Weissella cibaria wikim0187 having sarcopenia alleviating effect and use thereof

Abstract

The Weissella cibaria WIKIM0187 strain according to the present invention is a lactic acid bacterium isolated from kimchi, and exhibits an excellent sarcopenia alleviation effect, and thus can be variously used as a food composition, a pharmaceutical composition, an intestinal regulation composition, and a lactic acid bacterium starter, which comprise the strain as an active ingredient.

Description

Weissella cibaria WIKIM0187, which has the effect of improving sarcopenia, and its use

[0001] The present invention relates to a sarcopenia-improving composition containing Weissella cibaria WiKim0187 isolated from kimchi and containing the same.

[0002]

[0003] Sarcopenia is a disease in which muscle mass decreases with aging. When it occurs, it is a disease that not only reduces simple muscle mass, but also reduces exercise performance, immunity, and basal metabolic rate, and increases the incidence of chronic diseases such as diabetes, obesity, and cardiovascular disease, lowering the patient's survival rate. Also called muscle atrophy, it can be defined as a loss of size and mass of muscle cells and muscle tissue that occurs due to long-term inability to move due to aging, severe illness, nerve damage, etc., or due to nutritional disorders.

[0004] Muscles are broadly categorized into skeletal muscle, cardiac muscle, and visceral muscle. Of these, skeletal muscle is the most abundant tissue in the human body, accounting for 40-45% of body weight. Skeletal muscle is attached to bones via tendons and plays a role in generating bone movement and force. Skeletal muscle has the ability to regenerate and maintain itself depending on the environment, but this characteristic is lost with age. Consequently, as aging progresses, not only does muscle mass decrease, but muscle strength also decreases.

[0005] Muscle mass and strength peak around age 30, begin to decline in the 40s, and decline at a rate that doubles after age 70. Consequently, muscle mass decreases by approximately 50% between the ages of 20 and 90. Age-related muscle changes include a decrease in the number and size of muscle cells, changes in the ratio of muscle fiber types, decreased neuromuscular junction efficiency, intramuscular fat infiltration, decreased mitochondria and enzymes, decreased actin-myosin ratios within muscle fiber filaments, and delayed recovery from muscle damage.

[0006] Sarcopenia is caused by reduced physical activity, malnutrition, environmental factors, disease, inflammation, mitochondrial abnormalities, and hormonal changes. Genetic differences are also reported. Once sarcopenia develops, it rapidly worsens. When muscle loss due to sarcopenia becomes severe, the ability to store energy decreases, making one feel easily fatigued. The basal metabolic rate decreases, causing frequent weight fluctuations and easy weight gain. In other words, sarcopenia is a metabolic disease that directly causes muscle loss, which increases the risk of death due to decline and impairment of various bodily functions. In addition, it is also a cause of increased prevalence of metabolic diseases such as hypertension, diabetes, arthritis, obesity, and cancer due to decreased metabolism and weakened immunity.

[0007] International pharmaceutical companies and even domestic companies are developing and conducting Phase 2-4 clinical trials of biologics targeting myostatin and activin receptors to treat sarcopenia. However, these drugs have not shown significant efficacy in clinical trials or have caused adverse side effects. Currently, the market for treatments for age-related sarcopenia consists primarily of pain-relieving medications, exercise, hormone therapy, and dietary therapies. None of the proposed medications are currently available due to side effects or low efficacy.

[0008] The definition and diagnostic criteria for sarcopenia have recently been established, leading to growing interest in the condition. The rapid increase in sarcopenia patients due to aging is resulting in significant socioeconomic costs. Until a treatment is developed, prevention of sarcopenia is essential, and exercise and nutrition are the only effective treatments for improving it.

[0009] Therefore, various food materials should be developed and utilized to develop fundamental prevention and treatment methods for sarcopenia.

[0010] [Prior Art Literature]

[0011] [Patent Document]

[0012] (Patent Document 1) Domestic Registered Patent No. 10-2606636

[0013]

[0014] Accordingly, the inventors of the present invention have attempted to find a lactic acid bacteria strain that exhibits an effect of improving sarcopenia, and as a result, have isolated and identified a novel lactic acid bacteria strain, Weissella cibaria WiKim0187, thereby completing the present invention.

