WO2020091312A1 - Composition for preventing or treating alcoholic intestinal injury, containing probiotics as active ingredient - Google Patents

Abstract

The present invention pertains to a composition for preventing or treating alcoholic intestinal injury, the composition containing probiotics as an active ingredient. The composition of the present invention contains Lactobacillus bulgaricus CKDB001 (Accession Number: KCTC13669B), Lactobacillus helveticus CKDB001 (Accession Number: KCTC13670BP), Lactobacillus plantarum CKDB008 (Accession Number: KCTC13673BP), or Bifidobacterium bifidum CKDB001 (Accession Number: KCTC13114BP), which are novel isolated lactic acid strains, or a mixed strain thereof, has an excellent effect of reducing intestinal cell inflammation caused by alcoholic irritation, and can thus be usefully used as food or a therapeutic agent for preventing or alleviating alcoholic intestinal injury.

Description

Composition for preventing or treating alcoholic intestinal damage comprising probiotics as an active ingredient

This patent application claims priority to Korean Patent Application No. 10-2018-0131208 filed with the Korean Intellectual Property Office on October 30, 2018, and the disclosures of the patent application are incorporated herein by reference.

The present invention relates to a composition for preventing or treating alcoholic bowel damage comprising probiotics.

Alcohol (alcohol) is a favorite product that has been consumed for a long time to relieve stress or socialize. Small intake can help blood circulation and be beneficial to health, but chronic intake of excess can lead to liver disease such as hepatitis and cirrhosis. However, the liver is not the only organ affected by excessive alcohol intake. More than 80% of the alcohol consumed is absorbed into the body through the small intestine, so the effect of alcohol on the small intestine can never be overlooked.

The intestine is largely divided into a small intestine and a large intestine, and the small intestine can be further classified into a duodenum, a factory, and a ileum. The effect of alcohol is different for each part of the intestine. In the duodenum, bleeding and inflammatory reactions due to damage to the intestinal mucosa are most prominent, nutrient absorption is impaired in the plant, and contraction of villi in the ileum increases, resulting in diarrhea.

To prevent intestinal damage from alcohol intake above, abstinence is the best option, but eating foods that prevent intestinal damage can be a good alternative. However, most of the products sold on the market prevent liver damage, and products that prevent intestinal damage are not available. In addition, there are patents (KR10-1782848, KR10-1772810, KR10-2013-0036939, and KR10-1819026) for the prevention or treatment of alcoholic gastrointestinal diseases, all of which are inventions using natural extracts as active ingredients, and probiotics as active ingredients The present invention has not been developed yet. Therefore, the present inventors tried to find a functional probiotic strain that can prevent or treat alcoholic intestinal damage that can be applied as food and medicine.

The present inventors have made diligent research efforts to discover functional probiotic strains that can prevent or treat alcoholic bowel damage that can be applied as food and pharmaceutical products. As a result, a total of 530 types of lactic acid bacteria were first selected from 17 strains that confirmed safety, stability, and industrial potential, and then 12 types of lactic acid bacteria with excellent intestinal adhesion were selected. Among them, the cell survival rate due to alcohol was improved. Seven strains that can be selected were selected second. Finally, four strains that effectively lower the inflammatory cytokine expression increased due to alcohol were finally selected, and it was confirmed that the mixed strains of the four selected strains can effectively prevent inflammation of intestinal cells caused by alcohol than each single strain.

The four strains of the present invention are Lactobacillus bulgaricus CKDB001 (Accession No .: KCTC13669B), Lactobacillus helveticus CKDB001 (Accession No .: KCTC13670BP), Lactobacillus plantarum ) CKDB008 (Accession No .: KCTC13673BP), Bifidobacterium Bifidobacterium bifidum ) CKDB001 (accession number KCTC13114BP).

The present inventors confirmed that each of the selected four strains has excellent prevention or treatment effect against alcoholic intestinal damage, and in particular, these mixed strains can effectively prevent inflammation of intestinal cells caused by alcohol than each single strain. The present invention has been completed by clarifying.

Accordingly, an object of the present invention is to provide a method for preventing or treating alcoholic bowel damage, enteritis, or inflammatory bowel disease, comprising the step of administering to a subject a lactic acid bacteria composition comprising each selected single strain or a mixed strain thereof. Is to do.

Probiotics (Probiotics) is a microorganism that is beneficial to health when taken in an appropriate amount, and serves to protect the surface of the intestinal mucosa from various pathogens and foreign substances. Recently, various studies based on the hypothesis that probiotics will be involved in the protection of intestinal cells have been conducted as they show the effect of controlling intestinal mucosa defense and intestinal permeability in chronic inflammatory bowel disease.

The present inventors selected probiotics having an alcohol-protecting effect on epithelial cell lines for each organ of the small intestine, and developed a mixed strain of probiotics that can cover all damages of each small intestine.

In accordance with one aspect of the present invention, the invention provides a composition comprising a lactic acid bacteria Lactobacillus bacteria (Lactobacillus) in lactic acid bacteria, Bifidobacterium (Bifidobacterium) in lactic acid bacteria, or a mixture thereof.

According to an embodiment of the present invention, the Lactobacillus genus lactic acid bacteria are Lactobacillus bulgaricus , Lactobacillus helveticus , Lactobacillus helveticus , Lactobacillus plantarum , or mixtures thereof.

In a specific embodiment of the present invention, the Lactobacillus lactic acid bacteria are Lactobacillus bulgaricus CKDB001 (Accession No .: KCTC13669B), Lactobacillus helveticus CKDB001 (Accession No .: KCTC13670BP), Lactobacillus Lactobacillus plantarum CKDB008 (Accession No .: KCTC13673BP), or mixtures thereof.

According to another embodiment of the present invention, the lactic acid bacteria in the Bifidobacterium genus Bifidobacterium bifidum ), Bifidobacterium lactis , or mixtures thereof.

In a specific embodiment of the present invention, the lactic acid bacteria of the genus Bifidobacterium is Bifidobacterium bifidum CKDB001 (Accession No. KCTC13114BP).

