KR20230018758A - COMPOSITION FOR PREVENTING OR TREATING OBESITY OR NON-ALCOHOLIC FATTY LIVER COMPRISING Bifidobacterium animalis lactis MG741 - Google Patents

Abstract

The present invention relates to a composition for preventing or treating obesity or non-alcoholic fatty liver containing a Bifidobacterium animalis lactis MG741 strain. The Bifidobacterium animalis lactis MG741 strain isolated from feces of infants selected in the present invention is not only safe, exhibits properties suitable for probiotics, reduces body weight and body fat, and has an excellent effect of regulating the expression of proteins related to lipid metabolism. Therefore, the composition can be widely used in various fields for the prevention, alleviation and treatment of obesity or non-alcoholic fatty liver.

Description

Composition for prevention or treatment of obesity or non-alcoholic fatty liver containing Bifidobacterium animalis lactis MG741 strain

The present invention relates to a composition for preventing or treating obesity or non-alcoholic fatty liver containing Bifidobacterium animalis lactis MG741.

Lactic acid bacteria have special physiological activity and are generally regarded as safe bacteria (GRAS). Lactic acid bacteria are not only used in the production of various fermented foods, but are also widely used in dairy products and fermented fruit and vegetable products with functional and probiotic properties. Recently, as consumer demand for natural supplements that replace chemical supplements increases, they are emerging as alternatives. Probiotics are live microorganisms that have a very beneficial effect on the health of the host animal, improving the balance of the intestinal microflora and enhancing the absorption of nutrients. In addition, probiotics have the characteristics of showing the antibacterial action of pathogenic microorganisms in the intestinal environment. Probiotics include a variety of microorganisms, but the genera Lactobacillus and Bifidobacterium account for the most distribution.

Obesity is known to be a major cause of heart disease, cancer, arthritis, and diabetes, and although public awareness of obesity is increasing, the number of obese patients is continuously increasing. Obesity appears as the number of adipocytes increases and the lipid content of adipocytes increases as a result of adipogenesis. Adipocytes play a major role in synthesizing and storing excess calories as triglycerides, and as a result of adipocyte differentiation, the size and number of adipocytes increase and intracellular lipid accumulation is accelerated. In addition, obesity causes lipotoxicity and oxidative stress, and induces insulin resistance, which can cause hyperglycemia, which is a cause of various obesity-related diseases.

In addition, non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and is known to be closely related to type 2 diabetes, obesity and metabolic syndrome. Patients with non-alcoholic fatty liver disease are rapidly increasing in Korea due to westernized eating habits, obesity and an increase in the diabetic population. It is expected This is an average annual growth rate of 45%. However, the exact pathogenesis of non-alcoholic fatty liver disease has not been identified, the effects of drug treatments reported so far are not sufficient in the number of subjects studied, and there is no officially approved treatment system yet. Therefore, stability is secured even during long-term use, and there is a need for a new treatment that can exert a clear therapeutic effect on NAFLD.

However, conventional treatments have side effects such as heart disease, respiratory disease, and nervous system disease, as well as low durability of their efficacy, so it is necessary to develop more improved treatments, and products currently being developed have satisfactory therapeutic effects without side effects. As there is little, there is a demand for the development of new treatments for obesity and non-alcoholic fatty liver disease.

As a result of conducting research to obtain new functional probiotics, the inventors of the present invention confirmed that lactic acid bacteria isolated from feces of infants exhibit properties suitable for probiotics and can be effectively used for the treatment of obesity and non-alcoholic fat ducts, and the present invention completed.

Accordingly, an object of the present invention is to provide a composition for preventing or treating obesity or non-alcoholic fatty liver disease.

In order to achieve the above object, the present invention provides a Bifidobacterium animalis lactis MG741 strain.

In addition, the present invention provides a pharmaceutical composition for preventing or treating obesity or non-alcoholic fatty liver, comprising the strain, its culture medium or its cell-free supernatant.

In addition, the present invention provides a health functional food composition for preventing or improving obesity or non-alcoholic fatty liver, comprising the strain, its culture medium or its cell-free supernatant.

In addition, the present invention provides a feed additive composition for preventing or improving obesity or non-alcoholic fatty liver, including the strain, its culture medium or its cell-free supernatant.

