Composition containing specific components

Abstract

An object of the present invention is to provide a composition that exhibits the effect of promoting UCP1 expression and / or the effect of promoting brown adipocyte differentiation, which is free from side effects and can be taken over a long period of time. [Solution] The above object can be achieved by a composition containing at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine, and mint, and having the effect of promoting UCP1 expression and / or brown adipocyte differentiation.

Description

[Technical field]

[0001] The present invention relates to a composition which contains a specific component and has an effect of promoting UCP1 expression and / or promoting brown adipocyte differentiation. [Background technology]

[0002] Uncoupling proteins (UCPs) have the function of uncoupling oxidative phosphorylation in the inner mitochondrial membrane and dissipating energy as heat (see Non-Patent Document 1). In particular, UCP1 is specifically expressed in brown adipocytes, which are the site of heat production, and plays a role in converting neutral lipids into heat and directly consuming energy without going through ATP synthesis in the electron transport system. Furthermore, if the differentiation of precursor cells into brown adipocytes can be promoted, the expression level of UCP1 can be increased, making it possible to convert neutral lipids into heat. Therefore, if the expression level of UCP1 or brown adipocyte differentiation can be promoted, there is a possibility that obesity can be eliminated through the consumption of neutral lipids. Regarding such brown adipocyte differentiation promoters and UCP1 expression promoters, those containing catechins as active ingredients are described in Patent Document 1. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] JP 2014-65672 A [Non-patent literature]

[0004] [Non-Patent Document 1] Proceedings of the 124th Symposium of the Japanese Medical Association, p.62-70 Summary of the Invention [Problem to be solved by the invention]

[0005] The substance capable of promoting the expression level and activity of UCP1 described in Patent Document 1 contains a specific substance isolated and purified from a natural product as an active ingredient. However, since such a specific substance has a strong pharmacological action, care must be taken when taking it, and there is a risk of side effects caused by excessive intake.

[0006] The present invention has been made in consideration of the above circumstances, and an object of the invention is to provide a composition that exhibits an effect of promoting UCP1 expression and / or an effect of promoting brown adipocyte differentiation, which is free from side effects and can be taken for a long period of time and sustainedly. [Means for solving the problem]

[0007] The present inventors have conducted extensive research to solve the above problems, and have surprisingly found that the inclusion of a specific component provides a particularly significant UCP1 expression promoting effect and / or brown adipocyte differentiation promoting effect. The inventors have succeeded in creating compositions containing the specific component as compositions for promoting UCP1 expression, compositions for promoting metabolism, anti-obesity compositions, compositions for reducing body fat, and compositions for promoting thermogenesis. The present invention has been completed based on these findings and successful examples.

[0008] Therefore, according to the present invention, there is provided a cosmetic composition containing at least one ingredient selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint.

[0009] According to another aspect of the present invention, there is provided a composition for promoting UCP1 expression, which comprises at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint.

[0010] According to another aspect of the present invention, there is provided a composition for activating brown adipocytes, which comprises at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint.

[0011] According to another aspect of the present invention, there is provided a composition for promoting metabolism, which comprises at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint.

[0012] According to another aspect of the present invention, there is provided an anti-obesity composition comprising at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint.

[0013] According to another aspect of the present invention, there is provided a composition for reducing body fat, which comprises at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint.

[0014] According to another aspect of the present invention, there is provided a composition for promoting thermogenesis, which comprises at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint.

[0015] According to another aspect of the present invention, there is provided a composition comprising at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine, and mint. Effect of the Invention

[0016] The composition of the present invention has the effect of promoting UCP1 expression and / or the effect of promoting brown fat cell differentiation, and further, in relation to these effects, is expected to exhibit effects such as brown fat cell activation, metabolism promoting, anti-obesity, body fat reducing, thermogenesis promoting, UCP1 expression promoting, energy production promoting, neutral lipid lowering, sebum suppressing, and fat metabolism promoting.