[0015]

[0016] Accordingly, the purpose of the present invention is to provide Weissella cibaria WiKim0187 having the accession number KCCM13568P.

[0017] Another object of the present invention is to provide a food composition for improving sarcopenia, which comprises Weissella cibaria WiKim0187 (Accession No. KCCM13568P), a culture thereof, a pulverized product thereof, or an extract thereof as an active ingredient.

[0018] Another object of the present invention is to provide a pharmaceutical composition for improving sarcopenia, which comprises Weissella cibaria WiKim0187 (Accession No. KCCM13568P), a culture thereof, a lysate thereof, or an extract thereof as an active ingredient.

[0019] In addition, another object of the present invention is to provide a composition for a lactic acid bacteria starter for fermentation and a composition for a lactic acid bacteria starter for a fermentation comprising Weissella cibaria WiKim0187 (Accession No. KCCM13568P), a culture thereof, a pulverized product thereof or an extract thereof as an active ingredient.

[0020] In addition, another object of the present invention is to provide a method for improving or treating sarcopenia, which comprises administering a therapeutically effective amount of Weissella cibaria WiKim0187 (accession number KCCM13568P), a culture thereof, a lysate thereof, or an extract thereof to a subject in need thereof.

[0021] In addition, another object of the present invention is to provide a use of Weissella cibaria WiKim0187, accession number KCCM13568P, for improving sarcopenia.

[0022]

[0023] The present invention relates to a strain, Weissella cibaria WiKim0187, which is isolated from kimchi and has the effect of improving sarcopenia. The strain can be used in pharmaceutical compositions for preventing or treating sarcopenia, which contain the strain as an active ingredient, and can be used in functional foods such as starters for fermented foods.

[0024]

[0025] Figure 1 shows the 16S rRNA identification results of Weissella cibaria WiKim0187.

[0026] Figure 2 shows the results of hemolytic assay of Weissella cibaria WiKim0187.

[0027] Figure 3 shows the results of the forepaw grip strength test of Weissella cibaria WiKim0187 in an aging animal model of muscle loss.

[0028] Figure 4 shows the results of a forepaw grip strength test according to the intake of kimchi lactic acid bacteria in an aging animal model with muscle loss.

[0029] Figure 5 shows the results of DXA (Dual-energy X-ray absorptiometry) of Weissella cibaria WiKim0187 in a sarcopenic aging animal model.

[0030] Figure 6 shows the results of liver and hindlimb muscle tissue weights of Weissella cibaria WiKim0187 in a sarcopenic aging animal model.

[0031] Figure 7 shows the results of verifying the efficacy of improving muscle loss by confirming the expression of muscle loss marker mRNA in muscle tissue of Weissella cibaria WiKim0187.

[0032] Figure 8 shows the results of verifying the efficacy of improving muscle loss by confirming the expression of muscle differentiation marker mRNA in muscle tissue of Weissella cibaria WiKim0187.

[0033]

[0034] Hereinafter, the present invention will be described in more detail as follows.

[0035] The present invention relates to Weissella cibaria WiKim0187 and uses thereof.

[0036] The Weissella cibaria WiKim0187 strain of the present invention is a novel strain derived from kimchi, a fermented food of Korea.

[0037] The lactic acid bacteria strain isolated through the examples of the present invention was found to have a nucleic acid sequence of SEQ ID NO: 1 as a result of 16S rRNA base sequence analysis for identification and classification of microorganisms.

[0038] Accordingly, the microorganism of the present invention having the 16S rRNA base sequence of SEQ ID NO: 1 was named Weissella cibaria WiKim0187 (accession number KCCM13568P) and deposited at the Korea Microbiological Conservation Center on December 29, 2023 (domestic accession number KFCC11987P, international accession number KCCM13568P).

[0039] The Weissella cibaria WiKim0187 of the present invention is a gram-positive bacterium and a facultative anaerobe capable of growing under both aerobic and anaerobic conditions. It does not form spores, is not motile, and its cells take the form of rods.

[0040] In the following examples, Weissella cibaria WiKim0187 was selected and it was confirmed that Weissella cibaria WiKim0187 exhibited an excellent effect in improving sarcopenia.