In the most specific embodiment of the present invention, the lactic acid bacteria are Lactobacillus bulgaricus CKDB001 (Accession No .: KCTC13669BP), Lactobacillus helveticus CKDB001 (Accession No .: KCTC13670BP), Lactobacillus plantar ( Lactobacillus plantarum ) CKDB008 (Accession No .: KCTC13673BP), Bifidobacterium Bifidobacterium bifidum ) CKDB001 (Accession No. KCTC13114BP), or a mixed strain thereof.

The Lactobacillus bulgaricus CKDB001 strain of the present invention was deposited on October 23, 2018 at the Korea Institute of Biotechnology and Microbiology Resource Center (Korean Collection for Type Culture, KCTC), and was assigned a deposit number KCTC 13669BP.

In addition, the Lactobacillus helveticus CKDB001 strain of the present invention was deposited on October 23, 2018 at the Korea Institute of Biotechnology and Microbiology Resource Center (Korean Collection for Type Culture, KCTC), and was assigned a deposit number KCTC 13670BP.

In addition, the Lactobacillus plantarum CKDB008 strain of the present invention was deposited on October 23, 2018 at the Korea Institute of Biotechnology and Microbiology Resource Center (Korean Collection for Type Culture, KCTC), and was assigned a deposit number KCTC 13673BP.

Finally, the Bifidobacterium bifidum CKDB001 strain of the present invention was deposited on September 23, 2016 at the Korea Institute of Biotechnology and Microbiology Resource Center (Korean Collection for Type Culture, KCTC), and was assigned a deposit number KCTC13114BP.

The four strains were confirmed to be Gram positive, hemolytic, hemolytic, sporulation ability, and Catalase negative. The sugar availability of the strain was analyzed using the API 50 CHL kit (Table 1), and gene identification was performed by analyzing the 16s rDNA sequence for strain identification (Table 2).

Source		L. bulgaricusCKDB001	L. helveticusCKDB001	L. plantarumCKDB008	B.bifidumCKDB001
Temoin	0	+	+	+	-
Glycerol	One	-	-	-	-
Erythritol	2	-	-	-	-
D-Arabinose	3	-	-	-	-
L-Arabinose	4	-	-	+	-
D-Ribose	5	-	-	+	w
D-Xylose	6	-	-	-	-
L-Xylose	7	-	-	-	-
D-Adonitol	8	-	-	-	-
Methyl-βD-Xylopyranoside	9	-	-	-	-
D-Galactose	10	-	+	+	+
D-Glucose	11	+	+	+	+
D-Fructose	12	+	+	w	+
D-Mannose	13	+	+	+	-
L-Sorbose	14	-	-	-	-
L-Rhamnose	15	-	-	+	-
Dulcitol	16	-	-	-	-
Inositol	17	-	-	-	-
D-Mannitol	18	-	-	+	-
D-Sorbitol	19	-	-	-	-
Methyl-αD-Mannopyranoside	20	-	-	+	-
Methyl-αD-Glucopyranoside	21	-	-	-	-
N-Acetylglucosamine	22	+	+	+	+
Amygdaline	23	-	-	+	-
Arbutine	24	-	-	+	-
Esculine, citrate de fer	25	-	-	-	-
Salicine	26	-	-	+	-
D-Cellobiose	27	-	-	+	-
D-Maltose	28	w	+	+	-
D-Lactose	29	+	+	+	+
D-Melibiose	30	-	-	+	-
D-Saccharose	31	+	-	+	-
D-Trehalose	32	+	+	+	-
Inuline	33	-	-	-	-
D-Melezitose	34	-	-	+	-
D-Raffinose	35	-	-	+	-
Amidon	36	-	-	-	-
Glycogene	37	-	-	-	-
Xylitol	38	-	-	-	-
Gentiobiose	39	-	-	+	+
D-Turanose	40	-	-	+	-
D-Lyxose	41	-	-	-	-
D-Tagatose	42	-	-	-	-
D-Fucose	43	-	-	-	-
L-Fucose	44	-	-	-	-
D-Arabitol	45	-	-	+	-
L-Arabitol	46	-	-	-	-
Potassium GlucoNaTe	47	-	-	+	-
Potassium 2-CetoGluconate	48	-	-	-	-
Potassium 5-CetoGluconate	49	-	-	-	w

Strains	sequence
L. bulgaricus CKDB001	CTAGAGATAGGTGGTTCCCTTCGGGGACGCAGAGACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCTTTAGTTGCCATCATTAAGTTGGGCACTCTAAAGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGCAGTACAACGAGAAGCGAACCCGCGAGGGTAAGCGGATCTCTTAAAGCTGCTCTCAGTTCGGACTGCAGGCTGCAACTCGCCTGCACGAAGCTGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGAAGTCTGCAATGCCCAAAGTCGGTGAGATAACCTTTAT
L. helveticus CKDB001	GGAGTTCCCTTCGGGGACGCTAAGACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTTATTAGTTGCCAGCATTAAGTTGGGCACTCTAATGAGACTGCCGGTGATAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACAGTACAACGAGAAGCGAGCCTGCGAAGGCAAGCGAATCTCTGAAAGCTGTTCTCAGTTCGGACTGCAGTCTGCAACTCGACTGCACGAAGCTGGAATCGCTAGTAATCGCGGATCAGAACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGAAGTCTGCAATGCCCAAAGCCGGTGGCCTAACCTTCGGGAAGGAGCCGTCTAAGCAGGCAGATGACTGGGGTG
L. plantarum CKDB008	TAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTT
B. bifidum CKDB001	GTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGCCCCGTGTTGCCAGCACGTTATGGTGGGAACTCACGGGGGACCGCCGGGGTTAACTCGGAGGAAGGTGGGGATGACGTCAGATCATCATGCCCCTTACGTCCAGGGCTTCACGCATGCTACAATGGCCGGTACAGCGGGATGCGACATGGCGACATGGAGCGGATCCCTGAAAACCGGTCTCAGTTCGGATCGGAGCCTGCAACCCGGCTCCGTGAAGGCGGAGTCGCTAGTAATCGCGGATCAGCAACGCCGCGGTGAATGCGTTCCCGGGCCTTGTACACACCGCCCGTCAAGTCATGAAAGTGGGCAGCACCCGAAGCCGGTGGCCTAACCCCTTGTGGGATGGAGCCGTCTAAG

The Lactobacillus active ingredient of the composition Bulgaria kusu (Lactobacillus bulgaricus) CKDB001 (accession number: KCTC13669BP), Lactobacillus helveticus (Lactobacillus helveticus) CKDB001 (accession number: KCTC13670BP), Lactobacillus Planta column (Lactobacillus plantarum) CKDB008 (Accession No .: KCTC13673BP), Bifidobacterium Bifidobacterium bifidum ) CKDB001 (Accession No. KCTC13114BP) was selected as the strain that most effectively prevents intestinal damage caused by alcohol among a total of 17 probiotic strains. The lactic acid bacteria, which are active ingredients of the composition of the present invention, are Lactobacillus rhamnosus ( Lactobacillus rhamnosus) , which are commercially available strains that have the ability to adhere to small intestine (colon) and colon epithelial cells. GG strain, LGG), so it has very good gut adhesion regardless of the intestine.