[0017] The specific components used in the composition of the present invention have a proven track record of use in pharmaceuticals, cosmetics, etc., and therefore the composition of the present invention is highly safe. Thus, by virtue of the above-mentioned effects, the composition of the present invention is useful as a composition for promoting UCP1 expression, a composition for promoting metabolism, an anti-obesity composition, a composition for reducing body fat, and a composition for promoting thermogenesis, and is expected to be provided in a parenteral or oral form. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0018] The present invention will be described in detail below. The composition of the present invention has the effect of promoting UCP1 expression and / or the effect of promoting brown adipocyte differentiation, and contains at least any one of the following ingredients (hereinafter referred to as specific ingredients): hops, trehalose, aspartame, barley, ceramide, placenta, alanine, and mint.

[0019] The composition of the present invention may have either one of the effects of promoting UCP1 expression and the effect of promoting brown adipocyte differentiation, or may have both of these effects. In addition, the specific component may be any one of the specific components, or may be two or more components.

[0020] In addition to the effects of promoting UCP1 expression and promoting brown adipocyte differentiation, the composition of the present invention may also exhibit brown adipocyte activation, metabolism promoting, anti-obesity, body fat reducing, thermogenesis promoting, energy production promoting, neutral lipid lowering, sebum suppressing and fat metabolism promoting effects.

[0021] Because the composition of the present invention has the above-mentioned action, it can take the form of a composition for promoting UCP1 expression, a composition for promoting metabolism, an anti-obesity composition, a composition for reducing body fat, and a composition for promoting thermogenesis.

[0022] Hops are made from the common lupin ( Hops hopsThe hop plant may be any plant known as a hop plant, and is not particularly limited thereto. For example, the flowers (cones, seeds, fruits, leaves, stems, roots, and other parts of the hop plant may be used. It is preferable to use hop cones.

[0023] Mint is Mentha piperita ( Mint pepper The mint may be any part of the plant known as mint, including, for example, its flowers, seeds, fruits, leaves, stems, and roots. It is preferable to use mint leaves.

[0024] Barley is Hordeum vulgare ( Barley vulgar The barley extract is not particularly limited as long as it is known as a plant extract, and for example, its flowers, seeds, fruits, leaves, stems, roots, and other parts can be used. However, it is preferable to use barley leaves and stems.

[0025] The placenta is not particularly limited as long as it is a commonly known placenta, and examples thereof include animal placenta, which is an extract derived from the placenta of pigs, humans, cows, horses, sheep, etc., or a component thereof, marine placenta, which is an extract derived from the ovarian membrane of fish, etc., or a component thereof, and plant placenta, which is an extract derived from the germ of a plant, etc., or a component thereof. Animal placenta is preferred, and animal placenta derived from pigs is more preferred.

[0026] The ceramide is not particularly limited as long as it is a compound in which sphingosine and a fatty acid are bonded by an amide bond, and examples include ceramides derived from natural products and ceramides obtained by synthesis. Ceramides derived from natural products are preferred, and ceramides derived from rice, corn, konjac, Maitake mushroom, Pleurotus venus, soybean, wheat, beet, etc. are more preferred, and ceramide derived from rice (glucosylceramide) is even more preferred.

[0027] The trehalose is not particularly limited as long as it is a commonly known trehalose, and examples thereof include trehalose itself and trehalose derivatives such as its hydrates and optical isomers, with D-(+)-trehalose being preferred.

[0028] Aspartame is not particularly limited as long as it is aspartame as is commonly known, and examples thereof include aspartame itself and aspartame derivatives such as optical isomers thereof, and preferably aspartame having CAS registration number 22839-47-0.

[0029] The alanine is not particularly limited as long as it is a commonly known alanine, and examples thereof include alanine itself and derivatives thereof such as optical isomers, and preferably alanine having CAS Registry Number 338-69-2.