The Bifidobacterium animalis lactis MG741 strain isolated from feces of infants selected in the present invention is not only safe, but also exhibits properties suitable for probiotics, reduces body weight and body fat, and regulates the expression of proteins related to lipid metabolism. Since it is excellent, it can be widely used in various fields for the prevention, improvement, and treatment of obesity or non-alcoholic fatty liver.

1 is a diagram showing the results of body composition (A) and body fat mass (B) analysis in a mouse model.
Figure 2 is a diagram showing the results of weight gain (A) and epididymal fat (B) analysis in the mouse model.
Figure 3 is a diagram showing changes in endotoxin (A), total cholesterol (B), and LDL cholesterol (C) in serum of mice in a mouse model.
4 is a diagram showing changes in ALT (A) and AST (B) in serum in a mouse model.
Figure 5 is a diagram showing changes in liver tissue weight (A) and neutral lipids (B) in liver tissue in a mouse model.
Figure 6 is a diagram showing the results of measuring lipid metabolism-related protein expression changes (A) and SREBP1 (B), ChREBP (C), FAS (D), and ACC (E) expression levels in liver tissue of a mouse model.
Figure 7 is a diagram showing the blood leptin (A), fasting blood sugar (B), insulin (C) content and HOMA-IR (D) levels of the mouse model.
8 is a view showing the results of H&E staining and steatosis analysis using mouse model liver tissue (A), Oil red O staining and fat amount quantification results (B), and H&E staining and size analysis results of epididymal white fat (C).

Hereinafter, the present invention will be described in detail.

The present invention relates to Bifidobacterium animalis sub sp. lactis strain MG741.

The strain is a novel strain isolated from feces of infants, characterized in that it is a strain derived from the human body and is safe when ingested in the body, and in particular, since it was isolated from infants, it is characterized as a lactic acid bacterium that can be safely ingested by infants as well as adults.

The Bifidobacterium animalis sub sp. lactis MG741 strain of the present invention is a novel strain capable of preventing, treating or improving obesity or non-alcoholic fatty liver, and contains 16S rRNA represented by SEQ ID NO: 1 It is characterized in that it includes. The strain was deposited with the Korea Research Institute of Bioscience and Biotechnology on January 4, 2018, and was given accession number KCTC 13453BP.

The Bifidobacterium animalis lactis MG741 strain of the present invention has an excellent effect of reducing body weight and fat mass and regulating the expression of proteins related to lipid metabolism.

Accordingly, the present invention provides a composition for preventing, improving or treating obesity or non-alcoholic fatty liver, comprising the Bifidobacterium animalis lactis MG741 strain, its culture medium or its cell-free supernatant.

The composition includes a pharmaceutical composition, a food composition, a health functional food composition, a composition for adding feed, and the like.

More specifically, the present invention provides a pharmaceutical composition for preventing or treating obesity or non-alcoholic fatty liver, comprising the Bifidobacterium animalis lactis MG741 strain, its culture medium or its cell-free supernatant.

In the present invention, the "culture medium" is the entire strain, including the strain obtained by culturing the strain for a certain period of time in a medium capable of supplying nutrients so that the strain can grow and survive in vitro, its metabolites, extra nutrients, etc. It means a medium, but it is a concept that includes all of the supernatant from which the strain is removed after culturing the strain, and their extracts and fractions. The liquid from which the cells are removed from the culture medium is also called "supernatant" or "cell-free supernatant". It can be obtained by centrifuging to remove the sediment at the bottom and taking only the liquid at the top.

As used herein, the term "prevention" may refer to any action that suppresses or delays the onset of obesity-related diseases by administering the composition for preventing or treating obesity or non-alcoholic fatty liver disease according to the present invention to a subject.

In the present invention, the term "treatment" may refer to all actions to improve or benefit symptoms of related diseases by administering the composition according to the present invention to a subject suspected of having obesity or non-alcoholic fatty liver disease.

As used herein, the term "improvement" may refer to any activity that at least reduces a parameter associated with a condition to be treated, for example, the severity of a symptom.

In the present invention, the term "individual" may refer to all animals, including humans, who have or are likely to develop obesity or non-alcoholic fatty liver disease.

The composition of the present invention may further contain at least one known active ingredient having an anti-cancer effect on obesity or non-alcoholic fatty liver along with Bifidobacterium animalis lactis MG741 strain.

