WO2019088379A1 - 신규 유산균 및 이의 용도 - Google Patents
신규 유산균 및 이의 용도 Download PDFInfo
- Publication number
- WO2019088379A1 WO2019088379A1 PCT/KR2018/004590 KR2018004590W WO2019088379A1 WO 2019088379 A1 WO2019088379 A1 WO 2019088379A1 KR 2018004590 W KR2018004590 W KR 2018004590W WO 2019088379 A1 WO2019088379 A1 WO 2019088379A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- lactobacillus plantarum
- bifidobacterium longum
- lactobacillus
- lactic acid
- inflammatory disease
- Prior art date
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Images
Classifications
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
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- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
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- A61P3/00—Drugs for disorders of the metabolism
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A61K2035/11—Medicinal preparations comprising living procariotic cells
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
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- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
Definitions
- the present invention relates to a novel lactic acid bacterium, Lactobacillus plantarum and Bifidobacterium longum, and more particularly to a composition containing a novel lactic acid bacterium useful for the prevention and treatment of an inflammatory disease.
- Inflammation is one of a kind of defense reaction that appears in living tissue or organ for any external stimulus. Normally, the inflammatory reaction removes infected material from the outside and regenerates damaged tissue to regain its function. However, when the inflammatory reaction is continuously caused by the secretion of the inflammatory cytokine, a variety of inflammatory diseases are generated depending on the site of the inflammatory reaction.
- vaginitis a type of female inflammatory disease
- vaginal infectious disease caused by the proliferation of various anaerobic bacteria instead of bacteria normally present in the vagina.
- Most of the vaginitis is caused by bacterial vaginitis and fungal vaginitis.
- Typical cases of bacterial vaginosis are caused by the anaerobic strain Gardnerella vaginalis or Atopobium vaginae, It is mainly caused by Candida albicans (Sexually Transmitted Diseases, November 2006, vol 33, No. 11, 663-665).
- antibiotics are used for bacterial vaginitis and fungal vaginitis can be improved by using azole antifungals.
- a therapeutic agent has another problem that it is difficult to expect a fundamental therapeutic effect by causing another bacterial shift. Therefore, it is necessary to develop a therapeutic agent capable of inhibiting the inflammatory reaction so that normal lactic acid bacteria can be restored to their original state while eliminating vaginitis causing vaginosis.
- sufficient studies have not been made.
- the inventors of the present invention have found that a novel lactic acid bacterium isolated from kimchi and human feces inhibits the inflammatory reaction, inhibits the growth of vaginitis causing bacteria, At the same time, the present invention has been completed upon confirming that it is effective for the prevention and treatment of inflammatory diseases, specifically vaginitis, by showing the therapeutic effect of vaginitis.
- Another object of the present invention is to provide a composition for the prevention or treatment of an inflammatory disease comprising a novel lactic acid bacterium.
- Another object of the present invention is to provide a health functional food for preventing or ameliorating an inflammatory disease comprising a novel lactic acid bacterium.
- the present invention provides Lactobacillus plantarum NK3 (Depository Institution: Korean Microorganism Conservation Center, Date of Deposit: 2017.08.04, Accession Number: KCCM12089P).
- Lactobacillus plantarum NK3 of the present invention is a novel lactic acid bacteria of Lactobacillus plantarum isolated and identified from kimchi which is a traditional fermented food.
- the 16S rDNA nucleotide sequence for identification and classification of Lactobacillus plantarum NK3 of the present invention is shown in SEQ ID NO: 1 attached hereto.
- the Lactobacillus plantarum NK3 of the present invention may comprise the 16S rDNA of SEQ ID NO: 1.
- Lactobacillus plantarum NK3 of the present invention is gram-positive, and the cell type is bacillus.
- the physiological characteristics of the more specific Lactobacillus plantarum NK3 can be analyzed according to conventional methods in the art, and the results are shown in Table 2 below.
- Lactobacillus plantarum NK3 is a carbon source, which is selected from the group consisting of L-arabinose, D-ribose, D-galactose, D-glucose, D-fructose, D- mannose, mannitol, sorbitol, Mannoside, N-acetyl-glucosamine, amigalline, arbutin, escululin, salicin, cellobiose, maltose, lactose, melibiose, sucrose, trehalose, melizitose, gentiobiose, D-turanose and glucose You can use Nate.
- the present invention provides Bifidobacterium longum NK49 (Depository Institution: Korean Microorganism Conservation Center, Deposit date: 2017.08.04, Accession number: KCCM12088P) .
- Bifidobacterium longum NK49 of the present invention is a novel lactic acid bacterium of Bifidobacterium longum that has been isolated and identified from human feces.
- the 16S rDNA nucleotide sequence for identification and classification of Bifidobacterium longum NK49 of the present invention is shown in SEQ ID NO: 2 attached hereto.
- the Bifidobacterium longum NK49 of the present invention may comprise the 16S rDNA of SEQ ID NO: 2.
- Bifidobacterium longum NK49 As a result of analysis of the 16S rDNA sequence of SEQ ID NO: 2, 99% homology with the known Bifidobacterium longum isolates showed the highest molecular phylogenetic relationship with Bifidobacterium longum. Therefore, the lactic acid bacteria were identified as Bifidobacterium longum, named Bifidobacterium longum NK49, and deposited on August 4, 2017 (Accession No. KCCM12088P) at the Korean Microorganism Conservation Center.
- the Bifidobacterium longum NK49 of the present invention is a gram-positive bacterium, and the cell type is bacillus.
- the physiological characteristics of the more specific Bifidobacterium longum NK49 can be analyzed according to conventional methods in the art, and the results are shown in Table 3 below.
- Bifidobacterium longum sword NK49 is a carbon source which is a mixture of L-arabinose, D-ribose, D-xylose, D-galactose, D-glucose, D-fructose, mannitol, sorbitol, D-glucoside, esculine, salicin, maltose, lactose, melibiose, sucrose, raffinose and D-turanose can be used.
- the present invention provides a method for treating inflammatory diseases including Lactobacillus plantarum NK3 KCCM 12089P, Bifidobacterium longum NK49 KCCM 12088P or a mixture thereof Or a pharmaceutically acceptable salt thereof.
- the " Lactobacillus plantarum NK3 " of the present invention is the same as that described above.
- Lactobacillus plantarum NK3 contained in the pharmaceutical composition of the present invention may be a biomass of the present invention, a bacterium thereof, a culture thereof, a disruption thereof or an extract thereof, Lactobacillus plantarum NK3 can be used without limitation.
- the "Bifidobacterium longum NK49" of the present invention is the same as described above.
- Bifidobacterium longum NK49 contained in the pharmaceutical composition of the present invention may be a live cell, a bacterium thereof, a culture thereof, a lysate thereof, or an extract thereof, Lactobacillus plantarum NK3 can be used without limitation.
- " culture product " in the present invention means an object obtained by culturing a lactic acid bacterium in a known liquid medium or a solid medium, and the present invention includes a novel lactic acid bacterium.
