Lactic acid bacteria and use thereof

Abstract

To provide a new lactic acid bacterium and its use.SOLUTION: The features of the present invention for solving the above problems are as follows. It is characterized in that it can be used for various health-promoting applications using lactic acid bacteria Lacticaseibacillusparacaseisp. Shidare (accession number NITEABP-04138). Specifically, this lactic acid bacterium is characterized by exhibiting various effects such as promotion of production of IgA, IL-10, IL-12, and serotonin, normalization of stool shape and improvement of bowel movement, improvement of feeling of incomplete evacuation, adjustment of intestinal environment, immunoregulatory action, promotion of equol production, and further, promotion of collagen gene and hyaluronic acid gene expression, and EMT suppression.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a novel lactic acid bacterium and uses thereof. [Background technology]

[0002] Lactic acid bacteria derived from Somei-Yoshino cherry trees have been known to date as cherry-derived lactic acid bacteria, and are known to have the effect of promoting IL-12 production (Patent Document 1).

[0003] The body responds to infections by microorganisms such as bacteria and viruses, tumors, and cell damage with an immune response, which is regulated by direct or indirect interactions between immunocompetent cells. Cytokines such as interleukins and tumor necrosis factor α, produced by lymphocytes and macrophages, play an important role in regulating the immune response.

[0004] Currently, 35 types of cytokines belonging to the interleukin group are known, and the following effects have been confirmed for one of them, IL-12.

[0005] (1) It acts on NK cells and T cells to induce the production of interferon-γ and tumor necrosis factor-α, and activates macrophages. (2) Enhancement of the cytotoxic activity of NK cells and CD8+ T cells. (3) It acts synergistically with interleukin 2 to activate cytotoxic lymphocytes and induce lymphokine-activated killer cells. (4) Promotes differentiation of naive T cells into Th1 cells.

[0006] Due to these effects, IL-12 is expected to be useful in treating many diseases, including infections, tumors, allergies (atopic dermatitis, etc.), etc. Furthermore, it is known that IL-12 promotes interleukin-12 production, thereby reducing skin evaporation, thereby providing a skin barrier function (Patent Document 1).

[0007] On the other hand, IL-10 functions as an anti-inflammatory cytokine, and IL-12 acts to suppress excessive immune function after eliminating pathogens. Therefore, for the immune system to function normally, it is necessary to promote the production of both IL-10 and IL-12.

[0008] Equol, a metabolite of daidzein, a soy isoflavone, is a physiologically active substance that exhibits higher female hormone-like activity (i.e., estrogenic activity) than soy isoflavone itself. Equol has been reported to have effects such as alleviating menopausal symptoms, improving skin condition, and alleviating unpleasant symptoms associated with premenstrual syndrome (PMS) (e.g., Non-Patent Document 1, Non-Patent Document 2, and Patent Document 2).

[0009] Dysmenorrhea is characterized by pain and discomfort associated with menstruation, and is particularly prevalent among young women. In Japan, it is said that approximately 70-80% of menstruating women experience some kind of premenstrual symptoms. It is said that approximately 5.4% of women experience PMS so severe that it interferes with their daily lives (Non-Patent Document 3). Furthermore, over 90% of women report experiencing premenstrual symptoms such as abdominal bloating, headaches, and moodiness (Non-Patent Document 4). There are two types of dysmenorrhea: those with organic causes such as endometriosis or uterine fibroids (organic dysmenorrhea), and those without a clear cause (functional dysmenorrhea). Functional dysmenorrhea is particularly difficult to treat, and while NSAIDs are currently the primary painkillers used, they have yet to provide a fundamental solution. In recent years, it has become clear that the epithelial-mesenchymal transition (EMT) of endometrial epithelial cells is involved in the mechanism of dysmenorrhea, and this phenomenon is considered to be the primary cause of menstrual pain. In fact, a drug with anti-EMT activity (tranilast) has been confirmed to be effective, and pain relief has been reported. In the future, it is expected that treatment with safe ingredients with anti-EMT activity will contribute to the improvement of menstrual discomfort (Patent Document 3, Patent Document 4).

[0010] [Patent Document 1] Japanese Patent Publication No. 2023-040559 [Patent Document 2] International Patent Publication No. 2020-004568 [Patent Document 3] International Patent Publication No. 2010-147184 [Patent Document 4] International Patent Publication No. 2022-045211 [Non-Patent Document 1] Kenneth DR Setchelland Carlo Clerici, Equol:History, Chemistry, and Formation, The Journal of Nutrition, pp.1355S-1362S,2010 (doi: 10.3945 / jn.109.119776) [Non-patent document 2] 1. Shigeto Uchiyama et al., Journal of Intestinal Microbiology, 21(3), 217-220 (2007) [Non-patent document 3] Femtech Market Outlook CMC Publishing First edition published March 31, 2023 [Non-patent document 4] Winer, SA, Rapkin, AJ (2006). Premenstrual disorders: prevalence, etiology and impact. Journal of Reproductive Medicine; 51(4 Suppl):339-347. DISCLOSURE OF THE INVENTION [Problem to be solved by the invention]

[0011] Under these circumstances, the present inventors discovered that a novel lactic acid bacterium derived from weeping cherry tree (Prunus pendula) has intestinal regulating and immunomodulating effects, and thus completed the present invention. Furthermore, the present inventors have discovered that a novel lactic acid bacterium derived from weeping cherry tree (Prunus pendula) has the effect of promoting equol production, thereby completing the present invention. Furthermore, the present inventors discovered that a novel lactic acid bacterium derived from weeping cherry tree (Prunus pendula) has the ability to express collagen synthesis genes and hyaluronic acid synthesis genes, thereby completing the present invention. Furthermore, the present inventors have discovered that a novel lactic acid bacterium derived from weeping cherry tree (Prunus pendula) has EMT-inhibiting activity, thereby completing the present invention. That is, an object of the present invention is to provide novel lactic acid bacteria and uses thereof. [Means for solving the problem]

[0012] The features of the present invention for solving the above problems are as follows. 1. Lactic acid bacteria (Lacticaseibacillus paracasei sp. Shidare) ( Deposit number NITE ABP- 04138 ) Lactic acid bacteria. 2. An IgA production promoter containing the lactic acid bacteria shown in 1 above as an active ingredient. 3. An IL-10 production promoter containing the lactic acid bacteria described in 1 above as an active ingredient. 4. An IL-12 production promoter containing the lactic acid bacteria described in 1 above as an active ingredient. 5. A serotonin production promoter containing the lactic acid bacteria described in 1 above as an active ingredient. 6. A stool shape normalizing agent containing the lactic acid bacteria described in 1 above as an active ingredient. 7. A laxative containing the lactic acid bacteria described in 1 above as an active ingredient. 8. An agent for improving the feeling of incomplete bowel movements, containing the lactic acid bacteria described in 1 above as an active ingredient. 9. An intestinal regulator containing the lactic acid bacteria described in 1 above as an active ingredient. 10. An immunomodulator containing the lactic acid bacteria described in 1 above as an active ingredient. 11. An equol production promoter containing the lactic acid bacteria described in 1 above as an active ingredient. 12. A collagen synthesis gene expression promoter containing the lactic acid bacteria described in 1 above as an active ingredient. 13. A hyaluronic acid synthesis gene expression promoter containing the lactic acid bacteria described in 1 above as an active ingredient. 14. An EMT inhibitor containing the lactic acid bacteria described in 1 above as an active ingredient. 15. A preventive and therapeutic agent for functional dysmenorrhea, containing the lactic acid bacteria described in 1 above as an active ingredient. 16. A PMS preventive and therapeutic agent containing the lactic acid bacteria described in 1 above as an active ingredient. [Effects of the Invention]