[0030] Of the specific components, natural products may be processed products thereof. Examples of processed products include, but are not limited to, dried powders, shredded products and dried powders thereof, squeezed juice and dried powders thereof, extracts and dried powders thereof, etc. However, from the viewpoints of ease of processing, storage, transportation, etc., and versatility of use, it is preferable to use dried powders.

[0031] The method for obtaining dried natural products is not particularly limited, but for example, barley stalk and leaf powder can be produced by the method disclosed in Japanese Patent No. 3277181. That is, after harvesting barley stalks and leaves, they are washed with water to remove dirt and the like, drained, and then cut to an appropriate length. Then, as necessary, a blanching treatment such as hot water treatment or steam heat treatment is performed. The temperature and time of this treatment may be appropriately determined depending on the amount of barley stalks and leaves to be treated and the pH of the hot water. The blanched barley stalks and leaves are immediately cooled and dehydrated by centrifugation or the like. Then, they are dried so that the moisture content is 5 wt% or less. Barley stalk and leaf powder can be obtained by further undergoing a coarse grinding process, a heating process, a fine grinding process, and the like.

[0032] The specific ingredients are natural products, derivatives thereof, and synthetic products that have a long history of being ingested by humans and are therefore highly safe. Therefore, the composition of the present invention is highly practical and can be used for a variety of purposes in parenteral or oral forms, either as is or after processing.

[0033] The specific component may be one produced by a method generally known to those skilled in the art, or may be one that is available on the market.

[0034] The specific component exhibits a UCP1 expression promoting effect or a brown adipocyte differentiation promoting effect. In this specification, the "expression promoting effect" refers to, for example, in the case of a UCP1 expression promoting effect, at least one of the effects of promoting the expression amount of the UCP1 gene in a subject, increasing the translation amount of the UCP1 protein, and activating UCP1. In addition, the brown adipocyte activation effect refers to at least one of the effects of activating brown adipocytes and increasing the number of brown adipocytes.

[0035] Regarding the UCP1 expression promoting effect, as described in Non-Patent Document 1, it is known that the function of UCP1 is reduced in obese animals, that UCP1 is increased in animals that do not become obese even when they eat a lot, and that mice in which UCP1 expression has been artificially reduced become obese and mice with high UCP1 expression become thin. Therefore, an anti-obesity effect can be expected by promoting UCP1 expression, and a substance that actually has the effect of promoting UPC1 expression can promote lipolysis in white fat cells and at the same time activate UCP1, converting liberated fatty acids into heat, ultimately reducing body fat.

[0036] Thermogenesis promoting action refers to, for example, the action of promoting heat production from neutral lipids by brown fat cells. The neutral lipid lowering action refers to, for example, the action of decomposing neutral lipids to reduce the total amount of neutral lipids in the body. The sebum suppressing action refers to, for example, the action of suppressing the generation of sebum. The fat metabolism promoting action refers to, for example, the action of qualitatively converting fat cells and fat tissues that form an obese state into small normal fat cells. The body fat reducing action refers to, for example, the action of reducing body fat by converting free fatty acids into heat. The metabolism promoting action and the anti-obesity action refer to, for example, the action of promoting metabolism so as to reduce body fat and the action of suppressing obesity due to said action. The energy production promoting action refers to, for example, the action of producing energy through thermogenesis from neutral lipids by brown fat cells. Each action in this paragraph includes the action described in the respective meanings, but also includes those realized by other action mechanisms.

[0037] The anti-disease effect of an anti-disease composition such as the anti-obesity composition of the present invention means, for example, in the case of the anti-obesity effect of an anti-obesity composition, suppressing or delaying the onset of a current or future state considered to be obesity or obesity in a subject, or improving that state. Insulin resistance refers to a state in which the main function of insulin, the promotion of sugar absorption, is weakened in the liver, fat cells, skeletal muscles, etc. The anti-insulin resistance effect means suppressing or delaying the worsening of a value that is an index of current or future insulin resistance in a subject, as measured by the SSPG (steady-state plasma glucose) method or the like, or improving that value.