- the inflammatory diseases of the present invention can be used for the treatment and prophylaxis of arthritis, gout, hepatitis, asthma, obesity, keratitis, gastritis, enteritis, nephritis, colitis, diabetes, tuberculosis, bronchitis, pleurisy, peritonitis, spondylitis, pancreatitis, Inflammation, scleroderma, sepsis, burns, dermatitis, periodontitis and gingivitis.
- the inflammatory reaction was significantly inhibited in the treatment of Lactobacillus plantarum NK3 or Bifidobacterium longum NK49 with lipopolysaccharide, which is an inflammatory reaction inducer, in macrophages isolated from mice 1 and 2).
- the pharmaceutical composition comprising Lactobacillus plantarum NK3, Bifidobacterium longum NK49, or a mixture thereof can be effectively used for the prevention and treatment of inflammatory diseases.
- the inflammatory disease may be vaginitis.
- Lactobacillus plantarum NK3, Bifidobacterium longum NK49 or a mixture thereof exhibits a growth inhibitory effect in the vaginitis-causing bacterium, Gardnerella vaginalis and Atofibium pants, It was confirmed that the effect of suppressing the infection of Nerella vaginalis was excellent (Figs. 3 to 5).
- the pharmaceutical composition comprising Lactobacillus plantarum NK3, Bifidobacterium longum NK49, or a mixture thereof can be effectively used for the prevention and treatment of vaginitis, a specific inflammatory disease.
- the inflammatory disease may be colitis.
- administering Lactobacillus plantarum NK3, Bifidobacterium longum NK49, or a mixture thereof results in restoration of the length of the large intestine and reduction of the activity of myeloperoxidase , And the expression level and activity of cytokines, which are inflammatory indicators, were inhibited (Table 6). Accordingly, it has been confirmed that the pharmaceutical composition comprising Lactobacillus plantarum NK3, Bifidobacterium longum NK49, or a mixture thereof can be effectively used for the prevention and treatment of colitis, which is a specific inflammatory disease.
- compositions according to the present invention may be formulated into pharmaceutical formulations using methods well known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal.
- the pharmaceutical compositions according to the invention may additionally comprise a pharmaceutically acceptable carrier to the extent that the activity of the novel lactic acid bacteria is not inhibited.
- Such pharmaceutically acceptable carriers may be those conventionally used such as lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, But are not limited to, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
- the pharmaceutical composition of the present invention may contain diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and other pharmaceutically acceptable additives.
- the dosage of the pharmaceutical composition according to the present invention should be a pharmaceutically effective amount.
- a " pharmaceutically effective amount " means an amount sufficient to prevent or treat an inflammatory disease at a reasonable benefit / risk ratio applicable to medical treatment.
- Effective dose levels may be varied by those skilled in the art depending on factors such as the formulation method, the condition and weight of the patient, the sex, age, disease severity, form of the drug, route and duration of administration, excretion rate, have. Effective amounts may vary depending on the route of treatment, the use of excipients, and the likelihood of use with other agents, as will be appreciated by those skilled in the art.
- the composition of the present invention is generally administered to an adult at a dose of 0.0001 to 100 mg / kg per day, preferably 0.001 to 100 mg / kg per 1 kg of body weight per day .
- the dosage form is administered as described above, Lactobacillus plantarum NK3, Bifidobacterium longum NK49, or a mixture thereof of the present invention can be administered at 1 ⁇ 10 2 CFU / kg to 1 ⁇ 10 11 CFU / kg per day.
- the administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.
- the pharmaceutical composition of the present invention can be administered to mammals such as mice, livestock, humans, and the like through various routes.
- the pharmaceutical composition of the present invention may be administered orally or parenterally (for example, coated or intravenously, subcutaneously, intraperitoneally injected), but oral administration is preferred.
- oral administration For prevention and treatment of vaginitis, it can be administered into the vagina.
- Solid formulations for oral administration may include powders, granules, tablets, capsules, soft capsules, pills, and the like.
- liquid formulations for oral use include suspensions, solutions, emulsions, syrups and aerosols.
- formulations may be formulated in the form of sterilized aqueous solutions, liquid preparations, non-aqueous solutions, suspensions, emulsions, eye drops, ointments, syrups, suppositories, And preferably pharmaceutical compositions of creams, gels, patches, sprays, ointments, alerts, lotions, liniments, ointments, eye drops, pastes or cataplasms can be prepared and used , But is not limited thereto.
- the formulations for topical administration may be anhydrous or water-based, depending on the clinical formulation.
- the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like.
- the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.
- the present invention includes a step of administering to a subject a Lactobacillus plantarum NK3, a Bifidobacterium longum NK49, or a mixture thereof.
- the present invention provides a method for preventing or treating an inflammatory disease.
- " Lactobacillus plantarum NK3 ", " Bifidobacterium longum NK49 ", " administration ", and " inflammatory disease " and the like are the same as described above in the present invention.
- the subject refers to an animal, and typically can be a mammal that may exhibit beneficial effects in treatment with the novel lactic acid bacteria of the present invention.
- a preferred example of such a subject may include primates such as humans.
- Such subjects may also include all subjects with or at risk of having symptoms of inflammatory disease.
- the present invention provides a method for preventing or ameliorating an inflammatory disease comprising Lactobacillus plantarum NK3 KCCM12089P, Bifidobacterium longum NK49, KCCM12088P, or a mixture thereof, Provide food.
- " Lactobacillus plantarum NK3 ", " Bifidobacterium longum NK49 ", " administration ", and " inflammatory disease ", etc. of the present invention are the same as those described above.
- the health functional food is a food emphasizing the biological control function of food, and is added with added value so as to function and manifest to a specific purpose using physical, biochemical and biotechnological methods.
- These health functional food ingredients are designed and manufactured so that the body control functions related to regulation of body defense and body rhythm, prevention and recovery of disease are sufficiently exhibited to the living body, and food additives, And may contain raw materials.
- Lactobacillus plantarum NK3 or Bifidobacterium longum NK49 of the present invention When Lactobacillus plantarum NK3 or Bifidobacterium longum NK49 of the present invention is used as a health functional food (or a health functional beverage addition product), the novel lactic acid bacterium may be added as it is, or may be used in combination with other food or food ingredients, And the like.
- the mixing amount of Lactobacillus plantarum NK3 or Bifidobacterium longum NK49 can be suitably determined according to its use purpose (prevention, health or improvement, therapeutic treatment).
- the health functional food may contain flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate etc.), pectic acid and its salts, organic acids, A pH adjuster, a stabilizer, a preservative, a glycerin, an alcohol, a carbonating agent used in a carbonated drink, and the like.
- the health functional food of the present invention may contain pulp for the production of fruit and vegetable drinks. These components may be used singly or in combination, and the proportion of such additives is generally selected in the range of 0.001 to 50 parts by weight based on the total weight of the composition.