[0013] The lactic acid bacteria of the present invention have the effect of promoting IgA production, which contributes to improving intestinal immunity and regulating the intestinal function. The lactic acid bacteria of the present invention have the effect of promoting IL-10 production, making them useful as anti-inflammatory agents. The lactic acid bacteria of the present invention have the ability to promote Il-12 production. This activity (1) induces the production of interferon-γ and tumor necrosis factor-α by acting on NK cells and T cells, activates macrophages, enhances the cytotoxic activity of NK cells and CD8+ T cells, and synergizes with interleukin-2 to activate cytotoxic lymphocytes, induce lymphokine-activated killer cells, and promote the differentiation of naive T cells into Th1 cells. These findings make the bacteria effective in preventing and treating many diseases, including infectious diseases, tumors, and allergies (e.g., atopic dermatitis). As described above, the lactic acid bacteria of the present invention have the effect of promoting the production of both IL-10 and IL-12, and are therefore useful as immunomodulators. Furthermore, in human monitor tests, the lactic acid bacteria of the present invention have been shown to have the effects of promoting serotonin production, normalizing stool shape, improving bowel movements, and alleviating the feeling of incomplete evacuation, making them useful as anti-intestinal agents. Furthermore, the lactic acid bacteria of the present invention are useful as equol production promoters, and as such have the effects of alleviating menopausal symptoms, improving skin condition, and alleviating unpleasant symptoms caused by premenstrual syndrome (PMS). Furthermore, the lactic acid bacteria of the present invention have the effect of promoting the expression of collagen synthesis genes and hyaluronic acid synthesis genes, and as a result, the cherry blossom lactic acid bacteria have the effect of maintaining the moisture and elasticity of the skin. The lactic acid bacteria of the present invention are useful as EMT inhibitors, and are therefore useful as prophylactic and therapeutic agents for functional dysmenorrhea and PMS. [Brief explanation of the drawings]

[0014] [Figure 1] 1 is a graph showing the ability of the lactic acid bacteria of this example to induce IL-10 production using mouse splenocytes. [Figure 2] 1 is a graph showing the ability of the lactic acid bacteria of this example to induce IL-12 production using mouse splenocytes. [Figure 3] 1 is a graph showing the ability of the lactic acid bacteria of this example to induce IgA production using mouse splenocytes. [Figure 4] 1 is a graph showing the effect of the lactic acid bacterium of this example (Lacticaseibacillus paracasei sp. Shidare) on the amount of IgA in mouse feces. [Figure 5] 1 is a graph showing the effect of the lactic acid bacterium of this example (Lacticaseibacillus paracasei sp. Shidare) on the fecal volume of mice. [Figure 6] 1 is a graph showing the effect of the lactic acid bacterium of this example (Lacticaseibacillus paracasei sp. Shidare) on the amount of intestinal serotonin in mice. [Figure 7] 1 is a graph showing the effect of oral intake of the lactic acid bacteria of this example (Lacticaseibacillus paracasei sp. Shidare) on the frequency and amount of stool. [Figure 8] 1 is a graph showing the effect of oral intake of the lactic acid bacteria of this example (Lacticaseibacillus paracasei sp. Shidare) on stool shape score. [Figure 9] 1 is a graph showing the effect of oral intake of the lactic acid bacteria of an example (Lacticaseibacillus paracasei sp. Shidare) on the feeling of incomplete evacuation. [Figure 10]1 shows a phylogenetic tree of the lactic acid bacterium (Lacticaseibacillus paracasei sp. Shidare) of this example. [Figure 11] FIG. 1 is an explanatory diagram showing a method for identifying lactic acid bacteria (Lacticaseibacillus paracasei sp. Shidare) in this example. [Figure 12] 1 is a graph showing the effect of the lactic acid bacterium of this example (Lacticaseibacillus paracasei sp. Shidare) on equol levels in mouse blood. [Figure 13] 1 is a graph showing the results of evaluating the collagen and hyaluronic acid synthase gene expression effects in the lactic acid bacterium (Lacticaseibacillus paracasei sp. Shidare) of this example. [Figure 14] 1 is a graph showing the results of evaluating the EMT inhibitory effect of the lactic acid bacterium (Lacticaseibacillus paracasei sp. Shidare) of this example. DETAILED DESCRIPTION OF THE INVENTION

[0015] The present invention will be described in detail below. The present invention relates to a lactic acid bacterium (Lacticaseibacillus paracasei sp. Shidare) with accession number ( Deposit number NITE ABP-04138 ) is a lactic acid bacterium. The above lactic acid bacteria are derived from weeping cherry trees (Prunus pendula) and can be isolated and identified by the methods described in the Examples of this specification.

[0016] The lactic acid bacteria of the present invention can be used as an ingredient in various foods and beverages. Examples of foods and beverages include general foods such as confectioneries (gum, candy, caramel, chocolate, cookies, snacks, jelly, gummy candy, tablet candy, etc.), noodles (soba, udon, ramen, etc.), dairy products (milk, ice cream, yogurt, etc.), seasonings (miso, soy sauce, etc.), soups, beverages (juice, coffee, black tea, green tea, carbonated drinks, sports drinks, etc.), as well as health foods (tablets, capsules, etc.), and nutritional supplements (nutrition drinks, etc.). The agent of the present invention can be appropriately incorporated into these foods and beverages.

[0017] These foods and beverages can contain various ingredients depending on their type, such as glucose, fructose, sucrose, maltose, sorbitol, stevioside, corn syrup, lactose, citric acid, tartaric acid, malic acid, succinic acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid esters, polyglycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, gum arabic, carrageenan, casein, gelatin, pectin, agar, B vitamins, nicotinamide, calcium pantothenate, amino acids, calcium salts, colorants, flavorings, and preservatives.

[0018] When the agent of the present invention is applied to food and drink, the amount of active ingredient added is preferably 1 to 20 wt % in total relative to the food and drink, since the main purpose is disease prevention and health maintenance.

[0019] The lactic acid bacteria of the present invention may be used as a material for pharmaceuticals (including pharmaceuticals and quasi-drugs), which can be produced by appropriately blending the agent of the present invention with raw materials for pharmaceutical preparations. Examples of pharmaceutical raw materials that can be incorporated into the agent of the present invention include excipients (glucose, lactose, sucrose, sodium chloride, starch, calcium carbonate, kaolin, crystalline cellulose, cocoa butter, hardened vegetable oil, kaolin, talc, etc.), binders (distilled water, saline, ethanol water, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.), disintegrants (sodium alginate, agar, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, stearate monoglyceride, starch, lactose, gum arabic powder, gelatin, ethanol, etc.), disintegration inhibitors (sucrose, stearin, cocoa butter, hydrogenated oil, etc.), absorption enhancers (quaternary ammonium base, sodium lauryl sulfate, etc.), adsorbents (glycerin, starch, lactose, kaolin, bentonite, silicic acid, etc.), lubricants (purified talc, stearates, polyethylene glycol, etc.).