In addition, the lactic acid bacteria show a very high cell viability, similar to normal cells, as well as the commercial strain LGG strain despite ethanol treatment of small intestine (duodenum and ileum) epithelial cells.

In addition, the lactic acid bacteria significantly reduce the expression of inflammatory cytokines (IL-1β and TNFα) compared to the commercial strain LGG despite ethanol treatment of small intestine (duodenum and ileum) epithelial cells. Therefore, the lactic acid bacteria of the present invention shows a very good anti-inflammatory effect on the small intestine.

In particular, the four types of lactic acid bacteria deposited in the final selection and depository of the present invention have excellent intestinal adhesion ability, cell viability, and anti-inflammatory effect of each strain, but when used as a mixed strain mixed with them, the anti-inflammatory effect of each single strain It has characteristics in that it has a better anti-inflammatory effect (synergy effect). Therefore, the lactic acid bacteria of the present invention is more useful when used as a mixed strain.

The strain, or a culture thereof, which is an active ingredient of the compositions of the present invention, is a culture medium containing cells in addition to the cells isolated and / or purified from the strain, an extract of cells, a culture supernatant, a concentrate thereof, a concentrate, It is a dried product, and if necessary, a diluent, a diluent, and the like, and includes all of the state obtained by treating the culture medium and the culture.

The culture method, extraction method, separation method, concentration method, drying method, dilution method, etc. of the above-mentioned cell body are not particularly limited.

The medium for culturing the cells generally includes milk protein such as skim milk, whey, and casein, sugars, yeast extracts, etc., and various aerobic or anaerobic methods common to the culture method can be suitably used.

As the culture temperature, for example, 35 to 45 ° C is set, and during the culture, an alkali such as sodium hydroxide is used to use a neutralization culture method that maintains the pH of the medium from neutral to acidic, for example, about 5 to 6 pH. It might be. In addition to such a neutralization method, any culture method such as a batch culture method can be used. After cultivation, the culture or the supernatant may be concentrated, dried, or diluted, if necessary.

In addition, the supernatant and the cells of the culture may be separated using a centrifugation method or a membrane separation method, and the cells may be recovered in a concentrated state. In addition, the cells may be subjected to ultrasonic treatment or enzyme treatment to extract components in the cells, or the culture or supernatant, the cells or the extract may be dried. These can be used as an active ingredient of the composition of the present invention.

According to one embodiment of the present invention, lactic acid bacteria, which are active ingredients of the composition of the present invention, exhibit excellent cell viability and anti-inflammatory effects against the above-mentioned intestinal epithelial cell toxicity of alcohol. Therefore, the composition of the present invention has the purpose of preventing, improving or treating alcoholic bowel damage.

As demonstrated in the specific embodiment of the present invention, the lactic acid bacteria of the present invention exhibit the advantageous effects described above for various parts of the small intestine (duodenum, ileum) and large intestine. Thus, the “intestine” of the intestinal injury refers to the small intestine and / or large intestine, and more specifically the “small intestine” refers to the duodenum, plant, and / or ileum. Most specifically, the small intestine refers to the duodenum and / or ileum.

As demonstrated in a specific embodiment of the present invention, the lactic acid bacteria of the present invention significantly reduce the expression of inflammatory cytokines in the intestine. Therefore, the composition of the present invention has the purpose of preventing, improving or treating enteritis or inflammatory bowel disease.

In the present invention, the enteritis includes bacterial enteritis, viral enteritis, and alcoholic enteritis.

In addition, the inflammatory bowel disease in the present invention is selected from the group consisting of Crohn's disease, Behcet's disease, and ulcerative colitis.

The lactic acid bacteria composition of the present invention may be prepared as a food composition having the above-described use, or a pharmaceutical composition.

When the composition of the present invention is prepared as a food composition, as an active ingredient, as well as the lactic acid bacteria, it may include a component that is conventionally added in food production. The additive components include, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. Examples of the carbohydrate include monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, oligosaccharides, etc.) and polysaccharides (eg, dextrins, cyclodextrins, etc.). Sugars and sugar alcohols such as xylitol, sorbitol, erythritol, etc. As flavoring agents, natural flavoring agents (taumatin, stevia extracts (e.g. rebaudioside A, glycyrrhizine, etc.)) and synthetic flavoring agents (saccharin, aspar Tom).

For example, when the food composition of the present invention is prepared as a drink agent, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, jujube extract or licorice extract may be additionally included in addition to the above-mentioned strain as an active ingredient of the present invention. have.

The food composition of the present invention includes processing forms of all natural materials such as food, functional food, nutritional supplement, health food and food additives. Food compositions of this type can be prepared in various forms according to conventional methods known in the art.

For example, as a healthy food, the lactic acid bacteria themselves may be prepared in the form of tea, juice, and drink to be consumed or granulated, encapsulated, and powdered. In addition, food products include beverages (including alcoholic beverages), fruits and processed foods thereof (eg, canned fruits, canned foods, jams, marmalades, etc.), fish, meat, and processed foods (eg ham, sausage corn beef, etc.) ), Breads and noodles (e.g. udon, buckwheat noodles, ramen, spaghetti, macaroni, etc.), juice, various drinks, cookies, syrup, dairy products (e.g. yogurt, fermented milk, butter, cheese, etc.), edible vegetable oil, margarine, Vegetable protein, retort food, frozen food, various seasonings (eg, miso, soy sauce, sauce, etc.) can be prepared by adding the lactic acid bacteria of the present invention. In addition, in order to use the lactic acid bacteria of the present invention in the form of food additives, it may be prepared and used in the form of a powder or a concentrate.