- Foods to which the Lactobacillus plantarum NK3 or Bifidobacterium longum NK49 can be added include dairy products including sausages, meats, breads, chocolates, snacks, candies, confectionery, ramen, pizza, other noodles, gums, Products, soups, drinks, tea, drinks, alcoholic beverages and vitamin complexes.
- the liquid ingredient to be added in addition to the novel lactic acid bacteria may include various flavors or natural carbohydrates such as ordinary beverages as an additional ingredient.
- natural carbohydrates may be used in combination with monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose and polysaccharides such as dextrins and cyclodextrins and xylitol and sorbitol , Erythritol, and the like.
- monosaccharides such as glucose and fructose
- disaccharides such as maltose and sucrose
- polysaccharides such as dextrins and cyclodextrins and xylitol and sorbitol , Erythritol, and the like.
- the present invention relates to a pharmaceutical composition for the treatment of inflammatory diseases, comprising Lactobacillus plantarum NK3 KCCM12089P, Bifidobacterium longum NK49 KCCM12088P or a mixture thereof It provides usages.
- the present invention provides a composition comprising Lactobacillus plantarum NK3 KCCM12089P, Bifidobacterium longum NK49, KCCM12088P or mixtures thereof for use in the treatment of inflammatory diseases .
- the invention provides the use of Lactobacillus plantarum NK3 KCCM 12089P, Bifidobacterium longum NK49 KCCM 12088P or mixtures thereof for the treatment of inflammatory diseases.
- novel lactic acid bacteria Lactobacillus plantarum NK3, Bifidobacterium longum NK49 or a mixture thereof according to the present invention have an effect of inhibiting the inflammatory reaction. Accordingly, the novel lactic acid bacterium according to the present invention can be used as a composition for preventing or treating inflammatory diseases, and is particularly effective for the treatment and prevention of colitis and vaginitis.
- NF-kB p-p65 / p65
- NK3 Lactobacillus plantarum NK3
- Bifidobacterium longum NK49 novel lactic acid bacteria
- FIG. 2 is a graph showing the inhibitory effect of TNF- ⁇ on the macrophages of Lactobacillus plantarum NK3 and Bifidobacterium longum NK49, which are novel lactic acid bacteria.
- FIG. 3 is a graph showing the antimicrobial effect of Lactobacillus plantarum NK3, which is a novel lactic acid bacterium, and Gardnerella vaginalis, Bifidobacterium longum NK49.
- FIG. 4 is a graph showing the antimicrobial effect of Atobobium vaginae on Lactobacillus plantarum NK3 and Bifidobacterium longum NK49, which are new lactic acid bacteria.
- FIG. 5 is a graph showing the inhibitory effect on the infection of Gardnerella vaginalis of Lactobacillus plantarum NK3 and Bifidobacterium longum NK49, which are new lactic acid bacteria.
- G Only the guardian Nella trousers treated; GL5, GL6, GL7: treated with Gardnerella vaginalis and treated with 1 ⁇ 10 6 CFU / mL, 1 ⁇ 10 7 CFU / mL, and 1 ⁇ 10 8 CFU / mL of Lactobacillus plantarum, respectively; GB5, GB6, and GB7: treated with Gardnerella vaginalis and treated with 1 ⁇ 10 6 CFU / mL, 1 ⁇ 10 7 CFU / mL, and 1 ⁇ 10 8 CFU / mL of Bifidobacterium longum, respectively.
- FIG. 6 is a graph showing the inhibitory effect of the novel lactic acid bacteria Lactobacillus plantarum NK3, Bifidobacterium longum NK49 and mixtures thereof on the quality of the uterus and inflammation of the uterus.
- N normal group
- GC infected only with Gardnerella vaginalis
- oGL, oGB, oGM Experimental animals infected with Gardnerella vaginalis were infected with Lactobacillus plantarum, Bifidobacterium longagi Or a mixture thereof
- vGL, vGB, vGM Experimental animals infected with Gardnerella vaginalis were infected with Lactobacillus plantarum, Bifidobacterium longagi Or a mixture thereof is injected into the vagina. The same shall apply hereinafter.
- Figure 7 is a graph showing the inhibitory activity against the infected Gardnerella vaginalis of Lactobacillus plantarum NK3, Bifidobacterium longum NK49 and mixtures thereof, which is a novel lactic acid bacterium .
- FIG. 8 is a graph showing the inhibitory activity of myeloperoxidase activity of Lactobacillus plantarum NK3, Bifidobacterium longum NK49 and mixtures thereof, which is a novel lactic acid bacterium.
- FIG. 9 is a graph showing the inhibitory effect of TNF- ⁇ on Lactobacillus plantarum NK3, Bifidobacterium longum NK49, and a mixture thereof, which is a novel lactic acid bacterium.
- FIG. 10 is a graph showing the ability of the novel lactic acid bacteria, Lactobacillus plantarum NK3, Bifidobacterium longum NK49 and mixtures thereof to express IL-10.
- FIG. 11 is a graph showing the effect of Lactobacillus plantarum NK3, Bifidobacterium longum NK49, and the mixture thereof in the vaginal lactobacilli recovery effect of the novel lactic acid bacteria.
- Example 1 Isolation and identification of lactic acid bacteria
- Table 1 shows the strain names of the lactic acid bacteria.
- the lactic acid bacteria isolated from kimchi contained 5 kinds of Lactobacillus plantarum (Control Nos. 1 to 5 of Table 1), 5 kinds of Lactobacillus brevis (Control Nos. 6 to 10 of Table 1) , 5 kinds of Lactobacillus sakei (Control Nos. 11 to 15 of Table 1) and 5 kinds of Lactocacillus curvatus (Control Nos. 16 to 20 of Table 1).
- Lactobacillus isolated from human feces is classified into five types (Lactobacillus rhamnosus (control numbers 21 to 25 in Table 1), 5 kinds of Lactobacillus plantarum (control numbers 26 to 30 in Table 1) , 5 Lactobacillus reuteri (Control No. 31-35 in Table 1), 4 Lactobacillus johnsonii (Control No. 36-39 in Table 1), Lactobacillus mucosae
- Three types of Bifidobacterium adolescentis Control No. 43 to 45 in Table 1), five species of Bifidobacterium longum (Table 3) (Table 1, Control Number 40 to 42), Bifidobacterium adolescentis 1, Control No. 46-50).
- Control Number Strain name Control Number Strain name One Lactobacillus plantarum NK1 26 Lactobacillus plantarum NK26 2 Lactobacillus plantarum NK2 27 Lactobacillus plantarum NK27 3 Lactobacillus plantarum NK3 28 Lactobacillus plantarum NK28 4 Lactobacillus plantarum NK4 29 Lactobacillus plantarum NK29 5 Lactobacillus plantarum NK5 30 Lactobacillus plantarum NK30 6 Lactobacillus brevis NK6 31 Lactobacillus reuteri NK31 7 Lactobacillus brevis NK7 32 Lactobacillus reuteri NK32 8 Lactobacillus brevis NK8 33 Lactobacillus reuteri NK33 9 Lactobacillus brevis NK9 34 Lactobacillus reuteri NK34 10 Lactobacillus brevis NK10 35 Lactobacillus reuteri NK
- Lactobacillus plantarum NK3 (Accession No. KCCM12089P) was confirmed to be a gram-positive bacterium.