[0020] The lactic acid bacteria of the present invention can generally be administered orally in the form of tablets, pills, soft or hard capsules, fine granules, powders, granules, liquids, or the like.

[0021] The dosage may vary depending on the administration method, the condition, the age of the patient, etc., but typically, adults can be given 0.5 to 5000 mg of the active ingredient per day, and children can be given 0.5 to 3000 mg. The blending ratio of lactic acid bacteria can be changed as appropriate depending on the dosage form, but is usually about 0.3 to 15.0 wt% when administered orally or via mucosal absorption, and about 0.01 to 10 wt% when administered parenterally. Note that the dosage varies depending on various conditions, so in some cases a smaller dosage than the above is sufficient, and in other cases it may be necessary to administer a dosage exceeding the range.

[0022] The lactic acid bacteria of the present invention can also be expected to exert the above-mentioned effects when used as external skin preparations (including cosmetics, pharmaceuticals, and quasi-drugs). Examples of external skin preparations that can be formulated with the lactic acid bacteria of the present invention include emulsions, soaps, facial cleansers, bath additives, creams, emulsions, lotions, colognes, shaving creams, shaving lotions, cosmetic oils, suntan / sunscreen lotions, face powders, foundations, perfumes, masks, nail creams, enamel, enamel removers, eyebrow pencils, blushers, eye creams, eye shadows, mascara, eyeliners, lipsticks, lip balms, shampoos, rinses, hair dyes, dispersions, cleansers, etc. Examples of pharmaceutical or quasi-drug products that can be formulated with the lactic acid bacteria of the present invention include ointments, creams, external liquids, etc.

[0023] In addition to the lactic acid bacteria of the present invention, the above-mentioned topical skin preparations can contain, to the extent that the above-mentioned effects are not impaired, ingredients that are typically incorporated into topical skin preparations such as cosmetics and quasi-drugs, such as oils, higher alcohols, fatty acids, UV absorbers, powders, pigments, surfactants, polyhydric alcohols, sugars, polymers, physiologically active ingredients, solvents, antioxidants, fragrances, preservatives, etc. Examples are listed below, but the present invention is not limited to these examples.

[0024] (1) Examples of oil Ester-based oil phase ingredients: glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, butyl stearate, butyl myristate, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, diisopropyl adipate Isopropyl, Isoarachyl Neopentanoate, Caprylic / Capric Triglyceride, Trimethylolpropane Tri-2-Ethylhexanoate, Trimethylolpropane Triisostearate, Pentaerythritol Tetra-2-Ethylhexanoate, Cetyl Caprylate, Decyl Laurate, Hexyl Laurate, Decyl Myristate, Myristyl Myristate, Cetyl Myristate, Stearyl Stearate, Decyl Oleate, Cetyl Ricinoleate, Isostearyl Laurate, Isotridecyl Myristate, Isocetyl Myristate, Isostearate Myristate Tearyl, Isocetyl Palmitate, Isostearyl Palmitate, Octyl Stearate, Isocetyl Stearate, Isodecyl Oleate, Octyldodecyl Oleate, Octyldodecyl Linoleate, Isopropyl Isostearate, Cetostearyl 2-Ethylhexanoate, Stearyl 2-Ethylhexanoate, Hexyl Isostearate, Ethylene Glycol Dioctanoate, Ethylene Glycol Dioleate, Propylene Glycol Dicaprate, Propylene Glycol Dicaprylate, Propylene Glycol Dicaprylate, Dicaprin Neopentyl glycol acid, neopentyl glycol dioctanoate, glyceryl tricaprylate, glyceryl triundecylate, glyceryl triisopalmitate, glyceryl triisostearate, octyldodecyl neopentanoate, isostearyl octanoate, octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, octyldecyl isostearate, polyglycerin oleate, polyglycerin isostearate, dipropyl carbonate,Dialkyl carbonate (C12-18), triisocetyl citrate, triisoarachidyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, triethyl citrate, acetyltriethyl citrate, acetyltributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate propyl, dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phytosteryl isostearate, phytosteryl oleate, isocetyl 12-stearoylhydroxystearate, stearyl 12-stearoylhydroxystearate, isostearyl 12-stearoylhydroxystearate, etc. Hydrocarbon-based oil phase components: squalane, liquid paraffin, α-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybutene, microcrystalline wax, petrolatum, etc. Animal and vegetable oils and their hydrogenated oils, and naturally derived waxes: beef tallow, hydrogenated beef tallow, lard, hydrogenated lard, horse oil, hydrogenated horse oil, mink oil, orange roughy oil, fish oil, hydrogenated fish oil, egg yolk oil, and other animal oils and their hydrogenated oils, avocado oil, almond oil, olive oil, cocoa butter, kiwi seed oil, apricot kernel oil, kukui nut oil, sesame oil, wheat germ oil, rice germ oil, rice bran oil, safflower oil, shea butter, soybean oil, evening primrose oil, perilla oil, tea seed oil, camellia oil, corn oil, rapeseed oil, Examples of suitable oils include vegetable oils and their hydrogenated oils such as hydrogenated rapeseed oil, palm kernel oil, hydrogenated palm kernel oil, palm oil, hydrogenated palm oil, peanut oil, hydrogenated peanut oil, castor oil, hydrogenated castor oil, sunflower oil, grape seed oil, jojoba oil, hydrogenated jojoba oil, macadamia nut oil, medlar oil, cottonseed oil, hydrogenated cottonseed oil, coconut oil, and hydrogenated coconut oil; and waxes such as beeswax, high acid value beeswax, lanolin, reduced lanolin, hydrogenated lanolin, liquid lanolin, carnauba wax, and montan wax. Silicone-based oil phase components: dimethylpolysiloxane, methylphenylpolysiloxane, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, methylhydrogenpolysiloxane, polyether-modified organopolysiloxane, dimethylsiloxane-methylcetyloxysiloxane copolymer, dimethylsiloxane-methylstearoxysiloxane copolymer, alkyl-modified organopolysiloxane, terminal-modified organopolysiloxane, amino-modified silicone oil, amino-modified organopolysiloxane, dimethiconol, silicone gel, acrylic silicone, trimethylsiloxysilicate, silicone RTV rubber, etc. Fluorine-based oil phase components: perfluoropolyether, fluorine-modified organopolysiloxane, fluorinated pitch, fluorocarbon, fluoroalcohol, fluoroalkyl-polyoxyalkylene co-modified organopolysiloxane, etc.

