a remarkable rising trend in recent years.
At present, clinically, medicines aiming at UC mainly comprise salicylic acid medicines, glucocorticoid medicines and immunosuppressant. All three types of medicines can relieve UC to a certain extent, but all have disadvantages. Salicylic acid medicines can better inhibit prostaglandin synthesis and remove oxygen radicals so as to achieve the purpose of relieving inflammatory reaction, but can only relieve the inflammatory reaction in a short term and cannot realize radical treatment, and the salicylic acid western medicine for clinically treating UC is mainly sulfasalazine (SASP) and mainly aims at mild, moderate and chronic UC patients. In addition, salicylic acid drugs also have many side effects, such as causing gastrointestinal reactions, headache, reticulocyte increase, spermatozoa reduction, and rash, hepatotoxicity, leukopenia, anemia and the like caused by anaphylaxis, and the drugs also have antibacterial effect, are easy to cause flora disorder and enhance drug resistance. Glucocorticosteroids are the first choice for patients with severe or explosive UC, typical drugs such as betamethasone; however, glucocorticoid drugs cause side effects such as metabolic disturbance of the body and water retention, and can only be used as emergency drugs and cannot be taken for a long time. Immunosuppressive agents, such as cyclosporine, can inhibit UC by inhibiting the production of T-cell IL-2, affecting the progression of the immune response; however, immunosuppressant therapy has a large drug dependence and a long treatment period, is easy to cause renal toxicity and secondary infection, and can only be used as an auxiliary treatment means. Therefore, there is no safe and effective therapeutic agent for inflammation and related diseases, especially ulcerative enteritis.
Disclosure of Invention
The purpose of the present application is to provide a composition and its use.
The following technical scheme is adopted in the application:
one aspect of the present application discloses a composition comprising lactobacillus gasseri and/or a metabolite thereof, lactobacillus acidophilus and/or a metabolite thereof, and bifidobacterium pseudocatenulatum and/or a metabolite thereof.
The key point of the application lies in that the research finds that the combined use of lactobacillus gasseri, lactobacillus acidophilus and bifidobacterium pseudocatenulatum can prevent and treat inflammation and inflammation-related diseases, and particularly can effectively prevent and treat ulcerative enteritis; research shows that the prevention or treatment and prevention effects of the composition mainly comprise two aspects, on one hand, the lactobacillus gasseri, the lactobacillus acidophilus and the bifidobacterium pseudocatenulatum improve the intestinal microecology in vivo to form an ecological protection barrier consisting of beneficial bacteria, thereby having the prevention and treatment effects on inflammation or inflammation-related diseases; on the other hand, metabolites of Lactobacillus gasseri, Lactobacillus acidophilus and Bifidobacterium pseudocatenulatum serve as probiotic materials for the prevention and treatment of inflammation or inflammation-related diseases. Thus, the compositions of the present application are of significant utility in the prevention or treatment of inflammation or inflammation-related disorders, in particular in the prevention or treatment of ulcerative enteritis or related disorders thereof.
It should be noted that, in one implementation of the present application, the composition consisting of lactobacillus gasseri, lactobacillus acidophilus and bifidobacterium pseudocatenulatum improves the intestinal microecology in vivo through three bacteria to form an ecological protection barrier consisting of beneficial bacteria, thereby achieving the effect of preventing and treating ulcerative enteritis. It is understood that this improvement in the micro-ecology has not only preventive and therapeutic effects on ulcerative enteritis but also effects on other diseases associated with the micro-ecology, such as general enteritis or gastritis; thus, the compositions of the present application may be used to prevent or treat inflammation or inflammation-related disorders.
Preferably, lactobacillus gasseri is lactobacillus gasseri TF08-1 with the accession number GDMCC 60092, lactobacillus acidophilus is lactobacillus acidophilus AM13-1 with the accession number GDMCC 60091, and bifidobacterium pseudocatenulatum is bifidobacterium pseudocatenulatum TM12-14 with the accession number GDMCC 60089.
The key point of the application lies in that the research finds that the combined use of lactobacillus gasseri, lactobacillus acidophilus and bifidobacterium pseudocatenulatum can prevent and treat inflammation and inflammation-related diseases, and the three strains of lactobacillus gasseri TF08-1, lactobacillus acidophilus AM13-1 and bifidobacterium pseudocatenulatum TM12-14 have better combined use effect in the research process of the application, so that the three strains are respectively preserved. On the one hand, it is understood that, under the inventive idea of the present application, other strains of lactobacillus gasseri, lactobacillus acidophilus and bifidobacterium pseudocatenulatum can be used to achieve the effect even exceeding the effect of the three strains of the present application; on the other hand, in the case where the prevention or treatment effect on inflammation or inflammation-related diseases is required to be low, other strains of Lactobacillus gasseri, Lactobacillus acidophilus and Bifidobacterium pseudocatenulatum may be used.
Preferably, the compositions of the present application further comprise other probiotics and/or prebiotics.
It should be noted that the composition of the present application is characterized in that the combination of lactobacillus gasseri, lactobacillus acidophilus and bifidobacterium pseudocatenulatum can prevent and treat inflammation and inflammation-related diseases, and it can be understood that under the condition of not affecting the combined effect of lactobacillus gasseri, lactobacillus acidophilus and bifidobacterium pseudocatenulatum, other probiotics or prebiotics can be added, so that the composition of the present application has more functions or enhances the original effect thereof, and the probiotics or prebiotics can adopt probiotics or prebiotics reported in the existing research, and are not specifically limited herein.
Preferably, in one embodiment of the present application, the prebiotic in the composition of the present application is at least one selected from the group consisting of Fructooligosaccharide (FOS), Galactooligosaccharide (GOS), Xylooligosaccharide (XOS), lactulose oligosaccharide (lactulose), Soy Oligosaccharide (SOS), Inulin (Inulin) and oligosaccharide.
Preferably, the composition of the present application further comprises a substance that helps to maintain the viability of at least one of lactobacillus gasseri, lactobacillus acidophilus and bifidobacterium pseudocatenulatum.