[0041] The term "improvement" as used herein may mean any action that at least reduces a parameter associated with the condition being treated, for example, the severity of a symptom.

[0042] Meanwhile, safety must be ensured for the application of probiotic lactic acid bacteria to food. Accordingly, hemolytic toxicity of Weissella cibaria WiKim0187 was confirmed through a safety test. As a result of the hemolytic test, Weissella cibaria WiKim0187 of the present invention did not exhibit hemolysis.

[0043] Therefore, the Weissella cibaria WiKim0187 strain of the present invention can be utilized in various fields as a safe probiotic lactic acid bacterium for improving sarcopenia.

[0044] Accordingly, the present invention provides a food composition comprising Weissella cibaria WiKim0187, accession number KCCM13568P, a culture thereof, a pulverized product thereof, or an extract thereof as an active ingredient. The food composition may be in the form of a health functional food, a beverage, a bar, or the like.

[0045] Another aspect of the present invention is a health functional food composition for improving sarcopenia, comprising Weissella cibaria WiKim0187, accession number KCCM13568P, a culture thereof, a pulverized product thereof, or an extract thereof as an active ingredient.

[0046] The term "health functional food" in this specification means a food manufactured and processed using raw materials or ingredients with functionality useful to the human body in accordance with the Health Functional Food Act (Article 3, Paragraph 1), and "functionality" means obtaining a useful effect for health purposes, such as regulating nutrients or physiological actions for the structure and function of the human body (Article 2, Paragraph 2).

[0047] The food composition of the present invention can be used as a functional food or added to various foods. Foods that can be added to the composition of the present invention include, for example, beverages, vitamin complexes, and health supplements.

[0048] The food composition of the present invention may include ingredients commonly added during food manufacturing, such as proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. Examples of the carbohydrates mentioned above include monosaccharides such as glucose, fructose, etc.; disaccharides such as maltose, sucrose, oligosaccharides, etc.; and polysaccharides such as dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [thaumatin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (e.g., saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is manufactured into drinks and beverages, citric acid, high fructose corn syrup, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts, etc. can be additionally included.

[0049] The above food composition may additionally contain food additives, and its suitability as a “food additive” is determined by the specifications and standards for the relevant item in accordance with the general provisions and general test methods of the Food Additives Codex approved by the Ministry of Food and Drug Safety, unless otherwise specified.

[0050] Examples of items listed in the above "Food Additives Codex" include chemically synthesized products such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, and guar gum; and mixed preparations such as sodium L-glutamate preparations, alkaline agents for noodles, preservative preparations, and tar color preparations.

[0051] Foods containing the active ingredient of the present invention include confectionery such as bread, rice cakes, dried fruits, candies, chocolates, chewing gums, and jams; ice cream products such as ice cream, frozen desserts, and ice cream powders; dairy products such as milk, low-fat milk, lactose-hydrolyzed milk, processed milk, goat milk, fermented milk, buttermilk, condensed milk, milk cream, buttermilk, natural cheese, processed cheese, powdered milk, and whey; meat products such as processed meat products, processed egg products, and hamburgers; fish products such as processed fish products such as fish cakes, ham, sausage, and bacon; noodles such as ramen, dried noodles, fresh noodles, fried noodles, luxurious dried noodles, improved boiled noodles, frozen noodles, and pasta; beverages such as fruit drinks, vegetable drinks, carbonated drinks, soy milk, and yogurt, and mixed drinks; soy sauce, soybean paste, red pepper paste, soybean paste, cheonggukjang, and mixed sauce; Condiments such as vinegar, sauces, tomato ketchup, curry, dressings, margarine, shortening, and pizza are included, but are not limited to these.

[0052] In addition to the above, the composition of the present invention may include various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the composition of the present invention may include fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These ingredients may be used independently or in combination.

[0053] The beverage composition including the active ingredient of the present invention has no particular limitations on other ingredients, and may contain various flavoring agents or natural carbohydrates as additional ingredients, like conventional beverages. Examples of the above-mentioned natural carbohydrates include conventional sugars such as monosaccharides (e.g., glucose, fructose, etc.); disaccharides (e.g., maltose, sucrose, etc.); and polysaccharides (e.g., dextrin, cyclodextrin, etc.), and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (e.g., thaumatin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (e.g., saccharin, aspartame, etc.) can be advantageously used.