- the 16S rDNA of Lactobacillus plantarum NK3 was found to have the nucleotide sequence of SEQ ID NO: 1.
- the 16S rDNA sequence of Lactobacillus plantarum NK3 was compared with a BLAST search. As a result, a strain of Lactobacillus plantarum having the same 16S rDNA sequence was not found, and the known lactobacillus plantarum And it was confirmed that it was 99% homologous with 16S rDNA sequence of Lactobacillus plantarum strain.
- Bifidobacterium longum NK49 (Accession No. KCCM12088P) was confirmed to be a gram-positive bacterium.
- the 16S rDNA of Bifidobacterium longum NK49 was found to have the nucleotide sequence of SEQ ID NO: 2.
- a comparison of the 16S rDNA nucleotide sequence of Bifidobacterium longum NK49 with a BLAST search showed that the Bifidobacterium longum strain having the same 16S rDNA base sequence was not detected and that the known Bifidobacterium longum And 99% homology with the 16S rDNA sequence of the Bifidobacterium longum strain.
- Example 2 Comparison of activity of isolated lactic acid bacteria
- DPPH 2,2-Diphenyl-1-picrylhydrazyl
- a DPPH solution A suspension of lactic acid bacteria (1 ⁇ 10 6 CFU / ml) or a vitamin C solution (1 g / ml) was added to 0.1 ml of the DPPH solution and cultured at 37 ° C. for 20 minutes. The culture was centrifuged at 3000 rpm for 5 minutes to obtain a supernatant. Then, the antioxidant activity of the isolated lactic acid bacteria was calculated by measuring the absorbance of the supernatant at 517 nm. The antioxidant activity of each lactic acid bacterium is shown in Table 4 below.
- mice 2 ml of 4% thioglycolate sterilized in C57BL / 6 mice (male, 20-23 g at 6 weeks of age) was administered intraperitoneally. After 96 hours, the mice were anesthetized and 8 ml of RPMI 1640 medium was administered to the abdominal cavity of the mice. After 5 to 10 minutes, RPMI medium (macrophages) in the abdominal cavity of the mice was drawn, centrifuged at 1000 g for 10 minutes, and then washed twice with RPMI 1640 medium.
- the macrophages were spread on a 24-well plate at a rate of 0.5 ⁇ 10 6 per well and cultured in the same manner as in Example 1 except that the separated lactic acid bacterium (final treatment concentration: 1 ⁇ 10 4 cfu / ml, the same applies hereinafter) and lipopolysaccharide LPS) was treated for 2 or 24 hours before supernatants and cells were obtained.
- the obtained cells were homogenized in RIPA buffer (Gibco). (NF-kB), p-p65 (phosphor-NF-kB), and beta (beta) from the cells obtained by treating the amount of cytokine expression of TNF-a and IL- -actin was measured by an immunoblotting method.
- the level of expression of the inflammation index for each lactic acid bacterium is shown in Table 4 below.
- Caco2 cells, colon cancer cells, were distributed from a Korean cell line bank and cultured in RPMI 1640 medium for 48 hours. Then, Caco2 cells were dispensed in a 12-well plate at a dose of 2 x 10 6 per well. Each well was treated with 1 ⁇ g of LPS alone or with 1 ⁇ g of LPS and 1 ⁇ 10 4 CFU of lactic acid bacteria and cultured for 24 hours. Then, the cells cultured from each well were collected and the expression amount of ZO-1, a tight junction protein, was measured by immunoblotting. The expression levels of ZO-1 for each lactic acid bacterium are shown in Table 5 below.
- Lactobacillus plantarum NK3 and Bifidobacterium longum NK49 which are novel lactic acid bacteria among the isolated lactic acid bacteria, were evaluated for the expression of ZO-1, a close-coupled protein, (Table 4 and Table 5). The results are shown in Table 4 and Table 5, respectively. ≪ tb > < TABLE >
- Example 3 Measurement of inhibition of inflammatory response to macrophages
- mice 2 ml of 4% thioglycolate sterilized in C57BL / 6 mice (male, 20-23 g) was administered intraperitoneally. After 96 hours, the mice were anesthetized and 8 ml of RPMI 1640 medium was administered to the abdominal cavity of the mice. After 5 to 10 minutes, the RPMI medium (macrophages) in the abdominal cavity of the mice was drawn, centrifuged at 1000 g for 10 minutes, and washed twice with RPMI 1640 medium. The cells were cultured in a 24-well plate for 5 hours, and adhered cells were used as macrophages.
- the macrophages were spread in a number of 0.5 x 10 < 6 > per well, and cultured at 10 3 , 10 4, and 10 5 CFU / mL Lactobacillus plantarum NK3 and Bifidobacterium longum NK49, which are novel lactic acid bacteria, and lipopolysaccharide (LPS), an inflammatory reaction inducer, were treated for 90 minutes or 24 hours Post supernatant and cells.
- the obtained cells were homogenized in RIPA buffer (Gibco).
- the amount of expression of p65 (NF-kB), p-p65 (phosphor-NF-kB) and ⁇ -actin was measured by immunoblotting from the cells obtained by treatment for 90 minutes.
- the amount of TNF- ⁇ cytokine expressed in the culture supernatant after 24 hours of treatment was measured by ELISA kit (Ebioscience, San Diego, CA, USA).
- Example 4 Antimicrobial activity against vaginitis causing bacteria
- Lactobacillus plantarum NK3 and Bifidobacterium longum NK49 (1 ⁇ 10 6 , 1 ⁇ 10 7 , 1 ⁇ 10 8 CFU / mL) were added to the GAM medium to separate the new lactic acid bacteria Lactobacillus plantarum NK3 and Bifidobacterium longum NK49 (Gardnerella vaginalis) or Atopobium vaginae (1 x 10 6 CFU / mL) were transplanted.
- Yeast extract (1%), maltose (0.1%), glucose (0.1%) and horse serum (10%) were added to the BHI broth at 37 °C under anaerobic conditions And then cultured in the BHIS medium for 24 hours.
- the human cervical cancer cell line HeLa cells were cultured in RPMI 1640 medium (containing 10% heat-inactivated fetal calf serum) at 37 ° C, 5% CO 2 and 95% (1 ⁇ 10 5 , 1 ⁇ 10 6 , 1 ⁇ 10 7 CFU / mL) alone or in combination with Lactobacillus plantarum NK3 and Bifidobacterium longum NK49 (1 ⁇ 10 4 , 1 ⁇ 10 6 CFU / Gardnerella vaginalis (1 ⁇ 10 5 CFU / mL) was transplanted and the number of bacteria attached to HeLa cells after 24 hours was measured by qPCR.
- RPMI 1640 medium containing 10% heat-inactivated fetal calf serum
- mice were injected subcutaneously with 0.125 mg of ⁇ -estradiol 17-benzoate (Sigma, MO, USA) dissolved in olive oil.