It is understood that, in order to maintain the viability of lactobacillus gasseri, lactobacillus acidophilus and bifidobacterium pseudocatenulatum in the composition and ensure the efficacy thereof, various substances for maintaining the viability of the strains can be added into the composition, and the viable substances can be the viable substances reported in the prior research and are not particularly limited.
Preferably, in one implementation of the present application, the substance that helps to maintain the viability of at least one of lactobacillus gasseri, lactobacillus acidophilus and bifidobacterium pseudocatenulatum is selected from at least one of cysteine, glutathione, butyl hydroxy anisole, dibutyl methyl toluene, tocopherol, bamboo leaf antioxidant, D-erythorbic acid or its sodium salt, sodium ascorbate, calcium ascorbate, phospholipids, vitamin C and vitamin E.
Preferably, the composition of the present application further comprises a pharmaceutically or food acceptable carrier or adjuvant.
In one implementation of the present application, the prophylactic and therapeutic effects are achieved mainly by eating the composition of the present application. Therefore, the composition can also comprise pharmaceutically or food acceptable carriers or auxiliary materials for convenient use.
Preferably, the pharmaceutically or food acceptable carrier or excipient is at least one selected from glucose, lactose, sucrose, starch, mannitol, dextrin, fatty acid glyceride, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid ester, amino acid, gelatin, albumin, water and physiological saline.
In another aspect of the application, there is disclosed the use of a composition of the application for the preparation of a food, health product, food additive or medicament for the treatment or prevention of inflammation or inflammation-related disorder.
It is understood that the composition of the present application has effects of preventing and treating inflammation and inflammation-related diseases, and the composition of the present application may be prepared into various foods, health products, food additives or medicines for convenient use.
Preferably, the composition of the present application can be used for preparing food, health products, food additives or medicines for treating or preventing ulcerative enteritis or related diseases thereof.
In a further aspect of the present application is disclosed the use of a composition of the present application for the preparation of a food, nutraceutical, dietary supplement or pharmaceutical product for the control of weight loss in a mammal.
Among them, the weight loss of mammals, especially the weight loss of mammals due to inflammation.
Preferably, the inflammation is ulcerative enteritis, i.e., the control of weight loss in a mammal due to ulcerative enteritis.
In a further aspect of the present application is disclosed the use of a composition of the present application for the preparation of a food, nutraceutical, food additive or pharmaceutical product for reducing the disease activity index in a mammal.
In a further aspect of the present application is disclosed the use of a composition of the present application in the preparation of a food, nutraceutical, food additive or pharmaceutical product for improving intestinal disorders in a mammal.
It is noted that the composition of the present application can prevent and treat inflammation and inflammation-related diseases, and the key point is that the composition can control weight loss caused by inflammation and inflammation-related diseases, reduce disease activity index caused by inflammation and inflammation-related diseases, and improve intestinal lesions of mammals; therefore, the composition of the present application can also be used alone for preparing a food, health product, food additive or medicine for controlling the weight loss of mammals, reducing the disease activity index of mammals or improving intestinal lesions of mammals.
In yet another aspect, the present application discloses methods of treating or preventing inflammation or inflammation-related disorders using the compositions of the present application.
In yet another aspect of the present application, a method of controlling weight loss in a mammal using the composition of the present application is disclosed.
In yet another aspect of the present application, a method of reducing the disease activity index of a mammal using the composition of the present application is disclosed.
In yet another aspect of the present application, a method of ameliorating intestinal lesions in a mammal using the composition of the present application is disclosed.
In the above methods, the composition of the present application is administered to treat or prevent inflammation or inflammation-related diseases, control weight loss of a mammal, reduce disease activity index of a mammal, or improve intestinal disorders in a mammal.
In yet another aspect of the present application, a food product comprising the composition of the present application is disclosed.
Preferably, the food product is a lactic acid drink or a soy milk drink. The food of the present application is an edible product in any form in a broad sense, and is not limited to a lactic acid drink or a soy milk drink, and may be, for example, a fermented food or an animal feed.
It should be noted that the food product of the present application, which contains the composition of the present application, also has the effects of treating or preventing inflammation or inflammation-related diseases, controlling weight loss of mammals, reducing disease activity index of mammals, and improving intestinal lesions of mammals. It is understood that the food product of the present application, which is characterized by containing the composition of the present application, may be in a specific form, such as solid, liquid, etc., according to different food products or use requirements, and is not limited herein. In one implementation of the present application, the composition of the present application is mainly made into a common lactic acid beverage or soymilk beverage for convenient drinking; of course, solid food such as milk slices, cheese strips, etc. may be prepared, and are not particularly limited herein.
It should be noted that the amount of active bacteria or the intake amount of lactobacillus gasseri, lactobacillus acidophilus and bifidobacterium pseudocatenulatum in the food of the present application is not specifically limited, and in practical applications, the active bacteria or the intake amount can be flexibly selected according to actual conditions. Taking Lactobacillus gasseri TF08-1, Lactobacillus acidophilus AM13-1 and Bifidobacterium pseudocatenulatum TM12-14 as examples, the study of the present application showed that the daily intake of Lactobacillus gasseri TF08-1, Lactobacillus acidophilus AM13-1 and Bifidobacterium pseudocatenulatum TM12-14 was 109The cfu/mL composition is 0.2mL, has good treatment effect on ulcerative enteritis, and the active microbial inoculum amount can be used as reference dosage or reference intake amount of food, health care products, food additives or medicines.
In yet another aspect, the present application discloses a health product comprising the composition of the present application.
It should be noted that the health product of the present application, containing the composition of the present application, also has the effects of treating or preventing inflammation or inflammation-related diseases, controlling weight loss of mammals, reducing disease activity index of mammals and improving intestinal lesions of mammals.
In yet another aspect of the present application, a food additive comprising the composition of the present application is disclosed.
The composition of the present application can be administered in combination with a usual food material. For example, cereals including rice, flour, coarse cereals, and potatoes including potatoes, sweet potatoes, etc.; animal food including meat, poultry, fish, milk, eggs, etc.; legumes and products thereof, including soybeans and other dried legumes; vegetables and fruits including fresh beans, rhizomes, leafy vegetables, solanum melongena, etc.; pure heat energy food, including animal and vegetable oil, starch, edible sugar and wine, etc.; therefore, the composition can be independently used as a food additive or a modulator and added into various food materials for direct eating, and has the treatment or prevention effect on inflammation and related diseases thereof.