The pharmaceutical composition according to the present invention may contain a pharmaceutically effective amount of Bifidobacterium animalis lactis MG741 strain alone or may contain one or more pharmaceutically acceptable carriers, excipients or diluents.

The above "pharmaceutically effective amount" refers to an amount sufficient to exert an effect of preventing, improving or treating obesity or non-alcoholic fatty liver. The term "pharmaceutically acceptable" refers to a composition that is physiologically acceptable and does not usually cause allergic reactions such as gastrointestinal disorders and dizziness or similar reactions when administered to humans.

In addition, the pharmaceutical composition according to the present invention is formulated according to conventional methods into oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injection solutions. can Suitable formulations known in the art are preferably those disclosed in the literature (Remington's Pharmaceutical Science, recently, Mack Publishing Company, Easton PA). Carriers, excipients and diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginates, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate, mineral oil, and the like. When formulating the pharmaceutical composition, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations contain at least one excipient such as starch, calcium carbonate, sucrose, and lactose in the composition. , gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included. there is. Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents. As a base for the suppository, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogeratin and the like may be used.

The pharmaceutical composition of the present invention is the amount of active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human thought by a researcher, veterinarian, physician or other clinician, that is, the symptom of the disease or disorder being treated. It can be administered in a therapeutically effective amount that is an amount that induces remission. It is apparent to those skilled in the art that the therapeutically effective dosage and frequency of administration of the pharmaceutical composition of the present invention will vary depending on the desired effect. Therefore, the optimal dose to be administered can be easily determined by those skilled in the art, and depends on the type of disease, the severity of the disease, the content of the active ingredient and other ingredients contained in the composition, the type of formulation, and the age, weight, and general health of the patient. Condition, sex and diet, administration time, administration route and secretion rate of the composition, treatment period, can be adjusted according to various factors including drugs used simultaneously. For desirable effects, the pharmaceutical composition of the present invention may be administered in an amount of 1 to 10,000 mg/kg/day, preferably 1 to 200 mg/kg/day, and may be administered once a day or several times. It can also be administered in divided doses.

The pharmaceutical composition of the present invention can be administered to a subject by various routes. All modes of administration are contemplated, eg oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine intrathecal or intracerebrovascular injection.

In addition, the present invention provides a health functional food composition for preventing or improving obesity or non-alcoholic fatty liver, comprising the Bifidobacterium animalis lactis MG741 strain, its culture medium or its cell-free supernatant.

In the case of the health functional food composition of the present invention, the type of food is not particularly limited, and may include all foods in a conventional sense. Non-limiting examples of foods to which the substance can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea , drinks, alcoholic beverages, and vitamin complexes. When using the composition as a food additive, the composition may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to conventional methods.

In this specification, food means a natural product or processed product containing one or more nutrients, preferably means that it is in a state that can be eaten directly through a certain degree of processing process, and is a conventional meaning, The food composition is intended to include all foods, food additives, health functional foods and beverages.

Foods to which the composition of the present invention can be added include, for example, various foods, beverages, gum, candy, tea, vitamin complexes, and functional foods. In addition, in the present invention, the food includes special nutritious food (eg, formula milk, infant food, baby food, etc.), processed meat product, fish meat product, tofu, jelly, noodles (eg, ramen, noodles, etc.), health supplement food, seasoning food ( For example, soy sauce, soybean paste, red pepper paste, mixed soybean paste, etc.), sauces, confectionery (eg, snacks), dairy products (eg, fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (various types of kimchi, pickles, etc.), beverages (eg, fruit and vegetable beverages, soy milk, fermented beverages, ice cream, etc.), natural seasonings (eg, ramen soup, etc.), vitamin complexes, alcoholic beverages, alcoholic beverages, and other health supplements, but are not limited thereto. The food, beverage or food additive may be prepared by a conventional manufacturing method.

In the present invention, the term "health functional food" refers to a food manufactured and processed by using a specific ingredient as a raw material or by extracting, concentrating, refining, mixing, etc. a specific ingredient contained in a food raw material for the purpose of health supplement. It refers to food designed and processed so that it can sufficiently exert biological control functions such as biological defense, regulation of circadian rhythm, prevention and recovery of disease, etc. say what you can

When the Bifidobacterium animalis lactis MG741 strain according to the present invention is used as a health functional food, it can be added as it is or used together with other foods or food ingredients, which can be selected and used appropriately as needed.