[0025] (2) Examples of higher alcohols Examples include lauryl alcohol, myristyl alcohol, cetyl alcohol, stearyl alcohol, isostearyl alcohol, oleyl alcohol, behenyl alcohol, 2-ethylhexanol, hexadecyl alcohol, and octyldodecanol.

[0026] (3) Examples of fatty acids Examples of the acid include caprylic acid, capric acid, undecylenic acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, isostearic acid, oleic acid, linoleic acid, linolenic acid, arachidic acid, arachidonic acid, behenic acid, erucic acid, and 2-ethylhexanoic acid.

[0027] (4) Examples of UV absorbers Para-aminobenzoic acid, amyl para-aminobenzoate, ethyl dihydroxypropyl para-aminobenzoate, glyceryl para-aminobenzoate, ethyl para-aminobenzoate, octyl para-aminobenzoate, octyldimethyl para-aminobenzoate, ethylene glycol salicylate, octyl salicylate, triethanolamine salicylate, phenyl salicylate, butylphenyl salicylate, benzyl salicylate, homomenthyl salicylate, benzyl cinnamate, octyl para-methoxycinnamate, 2-ethylhexyl para-methoxycinnamate, glyceryl di-para-methoxycinnamate mono-2-ethylhexanoate, isopropyl para-methoxycinnamate, diethanolamine para-methoxyhydrocinnamate, diisopropyl and diisopropyl cinnamate mixture, urocanic acid, ethyl urocanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and its salts, di Examples of the hydroxymethoxybenzophenone include sodium dihydroxymethoxybenzophenone disulfonate, dihydroxybenzophenone, dihydroxydimethoxybenzophenone, hydroxyoctoxybenzophenone, tetrahydroxybenzophenone, butylmethoxydibenzoylmethane, 2,4,6-trianilino-p-(carbo-2-ethylhexyl-1-oxy)-1,3,5-triazine, 2-(2-hydroxy-5-methylphenyl)benzotriazole, methyl-O-aminobenzoate, 2-ethylhexyl-2-cyano-3,3-diphenylacrylate, phenylbenzimidazole sulfate, 3-(4-methylbenzylidene)camphor, isopropyldibenzoylmethane, 2-ethylhexyl 4-(3,4-dimethoxyphenylmethylene)-2,5-dioxo-1-imidazolidinepropionate, and polymer derivatives and silane derivatives thereof.

[0028] (5) Examples of powders and pigments Dyes such as Red No. 104, Red No. 201, Yellow No. 4, Blue No. 1, Black No. 401, lake dyes such as Yellow No. 4 AL Lake and Yellow No. 203 BA Lake, nylon powder, silk powder, urethane powder, Teflon (registered trademark) powder, silicone powder, polymethyl methacrylate powder, cellulose powder, starch, silicone elastomer spherical powder, polymers such as polyethylene powder, yellow iron oxide, red iron oxide, black iron oxide, chromium oxide, carbon black, colored pigments such as ultramarine and iron blue, zinc oxide, titanium oxide, acid Examples of suitable powders include white pigments such as cerium chloride, extender pigments such as talc, mica, sericite, kaolin, and plate-like barium sulfate, pearl pigments such as titanium dioxide, metal salts such as barium sulfate, calcium carbonate, magnesium carbonate, aluminum silicate, and magnesium silicate, inorganic powders such as silica and alumina, metal soaps such as aluminum stearate, magnesium stearate, zinc palmitate, zinc myristate, magnesium myristate, zinc laurate, and zinc undecylenate, bentonite, smectite, and boron nitride. There are no particular limitations on the shape (spherical, rod-like, needle-like, plate-like, irregular, flaky, spindle-like, etc.) and particle size of these powders. These powders may or may not have been previously surface-treated by conventionally known surface treatments, such as fluorine compound treatment, silicone treatment, silicone resin treatment, pendant treatment, silane coupling agent treatment, titanium coupling agent treatment, oil treatment, N-acylated lysine treatment, polyacrylic acid treatment, metal soap treatment, amino acid treatment, lecithin treatment, inorganic compound treatment, plasma treatment, mechanochemical treatment, etc.

[0029] (6) Examples of surfactants Anionic surfactants: fatty acid soaps, α-acylsulfonates, alkylsulfonates, alkylarylsulfonates, alkylnaphthalenesulfonates, alkyl sulfates, POE alkyl ether sulfates, alkylamide sulfates, alkyl phosphates, POE alkyl phosphates, alkylamide phosphates, alkyloylalkyl taurine salts, N-acylamino acid salts, POE alkyl ether carboxylates, alkyl sulfosuccinates, sodium alkylsulfoacetates, acylated hydrolyzed collagen peptide salts, perfluoroalkyl phosphate esters, etc. Cationic surfactants: alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, benzalkonium chloride, behenic acid amidopropyldimethylhydroxypropylammonium chloride, stearic acid diethylaminoethylamide, stearic acid dimethylaminopropylamide, lanolin derivative quaternary ammonium salts, etc. Amphoteric surfactants: Examples thereof include carboxybetaine type, amidobetaine type, sulfobetaine type, hydroxysulfobetaine type, amidosulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, and amidoamine type. Nonionic surfactants: Propylene glycol fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, sorbitan fatty acid esters, POE sorbitan fatty acid esters, POE sorbitan fatty acid esters, POE glycerin fatty acid esters, POE alkyl ethers, POE fatty acid esters, POE hydrogenated castor oil, POE castor oil, POE-POP copolymers, POE-POP alkyl ethers, polyether-modified silicone lauric acid alkanolamide, alkylamine oxide, hydrogenated soybean phospholipids, etc. Natural surfactants: Examples include lecithin, saponin, sugar-based surfactants, etc.

[0030] (7) Examples of polyhydric alcohols and sugars Examples of suitable sugars include ethylene glycol, diethylene glycol, polyethylene glycol, propylene glycol, dipropylene glycol, polypropylene glycol, glycerin, diglycerin, polyglycerin, 3-methyl-1,3-butanediol, 1,3-butylene glycol, sorbitol, mannitol, raffinose, erythritol, glucose, sucrose, fructose, xylitol, lactose, maltose, maltitol, trehalose, alkylated trehalose, mixed isomerized sugars, sulfated trehalose, pullulan, etc. Chemically modified versions of these sugars may also be used.