[0054] In another aspect, the present invention provides a pharmaceutical composition for preventing or treating sarcopenia, comprising Weissella cibaria WiKim0187, accession number KCCM13568P, a culture thereof, a lysate thereof, or an extract thereof as an active ingredient.

[0055] In the present invention, “sarcopenia” refers to a disease in which muscle protein breakdown occurs more than protein synthesis, resulting in a continuous decrease in muscle mass.

[0056] The term "prevention" used in the present invention may mean any act of inhibiting or delaying sarcopenia by administering to a subject a composition for preventing or improving sarcopenia according to the present invention.

[0057] The term “treatment” as used in the present invention may mean any act of improving or eliminating sarcopenia by administering the composition of the present invention.

[0058] When the composition according to the present invention is used as a pharmaceutical composition, the pharmaceutical composition of the present invention can be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the Weissella cibaria WiKim0187 of the above accession number KCCM13568P, and the adjuvant can be an excipient, a disintegrant, a sweetener, a binder, a coating agent, a swelling agent, a lubricant, a glidant, or a flavoring agent.

[0059] The above pharmaceutical composition may be preferably formulated as a pharmaceutical composition by including one or more pharmaceutically acceptable carriers in addition to the above-described effective ingredients for administration.

[0060] For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an orally acceptable, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, etc. In addition, if desired or necessary, suitable binders, lubricants, disintegrants, and coloring agents may also be included in the mixture. Suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tracheacanth, or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum, and the like. In the composition to be formulated as a liquid solution, acceptable pharmaceutical carriers include those that are sterile and biocompatible, such as saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and mixtures of one or more of these components. If necessary, other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate the composition into injectable formulations such as aqueous solutions, suspensions, and emulsions, pills, capsules, granules, or tablets.

[0061] In the present invention, a food composition containing the strain as an active ingredient may include a beverage such as fermented milk. Accordingly, the present invention provides a lactic acid bacteria starter for fermentation comprising Weissella cibaria WiKim0187, accession number KCCM13568P, or a culture thereof.

[0062] The term "lactic acid bacteria starter" as used herein refers to a starter that initiates fermentation by breaking down lactose in dairy products to produce lactic acid. Typically, it refers to culturing and propagating lactic acid bacteria stored as starter for the production of induced fermentation products, and in the present invention, it refers to culturing and propagating Weissella cibaria WiKim0187 before use.

[0063] In addition, the Weissella cibaria WiKim0187 of the present invention, accession number KCCM13568P, has the general intestinal function and immune enhancement effect of lactic acid bacteria.

[0064] Accordingly, the present invention provides a composition for the treatment of indigestion comprising, as an effective ingredient, Weissella cibaria WiKim0187, accession number KCCM13568P, a culture thereof, a pulverized product thereof, or an extract thereof.

[0065] The composition for the treatment of the present invention can be prepared and administered in various formulations and methods, and may be prepared by using Weissella cibaria WiKim0187 alone, deposited under the accession number KCCM13568P; a culture thereof; or a Weissella cibaria WiKim0187 strain in combination with Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus gasseri, Lactobacillus delbrueckii ssp. Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus helveticus, Lactobacillus fermentum, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus salivarius, Lactococcus lactis, Enterococcus faecium, Enterococcus faecalis, Streptococcus thermophilus, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum and Bifidobacterium animalis ssp. lactis.A mixture containing one or more probiotics selected from Bacillus lactis can be prepared and administered in the form of tablets, troches, capsules, elixirs, syrups, powders, suspensions, or granules by mixing the mixture with carriers and flavorings commonly used in the pharmaceutical field. The carriers may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, etc. The administration method may be oral, parenteral, or topical, but oral administration is preferred. In addition, the administration dosage may be appropriately selected depending on the absorption rate, inactivation rate, and excretion rate of the active ingredient in the body, and the age, sex, condition, etc. of the recipient.