Since the Bifidobacterium animalis lactis MG741 strain according to the present invention guarantees biosafety and shows an effect of increasing activity in proportion to the concentration, it is obvious that it can be used in an appropriate amount without being limited to a specific range.

In addition, there is no particular limitation on the type of health functional food in which the strain according to the present invention can be used. For example, ramen, other noodles, beverages, tea, drinks, alcoholic beverages, various soups, meat, sausages, bread, chocolate, candy, confectionery, pizza, chewing gum, dairy products including ice cream, or vitamin complexes.

In addition, the present invention provides a feed additive composition for preventing or improving obesity or non-alcoholic fatty liver, comprising a Bifidobacterium animalis lactis MG741 strain, its culture medium or its cell-free supernatant.

The feed additive of the present invention corresponds to supplementary feed under the Feed Management Act.

In the present invention, the term "feed" may refer to any natural or artificial diet, one meal, etc., or a component of the one meal meal, for or suitable for eating, ingesting, and digesting by an animal. The type of feed is not particularly limited, and feeds commonly used in the art may be used. Non-limiting examples of the feed include vegetable feeds such as grains, root fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal or grain by-products; Animal feed such as proteins, inorganic materials, oils, mineral oils, oils, single cell proteins, zooplankton, or food may be mentioned. These may be used alone or in combination of two or more.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are intended to illustrate the present invention by way of example, and the scope of the present invention is not limited to these examples.

Example 1. Identification of strain isolation from infant feces

Feces were collected from healthy Korean infants (2-15 months old). After mixing 9 mL of 0.85% saline solution with 1 g of feces and stirring for 3 minutes with a Stomacher (simple mixer, 3M), large particles or residues were removed through Whatman No.2 paper. After filtering, 1 mL was taken, diluted to 10 ⁸ with 0.85% saline dilution, poured into Raffinose-bifidobacterium (RB) agar, incubated at 37°C for 24 hours, and colonies were confirmed. The formed colonies were inoculated into MRS medium to which BCP was added to confirm whether yellow rings were formed, and then the colonies forming yellow rings were checked for Gram staining (positive), biochemical reaction (catalase negative), and morphology (rod). After that, the primary bacterial species was confirmed through the API 50 CHL kit.

Through the analysis of 16S rRNA among the primary isolated strains, a novel Bifidobacterium animalis sub sp. lactis (hereinafter referred to as Bifidobacterium animalis lactis), which was named MG741, prepared as a 20% glycerol stock and stored at -80°C. The 16S rRNA base sequence of the MG741 strain is shown in SEQ ID NO: 1.

Example 2. Evaluation of strain characteristics

In order to confirm the characteristics of the Bifidobacterium animalis lactis MG741 strain isolated and identified in Example 1, the enzyme activity was confirmed through the API ZYM Kit (Biomeriux, Co. France). The results are shown in Table 1.

Enzymes assayed for Substrate 741 Control (Negative) 0 Alkaline phosphatase 2-naphthyl phosphate One Esterase (C4) 2-naphthyl butyrate 0 Esterase Lipase (C8) 2-naphthyl caprylate 0 Lipase (C14) 2-naphthyl myristate 0 Leucine arylamidase L-leucyl-2-naphthylamide 4 Valine arylamidase L-valyl-2-naphthylamide 0 Crystine arylamidase L-cystyl-2-naphthylamide One Trypsin N-benzoyl-DL-arginine-2-naphthylamide 0 α-chymotrypsin N-glutaryl-phenylanine-2-naphthylamide One Acid phosphatase 2-naphtyl phosphate 0 Naphtol-AS-BI-phosphohydrolase Naphthol-AS-BI-phosphate 4 α-galactosidase 6-Br-2-naphthyl- α D-galactopyranoside 2 β-galactosidase 2-naphthyl- β D-galactopyranoside 3 β-glucuronidase Naphthol-AS-BI-β 0 α-glucosidase 2-naphthyl-αD-glucopyranoside 2 β-glucosidase 6-Br-2-naphthyl-β 0 N-acetyl-βglucosaminidase 1-naphthyl-N-acetyl-β 0 α-mannosidase 6-Br-2-naphthyl-αD-mannopyranoside 0 α-fucosidase 2-naphthyl-αL-fucopyranoside 0

As shown in Table 1, it was confirmed that the enzymes showing the enzyme activity score of 3 or higher in strain MG741 were Leucine arylamidase, Naphtol-AS-BI-phosphohydrolase and β-galactosidase.