In yet another aspect of the present application, a pharmaceutical product comprising the composition of the present application is disclosed.
Preferably, the pharmaceutical product is a tablet, granule, powder, enteric coated solution, solution or suspension.
The pharmaceutical product of the present application has an effect of treating or preventing inflammation and related diseases, because it contains the composition of the present application; the pharmaceutical product of the present application may be a composition of the present application alone or may be used in combination with other inflammatory drugs as long as the activity is not affected. It is to be understood that the drug of the present application may take various forms as long as the activity of each strain in the composition is not affected. The pharmaceutical composition of the present application may further comprise adjuvants commonly used in pharmaceutical compositions or dosage forms, such as stabilizers, wetting agents, emulsifiers, binders, isotonic agents, and the like.
The medicine can be administrated in any form of oral liquid, tablets, injection, orally disintegrating tablets, freeze-dried powder preparations or enteric-coated dosage forms. Enteric dosage forms, such as capsules or enteric tablets, are preferred so that the active ingredient of the drug, i.e., the microorganism, passes through the stomach smoothly without being destroyed by stomach acid. More preferably, the medicament of the present application may be formulated as enteric coated tablets for oral use.
The enteric formulation of the present application means a pharmaceutical formulation which is not disintegrated in gastric juice but is disintegrated and absorbed in intestinal juice, and the enteric formulation includes a capsule and an enteric tablet. Wherein the capsule is formed by encapsulating a powdered drug in a capsule shell which is acceptable for use with conventional medicaments; the enteric coated tablet is formed by coating a layer of enteric coating outside a common tablet medicament. The term "enteric coating" is abbreviated as "enteric coating" in the present application and includes all coatings which are approved for use with conventional drugs and which are not degraded by gastric acid but which are sufficiently decomposed in the small intestine to rapidly release the drug product of the present application. For example, the enteric coating of the present application can be maintained at 36-38 ℃ for more than 2 hours in synthetic gastric acid, e.g. HCl solution at pH 1, and preferably decomposed within 1.0 hour in synthetic intestinal fluid, e.g. buffer at pH 7.0.
Preferably, the enteric coating of the enteric coated tablet of the present application has a thickness of 5 to 100 μm, and preferably 20 to 80 μm. The enteric coating composition is selected from conventional materials known to those skilled in the art.
The content of active bacteria of each strain of the probiotic composition in the medicine of the application or the dosage of the medicine is not particularly limited, and in practical application, the content can be flexibly selected according to the health condition of an administration object. However, the studies of the present application showed that the daily intake of Lactobacillus gasseri TF08-1, Lactobacillus acidophilus AM13-1 and Bifidobacterium pseudocatenulatum TM12-14 all had concentrations of 1090.2mL of cfu/mL of probiotic composition has good treatment effect on ulcerative enteritis, and the dosage can be used as the content of active bacteria in the medicine or the reference of administration dosage.
The beneficial effect of this application lies in:
the composition provided by the application can have good treatment and prevention effects on inflammation, particularly ulcerative enteritis and related diseases thereof by combined use of lactobacillus gasseri, lactobacillus acidophilus and bifidobacterium pseudocatenulatum, and provides a novel composition which is safe, effective, small in toxic and side effects and not easy to generate resistance for treatment and prevention of inflammation and related diseases.
FIG. 1 is a Control group, a model group, and a VSL in an embodiment of the present application#The change curve of body weight of mice in group 3 and probiotic composition treatment groups;
FIG. 2 is a Control group, a model group, and a VSL in an embodiment of the present application#Profile of DAI index in mice of group 3 and probiotic composition treated groups.
The lactobacillus gasseri TF08-1 is preserved in the Guangdong province microorganism culture preservation center in 2016, 10 and 13 days, the preservation unit is the fifth building of the experimental building of the Mieli 100 province microorganism institute in Guangzhou city, Guangdong province, China, and the preservation number is GDMCC 60092.
Lactobacillus acidophilus AM13-1 was deposited in the microbial cultures Collection of Guangdong province in 2016, 10, 13 days, with the location of the culture unit being Wulou of the microbial institute's laboratory building, Mieli Zhonglu 100 province, Guangzhou, Guangdong province, China, and the number of the culture unit being GDMCC 60091.
Bifidobacterium pseudocatenulatum TM12-14 was deposited in Guangdong province microorganism culture collection center at 2016, 10, 13 days, with the location of the culture unit being Wulou of the Experimental building of microbial institute, Mieli Zhonglu, 100 province, Guangzhou, Guangdong province, China, and the deposition number being GDMCC 60089.
With the intensive study of intestinal microecology, the pathogenesis of ulcerative enteritis is closely related to the composition of intestinal microorganisms, the imbalance of intestinal bacteria is closely related to intestinal mucositis reaction, and the excessive proliferation of harmful bacteria can trigger inflammatory reaction to further induce the pathogenesis of ulcerative enteritis. Healthy human intestinal tracts contain a large number of beneficial bacteria, which form the first biological barrier of the intestinal tract.
Based on the above research and knowledge, the present application has developed and proposed a novel composition comprising lactobacillus gasseri and/or its metabolites, lactobacillus acidophilus and/or its metabolites, and bifidobacterium pseudocatenulatum and/or its metabolites. The composition not only has the functions of treating and preventing ulcerative enteritis, but also has the effect on other diseases related to the micro ecology, such as common enteritis, gastritis and the like; thus, the compositions of the present application can be used to prevent or treat inflammation or inflammation-related disorders.
In one implementation mode of the application, the composition consisting of lactobacillus gasseri TF08-1 with the accession number of GDMCC 60092, lactobacillus acidophilus AM13-1 with the accession number of GDMCC 60091 and bifidobacterium pseudocatenulatum TM12-14 with the accession number of GDMCC60089 has better treatment effect on ulcerative enteritis than VSL produced by Alfasigma company in America #3 composite probiotic preparation, can be used for preparing medicines for treating or preventing diseasesFood, health product, food additive or medicine for preventing inflammation and related diseases.