[0031] (8) Examples of polymers Acrylate / methacrylate copolymer (Plussize, manufactured by GOO Chemical Co., Ltd.), vinyl acetate / crotonic acid copolymer (Resin 28-1310, manufactured by NSC), vinyl acetate / crotonic acid / vinyl neodecanoate copolymer (28-2930, manufactured by NSC), methyl vinyl ether maleic acid half ester (Gantrez ES, manufactured by ISP), t-butyl acrylate / ethyl acrylate / methacrylic acid copolymer (Ruvimer, manufactured by BASF), vinylpyrrolidone / vinyl acetate / vinyl propionate copolymer (Ruviscol VAP, manufactured by BASF), vinyl acetate Vinyl acetate / crotonic acid copolymer (Rubyset CA, BASF), vinyl acetate / crotonic acid / vinyl pyrrolidone copolymer (Rubyset CAP, BASF), vinyl pyrrolidone / acrylate copolymer (Rubyflex, BASF), acrylate / acrylamide copolymer (Ultrahold, BASF), vinyl acetate / butyl maleate / isobornyl acrylate copolymer (Advantage, ISP), carboxyvinyl polymer (Carbopol, BFGoodrich), acrylic acid / alkyl methacrylate copolymer (Pemulen, BF Examples of such polymers include anionic polymers such as acetic acid amphoteric dialkylaminoethyl methacrylate polymer (Yukaformer, manufactured by Mitsubishi Chemical Corporation) and amphoteric polymers such as octylacrylamide acrylate / hydroxypropyl acrylate / butylaminoethyl methacrylate copolymer (AMPHOMER, manufactured by NSC Corporation); cationic polymers such as quaternized vinylpyrrolidone / dimethylaminoethyl methacrylate (GAFQUAT, manufactured by ISP Corporation) and methylvinylimidazolium chloride / vinylpyrrolidone copolymer (Rubycoat, manufactured by BASF Corporation); and nonionic polymers such as polyvinylpyrrolidone (Ruviscol K, manufactured by BASF Corporation), vinylpyrrolidone / vinyl acetate copolymer (Ruviscol VA, manufactured by BASF Corporation), vinylpyrrolidone / dimethylaminoethyl methacrylate copolymer (Copolymer 937, manufactured by ISP Corporation), and vinylcaprolactam / vinylpyrrolidone / dimethylaminoethyl methacrylate copolymer (Copolymer VC713, manufactured by ISP Corporation).Also suitable for use are naturally occurring polymeric compounds such as cellulose or derivatives thereof, keratin and collagen or derivatives thereof, calcium alginate, pullulan, agar, gelatin, tamarind seed polysaccharides, xanthan gum, carrageenan, high methoxyl pectin, low methoxyl pectin, guar gum, gum arabic, crystalline cellulose, arabinogalactan, karaya gum, tragacanth gum, alginic acid, albumin, casein, curdlan, gellan gum, and dextran.

[0032] (9) Examples of physiologically active ingredients Examples of physiologically active ingredients include substances that impart some kind of physiological activity to the skin when applied to the skin. Examples include whitening ingredients, immunostimulants, anti-aging agents, UV protection agents, slimming agents, tightening agents, antioxidants, hair growth agents, hair restoration agents, moisturizers, blood circulation promoters, antibacterial agents, disinfectants, drying agents, cooling agents, warming agents, vitamins, amino acids, wound healing promoters, irritation relievers, analgesics, cell activators, enzyme components, etc. Examples of suitable ingredients include angelica extract, avocado extract, hydrangea extract, althea extract, arnica extract, aloe extract, apricot extract, apricot kernel extract, ginkgo extract, fennel extract, turmeric extract, oolong tea extract, angelica tree extract, echinacea leaf extract, Scutellaria root extract, Phellodendron bark extract, Coptis japonica extract, barley extract, St. John's wort extract, white nettle extract, watercress extract, orange extract, Dried seawater, seaweed extract, hydrolyzed elastin, hydrolyzed wheat powder, hydrolyzed silk, chamomile extract, carrot extract, artemisia capillaris extract, licorice extract, kalk extract, kakyoku extract, cinchona extract, cucumber extract, guanosine, gardenia extract, kumazasa extract, sophora flavescens extract, walnut extract, grapefruit extract, clematis extract, chlorella extract, mulberry extract, gentian extract, black tea extract, yeast extract, burdock extract, Fermented rice bran extract, rice germ oil, comfrey extract, collagen, bilberry extract, Chinese radish extract, Bupleurum extract, umbilical cord extract, salvia extract, soapwort extract, bamboo extract, hawthorn extract, Japanese pepper extract, shiitake mushroom extract, rehmannia root extract, Lithospermum root extract, perilla extract, linden extract, meadowsweet extract, peony extract, calamus root extract, white birch extract, horsetail extract, ivy extract, hawthorn extract, se Sambucus extract, yarrow extract, peppermint extract, sage extract, mallow extract, cnidium extract, Swertia japonica extract, soybean extract, Chinese laurel extract, thyme extract, tea extract, clove extract, Imperata cylindrica extract, tangerine extract, angelica extract, calendula extract, peach kernel extract, spruce extract, Houttuynia cordata extract, tomato extract, natto extract, carrot extract, garlic extract, wild rose extract, hibiscus extract,Examples of such extracts include burdock root extract, parsley extract, honey, witch hazel extract, parietaria extract, burdock root extract, bisabolol, loquat extract, coltsfoot extract, butterbur stalk extract, poria cocos extract, butcher's broom extract, grape extract, propolis, loofah extract, safflower extract, peppermint extract, linden extract, peony extract, hop extract, pine extract, horse chestnut extract, skunk cabbage extract, soapberry extract, melissa extract, peach extract, cornflower extract, eucalyptus extract, saxifrage extract, coix seed extract, mugwort extract, lavender extract, apple extract, lettuce extract, lemon extract, astragalus extract, rose extract, rosemary extract, Roman chamomile extract, and royal jelly extract. In addition, it contains biopolymers such as deoxyribonucleic acid, mucopolysaccharides, sodium hyaluronate, sodium chondroitin sulfate, collagen, elastin, chitin, chitosan, and hydrolyzed eggshell membrane, moisturizing ingredients such as amino acids, hydrolyzed peptides, sodium lactate, urea, sodium pyrrolidone carboxylate, betaine, whey, and trimethylglycine, oily ingredients such as sphingolipids, ceramides, phytosphingosine, cholesterol, cholesterol derivatives, and phospholipids, ε-aminocaproic acid, glycyrrhizic acid, β-glycyrrhetinic acid, lysozyme chloride, and guaiazulene. Immune stimulants such as methicone and hydrocortisone, vitamins such as vitamin A, vitamin B2, vitamin B6, vitamin C, vitamin D, vitamin E, calcium pantothenate, biotin, nicotinamide, vitamin C ester, active ingredients such as allantoin, diisopropylamine dichloroacetate, 4-aminomethylcyclohexanecarboxylic acid, antioxidants such as tocopherol, carotenoids, flavonoids, tannins, lignans, saponins, cell activators such as alpha-hydroxy acids and beta-hydroxy acids, blood circulation promoters such as gamma-oryzanol and vitamin E derivatives, Wound healing agents such as retinol and retinol derivatives, skin whitening agents such as arbutin, kojic acid, placenta extract, sulfur, ellagic acid, linoleic acid, tranexamic acid, and glutathione, cepharanthine, licorice extract, capsicum tincture, hinokitiol, iodized garlic extract, pyridoxine hydrochloride, DL-α-tocopherol, DL-α-tocopherol acetate, nicotinic acid, nicotinic acid derivatives, calcium pantothenate, D-pantothenyl alcohol, acetylpantothenyl ethyl ether, biotin, allantoin, isopropyl methylphenol, estradiol Alcohol, ethinylestradiol, capronium chloride, benzalkonium chloride, diphenhydramine hydrochloride, tacanal, camphor, salicylic acid, nonylic acid vanillylamide, nonanoic acid vanillylamide, piroctone olamine, glyceryl pentadecanoate, L-menthol, mononitroguaiacol, resorcinol, gamma-aminobutyric acid, benzethonium chloride, mexiletine hydrochloride, auxin, female hormones, cantharides tincture, cyclosporine, zinc pyrithione, hydrocholorthisone, minoxidil, polyoxyethylene sorbitan monostearate, peppermint oil,Examples include hair growth agents such as Sasanishiki extract.