When the composition of the present invention is prepared as a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier included in the pharmaceutical composition of the present invention is commonly used in formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, but is not limited thereto It does not work.

The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention can be administered orally or parenterally, and is preferably applied by oral administration. The pharmaceutical composition of the present invention may be formulated in various oral or parenteral dosage forms, but is not limited thereto.

Suitable dosages of the pharmaceutical compositions of the invention vary by factors such as formulation method, mode of administration, patient's age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion, and response sensitivity, Usually, a skilled doctor can easily determine and prescribe a dose (pharmaceutical effective amount) effective for the desired treatment or prevention. According to a preferred embodiment of the invention, the daily dosage of the pharmaceutical composition of the invention is 0.0001-100 mg / kg. The term "pharmaceutical effective amount" as used herein means an amount sufficient to prevent or treat the aforementioned diseases.

As used herein, the term “treatment” refers to reduction, suppression, sedation, or eradication of a disease state, and “prevention” means to suppress the exacerbation of the disease state so that the disease state does not develop and includes the treatment.

Formulations for oral administration include, for example, tablets, pills, hard / soft capsules, liquids, suspensions, emulsifiers, and syrups. There are granules, elixirs, and the like, and these formulations may use one or more diluents or excipients, such as fillers, extenders, wetting agents, disintegrating agents, lubricants, binders, and surfactants, which are commonly used in addition to the active ingredients. As a disintegrating agent, agar, starch, alginic acid or its sodium salt, calcium phosphate monohydrogen anhydride, etc. may be used, and as a lubricant, silica, talc, stearic acid or its magnesium salt or calcium salt, polyethylene glycol, etc. may be used. , As the binder, magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidine, low-substituted hydroxypropylcellulose, and the like can be used. In addition, lactose, dextrose, sucrose, mannitol, sorbitol, and cellulose. Glycine or the like can be used as a diluent, and in some cases, commonly known boiling mixtures, absorbents, colorants, flavoring agents, sweeteners and the like can be used together.

The composition may be sterilized or contain preservatives, stabilizers, hydration or emulsification accelerators, salts for osmotic pressure control, adjuvants such as buffers, and other therapeutically useful substances, conventional methods of mixing, granulating or coating methods It can be formulated according to.

The pharmaceutical composition of the present invention is prepared in a unit dose form by formulating using a pharmaceutically acceptable carrier and / or excipient according to a method that can be easily carried out by those skilled in the art to which the present invention pertains. Or it can be manufactured by incorporating into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension, syrup or emulsion in an oil or aqueous medium, or may be in the form of ex-agent, powder, granule, tablet or capsule, and may further include a dispersant or stabilizer.

According to an aspect of the present invention, the present invention comprises the step of administering to the subject a composition comprising a lactic acid bacteria of the genus Lactobacillus (Lactobacillus), lactic acid bacteria of the genus Bifidobacterium, or mixtures thereof It provides a method of preventing or treating intestinal damage.

Alcoholic intestinal damage, which is the target disease of the treatment or prevention method of the present invention, is as defined in relation to the disease to be treated of the pharmaceutical composition.

In one embodiment of the invention the subject is a mammal or human.

According to another aspect of the present invention, the present invention comprises the step of administering to the subject a composition comprising a lactobacillus (Lactobacillus) genus lactic acid bacteria, Bifidobacterium genus lactic acid bacteria, or mixtures thereof, Provides a method for preventing or treating enteritis or inflammatory bowel disease.

Enteritis or inflammatory bowel disease, which is the target disease of the treatment or prevention method of the present invention, is as defined in relation to the target disease of the pharmaceutical composition.

The method for preventing or treating alcoholic intestinal damage, enteritis, or inflammatory bowel disease of the present invention includes a composition comprising the aforementioned Lactobacillus lactic acid bacteria, Bifidobacterium lactic acid bacteria, or mixtures thereof Since it is a method using the same active ingredient, the description thereof is omitted to avoid excessive complexity of the present specification for overlapping contents.

The features and advantages of the present invention are summarized as follows:

(a) The present invention provides a composition for preventing or treating alcoholic bowel damage comprising probiotics as an active ingredient. The compositions of the present invention relates to novel separation Lactobacillus own, Lactobacillus Bulgaria kusu (Lactobacillus bulgaricus) CKDB001 (accession number: KCTC13669B), Lactobacillus helveticus (Lactobacillus helveticus) CKDB001 (accession number: KCTC13670BP), Lactobacillus Planta column (Lactobacillus plantarum ) CKDB008 (Accession No .: KCTC13673BP), Bifidobacterium Bifidobacterium bifidum ) CKDB001 (Accession No. KCTC13114BP), or a mixed strain thereof.

(b) Since the present invention has an excellent effect of reducing inflammation of intestinal cells due to alcohol stimulation, it can be usefully used as a food or therapeutic agent for preventing and improving alcoholic bowel damage.

1A and 1B are diagrams showing the intestinal epithelial cell adhesion ability of the probiotic strain of the present invention.

Figure 2a is a diagram showing the protective effect of alcoholic cell damage to the duodenal cell line (HUTU-80) of the probiotic strain of the present invention.

Figure 2b is a diagram showing the protective effect of alcoholic cell damage to the ileum cell line (IEC-18) of the probiotic strain of the present invention.

Figure 3a is a diagram showing the inhibitory effect of mRNA expression of the inflammatory cytokine IL-1β in the duodenal cell line (HUTU-80) of the probiotic strain of the present invention.

Figure 3b is a diagram showing the inhibitory effect of mRNA expression of the inflammatory cytokine TNFα in the duodenal cell line (HUTU-80) of the probiotic strain of the present invention.

Figure 4a is a diagram showing the inhibitory effect of mRNA expression of the inflammatory cytokine IL-1β in the ileal cell line (IEC-18) of the probiotic strain of the present invention.