The present application will be described in further detail with reference to specific examples. The following examples are intended to be illustrative of the present application only and should not be construed as limiting the present application.
Example one
This example was carried out on the therapeutic effect of a probiotic composition of Lactobacillus gasseri TF08-1, Lactobacillus acidophilus AM13-1 and Bifidobacterium pseudocatenulatum TM12-14 on ulcerative enteritis, using a mouse model of sodium dextran sulfate (abbreviated as DSS) with molecular weight of 36000-50000 as a subject. The method comprises the following specific steps:
materials and methods
1. Strain culture and identification
1.1 Lactobacillus gasseri TF08-1
The Lactobacillus gasseri TF08-1 was isolated using PYG medium under anaerobic conditions at 37 ℃. TF08-1 has white colony with low bulge, nearly circular shape and wavy edge, and has colony diameter of about 1-2mm, and the microscopic shape of thallus is rod, gram positive, non-spore producing and flagellum. The strain is preserved in Guangdong province microorganism culture collection center with the preservation number of GDMCC 60092.
The specific separation and identification steps of the lactobacillus gasseri TF08-1 are as follows:
1.1.1 sample Collection
The isolated sample was obtained from a fecal sample from a 16-year-old healthy female volunteer who was resident in Shenzhen, Guangdong province. And the dietary and physical conditions of the volunteer were recorded in detail.
1.1.2 isolation and culture of the Strain
The preparation method comprises the following steps of preparing a separation culture medium in advance, wherein the culture medium is a PYG culture medium purchased from Kyork microorganism science and technology company and comprises the following specific components: 5g of peptone, 5g of trypticase, 10g of yeast powder, 5g of beef extract, 5g of glucose and K2HPO 4 2g，Tween 80 1mL，Cysteine-HCl·H 2O 0.5g, sodium sulfide 0.25g, heme 5mg, vitamin K 11 mu L, 40mL of inorganic salt solution, 1mg of Resazurin, 950mL of distilled water, pH 6.8-7.0, and sterilizing at 115 ℃ for 25 min. The solid medium was added with 1.5% agar and poured in an anaerobic cabinet. Each 1L of inorganic salt solution contains CaCl2·2H 2O 0.25g，MgSO 4·7H 2O 0.5g，K 2HPO 4 1g，KH 2PO 4 1g，NaHCO 3 10g，NaCl 2g。
Transferring the collected fresh feces sample to anaerobic chamber, suspending 0.2g feces in 1mL sterile Phosphate Buffer Solution (PBS), mixing well, performing gradient dilution, coating 100 μ L dilution solution, performing anaerobic culture at 37 deg.C for 3-4 days with anaerobic gas component N2：CO 2：H 2When the ratio is 90:5: 5. and selecting a single colony for streaking and purifying after the colony grows out of the plate to obtain a pure culture strain, and then carrying out identification and functional verification.
1.1.3 identification of 16S rDNA of Strain
The isolated strains were subjected to 16S rDNA identification to determine species classification information of the strains. Culturing the obtained isolated strain in a liquid PYG culture medium for 24h, centrifuging 1mL of bacterial liquid at 10000r/min for 5min, collecting thalli, extracting the genomic DNA of the strain, amplifying 16S rDNA by taking the genomic DNA as a template, and using a 16S rDNA universal primer.
The PCR amplification system of 16S rDNA is as follows: 10 XPCR buffer 3 u L, dNTP 2.5.5 uL, upstream primer 27F 0.5 uL, downstream primer 1492R 0.5 u L, Taq enzyme 0.3 uL, template 1 u L, ddH2O 18.2μL。
The amplification conditions for 16S rDNA were: pre-denaturation at 95 ℃ for 4min, then 30 cycles: denaturation at 95 ℃ for 30s, annealing at 57 ℃ for 40s, and extension at 72 ℃ for 1min for 30 s.
And purifying the PCR amplification product of the 16S rDNA, sequencing the PCR amplification product by 3730 to obtain a 16S rDNA sequence of the strain, and then performing comparison of databases of NCBI.
The upstream and downstream primers of the 16S rDNA universal primer in the test are respectively sequences shown in SEQ ID NO.1 and SEQ ID NO. 2. The result of the 16S rDNA sequence of the strain TF08-1 obtained by separation is the sequence shown in SEQ ID NO. 3. The NCBI blast comparison result shows that the TF08-1 strain isolated in this example has the highest homology with Lactobacillus gasseri and the similarity is 99.9%, so that the TF08-1 is judged to be Lactobacillus gasseri, named as Lactobacillus gasseri TF08-1, and is preserved.
SEQ ID NO.1：5’-AGAGTTTGATCATGGCTCAG-3’
SEQ ID NO.2：5’-TAGGGTTACCTTGTTACGACTT-3’
1.2 Lactobacillus acidophilus AM13-1
Lactobacillus acidophilus AM13-1 was isolated using PYG medium under anaerobic conditions at 37 ℃. The AM13-1 colony cultured in PYG medium for 2 days is white, convex, viscous, opaque, round, regular in edge, about 2-3mm in diameter, and the microscopic shape of the thallus is rod-shaped, gram-positive, and does not produce spores and flagella. The strain is preserved in Guangdong province microorganism culture collection center with the preservation number of GDMCC 60091.
The specific separation and identification steps of the lactobacillus acidophilus AM13-1 are as follows:
1.2.1 isolation culture
The separated sample is from a fecal sample of a healthy male in Shenzhen city, and the separation process of lactobacillus acidophilus AM13-1 is as follows:
(1) transferring the sample into an anaerobic box, taking about 0.2g of the sample, suspending the sample in 1mL of sterile PBS, fully and uniformly mixing, and then carrying out gradient dilution;
(2) 100 mu L of diluent is taken to be coated on a PYG medium plate, and the PYG medium plate is uniformly coated and placed in an anaerobic environment at 37 ℃ for culture, wherein the anaerobic gas composition is as follows: nitrogen gas: hydrogen gas: carbon dioxide 90:5: 5; the PYG medium was the same as "isolation culture of 1.1.2 Strain";
(3) culturing for 4 days, after bacterial colony grows on the flat plate, selecting single bacterial colony, carrying out streak purification, and carrying out anaerobic culture at 37 ℃;
(4) the pure single bacterium is preserved by glycerol and vacuum freeze drying.