[0033] (10) Examples of antioxidants Examples of antioxidants include sodium bisulfite, sodium sulfite, erythorbic acid, sodium erythorbate, dilauryl thiodipropionate, tocopherol, tolylbiguanide, nordihydroguaiaretic acid, parahydroxyanisole, butylhydroxyanisole, dibutylhydroxytoluene, ascorbyl stearate, ascorbyl palmitate, octyl gallate, propyl gallate, carotenoids, flavonoids, tannins, lignans, saponin, and plant extracts known to have antioxidant effects, such as apple extract and clove extract.

[0034] (11) Examples of solvents Examples include purified water, ethanol, lower alcohols, ethers, LPG, fluorocarbons, N-methylpyrrolidone, fluoroalcohols, volatile linear silicones, and next-generation chlorofluorocarbons. [Example]

[0035] Examples of the present invention are described below. Note that the examples shown below are provided to confirm the various actions and effects of the lactic acid bacteria of the present invention obtained by the present invention, and the scope of the present invention is not limited to these products and production methods.

[0036] Example: Method for isolating and identifying lactic acid bacteria The methods for isolating and identifying lactic acid bacteria are as follows. 1. Isolation method of lactic acid bacteria (accession number NITE ABP-04138) Weeping cherry blossom (Prunus pendula) petals were collected aseptically (using tweezers or gloves). They were then placed in a medium similar to that used in production (mainly yeast extract) and incubated overnight at 30°C and 37°C. The culture medium was then spread onto BCP-supplemented plate count agar (the purple medium turns yellow when acid is released, allowing the selection of acid-producing bacteria). The culture was then left to stand for 24 hours. Colonies that had turned yellow were then streaked onto BCP-supplemented plate count agar. Single colonies were then picked and cultured in a yeast extract-based liquid medium for 20 hours to confirm culturability. Selection was then performed based on turbidity (OD660 of approximately 4.0 or higher). Genomes were extracted from strains that passed the turbidity test, and lactic acid bacteria were selected based on 16s rRNA (700-800 bp from the front). 2. Identification method of isolated lactic acid bacteria A phylogenetic tree was drawn from the full-length 16s rRNA (1400-1500 bp) and used for identification. This phylogenetic tree is shown in Figure 10. Because this was L. casei, it was difficult to distinguish between L. casei and L. paracasei from the 16s rRNA, so a specific primer described in the reference was used for identification (Figure 11). As a result, it was identified as Lacticaseibacillus casei, named Lacticaseibacillus casei shidare, and the procedure for deposit was carried out (deposit number NITE ABP-04138). (References):Microbiology (Reading). 2017 Jul;163(7):950-960.

[0037] Test Example 1: Evaluation of IL-10 and IL-12 production promoting effect 1. Test Method Spleen cells were collected from BALB / cA mice (female, 10 weeks old) at a cell concentration of 2.5 × 10 6The samples were added to a final concentration of 1.0 μg / ml (IL-10 was 1.0 or 10 μg / ml) to a cell suspension prepared at 100 cells / ml (no addition was used as a negative control), and the cells were cultured at 37°C in a 5% CO2 environment for 24 hours for IL-12 and 96 hours for IL-10. The concentrations of each cytokine in the culture supernatant were then measured by ELISA. The results are shown in Figure 1 (IL-10) and Figure 2 (IL-12).

[0038] 2. Measurement results in Test Example 1 and effects of the embodiment From the above, it was confirmed that the lactic acid bacteria of this example (deposit number NITE ABP-04138) have an inhibitory effect on IL-12 and IL-10 production, and therefore the lactic acid bacteria of this example are useful as immunomodulators.

[0039] Test Example 2: Evaluation of IgA production promoting effect 1. Test Method Spleens were collected from BALB / cA mice (female, 10 weeks old) in a clean bench under as sterile conditions as possible. Cells were collected from the spleen using a cell strainer to obtain a cell concentration of 2.5 × 10 6 The cells were cultured at a final concentration of 1.0 or 10.0 μg / ml in RPMI 1640 medium (Wako) containing 10% FBS (Thermo Fisher Scientific) and Antibiotic Mixture (Thermo Fisher Scientific). The samples were added to the cell suspension to a final concentration of 1.0 or 10.0 μg / ml and cultured at 37°C in a 5% CO2 atmosphere for 9 days. The culture supernatant was then collected by centrifugation (1800 × g, 5 minutes), and the IgA concentration in the supernatant was measured by ELISA. The results are shown in Figure 3.

[0040] 2. Results and Effects of the Example in Test Example 2 3 shows that the lactic acid bacteria of this example (deposit number NITE ABP-04138) promote IgA production. This confirms that the lactic acid bacteria of this example promote IgA production and contribute to improving intestinal immunity and regulating the intestinal function.

[0041] Test Example 3: Evaluation of the effect on mouse fecal IgA levels 1. Test Method Since fecal IgA determination is widely used to evaluate the intestinal environment, fecal IgA was determined. BALB / c mice (female, 5 weeks old) were pre-fed for one week and then orally administered the lactic acid bacteria of the present invention at doses of 5 and 50 billion cells / kg / day for two weeks. After two weeks, feces were collected and the amount of IgA in the feces was quantified using an IgA Mouse Uncoated ELISA Kit (Invitrogen). The results are shown in Figure 4. 2. Results of Test Example 3 and Effects of the Examples As shown in Figure 4, it was revealed that the amount of IgA in feces was significantly increased in the group administered 50 billion cells / kg of the lactic acid bacteria of the example (deposit number NITE ABP-04138) compared to the control group. These results confirmed that ingestion of the lactic acid bacteria of this example (deposit number NITE ABP-04138) promotes IgA production and contributes to improving intestinal immunity and regulating the intestinal function.

[0042] Test Example 4: Effects on fecal volume and intestinal serotonin levels in mice 1. Test Method After one week of preliminary breeding, BALB / c mice (male, 7 weeks old) were orally administered the lactic acid bacteria of this example, the Shidare strain (accession number NITE ABP-04138), at 50 or 100 billion cells / kg / day for two weeks. After two weeks, feces were collected for 24 hours from each mouse, and the colon was also sampled. The collected colons were homogenized in RIPA Lysis buffer (ThermoFisher), and the amount of serotonin in the supernatant after centrifugation was quantified using a Serotonin ELISA Kit (Abnova). The results are shown in Figure 5 (fecal volume) and Figure 6 (serotonin volume).