Figure 4b is a diagram showing the inhibitory effect of mRNA expression of the inflammatory cytokine TNFα in the ileal cell line (IEC-18) of the probiotic strain of the present invention.

Figure 5a is a diagram showing the inhibitory effect of inflammatory cytokine expression in the ileal cell line (IEC-18) of the mixed strain of the present invention.

Figure 5b is a diagram showing the inhibitory effect of inflammatory cytokine expression in the duodenal cell line (HUTU-80) of the mixed strain of the present invention.

Hereinafter, the present invention will be described in more detail through examples. These examples are only intended to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Example

Throughout this specification, "%" used to indicate the concentration of a specific substance, unless otherwise specified, solids / solids (weight / weight)%, solids / liquids (weight / volume)%, and The liquid / liquid is (volume / volume)%.

1. Experimental materials

1.1 Isolation and screening of probiotic strains

1.1.1. Separation of new microorganisms

Probiotics used in this experiment was a strain isolated from Chong Kun Dang Bio (CKDBiO, Korea) and was isolated from feces, crude milk and other fermented foods. The collected sample was suspended in sterile anaerobic water, and after dilution of the decimal, it was spread on MRS (ManRogosaSharpe) solid medium and BL (Blood-liver) solid medium. After incubation in an anaerobic chamber maintained at 5% CO ₂ , 10% H ₂ , and 85% N ₂ atmospheric composition for 48 hours at 37 ° C., colonies having different forms were selected and purified. As a result of the experiment, a total of 530 lactic acid bacteria were isolated. Subsequently, the 530 strains of the isolated lactic acid bacteria were tested for hemolysis, whether gelatinase was produced, whether urease was produced, and whether bio-amine was produced. Was selected.

1.1.2 Comparative evaluation of digestive fluid (acidic pH and bile acid salt) resistance

The pH resistance and resistance to bile salts of each strain of lactic acid bacteria selected in 1.1.1 were compared and analyzed.

To confirm the resistance of the selected lactic acid bacteria to the acidic pH, an MRS or BL liquid medium adjusted to pH 2.5 using hydrochloric acid was prepared. After inoculating each of the strains with 1% in the medium, after diluting with 10% of anaerobic dilution (pH 6.2) in order to measure the initial number of bacteria, 1 ml was dispensed and plated using the same solid medium, followed by incubation at 37 ° C for 48 hours. . MRS liquid medium and BL liquid medium having a pH of 2.5 inoculated with the strain were cultured in a 37 ° C incubator for 2 hours, and then cultured in the same solid medium in the same manner as above. After colonization, colonies were counted to measure resistance to pH.

To confirm the bile resistance of the selected lactic acid bacteria, 0.3% oxgal (Sigma, USA) was added to the MRS or BL liquid medium using the method of Gilliland and Walker (1990), and 1% of the lactic acid bacteria after the second passage was completed. It was inoculated. As a control, the same amount of lactic acid bacteria was inoculated into the liquid medium of MRS or BL without addition of 0.3% oxgal. All samples were cultured in a 37 ° C. incubator and after 2 hours, cultured in each lactic acid bacteria growth solid medium to measure the number of microorganisms according to a conventional resistance measurement method (Equation 1).

Equation 1

Survival rate (%) = [(CFU / ml _120min ) / (CFU / ml _0min )] x100

As a result of the experiment, a total of 128 weeks were selected by selecting the top 30% of the strains having high acid resistance and bile resistance for each species.

1.1.3 Preparation of Raw Lactic Acid Bacteria

For the production of high-concentration powder of the selected 128-week lactic acid bacteria, the culture medium was cultured by setting the optimized growth medium and culture conditions for each strain, and then the cell recovery and freeze-drying process was performed in a conventional method (60 ° C freezer after centrifugation). After freezing, lyophilization was performed under operating conditions between 0 ° C and 45 ° C). After recovering the cells, 5-50% (volume / weight)% of maltodextrin or 5-50% (volume / weight) of trehalose or 5-50% (volume / weight) of cellulose is added to the concentrate compared to the concentrate, and then lyophilized and pulverized to prepare lactic acid bacteria raw material. Did. As a result, a total of 17 strains (Table 3) with the highest probability of industrialization due to the long-term storage stability and the highest original survival rate by species were selected. In the experiment, the powdered strain was suspended in an animal cell culture medium and used.

List of probiotic strains used in the study

Serial number	Strain name	Raw survival rate (%)	Abbreviation
One	Lactobacillus helveticus CKDB001	60	LH
2	Lactobacillus gasseri CKDB026	50	LG
3	Lactobacillus reuteri CKDB016	67	LU
4	Lactobacillus plantarum CKDB008	82	LP
5	Lactobacillus acidophilus CKDB007	29	LA
6	Lactobacillus bulgaricus CKDB001	72	LB
7	Lactobacillus casei CKDB007	75	LC
8	Lactobacillus fermentum CKDB004	74	LF
9	Lactobacillus rhamnosus CKDB	80	LR
10	Lactobacillus salivarius CKDB001	75	LS
11	Lactobacillus paracasei CKDB005	68	LPARA
12	Lactococcus lactis CKDB007	81	LCL
13	Streptococcus thermophilus CKDB021	70	ST
14	Bifidobacterium lactis CKDB005	35	BL
15	Bifidobacterium longum CKDB004	40	BO
16	Bifidobacterium bifidum CKDB001	27	BF
17	Bifidobacterium breve CKDB002	38	BB

1.2 Cell culture The cells of HUTU-80 (duodenal epithelial cell line, Human), IEC-18 (colon epithelial cell line, Rat), and HT-29 (colon epithelial cell line, Human) are distributed from the Korea Cell Line Bank (KCLB, Seoul, Korea). HUTU-80 and IEC-18 used Dulbeco's Modified Eagle's Media (DMEM) medium, HT-29 used Rowell Park Memorial Institue (RPMI) 1640 medium, and Penicillin / Streptomycin (P / S) for each medium. Incubation was performed at 37 ° C and 5% CO ₂ using a culture solution to which 1% and Fetal Bovine Serum (FBS) 10% were added.

1.3 Statistical Processing

The significance difference test for the experimental results was analyzed by variance analysis (Anova: analysis of variance) in the Prism software according to the multiple verification method at the p <0.05 level.