1.2.2 identification of 16S rDNA of AM13-1
Extracting genome DNA, performing 16S rDNA amplification by using the DNA as a template, adopting a 16S rDNA universal primer, performing pre-denaturation at 95 ℃ for 4min under the amplification condition, and then performing 30 cycles: denaturation at 95 ℃ for 30s, annealing at 57 ℃ for 40s, and extension at 72 ℃ for 1min for 30 s. And purifying the amplified PCR product, sequencing the PCR product by 3730 to obtain a 16S rDNA full-length sequence of AM13-1, and aligning the 16S rDNA sequence of AF13-1 in a database of NCBI.
The 16S rDNA universal primer and the PCR amplification system of the test are the same as the 16S rDNA identification of the 1.1.3 strain. The result of the 16S rDNA sequence of the strain AM13-1 obtained by separation is the sequence shown in SEQ ID NO. 4. The NCBI blast comparison result shows that the AM13-1 strain isolated in the example has the highest homology with Lactobacillus acidophilus and the similarity is 100%, so that AM13-1 is judged to be Lactobacillus acidophilus and named as Lactobacillus acidophilus AM13-1, and the Lactobacillus acidophilus strain is preserved.
1.3 Bifidobacterium pseudocatenulatum TM12-14
Bifidobacterium pseudocatenulatum TM12-14 was isolated using PYG medium under anaerobic conditions at 37 ℃. The TM12-14 colonies cultured in PYG medium for 2 days are white, convex, round, and neat in edge, the diameter of the colonies is about 1-2mm, the microscopic morphology of the thalli is in a branched rod shape, gram staining is positive, and no spore or flagellum is produced. The strain is preserved in Guangdong province microorganism culture collection center with the preservation number of GDMCC 60089.
The specific separation and identification steps of the bifidobacterium pseudocatenulatum TM12-14 are as follows:
1.3.1 sample Collection
The samples were isolated from faeces of a 14 year old healthy male, collected into sterile sample tubes and brought back to the laboratory within 1h for sorting.
1.3.2 isolation and purification of Bifidobacterium pseudocatenulatum
Immediately transferring the collected fresh sample to an anaerobic operation box, putting 0.2g of the sample into 1mL of sterile PBS, fully shaking and uniformly mixing, then carrying out gradient dilution coating, adopting a PYG medium plate as a culture medium, carrying out anaerobic culture at 37 ℃, wherein the anaerobic gas component is N2：CO 2：H 2＝90：5：5。After 3 days of culture, single colony is selected for streaking and purification, and pure culture of each single strain is obtained. The PYG medium was the same as that used in the isolation culture of the 1.1.2 strain.
1.3.3 Strain preservation
The obtained pure culture strain was cultured to a concentration of about 109cfu/mL, 400 μ L of 40% glycerol was added to 400 μ L of the bacterial solution to make the glycerol concentration reach 20%, and then the mixture was stored at-80 ℃ and ultra-low temperature.
1.3.416S rDNA identification
Culturing the obtained separated strain in a liquid PYG culture medium for 24h, centrifuging 1mL of bacterial liquid at 10000r/min for 5min, collecting the thallus, and extracting the genome DNA. The genome DNA is used as a template, and a 16S rDNA universal primer is used for PCR amplification, wherein the PCR amplification primer, the system and the conditions are the same as those of the 16S rDNA identification of the 1.1.3 strain.
And carrying out electrophoresis detection, purification and 3730 sequencing on the obtained 16S rDNA amplification product to obtain a 16S rDNA sequence of the strain, and then carrying out comparison of an NCBI database.
The sequencing result shows that the length of the 16S rDNA of the strain TM12-14 is 1400bp, and the sequence is shown as SEQ ID NO. 5. NCBI blast alignment results show that the strain TM12-14 has the highest homology with Bifidobacterium pseudocatenulatum, is identified as Bifidobacterium pseudocatenulatum, is named as Bifidobacterium pseudocatenulatum TM12-14, and is deposited.
2. Mouse model
The mouse models selected in this example were: DSS (Dextran Sulfate sodium Sulfate, Na, molecular weight 36000-50000) induced ulcerative enteritis mouse model.
Specifically, 48 mice were selected from the group of mice purchased from the center of Hubei medical laboratory animals, all of which were 8 weeks old and 20 g. + -. 2g in weight, and were bred in the environment of a SPF-rated rat house. The 48 mice were randomly divided into 4 groups of 12 mice each for subsequent testing.
DSS molding: and (3) continuously drinking 0.15% DSS for seven days to obtain the ulcerative enteritis mouse model.
3. Test method
The 48 mice were randomly divided into 4 groups of 12 mice each, 4 groups being normal (i.e., control), model, probiotic composition treatment and VSL, respectively#3 treatment groups, the specific treatment modalities for each group were as follows:
normal group: the mice were fed with normal feed and each mouse was gavaged with 0.2mL of PBS buffer per day.
Model group: feeding with the same feed, and performing DSS (direct sequence spread spectrum) molding: adding DSS into the drinking water of the mice, wherein the addition amount of the DSS is 0.15 percent of the final concentration, feeding for seven days, and feeding each mouse with 0.2mL of PBS buffer solution every day.
Probiotic composition treatment group: feeding with the same feed, feeding each mouse with 0.2mL of probiotic composition bacteria liquid 3 days before DSS molding, and then performing DSS molding: adding DSS into drinking water of mice, wherein the addition amount of the DSS is 0.15 percent of the final concentration, feeding for seven days, and feeding each mouse with 0.2mL of probiotic composition bacterial liquid per day.
The probiotic composition bacterial liquid is prepared by the following method:
culturing Lactobacillus gasseri TF08-1, Lactobacillus acidophilus AM13-1 and Bifidobacterium pseudocatenulatum TM12-14 for 24 hr, centrifuging, collecting thallus, suspending with PBS, and adjusting the concentration to 109cfu/mL, mixing the three strains according to the ratio of 1:1:1 to prepare the probiotic composition bacterial liquid.