[0066] The amount of Weissella cibaria WiKim0187 included in the composition according to the present invention is about 10 per use. 6 10 inland 12 cfu / g, for example 10 7 10 inland 11 cfu / g, 10 8 10 inland 10 cfu / g. When administering strains, it is preferable to administer them in a live state, and they can be administered in a killed or attenuated state before ingestion. In addition, when manufacturing using culture supernatants, etc., an additional sterilization process through heat treatment may be performed. The amount of strains and daily intake required to achieve minimum efficacy may vary depending on the physical or health condition of the ingester, but is generally about 10 6 10 inland 12 cfu / g, for example 10 7 10 inland 11 cfu / g, 10 8 10 inland 10 It could be cfu / g.

[0067] In addition, the present invention provides a method for treating sarcopenia, which comprises administering to a subject in need thereof a therapeutically effective amount of Weissella cibaria WiKim0187 (accession number KCCM13568P), a culture thereof, a lysate thereof, or an extract thereof.

[0068] The term “subject” as used herein refers to a mammal that is the subject of treatment, observation or experiment, preferably a human.

[0069] The term “therapeutically effective amount” as used herein means the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue, animal, or human, as conceived by a researcher, veterinarian, physician, or other clinician, including an amount that induces alleviation of symptoms of a disease or disorder being treated. It will be apparent to those skilled in the art that the therapeutically effective amount and frequency of administration of the active ingredient of the present invention will vary depending on the desired effect. Therefore, the optimal dosage to be administered can be readily determined by those skilled in the art, and can be adjusted according to various factors, including 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 patient’s age, weight, general health, sex, and diet, the time of administration, the route of administration, and the excretion rate of the composition, the treatment period, and concurrently used drugs.

[0070] In the treatment method of the present invention, for adults, it is preferable to administer Weissella cibaria WiKim0187, its culture, its lysate or its extract once or several times a day at a dose of 0.01 mg / kg to 200 mg / kg.

[0071] In the treatment method of the present invention, a composition comprising the Weissella cibaria WiKim0187 of the present invention, its culture, its lysate or its extract as an active ingredient can be administered in a conventional manner via oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes.

[0072] The present invention may include the use of Weissella cibaria WiKim0187, accession number KCCM13568P, for improving sarcopenia.

[0073]

[0074] The advantages and features of the present invention, and the methods for achieving them, will become clearer with reference to the embodiments described in detail below. However, the present invention is not limited to the embodiments disclosed below, but may be implemented in various different forms. These embodiments are provided solely to ensure that the disclosure of the present invention is complete and to fully inform those skilled in the art of the scope of the invention. The present invention is defined solely by the scope of the claims.

[0075]

[0076] [Example]

[0077] Manufacturing Example 1: Isolation and identification of Weissella cibaria WiKim0187

[0078] The inventors isolated kimchi lactic acid bacteria from cabbage kimchi. The kimchi sample was crushed, and the kimchi extract was spread on MRS agar medium (Difco) and cultured at 30°C for at least 24 hours. Single colonies formed after culture were subcultured to select single colonies, and the selected single colonies were finally identified through 16S RNA sequencing.

[0079] As a result of phylogenetic tree microorganism analysis through analysis of the base sequence of 16S rRNA of the selected strain, the microorganism of the present invention was identified as having the 16S rDNA base sequence of SEQ ID No: 1, and was named Weissella cibaria WiKim0187 and deposited at the Korea Microbiological Conservation Center on December 29, 2023 (domestic accession number KFCC11987P, international accession number KCCM13568P) [Fig. 1].

[0080] SEQ ID No: 1

[0081] CCTTCGGGGACAAGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTACTAGTTGCCAGCATTCAGTTGGGCACTCTAGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGCGTATACA ACGAGTTGCCAACCCGCGAGGGTGAGCTAATCTCTTAAAGTACGTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGCCGGTGGGGAACTTCGGAGCCGCCTCAAGGCGATGGTAA

[0082] The general mycological characteristics of Weissella cibariae WiKim0187 are that it is a Gram-positive bacterium, a facultative anaerobe capable of growth under both aerobic and anaerobic conditions, non-spore forming, non-motile, and has a rod-shaped cell structure. Additional characteristics were observed through experiments using the API 50 CH kit for identification of lactic acid bacteria from Biomerieux, and the results are shown in Table 1.