- Gardnerella vaginalis (1 x 10 8 CFU / mouse) was transplanted into the vagina of the mice.
- the experiment was carried out after 7 days of lactic acid bacteria administration (14 days after transplantation) and 24 hours after sacrifice of animal model.
- Gardnerella vaginalis was transplanted into the vagina and inflammation accompanied by vaginal and uterine edema occurred.
- oral or vaginal administration of the novel lactic acid bacteria, Lactobacillus plantarum NK3, Bifidobacterium longum NK49, or a mixture thereof significantly reduced the appearance and inflammation of the uterus (Fig. 6).
- vagina was washed with sterilized physiological saline (0.5 ml) at 24 hours and 48 hours.
- the washed vaginal washes were separated by Qaigen DNA purification kit, and lactobacillus and Gardnerella vaginalis strains were quantitated by PCR.
- MPO myeloperoxidase
- Lysis buffer (20 mM HEPES, 1.5 mM MgCl 2, 0.4 mM NaCl, 1 mM EDTA, 1 mM dithiothreitol, 0.5 mM phenyl methyl sulfonyl fluoride, 20 ⁇ g / mL trypsin inhibitor, 1 % nonidet p-40, 20% glycerol) was added and homogenized. Proteins were quantitated by the Bradford assay and samples were prepared to contain 50 ⁇ g / ⁇ L of protein.
- an electrophoresis sample buffer (Laemmli sample buffer 950 ⁇ L + ⁇ Me 50 ⁇ L) was added to the sample, followed by denaturation at 100 ° C. for 3 minutes and electrophoresis (150 V, 30 mA) was performed. The electrophoresed gel was transferred to the membrane at 30V for 2 hours. 15 ml of the membrane was added to 5% skim milk / PBS-T (0.05% tween 20 / PBS) and shaken for 1 hour.
- an electrophoresis sample buffer (Laemmli sample buffer 950 ⁇ L + ⁇ Me 50 ⁇ L) was added to the sample, followed by denaturation at 100 ° C. for 3 minutes and electrophoresis (150 V, 30 mA) was performed. The electrophoresed gel was transferred to the membrane at 30V for 2 hours. 15 ml of the membrane was added to 5% skim milk / PBS-T (0.05% tween 20 / PBS) and shaken
- the group infected with Gardnerella vaginalis showed an increase in TNF- ⁇ expression and a decrease in IL-10 expression in vaginal tissue.
- TNF- ⁇ expression is suppressed and the expression of IL-10 is suppressed in the group administered with oral or vaginal administration of Lactobacillus plantarum NK3, Bifidobacterium longum NK49 or a mixture thereof (Figs. 9 and 10).
- mice were washed twice in vagina with 0.5 ml of sterile physiological saline. The washed vaginal washes were collected and centrifuged (10,000 g, 10 min). The precipitate was extracted with a bacterial genomic DNA extraction kit (QIAGEN DNeasy Feces kit; Qiagen, Hilden, Germany). 10 ng of the extracted DNA was subjected to DNA polymerase activation (95 ° C., 30 sec), denaturation (95 ° C., 5 sec) and amplification (63 ° C., 30 sec) in the SYBER premix with the following primers in a Qiagen thermal cycler PCR was repeatedly performed.
- DNA polymerase activation 95 ° C., 30 sec
- denaturation 95 ° C., 5 sec
- amplification 63 ° C., 30 sec
- Lactobacilli forward (5'-3 ') CTC AAA ACT AAA CAA AGT TTC (SEQ ID NO: 3);
- Lactobacilli reverse (5'-3 ') CTT GTA CAC ACC GCC CGT (SEQ ID NO: 4);
- Control 16S rDNA reverse (5'-3 ') AAG GAG GTG WTC CAR CC (SEQ ID NO: 6).
- Lactobacillus plantarum NK3, Bifidobacterium longum NK49, or a mixture thereof was significantly reduced when the plant was infected with Gardnerella vaginalis.
- lactobacilli increased (FIG. 11). This means that vaginitis induced by Gardnerella vaginalis and reduced lactobacilli have been restored by the administration of the lactic acid bacteria to restore the acidic conditions.
- Example 5 Effect of lactic acid bacteria on colitis in an animal model
- mice Male, 21-23g, 6 weeks old were used for 6 weeks in each group, and the mice were adapted to the laboratory for 1 week.
- One group was used as a normal group, and the other group mice were induced with 2,4,6-trinitrobenzenesulfonic acid (TNBS) as colitis.
- TNBS 2,4,6-trinitrobenzenesulfonic acid
- animals were anesthetized with ether and mixed with 50% ethanol.
- the TNBS solution was injected into the large intestine through the anus using a 1 ml round syringe and kept in the vertical for 30 seconds to induce inflammation Respectively.
- 0.1 ml of physiological saline was orally administered to the normal group.
- a 1% dextrose solution of a lactic acid bacterium suspension was orally administered to the experimental animals in the normal group instead of the novel lactic acid bacteria.
- the experimental animals of the positive control group were orally administered sulfasalazine, a colitis treating drug, in an amount of 50 mg / kg instead of the novel lactic acid bacteria.
- Western blotting was used to measure inflammatory response indicators such as p-p65, p65, COX-2 and ⁇ -actin. Specifically, 50 ⁇ g of the supernatant obtained in the same manner as in the measurement of the activity of Myeloperoxidase (MPO) was subjected to immunoblotting. The expression level of cytokines (IL-17, TNF- ⁇ ) was measured using an ELISA kit.
- MPO Myeloperoxidase
- the inventors of the present invention deposited Lactobacillus plantarum NK3 on KCCM12089P on August 4, 2017, at the Korea Microorganism Conservation Center (address: 45 Yurim Building, 2 Gagil in Haejeon, Seoul, Korea) I got a security number.
- KCCM12088P has been given the accession number.