2. of the present invention Probiotic  Intestinal epithelial cells Adhesion  Measure

In the intestine, numerous intestinal bacteria are inhabited by competing and interacting with each other. Probiotics in these intestines are essential for the ability to attach the intestines in order to exert such functions as suppressing harmful bacteria, strengthening barrier functions, and regulating immune functions. Therefore, the present inventors measured the adhesion capacity to the intestinal epithelial cells (IEC-18) and the colon epithelial cells (HT-29) in order to confirm the intestinal adhesion ability of the probiotic strains of the present invention.

2.1 Measurement of adhesion of probiotic strains to ileal epithelial cell line (IEC-18)

IEC-18 was dispensed into a 12-well cell culture dish at a concentration of 1 x 10 ⁵ cells / mL and cultured in a cell incubator (37 ° C for 5 days, 5% CO ₂ ) until a single layer. The probiotic strain was diluted with 1Х10 ⁹ CFU / mL in DMEM without antibiotics. When IEC-18 achieves confluency, 1 mL of each fungus solution is dispensed into the cells and incubated in a cell incubator for 90 minutes. Thereafter, the cells were washed twice with PBS and treated with Trypsin-EDTA to remove the cells. After diluting the separated cells and inoculating them on an MRS agar plate, the number of viable cells is measured after incubation at 37 ° C. for 18 hours. As a comparative group, a commercial strain Lactobacillus rhamnosus GG was used, and the intestinal adhesion rate was calculated as shown in Equation 2.

Equation 2

Intestinal adhesion rate (%) = (viable cell count / initial viable cell count after 90 min incubation) x 100

As a result of the experiment, among 17 strains of CKDBiO, B. bifidum is the strain that has superior intestinal adhesion ability than IEC L. GG (0.02%) in IEC-18. CKDB001 (0.18%), L. plantarum CKDB008 (0.11%), S. thermophilus CKDB021 (0.08%), L. salivarius CKDB001 (0.05%), L. bulgaricus CKDB001 (0.04%), L. helveticus CKDB001 (0.02%), and L. fermentum Seven types of CKDB004 (0.02%) were identified (FIG. 1A).

2.2 Measurement of adhesion of probiotic strain to colon epithelial cell line (HT-29)

HT-29 was dispensed in a 12 well cell culture dish at a concentration of 1 x 10 ⁵ cells / mL and cultured in a cell culture medium (37 ° C., 5% CO _{2 for} about 5 days) until a monolayer. Each probiotic strain was diluted with 1Х10 ⁹ CFU / mL in RPMI 1640 without antibiotics. When HT-29 formed confluency, 1 mL of each fungus solution was dispensed into cells and cultured in a cell incubator for 90 minutes. Thereafter, the cells were washed twice with PBS and treated with Trypsin-EDTA to remove the cells. The detached cells were serially diluted and inoculated onto an MRS agar plate, and the number of viable cells was measured after incubation at 37 ° C. for 18 hours. As a comparative group, a commercial strain, Lactobacillus rhamnosus GG, was used, and the intestinal adhesion rate was calculated as shown in Equation 2 in the same manner as in 2.1 above.

Equation 2

Intestinal adhesion rate (%) = (viable cell count / initial viable cell count after 90 min incubation) x 100

In HT-29, the strain with superior intestinal adhesion ability than LGG (2.23%) was L. helveticus CKDB001 (8.17%), L. reuteri CKDB016 (4.88%), L. plantarum CKDB008 (4.41%), L. acidophilus CKDB007 (3.43%), L. bulgaricus CKDB001 (3.39%), L. casei 9 types of CKDB007 (3.32%), B. lactis CKD005 (3.22%), B. longum CKD004 (3.11%), and L. fermentum CKD004 (2.37%) were identified (FIG. 1B).

2.3. Sintering

Considering the strain having excellent intestinal adhesion ability to both cell lines, a total of 12 out of 17 species were selected. Among them, BBF and LH were found to have significantly superior intestinal adhesion ability compared to LGG, which is well known as a strain having excellent intestinal adhesion ability.

3. The present invention Probiotic  Protective effect of alcoholic intestinal cell damage-Cell viability measurement ( MTT  assay)

MTT assay was performed to evaluate the toxicity of alcohol to intestinal epithelial cells and the prophylactic effect of probiotics. The MTT assay was conducted with 12 strains that were superior to LGG in the previous intestinal adhesion test.

3.1 Experimental method

First, HUTU-80 and IEC-18 were dispensed in a 96-well cell culture dish at a concentration of 1Х10 ⁵ cell / mL and cultured in a cell culture medium (37 ° C, 5% CO ₂ ) until a single layer. The probiotic strain was diluted with 1Х10 ⁶ CFU / mL in cell medium MEM without antibiotics. When the cells were confluency, 200 μL of each fungus was dispensed into the cells and cultured in a cell incubator for 90 minutes. Thereafter, the cells were washed twice with PBS, and cell culture medium containing ethanol was dispensed and reacted for 90 minutes. After the reaction was completed, the medium was removed, and then the MTT solution was added, incubated for 3 hours in a cell incubator, and then Dimethyl sulfoxide (DMSO) was added to terminate the reaction. DMSO was reacted for 10 minutes and cell viability was calculated by measuring absorbance at 540 nm. The control group was treated only with ethanol, and the comparison group used a commercial strain, Lactobacillus rhamnosus GG. Cell viability was calculated as shown in Equation 3.

Equation 3

Cell viability (%) = (Test group absorbance value / Control absorbance value) x 100

3.2 Protective effect on duodenal epithelial cell line (HUTU-80)

In the case of the duodenal epithelial cell line (HUTU-80), the cell viability was reduced to 33% compared to the normal group in the control group with only ethanol. When the comparative bacteria LGG was pretreated, the cell viability increased to 82%. Two of the 12 strains that showed higher cell viability than LGG. That is, L. plantarum CKDB008 is 95%, L. salivarius CKDB001 showed 88% cell viability and showed a significant difference from the control group as well as cell viability similar to normal cells (Fig. 2A).