Next, in order to confirm the carbohydrate fermentation profile, API 50CHL kit (Biomeriux, France) was used. The results are shown in Table 2.

Substrate MG741 Substrate MG741 Control (Negative) - Esculin + Glycerol - Salicin + Erythritol - D-cellobiose - D-arabinose - D-maltose + L-arabinose + D-lactose + D-ribose + D-melibose + D-xylose + D-sucrose + L-xylose - D-trehalose - D-adonitol - Inulin - Methyl-β D-xylopyranoside - D-melezitose - D-galactose + D-raffinose + D-glucose + Starch + D-fructose - Glycogen - D-mannose + Xylitol - L-sorbose + Gentiobiose + L-rhamnose - D-turanose - Dulcitol - D-lyxose + Inositol - D-tagatose - D-mannitol - D-fucose - D-sorbitol - L-fucose - Methyl-α D-Mannoside - D-arabitol - Methyl-α D-glucoside + L-arabirol - N-acethyl-glucosamine - Gluconate - Amygdalin + 2-keto-gluconate - Arbutin + 5-keto-gluconate -

As shown in Table 2, strain MG741 is L-arabinose, D-ribose, D-xylose, D-galactose, D-glucose, D-mannose, D-sorbose, methyl-alpha-D-glucoside, amygdalin , arbutin, esculin, salicin, D-maltose, D-lactose, D-melibiose, D-sucrose, D-raffinose, starch, gentiobiose, and D-lyxose. Confirmed.

Example 3. Obesity and non-alcoholic activity evaluation method

3-1. Obesity and non-alcoholic fatty liver model construction

Forty-five 6-week-old mice (C57BL/6J, male) were distributed from the central laboratory and allowed to acclimatize for one week. Thereafter, the mice were divided into normal diet group (ND), experimental control group (High fat diet group, HFD), and sample group (MG741, MD6), and each substance was orally administered for 12 weeks. In the case of the sample group, the Bifidobacterium animalis lactis MG741 strain isolated in Example 1 was dissolved in physiological saline and administered in an amount of 10 ⁶ CFU. Obesity and non-alcoholic fatty liver were induced in all groups except the normal group by providing a high-fat diet. Body weight and feed intake were measured at intervals of one week from the start of the experiment.

3-2. Serological analysis

After 12 weeks of induction of obesity and non-alcoholic fatty liver according to Example 3-1, the experimental animals were fasted for 18 hours, anesthetized, blood was collected from the abdominal vena cava, and serum was separated by centrifugation at 3,000 rpm for 15 minutes. The separated serum was stored in a -70°C freezer until analysis.

The content of aminotransferase (ALT) and aspartic acid aminotransferase (AST) in serum was determined by Cell Biolabs, Inc. (Beverly, MA, USA), and the total cholesterol, high density lipoprotein (HDL) cholesterol, and triglyceride content in the liver were measured by purchasing a kit from Abcam (Cambridge, MA, USA). The contents of endotoxin (Enotoxin) and insulin (Insulin) were measured by purchasing a kit from Thermo fisher Waltham, MA, USA). HOMA-IR was calculated as fasting blood glucose × insulin concentration ÷ 405.

3-3. Protein expression analysis through western blot

For the analysis of obesity and non-alcoholic fatty liver biomarkers, a lysis buffer containing a protease inhibitor was added to mouse liver tissue, and then homogenized and centrifuged at 10,000 × g for 5 minutes to obtain a supernatant. After quantifying the protein, the same amount of protein was electrophoresed using 10% SDS-polyacrylamide gel (Bio-Rad, CA, USA) and then transferred to a PVDF membrane (Bio-Rad, CA, USA). After blocking non-specific binding sites in 5% skim milk (0.1% Tween 20 containing PBS, PBST) solution, primary antibodies (anti-β-actin, anti-SREBP1c, anti-ACC, anti-FAS, anti -ChREBP, Abcam) at 4°C overnight, incubated with secondary antibody (anti-rabbit IgG or anti-mouse IgG linked with horseradish peroxidase, Cell signaling Technology) for 1 hour at room temperature, streptavidin-horseradish peroxidase and It was detected using a ChemiDoc XRS device (Bio-Rad, USA).