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Abstract
Description
관리번호 | 균주명 | 관리번호 | 균주명 |
1 | Lactobacillus plantarum NK1 | 26 | Lactobacillus plantarum NK26 |
2 | Lactobacillus plantarum NK2 | 27 | Lactobacillus plantarum NK27 |
3 | Lactobacillus plantarum NK3 | 28 | Lactobacillus plantarum NK28 |
4 | Lactobacillus plantarum NK4 | 29 | Lactobacillus plantarum NK29 |
5 | Lactobacillus plantarum NK5 | 30 | Lactobacillus plantarum NK30 |
6 | Lactobacillus brevis NK6 | 31 | Lactobacillus reuteri NK31 |
7 | Lactobacillus brevis NK7 | 32 | Lactobacillus reuteri NK32 |
8 | Lactobacillus brevis NK8 | 33 | Lactobacillus reuteri NK33 |
9 | Lactobacillus brevis NK9 | 34 | Lactobacillus reuteri NK34 |
10 | Lactobacillus brevis NK10 | 35 | Lactobacillus reuteri NK35 |
11 | Lactobacillus sakei NK11 | 36 | Lactobacillus johnsonii NK36 |
12 | Lactobacillus sakei NK12 | 37 | Lactobacillus johnsonii NK37 |
13 | Lactobacillus sakei NK13 | 38 | Lactobacillus johnsonii NK38 |
14 | Lactobacillus sakei NK14 | 39 | Lactobacillus johnsonii NK39 |
15 | Lactobacillus sakei NK15 | 40 | Lactobacillus mucosae NK40 |
16 | Lactobacillus curvatus NK16 | 41 | Lactobacillus mucosae NK41 |
17 | Lactobacillus curvatus NK17 | 42 | Lactobacillus mucosae NK42 |
18 | Lactobacillus curvatus NK18 | 43 | Bifidobacterium adolescentis NK43 |
19 | Lactobacillus curvatus NK19 | 44 | Bifidobacterium adolescentis NK44 |
20 | Lactobacillus curvatus NK20 | 45 | Bifidobacterium adolescentis NK45 |
21 | Lactobacillus rhamnosus NK21 | 46 | Bifidobacterium longum NK46 |
22 | Lactobacillus rhamnosus NK22 | 47 | Bifidobacterium longum NK47 |
23 | Lactobacillus rhamnosus NK23 | 48 | Bifidobacterium longum NK48 |
24 | Lactobacillus rhamnosus NK24 | 49 | Bifidobacterium longum NK49 |
25 | Lactobacillus rhamnosus NK25 | 50 | Bifidobacterium longum NK50 |
탄소원 | NK3 | 탄소원 | NK3 |
CONTROL | - | 에스큘린 | + |
글리세롤 | - | 살리신 | + |
에리쓰리톨 | - | 셀로비오스 | + |
D-아라비노오스 | - | 말토오스 | + |
L-아라비노오스 | + | 락토오스 | + |
D-리보오스 | + | 멜리비오스 | + |
D-자일로오스 | - | 수크로오스 | + |
L-자일로오스 | - | 트레할로오스 | + |
D-아도니톨 | - | 이뉼린 | - |
메틸-BD-자일로피라노사이드Methyl-BD-Xylopyranoside | - | 멜리지토오스 | + |
D-갈락토오스 | + | 라피노오스 | - |
D-글루코오스 | + | 전분(Starch) | - |
D-프럭토오스 | + | 글리코겐 | - |
D-만노오스 | + | 자일리톨 | - |
L-소르보오스 | - | 겐티오비오스 | + |
람노서스 | - | D-투라노오스 | + |
둘시톨 | - | D-릭소오스 | - |
이노시톨 | - | D-타가토오스 | - |
만니톨 | + | D-푸코오스 | - |
솔비톨 | + | L-푸코오스 | - |
α-메틸-D-만노사이드 | ± | D-아라비톨 | - |
α-메틸-D-글루코사이드 | - | L-아라비톨 | - |
N-아세틸-글루코사민 | + | 글루코네이트 | ± |
아미그달린 | + | 2-케토-글루코네이트 | - |
알부틴 | + | 5-케토-글루코네이트 | - |
탄소원 | NK49 | 탄소원 | NK49 |
CONTROL | - | 에스큘린 | + |
글리세롤 | - | 살리신 | + |
에리쓰리톨 | - | 셀로비오스 | - |
D-아라비노오스 | - | 말토오스 | + |
L-아라비노오스 | + | 락토오스 | + |
D-리보오스 | + | 멜리비오스 | + |
D-자일로오스 | ± | 수크로오스 | + |
L-자일로오스 | - | 트레할로오스 | - |
D-아도니톨 | - | 이뉼린 | - |
메틸-BD-자일로피라노사이드Methyl-BD-Xylopyranoside | - | 멜리지토오스 | - |
D-갈락토오스 | + | 라피노오스 | + |
D-글루코오스 | + | 전분(Starch) | - |
D-프럭토오스 | + | 글리코겐 | - |
D-만노오스 | - | 자일리톨 | - |
L-소르보오스 | - | 겐티오비오스 | - |
람노서스 | - | D-투라노오스 | ± |
둘시톨 | - | D-릭소오스 | - |
이노시톨 | - | D-타가토오스 | - |
만니톨 | + | D-푸코오스 | - |
솔비톨 | + | L-푸코오스 | - |
α-메틸-D-만노사이드 | - | D-아라비톨 | - |
α-메틸-D-글루코사이드 | ± | L-아라비톨 | - |
N-아세틸-글루코사민 | - | 글루코네이트 | - |
아미그달린 | - | 2-케토-글루코네이트 | - |
알부틴 | - | 5-케토-글루코네이트 | - |
관리번호 | 균주명 | 항산화 활성 | TNF-α억제능 | IL-10발현증가 | NF-kB억제능 |
1 | Lactobacillus plantarum NK1 | + | + | + | + |
2 | Lactobacillus plantarum NK2 | + | ++ | + | + |
3 | Lactobacillus plantarum NK3 | +++ | +++ | +++ | +++ |
4 | Lactobacillus plantarum NK4 | + | + | + | ++ |
5 | Lactobacillus plantarum NK5 | ++ | ++ | ++ | ++ |
6 | Lactobacillus brevis NK6 | + | + | + | + |
7 | Lactobacillus brevis NK7 | + | + | + | + |
8 | Lactobacillus brevis NK8 | + | + | + | + |
9 | Lactobacillus brevis NK9 | + | + | + | + |
10 | Lactobacillus brevis NK10 | - | + | + | + |
11 | Lactobacillus sakei NK11 | + | + | + | + |
12 | Lactobacillus sakei NK12 | - | ++ | + | + |
13 | Lactobacillus sakei NK13 | - | ++ | ++ | + |
14 | Lactobacillus sakei NK14 | - | + | + | + |
15 | Lactobacillus sakei NK15 | + | + | + | + |
16 | Lactobacillus curvatus NK16 | + | + | + | + |
17 | Lactobacillus curvatus NK17 | + | + | + | + |
18 | Lactobacillus curvatus NK18 | + | + | + | + |
19 | Lactobacillus curvatus NK19 | + | + | + | + |
20 | Lactobacillus curvatus NK20 | + | + | + | + |
21 | Lactobacillus rhamnosus NK21 | + | + | + | + |
22 | Lactobacillus rhamnosus NK22 | + | + | + | + |
23 | Lactobacillus rhamnosus NK23 | + | + | + | + |
24 | Lactobacillus rhamnosus NK24 | ++ | + | + | + |
25 | Lactobacillus rhamnosus NK25 | ++ | ++ | ++ | ++ |