[0043] 2. Results and Effects of the Examples in Test Example 4 As shown in Figure 5, a tendency for fecal mass to increase was confirmed in the group administered 100 billion cells / kg of the lactic acid bacteria of this example (deposit number NITE ABP-04138).Furthermore, as shown in Figure 6, a significant increase in intestinal serotonin levels was confirmed in the group administered 100 billion cells / kg of the lactic acid bacteria of this example. These results confirmed that ingestion of the lactic acid bacteria of this example (deposit number NITE ABP-04138) promotes intestinal serotonin production, activates intestinal peristalsis, and promotes bowel movements.

[0044] Test Example 5: Effect of oral intake of cherry lactic acid bacteria (Lacticaseibacillus paracasei sp. shidare) (Deposit No. NITE ABP-04138) on bowel movements and stool shape in humans 1. Test Method Twenty-eight Japanese adult men and women (12 men, 16 women, mean age: 39.9 ± 11.6 years) were divided into three groups: a placebo group, a 50 billion / day group, and a 100 billion / day group, with an equal gender ratio and average age. The test substances were hard capsules containing the lactic acid bacteria of this example (accession number NITE ABP-04138) and starch hydrolysate at the respective daily intake amounts, and a placebo capsule containing only starch hydrolysate. The ingestion period was two weeks, with the above capsules taken once daily. Evaluation items included weekly stool volume and frequency, stool shape score according to the Bristol Scale, odor, and a questionnaire regarding residual evacuation. Stool volume was visually measured in terms of the number of ping-pong balls it represented. The Bristol scale was rated as follows: 1: hard stool, 2: hard stool, 3: slightly hard stool, 4: normal stool, 5: slightly soft stool, 6: muddy stool, and 7: watery stool. The subjects were asked to observe the shape of their stool at the time of defecation. The odor questionnaire was rated as 1: bad smell, 2: sour smell, 3: hardly noticeable, and 4: not noticeable at all. The feeling of incomplete bowel movement was rated as 1: no bowel movement in a day, 2: feeling of incomplete bowel movement, 3: mostly clear, and 4: clear. The results are shown in Figure 7 (number of bowel movements and amount), Figure 8 (stool shape score), and Figure 9 (feeling of incomplete bowel movement).

[0045] 2. Results and Effects of the Examples in Test Example 4 As shown in Figure 7, a tendency for an increase in bowel movement frequency was confirmed in the 50 billion / day group of the lactic acid bacteria of this example (deposit number NITE ABP-04138), and a significant increase in stool volume was confirmed in both the 50 billion / day and 100 billion / day groups compared to the placebo group. Furthermore, as shown in Figure 8, in the 100 billion / day group, a significant improvement in stool shape score was confirmed in the second week compared to the first week of intake. This confirmed that stool shape improves by taking the lactic acid bacteria of this example (deposit number NITE ABP-04138). Furthermore, as shown in Figure 8, the questionnaire results confirmed that the feeling of incomplete bowel movement tended to improve in the second week compared to the first week in the 50 billion / day group. This confirmed that taking the lactic acid bacteria of this example (deposit number NITE ABP-04138) improved the feeling of incomplete bowel movement. From the above results, it was revealed that continuous oral intake of the lactic acid bacteria of this example improves bowel movements and normalizes the shape of stools.

[0046] Test Example 6: Evaluation of equol production promoting effect 1. Test Method After one week of preliminary breeding, ddY mice (female, 8 weeks old) were orally administered the lactic acid bacteria of this example (deposit number NITE ABP-04138) at 500 and 100 billion cells / kg / day for two weeks. Concurrently with the administration of the samples, all groups, including the control group, were orally administered daidzein at 5 mg / kg / day. After two weeks, blood samples were collected, and serum was separated from the collected blood. The equol concentration in the serum was quantified using an Equol ELISA Kit (Healthcare Systems). The results are shown in Figure 12.

[0047] 2. Results and Effects of Examples in Test Example 6 As shown in FIG. 12, the serum equol concentration tended to increase in a concentration-dependent manner in the group administered with the lactic acid bacteria of this example (deposit number NITE ABP-04138) compared to the control group. These results confirmed that ingestion of the lactic acid bacteria of this example (deposit number NITE ABP-04138) promotes equol production.

[0048] Test Example 7: Evaluation of collagen synthase gene (COL1A1) and hyaluronic acid synthase gene (HAS2) expression promoting effect 1. Experimental Method (1) Method for culturing human dermal fibroblast cells (NHDF cells) Human dermal fibroblasts purchased from Kurabo Co., Ltd. were cultured and used for the experiment. The medium used was Dulbecco's Modified Eagles Medium (DMEM containing 1000 mg / L glucose) supplemented with 10 (v / v)% FBS (fetal bovine serum), 100 units / mL penicillin G, and 100 μg / mL streptomycin. Cells were cultured in 75 cm 2The culture was carried out in a culture flask at 37°C in the presence of 5% CO2. The cultured cells were washed twice with PBS (-) and then detached from the flask with a phenol red-containing 0.25 w / v% trypsin-1 mM EDTA·4Na solution for use in the experiment. (2) Evaluation method Method for evaluating the promotion of collagen (COL1A1) and hyaluronic acid (HAS2) synthase gene expression using NHDF cells NHDF cell suspension (1.6 × 10 5 Cells / mL) were seeded into a 12-well plate (1 mL / well) and cultured for 24 hours. After culture, the medium was replaced with medium containing the test substance. After 24 hours of culture, RNA was extracted from the cells according to standard methods, and the expression level of each gene was confirmed by RT-PCR.

[0049] 2. Results and Effects of the Example in Test Example 7 The effects of the lactic acid bacteria of this example (deposit number NITE ABP-04138) on the expression of various genes are shown in Figure 13. As shown in Figure 13, the addition of 10 μg / ml of Sakura lactic acid bacteria to NHDF cells significantly promoted the expression of the collagen synthase gene (COL1A1). Furthermore, the addition of 1 μg / ml of Sakura lactic acid bacteria significantly promoted the expression of the hyaluronic acid synthase gene (HAS2). This confirmed that Sakura lactic acid bacteria promotes the synthesis of collagen and hyaluronic acid. These results confirm that cherry blossom lactic acid bacteria have the effect of maintaining skin moisture and elasticity.

[0050] Test Example 8: Evaluation of EMT inhibitory effect 1. Experimental Method Immortalized human retinal pigment epithelial cell line ARPE-19 was cultured in a 96-well glass plate at 1 × 10 4Cells were seeded per well and cultured for 5 days at 37°C in a 5% CO2 environment. DMEM-F12 medium served as the control. EMT induction medium was prepared by adding TGF-β2 (5 ng / ml) and TNF-α (100 ng / ml) to DMEM-F12 medium. Pre-prepared 3 mg / ml, 1 mg / ml, and 0.3 mg / ml samples were diluted 1000-fold with EMT induction medium to form 3 μg / ml, 1 μg / ml, and 0.3 μg / ml test samples, respectively. A blank was prepared by diluting the sample solvent 1000-fold. The original medium was replaced with the control, EMT induction medium, and test sample, and the RPE cells were cultured at 37°C in a 5% CO2 environment. After 48 hours, the plates were removed from the incubator, washed twice with PBS, and fixed with 4% paraformaldehyde for 30 minutes at room temperature. After washing twice with PBS and blocking for 1 hour, the cells were stained for F-actin (Alexa fluor 568 phalloidin, Invitrogen) and nuclei (Hoechst 33342, Invitrogen) for 1 hour. Immediately after washing with PBS, five images per well were taken at 10x magnification using a fluorescence microscope (LAS AF with DMI6000B, Leica) at the top, bottom, center, left, and right positions. The captured fluorescent images were quantified using Image-J. The EMT inhibition rate was calculated using the following formula. The results are shown in Figure 14.

number

[0051] 2. Results and Effects of the Example in Test Example 8 As shown in Figure 14, it was confirmed that the addition of cherry lactic acid bacteria increased the EMT inhibition rate in a concentration-dependent manner. These results confirmed that Sakura lactic acid bacteria suppress epithelial EMT, support the health of the uterine epithelium, and have preventive and therapeutic effects on functional dysmenorrhea, thereby suppressing and improving discomfort such as menstrual pain.