[0083] [Table 1]

[0084]

[0085]

[0086] Example 1: Confirmation of hemolytic activity of the WiKim0187 strain

[0087] For the above-mentioned Weissella cibaria WiKim0187 to be used as a food ingredient or additive, it must be safe, free of hemolytic toxicity to the human body. Therefore, the hemolytic activity of Weissella cibaria WiKim0187 was tested to determine whether it could induce the destruction or decomposition of red blood cells.

[0088] The above hemolytic activity was confirmed by smearing Weissella cibaria WiKim0187 on blood solid medium supplemented with 5% sheep blood (KisanBio, Korea), culturing at 30°C for 48 hours, and determining hemolytic activity by examining the formation of a transparent ring around the cells. The results are shown in Fig. 2.

[0089] As shown in Fig. 2, the above-mentioned Weissella cibaria WiKim0187 was confirmed not to induce a hemolytic reaction as no clear ring formed around the cell due to destruction of red blood cells in the hemolytic test.

[0090] Therefore, the above-mentioned Weissella cibaria WiKim0187 was confirmed to have no harmful effects on the human body, such as hemolysis, and thus was confirmed to be safe when applied to food.

[0091]

[0092] Manufacturing Example 2: Preparation of freeze-dried product of Weissella cibaria WiKim0187 strain culture solution

[0093] For the freeze-drying of the strain Weissella cibaria WiKim0187, the culture medium was prepared with the following composition. Sugar, Peptone, Yeast extract, Na-acetate, Trisodium citrate, Ammonium sulfate, KH2PO4, MnSO4, and MgSO4 were added, and Weissella cibaria WiKim0187 was cultured at pH 6.0, 32℃, for 12 hours, and freeze-dried. The freeze-drying protectants included trehalose, lactose, H powder, and alginate. The freeze-dried product without Weissella cibaria WiKim0187 was weighed the same as the freeze-dried product with Weissella cibaria WiKim0187, suspended in DPBS, and administered orally daily. Weissella cibaria WiKim0187 was 1 × 10 9 It was administered daily at a concentration of CFU / 0.2 mL.

[0094]

[0095] Manufacturing Example 3: Manufacturing of a group receiving Weissella cibaria WiKim0187 in a sarcopenic aging mouse model.

[0096] In order to confirm the muscle-loss improvement effect of the present invention's Weissella cibaria WiKim0187, male C57BL / 6, 8-month-old aged mice (sarcopenic aging mouse model) were used. The aged mice were administered 1×10 of the excipient or the lyophilized product of Weissella cibaria WiKim0187 for 4 months. 9 Oral administration was performed at a concentration of CFU / 0.2 mL (Weissella cibaria WiKim0187 intake group).

[0097]

[0098] Example 2: Verification of the muscle strength-improving effect of the Weissella cibaria WiKim0187 strain in a sarcopenic aging mouse model.

[0099] 1) Forelimb grip test

[0100] Waysella Sibaria WiKim0187 1×10 9 The mice were administered orally daily at a concentration of 1 CFU / 0.2 mL, and forepaw grip strength was measured monthly at one-month intervals. Forepaw grip strength was evaluated using a mouse behavior analysis device (UgoBasile, Italy), and the forepaw grip strength was evaluated by dividing the average value by body weight after excluding the highest or lowest value, and measuring the forepaw grip strength five times in three steps.

[0101] As shown in Fig. 3, the forefoot grip strength and forefoot grip strength according to body weight of the group that consumed Weissella cibaria WiKim0187 (Weissella cibaria WiKim0187 consumption group) were higher than those of the group that consumed the excipient.

[0102] As shown in Fig. 4, the group that consumed the excipient showed a decrease in forefoot grip strength over time, whereas the group that consumed Weissella cibaria WiKim0187 showed almost maintained forefoot grip strength, indicating that Weissella cibaria WiKim0187 has the effect of improving muscle strength and preventing sarcopenia due to aging.

[0103]

[0104] Example 3: Verification of the inhibition of muscle loss by the Weissella cibaria WiKim0187 strain in a sarcopenic aging mouse model.