3.3 Protective effect on ileal epithelial cell line (IEC-18)

In the case of the ileal epithelial cell line (IEC-18), the cell viability was reduced to 59% compared to the normal group in the control group with only ethanol. When the comparative bacteria LGG was pretreated, the cell viability increased to 77%. Among the 12 strains, 5 cells showed higher cell viability than LGG. In other words, B. bifidum CKDB001 , L. bulgaricus CKDB001 , L. fermentum CKDB004 , L. helveticus CKDB001 , and L. acidophilus CKDB007 showed a cell survival rate of 80% or more and showed a significant difference from the control group (FIG. 2B).

4. The present invention Probiotic  Inhibitory effect of inflammatory cytokines in the intestine

RT-PCR was performed to confirm that the probiotic strain of the present invention inhibits mRNA expression of inflammatory cytokines induced by alcohol. Confirmation of the inflammatory cytokine mRNA shedding amount was conducted with seven strains that were superior to LGG in the previous intestinal adhesion test and MTT assay.

4.1 Experimental method

First, divide the duodenal epithelial cell line (HUTU-80) and ileal epithelial cell line (IEC-18) into a 6-well cell culture dish at a concentration of 1Х10 ⁵ cells / mL, and then incubate until a single layer (37 ℃, 5% CO _2). ). Probiotics were diluted in 1MEM ¹⁰ CFU / mL in DMEM without antibiotics. When the cells are confluent, each fungus solution is dispensed into cells by 3 mL and cultured in a cell incubator for 90 minutes. Thereafter, the cells were washed twice with PBS, and cell culture medium containing ethanol was dispensed and reacted for 90 minutes. After the reaction, the cells were removed by treatment with Trypsin-EDTA. Thereafter, RNA was extracted from each cell line and cDNA was synthesized. And the mRNA expression level of inflammatory cytokines was confirmed through RT-PCR. The genomic base sequence used for PCR analysis is described in Table 4.

Genome sequence used for PCR analysis

Cell line	dielectric	Forward	Reverse
HUTU-80 (Human)	GAPDH	ACCCACTCCTCCACCTTTGA	CTGTTGCTGTAGCCAAATTCGT
	IL-1β	CCGACCACCACTACAGCAAG	GGGCAGGGAACCAGCATCTT
	TNFα	CTGGGCAGGTCTACTTTGGG	CTGGAGGCCCCAGTTTGAAT
IEC-18 (Rat)	GAPDH	AGTGCCAGCCTCGTCTCATA	GATGGTGATGGGTTTCCCGT
	IL-1β	CACCTCTCAAGCAGAGCACAG	GGGTTCCATGGTGAAGTCAAC
	TNFα	GGCTTTCGGAACTCACTGGA	CCCGTAGGGCGATTACAGTC

4.2 Effect of reducing inflammatory cytokine mRNA expression in duodenal epithelial cell line (HUTU-80)

In the duodenal epithelial cell line, the expression level of IL-1β mRNA was increased by 385% and the expression of TNFα by 357% compared to normal in the ethanol-only control group. When the comparative bacteria LGG were pretreated, IL-1β decreased to 235% and TNFα to 257%. B. bifidum is the strain that has lower IL-1β expression than LGG It was reduced to 155% with one CKDB001. B. bifidum was the strain that lowered TNFα expression level than LGG. CKDB001, L. plantarum Two types of CKDB008 were reduced to 130% and 142%, respectively. Therefore, two of the seven candidate strains were found to be able to reduce the inflammatory response of duodenal cells caused by alcohol more effectively than the comparative group LGG (FIGS. 3A and 3B).

4.3 Effect of reducing inflammatory cytokine mRNA expression in ileal epithelial cell line (IEC-18)

In the case of the control group that responded only to ethanol in the duodenal epithelial cell line, IL-1β mRNA expression was increased by 320% and TNFα expression was increased by 289% compared to normal. When pre-treated with the comparative bacteria LGG, IL-1β decreased to 201% and TNFα to 223%. L. bulgaricus is a strain that has lower IL-1β expression than LGG. CKDB001 , L. helveticus CKDB001 , L. plantarum It was reduced to 145%, 166%, and 182%, respectively, with three types of CKDB008. Strains with lower TNFα expression than LGG were L. helveticus It was reduced to 191% with one CKDB001. Therefore, three of the seven candidate strains were found to be able to reduce the inflammatory response of ileal epithelial cells due to alcohol more effectively than the comparative group LGG (FIGS. 4A and 4B).

4.4 Sintering

B. bifidum In the case of CKDB001, the inflammatory cytokine expression could be effectively reduced in the duodenal cell line, but the effect was relatively insignificant in the ileal cell line. L. helveticus CKDB001 showed the opposite trend. Therefore, when these strains are mixed, it is considered that they can complement each other and exhibit anti-inflammatory effects across all small intestine sites.

5. Verification of the effect of inhibiting the expression of inflammatory cytokines in the intestine of the mixed strain of the present invention

The present inventors, in order to confirm that the strain mixture effective for damage to each small intestine site selected in the above 4 is more effective than a single strain, mixed strain of the present invention (4 excellent strains for inhibiting the expression level of inflammatory cytokine mRNA: B. bifidum CKDB001, L. helveticus CKDB001, L. plantarum CKDB008, L. bulgaricus CKDB001) was tested to inhibit the expression of inflammatory cytokines in the intestine. The experimental method was carried out in the same manner as in the above 4, except that the mixed strains of the above 4 were set as the experimental group.

5.1 Effect of reducing the expression level of inflammatory cytokine mRNA in ileal epithelial cell line (IEC-18) of mixed strains

In the control group that reacted with only 5% ethanol in the ileum cell line, the expression level of IL-1β mRNA increased to 481% compared to normal, and decreased to 220% when pretreated with the comparative bacteria LGG. L. helveticus CKDB001, L. plantarum CKDB008 and L. bulgaricus CKDB001 lowered IL-1β mRNA expression levels to 110%, 202%, and 132%, respectively, and were found to be more effective than LGG. IL-4 β mRNA expression was 103 when pretreated by mixing four bacteria. It was significantly lowered to%, showing a similar degree to Normal.