3-4 Histological analysis

For histological analysis, hematoxylin and eosin (H&E) staining and Oil Red O (ORO) staining were performed. For H&E staining, liver and white fat were fixed in 4% formaldehyde and paraffinized. Tissues were then cut and stained with H&E. In addition, for ORO staining, liver tissue was frozen in OCT solution and cut. After that, the sections were fixed in 4% formaldehyde and then stained with ORO. All slides were scanned and analyzed using a pannoramic 250 Flash III slide scanner (3DHistech, Ltd., Budapest, Hungary) and CaseViewer software (3DHistech, Ltd).

3-5. statistical processing

The experimental results were expressed as mean ± standard error. Statistical analysis between the experimental groups was performed by one-way analysis of variance (ANOVA) using SPSS Statistics 20 software (IBM Inc, Chicago, IL, USA). For the items, statistical significance was tested at the level of p<0.05 by Tukey's multiple range test.

Example 4. Verification of treatment effect on obesity and non-alcoholic fatty liver

4-1. Effects on body weight and fat mass

In order to confirm the effect of the MG741 strain according to the present invention on body weight and fat mass, body fat mass was measured using a body composition analyzer. In addition, the body weight was measured during the experiment period (8 weeks), and after the experiment was completed, the white epididymal adipose tissue was removed and the weight measurement result was confirmed. The results are shown in Figures 1 and 2.

As shown in FIG. 1, as a result of body fat mass measurement, it was confirmed that the control group showed an increase in body fat mass by 111.2% compared to the normal group, whereas the MG741 administration group (MD6) showed a decrease in body fat mass by about 20.7% compared to the experimental control group.

In addition, as shown in FIG. 2, during the experimental period, the weight of the normal group increased by 6.6 g, while the experimental control group increased by 24.2 g, and the MG741-administered group (MD6) showed a decrease in body weight by about 22.4% compared to the experimental control group. In addition, the amount of fat extracted from the experimental animals increased in the experimental control group compared to the normal group, while the MD6-administered group decreased by 14.5% compared to the experimental control group.

Through the above experimental results, it was confirmed that the MG741 strain of the present invention has an excellent anti-obesity effect of inhibiting weight gain and reducing body fat mass and weight of adipose tissue.

4-2. Effects on serum endotoxins and lipid concentrations

Serum endotoxin and lipid concentrations were measured, and the results are shown in FIG. 3 .

As shown in Figure 3A, compared to the normal group, the experimental control group increased the amount of endotoxin by 164.0%, but it was confirmed that the MG741 administration group (MD6) decreased by 57.4% compared to the experimental control group.

In addition, as a result of measuring the concentration of total cholesterol and LDL cholesterol, which are greatly affected by the high-fat diet, as shown in FIGS. 3B and 3C, total cholesterol increased by 108.5% in the control group compared to the normal group, but the MG741 administration group MD6) was confirmed to be reduced by 15.9% compared to the experimental control group. In addition, LDL cholesterol increased by 198.2% in the control group compared to the normal group, but decreased by 31.1% in the MG741-administered group (MD6).

Through this, it was confirmed that the MG741 strain according to the present invention can reduce inflammation with anti-obesity effect by reducing the concentration of endotoxin in serum, help to strengthen immunity, and have excellent cholesterol reduction effect.

4-3. Effect on the concentration of enzymes related to liver function in serum

In order to observe changes in enzymes related to liver function in the blood, ALT and AST contents of serum were measured. The results are shown in FIG. 4 .

As shown in Figure 4, the ALT level in the serum was 554.3% higher in the control group than in the normal group, and decreased by 49.6% in the MG741 administration group (MD6) compared to the control group. In addition, the AST level in the serum was 145.3% higher in the control group than in the normal group, and was reduced by 36.0% in the MG741-administered group (MD6) compared to the control group.

Through this, it was confirmed that the concentration of liver function-related enzymes in the serum was decreased in the MG741 administration group, thus showing the effect of improving non-alcoholic fatty liver.