26 | Lactobacillus plantarum NK26 | + | + | + | + |
27 | Lactobacillus plantarum NK27 | + | + | + | + |
28 | Lactobacillus plantarum NK28 | + | + | + | + |
29 | Lactobacillus plantarum NK29 | + | + | + | + |
30 | Lactobacillus plantarum NK30 | + | + | + | + |
31 | Lactobacillus reuteri NK31 | + | + | + | + |
32 | Lactobacillus reuteri NK32 | ++ | ++ | ++ | ++ |
33 | Lactobacillus reuteri NK33 | +++ | ++ | ++ | ++ |
34 | Lactobacillus reuteri NK34 | + | + | + | + |
35 | Lactobacillus reuteri NK35 | + | + | + | + |
36 | Lactobacillus johnsonii NK36 | ++ | ++ | ++ | + |
37 | Lactobacillus johnsonii NK37 | ++ | ++ | ++ | ++ |
38 | Lactobacillus johnsonii NK38 | + | + | + | + |
39 | Lactobacillus johnsonii NK39 | + | + | + | + |
40 | Lactobacillus mucosae NK40 | ++ | ++ | ++ | ++ |
41 | Lactobacillus mucosae NK41 | ++ | ++ | +++ | +++ |
42 | Lactobacillus mucosae NK42 | + | + | + | + |
43 | Bifidobacterium adolescentis NK43 | + | + | + | + |
44 | Bifidobacterium adolescentis NK44 | ++ | ++ | ++ | ++ |
45 | Bifidobacterium adolescentis NK45 | + | + | + | + |
46 | Bifidobacterium longum NK46 | +++ | ++ | +++ | +++ |
47 | Bifidobacterium longum NK47 | ++ | ++ | ++ | ++ |
48 | Bifidobacterium longum NK48 | + | + | + | + |
49 | Bifidobacterium longum NK49 | +++ | +++ | +++ | +++ |
50 | Bifidobacterium longum NK50 | + | + | + | + |
관리번호 | 균주명 | ZO-1발현증가 |
1 | Lactobacillus plantarum NK1 | + |
2 | Lactobacillus plantarum NK2 | + |
3 | Lactobacillus plantarum NK3 | ++ |
4 | Lactobacillus plantarum NK4 | + |
5 | Lactobacillus plantarum NK5 | ++ |
6 | Lactobacillus brevis NK6 | + |
7 | Lactobacillus brevis NK7 | + |
8 | Lactobacillus brevis NK8 | + |
9 | Lactobacillus brevis NK9 | + |
10 | Lactobacillus brevis NK10 | - |
11 | Lactobacillus sakei NK11 | + |
12 | Lactobacillus sakei NK12 | - |
13 | Lactobacillus sakei NK13 | + |
14 | Lactobacillus sakei NK14 | + |
15 | Lactobacillus sakei NK15 | + |
16 | Lactobacillus curvatus NK16 | + |
17 | Lactobacillus curvatus NK17 | + |
18 | Lactobacillus curvatus NK18 | + |
19 | Lactobacillus curvatus NK19 | + |
20 | Lactobacillus curvatus NK20 | + |
21 | Lactobacillus rhamnosus NK21 | + |
22 | Lactobacillus rhamnosus NK22 | + |
23 | Lactobacillus rhamnosus NK23 | + |
24 | Lactobacillus rhamnosus NK24 | ++ |
25 | Lactobacillus rhamnosus NK25 | + |
26 | Lactobacillus plantarum NK26 | + |
27 | Lactobacillus plantarum NK27 | + |
28 | Lactobacillus plantarum NK28 | + |
29 | Lactobacillus plantarum NK29 | + |
30 | Lactobacillus plantarum NK30 | + |
31 | Lactobacillus reuteri NK31 | + |
32 | Lactobacillus reuteri NK32 | ++ |
33 | Lactobacillus reuteri NK33 | ++ |
34 | Lactobacillus reuteri NK34 | + |
35 | Lactobacillus reuteri NK35 | + |
36 | Lactobacillus johnsonii NK36 | + |
37 | Lactobacillus johnsonii NK37 | ++ |
38 | Lactobacillus johnsonii NK38 | + |
39 | Lactobacillus johnsonii NK39 | + |
40 | Lactobacillus mucosae NK40 | ++ |
41 | Lactobacillus mucosae NK41 | ++ |
42 | Lactobacillus mucosae NK42 | + |
43 | Bifidobacterium adolescentis NK43 | + |
44 | Bifidobacterium adolescentis NK44 | ++ |
45 | Bifidobacterium adolescentis NK45 | + |
46 | Bifidobacterium longum NK46 | ++ |
47 | Bifidobacterium longum NK47 | + |
48 | Bifidobacterium longum NK48 | + |
49 | Bifidobacterium longum NK49 | + |
50 | Bifidobacterium longum NK50 | + |
실험군 | 체중변화g | 대장길이 cm | MPO 활성μU/㎎ | TNF-αpg/mg | IL-17pg/mg | NF-kB활성p-p65/p65 | COX-2활성 |
정상군 | 0.9 | 6.5 | 0.21 | 35 | 17 | 0.12 | 0.23 |
유도군 | -1.9 | 4.4 | 1.86 | 265 | 89 | 0.32 | 0.52 |
LP NK3 | -0.9 | 5.2 | 1.12 | 89 | 45 | 0.23 | 0.35 |
BL NK49 | -0.7 | 5.4 | 0.87 | 95 | 38 | 0.22 | 0.36 |
혼합물 | -0.7 | 5.5 | 0.82 | 88 | 35 | 0.19 | 0.29 |
양성대조군 | -1.0 | 5.1 | 1.26 | 105 | 47 | 0.25 | 0.42 |
Claims (20)
- 락토바실러스 플란타럼 NK3 (Lactobacillus plantarum NK3) KCCM12089P.
- 비피도박테리움 롱검 NK49 (Bifidobacterium longum NK49) KCCM12088P.
- 제1항에 있어서, 상기 락토바실러스 플란타럼 NK3 (Lactobacillus plantarum NK3) KCCM12089P는 서열번호 1의 16S rDNA 염기서열을 포함하는, 락토바실러스 플란타럼 NK3 (Lactobacillus plantarum NK3) KCCM12089P.
- 제2항에 있어서, 상기 비피도박테리움 롱검 NK49 (Bifidobacterium longum NK49) KCCM12088P은 서열번호 2의 16S rDNA 염기서열을 포함하는, 비피도박테리움 롱검 NK49 (Bifidobacterium longum NK49) KCCM12088P.
- 락토바실러스 플란타럼 NK3 (Lactobacillus plantarum NK3) KCCM12089P, 비피도박테리움 롱검 NK49 (Bifidobacterium longum NK49) KCCM12088P 또는 이들의 혼합물을 포함하는 염증 질환의 예방 또는 치료용 약학 조성물.
- 제5항에 있어서, 상기 락토바실러스 플란타럼 NK3 (Lactobacillus plantarum NK3) KCCM12089P은 이의 생균체, 이의 사균체, 이의 배양물, 이의 파쇄물 또는 이의 추출물인, 염증 질환의 예방 또는 치료용 약학 조성물.
- 제5항에 있어서, 상기 비피도박테리움 롱검 NK49 (Bifidobacterium longum NK49) KCCM12088P은 이의 생균체, 이의 사균체, 이의 배양물, 이의 파쇄물 또는 이의 추출물인, 염증 질환의 예방 또는 치료용 약학 조성물.