The weight, diet and drinking conditions of the mice were recorded every day after the DSS modeling, while the fecal character and fecal occult blood condition of the mice were observed, and disease activity index (abbreviated DAI) of the mice was calculated on days 1, 3, 5 and 7, respectively, with the DAI scoring criteria detailed in table 1. After the experiment was completed, the mice were sacrificed and all mice were bled, decapped, colons were removed, photographed, weighed, and the colons length was measured. Colonic tissue was stored in a-80 ℃ freezer and paraformaldehyde. Wherein the recording time of the normal group is the same as the DSS molding.
TABLE 1 DAI index rating Table
In Table 1, "stool shape", "normal" stool means formed stool, "loose" stool means pasty, semi-formed stool not adhering to the anus, and "loose" means watery stool adhering to the anus. In the aspect of fecal occult blood/weak ocular bloody stool, the term "normal" means that the blood of the mouse stool is negative; "macroscopic bloody stool" means that red or brown bloody stool can be directly observed by the naked eye; "occult blood positive" means an unobvious visual stool that is positive for stool blood using tetramethylbenzidine. The DAI index is equal to the sum of the three integrals of body weight, "stool behavior" and "fecal occult blood/weak ocular bloody stool".
Second, results and analysis
1. Weight change
The body weights of the mice on day 1, day 3, day 5 and day 7 were counted, and the average body weights of the mice in each group are shown in table 2 and fig. 1.
TABLE 2 mean body weight of the respective mice
| Grouping | Day 1 (g) | Day 3 (g) | Day 5 (g) | Day 7 (g) |
| Control | 22.32±0.45 | 23.73±0.64 | 24.98±0.96 | 25.52±1.26 |
| Model set | 22.41±0.52 | 21.65±0.71* | 20.02±1.26* | 18.21±1.57** |
| Probiotics | 22.38±0.61 | 21.97±0.85 | 21.47±1.35 | 21.32±1.49 ▲ |
| |
22.54±0.71 | 21.87±0.92 | 21.32±1.48 | 20.01±1.66 ▲ |
In Table 2, "Control" refers to the normal group, "probiotic", i.e., probiotic composition treatment group, "VSL#3' is VSL#And 3, treatment groups. "" indicates that the body weight of mice in the model group differed from the body weight of mice in the normal group by a significant level P<0.05, ". star" means that the body weight of mice in the model group differed from that in the normal group by a significant level P<0.01, ". tangle-solidup" means "probiotic" and "VSL #3 "significant level of difference P between two groups of mice compared to the model group<0.05。
The results in Table 2 and FIG. 1 show that the body weight of the mice in the normal group tended to increase slowly, and the mice induced by DSS, i.e., the model group, "probiotic" group and "VSL#In the 3' group, the body weight of the three groups was continuously decreased, and the body weight decrease started to be significant on the 3 rd day (i.e.. about.P) in the model group compared with the control group<0.05), day 7, the difference between the two was more pronounced (i.e. P)<0.01). And probiotic compositions and VSLs #3 can slow down the weight loss of UC mice, and on day 7, the probiotics and the VSL #3 "weight loss control in these two groups of mice was significant relative to the model group (i.e.P. tangle-solidup)<0.05). Probiotics compositions and VSLs are described#3 can control the weight loss caused by UC. Also, the body weight of the mice (i.e., "probiotics") in the probiotic composition group was slightly higher than the VSL at day 7#3, the effect of the probiotic composition on controlling the body weight reduction of UC mice is slightly better than that of VSL #3。
Changes in DAI
DSS-induced ulcerative enteritis mice induced changes in DAI index due to weight loss, stool traits and changes in hematochezia status, and statistics of mouse DAI index at day 1, day 3, day 5 and day 7 are shown in table 3 and figure 2. In table 3, the DAI of each group of mice was averaged for each group of mice.
TABLE 3 mouse DAI values
| | Day | 1 | |
|
|
| Control | 1.1±0.5 | 1.1±0.7 | 1.2±0.8 | 1.3±0.8 | |
| Model set | 1.1±0.5 | 3.6±1.1* | 7.2±1.6** | 9.4±2.0** | |
| Probiotics | 1.2±0.4 | 3.3±1.2 | 6.0±1.5 ▲ | 6.5±1.8 ▲ | |
| |
1.1±0.4 | 3.4±1.3 | 6.6±1.6 | 7.8±1.9 ▲ |
In Table 3, "Control" refers to the normal group, "probiotic" i.e., probiotic composition treatment group, "VSL#3' is VSL#And 3, treatment groups. "" indicates that the model group differed significantly from the normal group mouse DAI index by a significant level P<0.05, ". x" means that the model group differed from the mouse DAI index of the normal group by a significant level P<0.01, ". tangle-solidup" means "probiotic" and "VSL #3 "significant level of difference P between two groups of mice compared to the model group<0.05。
Table 3 and figure 2 data show that the DAI in normal group mice remained essentially flat, while the model, probiotic and VSL groups were maintained with DSS induction#The DAI was gradually increased in 3 mice, and the DAI of the model group mice became initially significant on day 3 (i.e., P) compared to the control group<0.05), day 7 mice in the model group reached the highest level of DAI (i.e., P relative to control group)<0.01). Probiotic intervention can control the increase of DAI, and the DAI value of the probiotic mice on 5 days and 7 days is remarkably controlled relative to the model group (namely P)<0.05) and the DAI of the probiotic intervention group mice was slightly lower than VSL # 3 at day 7, indicating that the probiotic composition of the present example is more effective in controlling the DAI rise in UC mice than VSL # 3.
3. Changes in colon Length
The colon tissue of the UC model mouse is altered mainly because of the occurrence of ulcers and inflammation resulting in a shortening of the colon tissue, and after the treatment is finished, the colon length of the mouse measured by dissection is shown in table 4.