In the case of TNFα, the expression level increased to 343% compared to normal when treated with 5% ethanol, and decreased to 150% when pretreated with the comparative bacteria LGG. L. helveticus It was found that CKDB001 lowered the TNFα mRNA expression level to 104% and was more effective than LGG. When 4 bacteria were mixed and pretreated, it was lowered to 105%, and the effect of suppressing TNFα mRNA expression was found to be very excellent (FIG. 5A).

5.2 Effect of reducing the expression level of inflammatory cytokine mRNA in the duodenal epithelial cell line (HUTU-80) of the mixed strain

In the control group in which only 2.5% ethanol was reacted in the duodenal cell line, the expression level of IL-1β mRNA increased to 427% compared to normal, and decreased to 217% when pretreated with the comparative group LGG. B. bifidum CKDB001, L. plantarum CKDB008 lowered IL-1β mRNA expression to 168% and 212%, respectively, and was found to be more effective than LGG. When pre-treated by mixing four bacteria, IL-1β mRNA expression was significantly lowered to 150%, synergistic. Appeared.

In the case of TNFα, expression was increased to 411% compared to normal when treated with 2.5% ethanol, and decreased to 266% when pretreated with the comparative group LGG. B. bifidum CKDB001, L. plantarum CKDB008 lowered the mRNA expression levels to 143% and 150%, respectively, and was found to be more effective than LGG, and when 4 bacteria were mixed and pretreated, the TNFα mRNA expression level was significantly lower to 138%, which was the most effective (FIG. 5B). .

5.3 Sintering

The present inventors confirmed from the above results that the mixed strain of the present invention is more effective in suppressing inflammatory cytokine expression compared to a single strain in both ileum cell line and duodenal cell line.

6. Conclusion

In order to develop a probiotic mixture that effectively prevents intestinal damage caused by alcohol, each intestine is subdivided to verify the efficacy of the strain for each site. 17 strains were selected from 530 types of lactic acid bacteria, which confirmed safety, stability, and industrial potential. After that, 12 species with excellent intestinal adhesion were selected, and among them, 7 strains capable of improving the cell viability due to alcohol were selected second.

Finally, four strains that effectively lower the inflammatory cytokine expression increased due to alcohol were finally selected, and it was confirmed that the mixed strains of the four selected strains can effectively prevent inflammation of intestinal cells caused by alcohol than each single strain. This mixed strain can be said to be an excavation in that it exhibits an excellent effect than LGG bacteria, which has been proven to be effective against intestinal adhesion and anti-inflammatory properties.

In addition, most of the existing studies are studies to prevent and treat liver damage caused by alcohol, and a routine method of measuring blood alcohol concentration, inflammatory response in the liver, and histological results was used. In this study, it is significant in that research was conducted to prevent alcoholic intestinal damage as well as all of the intestinal tract by confirming the degree of damage and relief of the inflammatory response.

[Accession Number]

Depository name: Korea Institute of Biotechnology

Accession number: KCTC13669BP

Date of Deposit: 20181023

Depository name: Korea Institute of Biotechnology

Accession number: KCTC13670BP

Date of Deposit: 20181023

Depository name: Korea Institute of Biotechnology

Accession number: KCTC13673BP

Date of Deposit: 20181023

Depository name: Korea Institute of Biotechnology

Accession number: KCTC13114BP

Date of accession: 20160923

[Correction by Article 91 of the Rules 10.12.2019]

[Correction by Article 91 of the Rules 10.12.2019]

[Correction by Article 91 of the Rules 10.12.2019]

[Correction by Article 91 of the Rules 10.12.2019]

Claims (14)

1. Lactobacillus ( Lactobacillus ) genus lactic acid bacteria, Bifidobacterium ( Bifidobacterium ) genus lactic acid bacteria, or a composition comprising a mixture thereof.
2. The method of claim 1, wherein the Lactobacillus lactic acid bacteria are Lactobacillus bulgaricus , Lactobacillus helveticus , Lactobacillus helveticus , Lactobacillus plantarum , or a mixture thereof.
3. The method of claim 1, wherein the Lactobacillus lactic acid bacteria are Lactobacillus bulgaricus CKDB001 (Accession No .: KCTC13669B), Lactobacillus helveticus CKDB001 (Accession No .: KCTC13670BP), Lactobacillus ( Lactobacillus plantarum ) CKDB008 (Accession No .: KCTC13673BP), or a mixture thereof, a lactic acid bacteria composition.
4. The composition of claim 1, wherein the lactic acid bacteria of the genus Bifidobacterium is Bifidobacterium bifidum .
5. According to claim 1, wherein the Bifidobacterium lactic acid bacteria are Bifidobacterium bifidum ( Bifidobacterium bifidum ) CKDB001 (Accession No. KCTC13114BP), lactic acid bacteria composition.
6. The method of claim 1, wherein the composition is Lactobacillus bulgaricus ( Lactobacillus bulgaricus ) CKDB001 (Accession No .: KCTC13669B), Lactobacillus helveticus ( Lactobacillus helveticus ) CKDB001 (Accession No .: KCTC13670BP), Lactobacillus plantarum ) CKDB008 (Accession No .: KCTC13673BP), Bifidobacterium Bifidobacterium bifidum ) CKDB001 (Accession No. KCTC13114BP), or a lactic acid bacteria composition comprising a mixed strain thereof.
7. According to claim 1, The composition is for preventing or treating alcoholic bowel damage, lactic acid bacteria composition.
8. The composition of claim 7, wherein the intestine is a small intestine.
9. The composition of claim 1, wherein the composition is for the prevention or treatment of enteritis or inflammatory bowel disease.
10. The composition of claim 9, wherein the inflammatory bowel disease is selected from the group consisting of Crohn's disease, Behcet's disease, and ulcerative colitis.
11. A method of preventing or treating alcoholic intestinal injury, comprising administering to a subject a composition comprising a lactobacillus genus of claim 1, a lactobacillus genus of Bactidobacterium, or a mixture thereof.
12. The method of claim 11, wherein the intestine is a small intestine.
13. A method of preventing or treating enteritis, or inflammatory bowel disease, comprising administering to a subject a composition comprising a lactobacillus genus of claim 1, a lactic acid bacterium of Bifidobacterium, or a mixture thereof. .
14. The method of claim 13, wherein the inflammatory bowel disease is selected from the group consisting of Crohn's disease, Behcet's disease, and ulcerative colitis.

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