- 제5항에 있어서, 상기 염증 질환은 관절염, 통풍, 간염, 천식, 비만, 각막염, 위염, 장염, 신장염, 대장염, 당뇨, 결핵, 기관지염, 흉막염, 복막염, 척추염, 췌장염, 염증 통증, 요도염, 방광염, 질염, 동맥경화증, 패혈증, 화상, 피부염, 치주염 및 치은염을 포함하는 군에서 선택되는 어느 하나 이상인, 염증 질환의 예방 또는 치료용 약학 조성물.
- 제5항에 있어서, 상기 염증 질환은 대장염 또는 질염인, 염증 질환의 예방 또는 치료용 약학 조성물.
- 락토바실러스 플란타럼 NK3 (Lactobacillus plantarum NK3) KCCM12089P, 비피도박테리움 롱검 NK49 (Bifidobacterium longum NK49) KCCM12088P 또는 이들의 혼합물을 포함하는 염증 질환의 예방 또는 개선용 건강기능 식품.
- 제5항에 있어서, 상기 락토바실러스 플란타럼 NK3 (Lactobacillus plantarum NK3) KCCM12089P은 이의 생균체, 이의 사균체, 이의 배양물, 이의 파쇄물 또는 이의 추출물인, 염증 질환의 예방 또는 개선용 건강기능 식품.
- 제10항에 있어서, 상기 비피도박테리움 롱검 NK49 (Bifidobacterium longum NK49) KCCM12088P은 이의 생균체, 이의 사균체, 이의 배양물, 이의 파쇄물 또는 이의 추출물인, 염증 질환의 예방 또는 개선용 건강기능 식품.
- 제10항에 있어서, 상기 염증 질환은 관절염, 통풍, 간염, 천식, 비만, 각막염, 위염, 장염, 신장염, 대장염, 당뇨, 결핵, 기관지염, 흉막염, 복막염, 척추염, 췌장염, 염증 통증, 요도염, 방광염, 질염, 동맥경화증, 패혈증, 화상, 피부염, 치주염 및 치은염을 포함하는 군에서 선택되는 어느 하나 이상인, 염증 질환의 예방 또는 개선용 건강기능 식품.
- 제10항에 있어서, 상기 염증 질환은 대장염 또는 질염인, 염증 질환의 예방 또는 개선용 건강기능 식품.
- 락토바실러스 플란타럼 NK3(Lactobacillus plantarum NK3) KCCM12089P, 비피도박테리움 롱검 NK49(Bifidobacterium longum NK49) KCCM12088P 또는 이들의 혼합물을 대상체에 투여하는 단계를 포함하는 염증 질환의 예방 또는 치료방법.
- 제15항에 있어서, 상기 염증 질환은 관절염, 통풍, 간염, 천식, 비만, 각막염, 위염, 장염, 신장염, 대장염, 당뇨, 결핵, 기관지염, 흉막염, 복막염, 척추염, 췌장염, 염증 통증, 요도염, 방광염, 질염, 동맥경화증, 패혈증, 화상, 피부염, 치주염 및 치은염을 포함하는 군에서 선택되는 어느 하나 이상인, 염증 질환의 예방 또는 치료방법.
- 염증 질환의 치료를 위한 약제의 제조를 위한 락토바실러스 플란타럼 NK3 (Lactobacillus plantarum NK3) KCCM12089P, 비피도박테리움 롱검 NK49 (Bifidobacterium longum NK49) KCCM12088P 또는 이들의 혼합물의 용도.
- 제17항에 있어서, 상기 염증 질환은 관절염, 통풍, 간염, 천식, 비만, 각막염, 위염, 장염, 신장염, 대장염, 당뇨, 결핵, 기관지염, 흉막염, 복막염, 척추염, 췌장염, 염증 통증, 요도염, 방광염, 질염, 동맥경화증, 패혈증, 화상, 피부염, 치주염 및 치은염을 포함하는 군에서 선택되는 어느 하나 이상인, 용도.
- 염증 질환의 치료에 사용하기 위한 락토바실러스 플란타럼 NK3 (Lactobacillus plantarum NK3) KCCM12089P, 비피도박테리움 롱검 NK49 (Bifidobacterium longum NK49) KCCM12088P 또는 이들의 혼합물을 포함하는 조성물.
- 제19항에 있어서, 상기 염증 질환은 관절염, 통풍, 간염, 천식, 비만, 각막염, 위염, 장염, 신장염, 대장염, 당뇨, 결핵, 기관지염, 흉막염, 복막염, 척추염, 췌장염, 염증 통증, 요도염, 방광염, 질염, 동맥경화증, 패혈증, 화상, 피부염, 치주염 및 치은염을 포함하는 군에서 선택되는 어느 하나 이상인, 조성물.
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EP4265297A1 (en) * | 2020-12-16 | 2023-10-25 | Meiji Co., Ltd | Composition for improving inflammation of brain tissue |
KR102533469B1 (ko) * | 2021-01-05 | 2023-05-17 | 연세대학교 산학협력단 | 염증성 장 질환의 예방 또는 치료용 조성물 |
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KR101784847B1 (ko) * | 2016-05-10 | 2017-10-13 | 주식회사 하우동천 | 질염 예방 및 치료용 유산균 함유 조성물 및 이의 용도 |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
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WO2022192755A1 (en) * | 2021-03-12 | 2022-09-15 | MarvelBiome, Inc. | Methods and uses of microbiome compositions, components, or metabolites for treating eye disorders |
US20240066075A1 (en) * | 2021-03-12 | 2024-02-29 | MarvelBiome, Inc. | Methods and uses of microbiome compositions, components, or metabolites for treating eye disorders |
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US12036253B2 (en) | 2024-07-16 |
US20230181656A1 (en) | 2023-06-15 |
BR112021008358A2 (pt) | 2021-08-03 |
US11554146B2 (en) | 2023-01-17 |
EP3875576A1 (en) | 2021-09-08 |
JP2021501586A (ja) | 2021-01-21 |
AU2018358456B2 (en) | 2021-10-28 |
US20210260140A1 (en) | 2021-08-26 |
KR20190050294A (ko) | 2019-05-10 |
CA3081210A1 (en) | 2019-05-09 |
AU2021269282B2 (en) | 2023-08-17 |
AU2018358456A1 (en) | 2020-05-28 |
SG11202104408UA (en) | 2021-05-28 |
US20230181657A1 (en) | 2023-06-15 |
JP6964191B2 (ja) | 2021-11-10 |
CA3081210C (en) | 2023-04-11 |
EP3875576A4 (en) | 2022-03-02 |
KR102173547B1 (ko) | 2020-11-03 |
KR102151666B1 (ko) | 2020-09-04 |
AU2021269282A1 (en) | 2021-12-09 |
CN113302279B (zh) | 2024-07-30 |
WO2020091166A1 (ko) | 2020-05-07 |
KR20200104848A (ko) | 2020-09-04 |
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