[0052] The lactic acid bacteria of the present invention ( Deposit number NITE ABP-04138 The following formulation examples are not intended to limit the scope of the present invention. Formulation example 1: Chewing gum Sugar 53.0wt% Gum base 20.0 Glucose 10.0 Starch syrup 16.0 Fragrance 0.5 Lactic acid bacteria of the present invention 0.5 100.0wt%

[0053] Mixing example 2: Gummies Reduced starch syrup 40.0wt% Granulated sugar 20.0 Glucose 20.0 Gelatin 4.7 Wednesday 9.68 Yuzu juice 4.0 Yuzu flavor 0.6 Dye 0.02 Lactic acid bacteria of the present invention 1.0 100.0wt%

[0054] Mixing example 3: Candy Sugar 50.0wt% Starch syrup 33.0 Wednesday 14.4 organic acid 2.0 Fragrance 0.2 Lactic acid bacteria of the present invention 0.4 100.0wt%

[0055] Blending example 4: Yogurt (hard / soft) Milk 41.5wt% Skim milk powder 5.8 Sugar 8.0 Agar 0.15 Gelatin 0.1 Lactic acid bacteria 0.005 Lactic acid bacteria of the present invention 0.4 fragrance trace amount water residue 100.0wt%

[0056] Formulation example 5: Soft drink High fructose corn syrup 30.0wt% Emulsifier 0.5 Lactic acid bacteria of the present invention 0.3 Fragrance (appropriate amount) Purified water remainder 100.0wt%

[0057] Formulation example 6: Tablet confectionery Sugar 76.4wt% Glucose 19.0 Sucrose fatty acid ester 0.2 Lactic acid bacteria of the present invention 0.5 Purified water 3.9 100.0wt%

[0058] Formulation example 7: Soft capsule Brown rice germ oil 47.0wt% Yuzu seed oil 40.0 Emulsifier 12.0 Lactic acid bacteria of the present invention 1.0 100.0wt%

[0059] Formulation example 8: Tablets Lactose 54.0wt% Microcrystalline cellulose 30.0 Starch decomposition product 10.0 Glycerin fatty acid ester 5.0 Lactic acid bacteria of the present invention 1.0 100.0wt%

[0060] Formulation example 9: Cosmetic cream Squalane 20.0wt% Beeswax 5.0 Refined Jojoba Oil 5.0 Glycerin 5.0 Glycerin Monostearate 2.0 Polyoxyethylene (20) sorbitan Monostearate 2.0 Lactic acid bacteria of the present invention 2.0 Preservatives (appropriate amount) Fragrance (appropriate amount) Purified water remainder 100.0wt%

[0061] Formulation example 10: Lotion Ethanol 5.0wt% Glycerin 2.0 1,3-butylene glycol 2.0 Polyethylene oleyl ether 0.5 Sodium citrate 0.1 Citric acid 0.1 Lactic acid bacteria of the present invention 0.1 Purified water remainder 100.0wt%

[0062] Formulation example 11: Body gel Macadamia nut oil 2.0wt% Octyldodecyl myristate 10.0 Methylphenylpolysiloxane 5.0 Behenyl Alcohol 3.0 Stearic Acid 3.0 Batyl alcohol 1.0 Glyceryl Monostearate 1.0 Polyoxyethylene sorbitol tetraoleate 2.0 Hydrogenated soybean phospholipid 1.0 Ceramide 0.1 Retinol Palmitate 0.1 Preservatives (appropriate amount) Centella asiatica extract 1.0 Lactic acid bacteria of the present invention 1.0 1,3-butylene glycol 5.0 Purified water remainder 100.0wt%

[0063] Formulation example 12: Emulsion Squalane 4.0wt% Vaseline 2.5 Cetyl alcohol 2.0 Glycerin 2.0 Lipophilic Glyceryl Monostearate 1.0 Stearic Acid 1.0 L-Arginine 1.0 Lactic acid bacteria of the present invention 0.5 Potassium hydroxide 0.1 fragrance trace amount Purified water remainder 100.0wt%

[0064] Formulation example 13: Bath additive (liquid) Propylene glycol 50.0wt% Ethanol 20.0 Sodium sulfate 5.0 Lactic acid bacteria of the present invention 0.5 Lanolin 0.5 Avocado oil 0.5 Dye 1.5 Fragrance 22.0 100.0wt% [Industrial Applicability]

[0065] As described above, the present invention can provide novel lactic acid bacteria and uses thereof.

Claims

1. A lactic acid bacterium represented by Lacticaseibacillus paracasei sp. Shidare (deposit number NITE ABP-04138).

2. An IgA production promoter comprising the lactic acid bacterium according to claim 1 as an active ingredient.

3. An IL-10 production promoter comprising the lactic acid bacterium according to claim 1 as an active ingredient.

4. An IL-12 production promoter comprising the lactic acid bacterium according to claim 1 as an active ingredient.

5. A serotonin production promoter comprising the lactic acid bacterium according to claim 1 as an active ingredient.

6. A stool shape normalizing agent containing the lactic acid bacterium according to claim 1 as an active ingredient.

7. A laxative containing the lactic acid bacterium according to claim 1 as an active ingredient.

8. An agent for improving the feeling of incomplete evacuation, comprising the lactic acid bacterium according to claim 1 as an active ingredient.

9. 10. An intestinal regulator comprising the lactic acid bacterium according to claim 1 as an active ingredient.

10. An immunomodulator containing the lactic acid bacteria of claim 1 as an active ingredient.

11. An equol production promoter comprising the lactic acid bacterium according to claim 1 as an active ingredient.

12. A collagen synthesis gene expression promoter comprising the lactic acid bacterium according to claim 1 as an active ingredient.

13. A hyaluronic acid synthesis gene expression promoter containing the lactic acid bacterium according to claim 1 as an active ingredient.

14. An EMT inhibitor containing the lactic acid bacterium according to claim 1 as an active ingredient.

15. A preventive and therapeutic agent for functional dysmenorrhea, comprising the lactic acid bacterium according to claim 1 as an active ingredient.

16. A PMS preventive and therapeutic agent containing the lactic acid bacteria of claim 1 as an active ingredient.

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