4-4. Effects on liver tissue

Liver weight and triglyceride (TG) were measured. The results are shown in FIG. 5 .

As shown in Figure 5, it was confirmed that the weight of liver tissue increased by 145.9% in the control group compared to the normal group and decreased by 21.7% in the MG741 administration group compared to the experimental control group. In addition, the neutral lipid concentration in the liver was increased by 156.0% in the experimental control group compared to the normal group, and it was confirmed that the MG741 administration group decreased by 35.0% compared to the experimental control group.

Through this, it was confirmed that the concentration of neutral lipids in the liver was decreased in the MG741 administration group, thus showing the effect of improving non-alcoholic fatty liver.

4-5. Effects on protein changes related to liver lipid metabolism

Lipid metabolism-related proteins sterol regulatory element binding protein-1 (SREBP1), carbohydrate response element binding protein (ChREBP), acetyl coenzyme A carboxylase Expression changes of acetyl CoA carboxylase (ACC) and fatty acid synthase (FAS) were confirmed. The results are shown in FIG. 6 .

As shown in FIG. 6, sterol regulatory element binding protein-1 (SREBP1), a transcription factor that regulates fat metabolism, and carbohydrate response element binding protein, a protein that promotes fat synthesis, , ChREBP) decreased by 26.8% and 20.2%, respectively, in the MG741 CFU-administered group (MD6) compared to the control group. In addition, it was confirmed that the expression of ACC and FAS, which are enzymes involved in fatty acid synthesis, decreased by 46.2% and 47.0% in the MG741 administration group compared to the control group.

Through this, it was confirmed that the MG741 strain of the present invention exhibits an improvement and therapeutic effect on non-alcoholic fatty liver through the regulation of expression of proteins related to lipid metabolism.

4-6. Effects on leptin and insulin resistance

Blood leptin content, fasting blood glucose, insulin and HOMA-IR were measured. The results are shown in FIG. 7 .

As shown in Figure 7, the content of leptin in the blood was increased by 1927.0% in the control group compared to the normal group, and it was confirmed that the MG741 administration group decreased by 37.3% compared to the experimental control group. In addition, fasting blood glucose increased by 185.5% in the control group compared to the normal group, and decreased by 18.9% in the MG741-administered group compared to the experimental control group. In addition, blood insulin concentration increased by 775.4% in the experimental control group compared to the normal group, and decreased by 61.2% in the MG741 administration group compared to the experimental control group. Finally, insulin resistance increased by 1438.7% in the control group compared to the normal group, and decreased by 68.5% in the MG741 administration group compared to the experimental control group.

Based on the above results, it was confirmed that the MG741 strain according to the present invention exhibits excellent effects in the treatment of non-alcoholic fatty liver disease by reducing the concentration of leptin in blood and reducing fasting blood glucose and insulin concentrations to improve insulin resistance.

4-7. Effects on liver and adipose tissue

H&E and ORO staining were performed to confirm the effects of long-term high-fat diet on liver and adipose tissue. The results are shown in FIG. 8 .

As shown in FIG. 8, as a result of H&E staining of the liver tissue, it was confirmed that steatosis was severely induced in the liver tissue in the control group compared to the normal group (score 0) (score 2.67), and the MG741 administration group was the experimental control group. It showed a state of 61.0% decrease compared to that. In addition, as a result of confirming the degree of fat cell staining through ORO staining of liver tissue, it was confirmed that the stained range increased by 900.6% in the control group compared to the normal group, and the MG741 administration group showed a 70.3% decrease compared to the experimental control group.

Next, as a result of confirming the expansion of adipose tissue through H&E staining, it increased by 367.0% in the control group compared to the normal group, and the size of the MG741 administration group decreased by 9.7% compared to the experimental control group.

Through the above experiments, it was confirmed that the accumulation of fat in liver tissue and the expansion of adipose tissue were significantly reduced through the administration of the MG741 strain according to the present invention, and the MG741 strain according to the present invention has an excellent effect on the treatment of non-alcoholic fatty liver and obesity confirmed that it will appear.

Through the above experiments, the Bifidobacterium animalis lactis MG741 strain, which showed excellent anti-obesity and non-alcoholic fatty liver treatment effects, was deposited with the Korea Research Institute of Bioscience and Biotechnology on January 4, 2018, and was given accession number KCTC 13453BP .