TABLE 4 Colon Length in mice
| Grouping | Colon Length (cm) |
| Control | 8.38±0.49 |
| Model set | 5.02±0.87** |
| |
6.30±0.67 ▲ |
| Probiotics | 6.63±0.71 ▲ |
In Table 4, "Control" refers to the normal group, "probiotic", i.e., probiotic composition treatment group, "VSL#3' is VSL#And 3, treatment groups. "" indicates that the colon length of the mice in the model group is different from that of the mice in the normal group by a significant level P<0.01, ". tangle-solidup" means "probiotic" and "VSL#3' two groups of mice have a significant difference in colon length P relative to the model group of mice<0.05。
The results in table 4 show that colon tissue shortening was more severe in the model group and very significant (. about.P.) compared to the control group in mice after 7 days of DSS induction<0.01). And probiotic compositions and VSLs #3 can significantly control the shortening of the colon of the mouse, and obtains significant control (P) relative to a model group<0.05). The colon length ratio VSL of the probiotic composition group mice can be found through the data in the table#The long colon length of 3 groups of mice can indicate that the probiotic composition has stronger capability of controlling the colon shortening of UC mice than VSL #3。
The results in tables 2 to 4 and fig. 1 and 2 show that the probiotic composition of the present example has therapeutic and prophylactic effects on ulcerative enteritis and the therapeutic effect is slightly superior to the existing VSL #3, and (3) preparing the product.
Example two
In this example, the composition demonstrated to have therapeutic and preventive effects on ulcerative enteritis in the first example was prepared into a common food as follows:
mixing milk, vitamin C and white sugar with cultured Lactobacillus gasseri TF08-1, Lactobacillus acidophilus AM13-1 and Bifidobacterium pseudocatenulatum TM12-14 according to the formula shown in Table 5, and making into food with therapeutic and prophylactic effects on ulcerative enteritis.
TABLE 5 food formulation with probiotic composition
| Raw materials | Mass percent (%) |
| Lactobacillus gasseri TF08-1 | 0.15 |
| Lactobacillus acidophilus AM13-1 | 0.15 |
| Bifidobacterium pseudocatenulatum TM12-14 | 0.15 |
| Milk | 90.0 |
| White sugar | 9.0 |
| Vitamin C | 0.55 |
Mixing milk and white sugar according to the formula of Table 5, stirring, preheating, homogenizing under 20Mpa, sterilizing at 90 deg.C for 5-10 min, cooling to 40-43 deg.C, adding protective agent (i.e. vitamin C), inoculating 1-100 × 106cfu/g of mixed probiotic food composition of three kinds of mixed probiotic bacteria, namely Lactobacillus gasseri TF08-1, Lactobacillus acidophilus AM13-1 and Bifidobacterium pseudocatenulatum TM 12-14.
The milk product of this example was added to the DSS-molded mouse feed and fed and tested according to the probiotic composition treatment group of example one, except that the milk product of this example was added to the feed only, and no additional gavage probiotic composition bacterial liquid was added. The detection result shows that the milk product of the embodiment can also control the weight reduction of UC mice, reduce the disease activity index DAI of the mice, improve intestinal lesions and has the treatment and prevention effects on ulcerative enteritis.
EXAMPLE III
In this example, a probiotic composition, which demonstrates the therapeutic and prophylactic effects of ulcerative enteritis as described in example one, was formulated as a medicament for the treatment of ulcerative enteritis, as shown in table 6:
TABLE 6 pharmaceutical formulations containing probiotic compositions
| Raw materials | Mass percent (%) |
| Lactobacillus gasseri TF08-1 | 0.5％ |
| Lactobacillus acidophilus AM13-1 | 0.5％ |
| Bifidobacterium pseudocatenulatum TM12-14 | 0.5％ |
| Lactose | 2.0％ |
| Yeast powder | 2.0％ |
| Peptone | 1.0％ |
| Purified water | 93％ |
| Vitamin C | 0.5％ |
Mixing lactose, yeast powder and peptone with purified water at the ratio of Table 6, preheating to 60-65 deg.C, homogenizing under 20Mpa, sterilizing at 90 deg.C for 20-30 min, cooling to 36-38 deg.C, mixing with protective agent (i.e. vitamin C), and respectively adding into the mixture 1-50 × 106cfu/mL Lactobacillus gasseri TF08-1, Lactobacillus acidophilus AM13-1 and Bifidobacterium pseudocatenulatum TM12-14 viable bacteria, fermenting at 36-38 ℃ until the pH value is 6.0, centrifuging, and freeze-drying until the water content is less than 3%, thus preparing the composite probiotic freeze-dried product. Weighing 0.5 g of freeze-dried substance, mixing with maltodextrin in equal amount, and encapsulating to obtain the product containing Lactobacillus gasseri gThe three strains of the asseri TF08-1, Lactobacillus acidophilus AM13-1 and Bifidobacterium pseudocatenulatum (Bifidobacterium pseudocatenulatum) 12-14 are combined with probiotics to form a capsule pharmaceutical composition.
The probiotic composition bacterial liquid in the probiotic composition treatment group of the example one was replaced with the capsule pharmaceutical composition of the example one, and the capsule pharmaceutical composition of the example was intragastrically administered in the same manner as in the example one, one capsule per day, and tested in the same manner as in the example one. The results show that the capsule pharmaceutical composition of the embodiment can also control the weight loss of UC mice, reduce the disease activity index DAI of the mice, improve intestinal lesions and has the treatment and prevention effects on ulcerative enteritis.
Example four
Preparation method of medicine for treating ulcerative enteritis (UC)
1. Preparing bacterial liquid: will be 1 × 109The method comprises the steps of respectively carrying out anaerobic culture on cfu/mL Lactobacillus gasseri TF08-1, Lactobacillus acidophilus AM13-1 and Bifidobacterium pseudocatenulatum TM12-14, wherein the anaerobic culture medium adopts PYG culture medium, carrying out anaerobic fermentation at 37 ℃ for 2-3 days, and then mixing in equal proportion to obtain the final concentration of about 1 × 109cfu/mL。
2. Preparation of growth factors: mixing skimmed milk and casein, centrifuging, and ultrafiltering to obtain crude extract of milk growth factor containing vitamins, purine, and pyrimidine.