[0105] 1) Dual-energy X-ray absorptiometry

[0106] The Weissella cibaria WiKim0187 intake group and the vehicle group were measured using a small animal Dual-energy X-ray absorptiometry device (iNSiGHT VET DXA, OsteoSys, Korea).

[0107] As shown in Figure 5, the group that consumed Weissella cibaria WiKim0187 showed significantly higher muscle mass compared to the group that consumed the excipient, indicating that Weissella cibaria WiKim0187 had the effect of suppressing muscle loss due to aging.

[0108] 2) Measurement of liver and hind leg muscle tissue weight

[0109] When the animals were sacrificed after orally administering the Weissella cibaria WiKim0187 intake group and the vehicle group for 4 months, the liver, tibialis anterior, gastrocneminus, and quadriceps were separated and weighed.

[0110] As shown in Figure 6, the weight of the liver and hind leg muscle tissue was measured to be higher than that of the excipient group, indicating that Weissella cibaria WiKim0187 has the effect of suppressing muscle loss due to aging.

[0111]

[0112] Example 4: Verification of the efficacy of the Weissella cibaria WiKim0187 strain in improving muscle loss in a sarcopenic aging mouse model.

[0113] 1) Confirmation of decreased expression of muscle tissue sarcopenia marker genes (MuRF-1, Atrogin-1)

[0114] To verify the efficacy of Weissella cibaria WiKim0187 in improving muscle loss, muscle tissues from aged mice orally administered for 4 months were collected and the gene expression of skeletal muscle atrophy factors Muscle RING finger 1 (MuRF-1) and Atrogin-1 was measured.

[0115] As shown in Fig. 7, in the gastrocnemius muscle tissue, the MuRF-1 gene and Atrogin-1 gene showed significantly lower expression levels in the Weissella cibaria WiKim0187 group than in the vehicle group. In addition, in the tibialis anterior muscle tissue, the expression of the Atrogin-1 gene was measured to be significantly lower in the Weissella cibaria WiKim0187 group, indicating that Weissella cibaria WiKim0187 showed the efficacy of improving sarcopenia by lowering the expression of skeletal muscle atrophy factors.

[0116] 2) Confirmation of increased expression of muscle differentiation marker genes (MyD, Myogenin) in muscle tissue

[0117] To verify the muscle function-enhancing efficacy of Weissella cibaria WiKim0187, muscle tissues from aged mice orally administered for 4 months were collected and the gene expression of muscle protein synthesis factors MyoD and Myogenin was measured.

[0118] As shown in Fig. 8, the expression levels of MyoD and Myogenin genes in the quadriceps femoris were higher than in the excipient group, indicating that Weissella cibaria WiKim0187 was involved in protein synthesis in the quadriceps femoris and had an effect of improving muscle loss.

[0119]

Claims

1. Weissella cibaria WiKim0187 (Accession number KCCM13568P).

2. In paragraph 1, A strain that has the effect of alleviating sarcopenia.

3. In paragraph 1, A strain having 16S rRNA represented by SEQ ID NO:

1.

4. A food composition for improving sarcopenia, comprising Weissella cibaria WiKim0187 (Accession No. KCCM13568P), a culture thereof, a pulverized product thereof, or an extract thereof as an active ingredient.

5. In paragraph 4, The above food is a composition that is a health functional food.

6. In paragraph 4, The above food is a beverage, bar, or fermented milk composition.

7. A pharmaceutical composition for preventing or treating sarcopenia, comprising Weissella cibaria WiKim0187 (Accession No. KCCM13568P), a culture thereof, a lysate thereof, or an extract thereof as an active ingredient.

8. A composition for a laxative comprising Weissella cibaria WiKim0187 (Accession No. KCCM13568P), a culture thereof, a pulverized product thereof, or an extract thereof as an active ingredient.

9. A lactic acid bacteria starter for fermentation comprising Weissella cibaria WiKim0187 (accession number KCCM13568P) or a culture thereof as an active ingredient.

10. A method for improving or treating sarcopenia, comprising administering to a subject in need thereof a therapeutically effective amount of Weissella cibaria WiKim0187 (accession number KCCM13568P), a culture thereof, a lysate thereof, or an extract thereof.

11. Use of Weissella cibaria WiKim0187, accession number KCCM13568P, for improving sarcopenia.

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