Korea Research Institute of Bioscience and Biotechnology KCTC13453BP 20180104

<110> MEDIOGEN Co.,Ltd. KOREA FOOD RESEARCH INSTITUTE <120> COMPOSITION FOR PREVENTING OR TREATING OBESITY OR NON-ALCOHOLIC FATTY LIVER COMPRISING Bifidobacterium animalis lactis MG741 <130> 1-43 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1404 <212> DNA <213> unknown <220> <223> Bifidobacterium animalis lactis MG741 16S rRNA <400> 1 agtcgaacgg gatccctggc agcttgctgt cggggtgaga gtggcgaacg ggtgagtaat 60 gcgtgaccaa cctgccctgt gcaccggaat agctcctgga aacgggtggt aataccggat 120 gctccgctcc atcgcatggt ggggtgggaa atgcttttgc ggcatgggat ggggtcgcgt 180 cctatcagct tgttggcggg gtgatggccc accaaggcgt tgacgggtag ccggcctgag 240 agggtgaccg gccacattgg gactgagata cggcccagac tcctacggga ggcagcagtg 300 gggaatattg cacaatgggc gcaagcctga tgcagcgacg ccgcgtgcgg gatggaggcc 360 ttcgggttgt aaaccgcttt tgttcaaggg caaggcacgg tttcggccgt gttgagtgga 420 ttgttcgaat aagcaccggc taactacgtg ccagcagccg cggtaatacg tagggtgcga 480 gcgttatccg gatttattgg gcgtaaaggg ctcgtaggcg gttcgtcgcg tccggtgtga 540 aagtccatcg cctaacggtg gatctgcgcc gggtacgggc gggctggagt gcggtagggg 600 agactggaat tcccggtgta acggtggaat gtgtagatat cgggaagaac accaatggcg 660 aaggcaggtc tctgggccgt cactgacgct gaggagcgaa agcgtgggga gcgaacagga 720 ttagataccc tggtagtcca cgccgtaaac ggtggatgct ggatgtgggg ccctttccac 780 gggtccccgtg tcggagccaa cgcgttaagc atcccgcctg gggagtacgg ccgcaaggct 840 aaaactcaaa gaaattgacg ggggcccgca caagcggcgg agcatgcgga ttaattcgat 900 gcaacgcgaa gaaccttacc tgggcttgac atgtgccgga tcgccgtgga gacacggttt 960 cccttcgggg ccggttcaca ggtggtgcat ggtcgtcgtc agctcgtgtc gtgagatgtt 1020 gggttaagtc ccgcaacgag cgcaaccctc gccgcatgtt gccagcgggt gatgccggga 1080 actcatgtgg gaccgccggg gtcaactcgg aggaaggtgg ggatgacgtc agatcatcat 1140 gccccttacg tccagggctt cacgcatgct acaatggccg gtacaacgcg gtgcgacacg 1200 gtgacgtggg gcggatcgct gaaaaccggt ctcagttcgg atcgcagtct gcaactcgac 1260 tgcgtgaagg cggagtcgct agtaatcgcg gatcagcaac gccgcggtga atgcgttccc 1320 gggccttgta cacaccgccc gtcaagtcat gaaagtgggt agcactccga agccggtggc 1380 ccgacccttg tggggggagc cgtc 1404

Claims (6)

Accession number KCTC13453BP, Bifidobacterium animalis lactis MG741 strain.
According to claim 1, wherein the strain is isolated from feces of infants, Bifidobacterium animalis lactis MG741 strain.
According to claim 1, wherein the strain has a 16S rRNA represented by SEQ ID NO: 1, Bifidobacterium animalis lactis MG741 strain.
A pharmaceutical composition for preventing or treating obesity or non-alcoholic fatty liver, comprising the strain according to any one of claims 1 to 3, its culture medium or its cell-free supernatant.
A functional food composition for preventing or improving obesity or non-alcoholic fatty liver, comprising the strain according to any one of claims 1 to 3, its culture medium or its cell-free supernatant.
A composition for preventing obesity or non-alcoholic fatty liver or adding feed, comprising the strain according to any one of claims 1 to 3, its culture medium or its cell-free supernatant.

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