3. Preparation of a medicament formulation: adding 5 volumes of growth factor and 1 volume of protective agent (i.e. vitamin C) into 100 volumes of fermented bacteria liquid mixture, stirring thoroughly, adding starch adjuvant such as maltodextrin, and making into pharmaceutical dosage form.
The pharmaceutical dosage form can be further made into slurry, powder or granule.
The drug formulation of the present example was directly added to the mice feed for DSS modeling, and the feeding and testing were performed according to the probiotic composition treatment group of the first example, except that the drug formulation of the present example was only added to the feed, and no additional gavage probiotic composition bacterial liquid was added. The detection result shows that the drug product of the embodiment can also control the weight loss of UC mice, reduce the disease activity index DAI of the mice, improve intestinal lesions and has the treatment and prevention effects on ulcerative enteritis.
The above examples demonstrate that the combined use of Lactobacillus gasseri, Lactobacillus acidophilus and Bifidobacterium pseudocatenulatum can treat and prevent ulcerative enteritis; moreover, the composition of the three bacteria can be prepared into various foods or medicines for use; of course, it is understood that the composition of the three bacteria can be made into various foods and medicines, and also made into various health products or food additives.
In addition, studies have shown that the therapeutic effect of the three bacteria is largely based on the improvement of the micro-ecology, which has therapeutic and preventive effects not only on ulcerative enteritis but also on other diseases associated with the micro-ecology, such as common enteritis, gastritis, etc.; thus, the compositions of the present application may be used for the prevention or treatment of inflammation or inflammation-related disorders, in particular various inflammatory bowel diseases, gastritis.
The foregoing is a more detailed description of the present application in connection with specific embodiments thereof, and it is not intended that the present application be limited to the specific embodiments thereof. For those skilled in the art to which the present application pertains, several simple deductions or substitutions may be made without departing from the concept of the present application, and all should be considered as belonging to the protection scope of the present application.
Claims (25)
- A composition characterized by: the composition comprises lactobacillus gasseri and/or metabolites thereof, lactobacillus acidophilus and/or metabolites thereof, and bifidobacterium pseudocatenulatum and/or metabolites thereof.
- The composition of claim 1, wherein: the lactobacillus gasseri is lactobacillus gasseri TF08-1 with a deposit number GDMCC 60092, the lactobacillus acidophilus is lactobacillus acidophilus AM13-1 with a deposit number GDMCC 60091, and the bifidobacterium pseudocatenulatum is bifidobacterium pseudocatenulatum TM12-14 with a deposit number GDMCC 60089.
- The composition according to claim 1 or 2, characterized in that: the composition further comprises other probiotics and/or prebiotics; preferably, the prebiotic is selected from at least one of fructooligosaccharide, galactooligosaccharide, xylooligosaccharide, lactulose oligosaccharide, soy oligosaccharide, inulin and oligosaccharide.
- The composition according to claim 1 or 2, characterized in that: the composition further comprises a substance that helps to maintain viability of at least one of lactobacillus gasseri, lactobacillus acidophilus and bifidobacterium pseudocatenulatum.
- The composition of claim 4, wherein: the substance for maintaining the activity of at least one of Lactobacillus gasseri, Lactobacillus acidophilus and Bifidobacterium pseudocatenulatum is at least one selected from cysteine, glutathione, butylated hydroxyanisole, dibutyl methyl toluene, tocopherol, bamboo leaf antioxidant, D-isoascorbic acid or its sodium salt, sodium ascorbate, calcium ascorbate, phospholipid, vitamin C and vitamin E.
- The composition according to claim 1 or 2, characterized in that: the composition also comprises a pharmaceutically or food acceptable carrier or auxiliary material.
- The composition of claim 6, wherein: the pharmaceutically or food acceptable carrier or adjuvant is at least one selected from glucose, lactose, sucrose, starch, mannitol, dextrin, fatty glyceride, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid ester, amino acid, gelatin, albumin, water and physiological saline.
- Use of a composition according to any one of claims 1 to 7 for the preparation of a food, health product, food additive or pharmaceutical product for the treatment or prevention of inflammation or inflammation-related disorders.
- Use according to claim 8, characterized in that: the inflammation is ulcerative enteritis.
- Use of a composition according to any one of claims 1 to 7 for the preparation of a food, nutraceutical, dietary supplement or pharmaceutical product for controlling weight loss in a mammal.
- Use according to claim 10, characterized in that: the weight loss of the mammal refers to the weight loss of the mammal caused by inflammation; preferably, the inflammation is ulcerative enteritis.
- Use of a composition according to any one of claims 1 to 7 for the preparation of a food, nutraceutical, food additive or pharmaceutical product for reducing the disease activity index in a mammal.
- Use of a composition according to any one of claims 1 to 7 in the manufacture of a food, nutraceutical, food additive or pharmaceutical product for improving intestinal pathologies in a mammal.
- A method of treating or preventing inflammation or inflammation-related disorder using the composition of any one of claims 1-7.
- A method of controlling weight loss in a mammal using a composition according to any one of claims 1 to 7.
- A method of reducing the disease activity index of a mammal using the composition of any one of claims 1-7.
- A method of ameliorating intestinal lesions in a mammal using the composition of any one of claims 1 to 7.
- A food product characterized by: the food product comprising the composition of any one of claims 1 to 7.
- The food product of claim 18, wherein: the food is a lactic acid beverage or a soybean milk beverage.
- A health product is characterized in that: the health product contains the composition of any one of claims 1 to 7.
- A food additive characterized by: the food additive comprises the composition of any one of claims 1 to 7.
- A pharmaceutical product characterized by: the pharmaceutical product contains the composition according to any one of claims 1 to 7.
- The pharmaceutical product of claim 22, wherein: the medicine is tablet, granule, powder, enteric-coated preparation, solution or suspension.
- The pharmaceutical product of claim 22, wherein: the medicine is an enteric solvent which is a capsule or an enteric tablet; preferably, the medicament is an enteric coated tablet.
- The pharmaceutical product of claim 24, wherein: the enteric coating thickness of the enteric-coated tablet is 5-100 μm, preferably, the thickness is 20-80 μm.
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