[0038] Obesity is known to cause various diseases, such as type 2 diabetes, hypertension, and hyperlipidemia. Furthermore, these diseases may lead to stroke and ischemic heart disease. In an obese state, fat cells are enlarged, TNF-α and free fatty acids (FFA) are secreted, which inhibit the uptake of sugar in insulin target cells such as muscle cells and liver cells, and suppress the secretion of adiponectin, which promotes insulin action, causing a decrease in insulin sensitivity at the cellular level, resulting in insulin resistance. The above diseases caused by obesity are considered to be a series of metabolic abnormalities based on insulin resistance. Therefore, by using the composition of the present invention, it is possible to prevent or improve obesity and diseases associated with obesity, such as type 2 diabetes, hypertension, and hyperlipidemia.

[0039] Here, prevention and amelioration of type 2 diabetes refers to suppressing or delaying the onset of a diabetic state or borderline state, or moving from such a state closer to a state called the normal range. Prevention and amelioration of hypertension refers to suppressing or delaying the onset of a hypertensive state or borderline state, or moving from such a state closer to a state called the normal range. Prevention and amelioration of hyperlipidemia refers to suppressing or delaying the onset of a hyperlipidemic state or borderline state, or moving from such a state closer to a state called the normal range.

[0040] The method for producing the composition of the present invention is not particularly limited, and for example, a solid composition can be obtained by incorporating a specific component at room temperature or under heating. Alternatively, this solid composition can be dissolved in a solvent such as water to obtain a liquid composition. Furthermore, a liquid composition can be obtained by adding and mixing other solid components to a liquid product of a specific component. The obtained liquid composition may be powdered through a drying process. Examples of the drying process in this case include, but are not limited to, spray drying and freeze drying.

[0041] The composition of the present invention can be used as it is or mixed with other components depending on the application. In this way, the composition of the present invention can contain various ingredients other than the specific ingredients as long as the object of the present invention can be achieved.

[0042] For example, the composition of the present invention may further contain additives used in normal processing, such as excipients, bulking agents, binders, thickeners, emulsifiers, colorants, flavors, flavor oils, etc. The amount of additive used is not particularly limited as long as it does not interfere with the solution of the problems of the present invention, and may be appropriately adjusted.

[0043] The composition of the present invention is not particularly limited as long as it is in a commonly used form, and may take any of the following forms: liquid, lotion, mousse, gel, emulsion, suspension, cream, ointment, sheet, aerosol, spray, stick, powder, grain, granule, tablet, rod, plate, block, solid, paste, capsule, caplet, etc.

[0044] The composition of the present invention can be used as a non-oral composition, for example, in a form suitable for cosmetics. That is, one aspect of the composition of the present invention is a cosmetic composition. For example, the composition of the present invention can be processed into various forms such as lotions, emulsions, gels, creams, and ointments, either as is or mixed with additives that are usually used in the processing of cosmetics. Specifically, it can be used as a lotion, cosmetic cream, emulsion, cream, pack, hair tonic, hair cream, shampoo, hair rinse, treatment, face wash, foundation, hair growth agent, aqueous ointment, spray, etc.

[0045] The content of the specific component contained in the composition of the present invention is not particularly limited as long as the effect of promoting UCP1 expression and / or brown adipocyte differentiation is observed; for oral compositions, the content is, for example, 0.0001 wt% or more, preferably 0.001 wt% or more, relative to the total composition, and for non-oral compositions such as cosmetics, the content is, for example, 0.00001 wt% or more, preferably 0.0001 wt% or more.

[0046] A specific embodiment of the composition of the present invention is, for example, a composition containing any one of the specific components in an amount of 0.001 to 100 wt% relative to the entire composition. Among these, a composition containing the specific component in an amount of 0.01 to 80 wt% is more preferred.

[0047] Another specific embodiment of the composition of the present invention is a composition containing any two of the specific components in a total amount of 10 to 100 wt% relative to the entire composition, including a composition containing one of the two components in a ratio of 10 to 90 wt% and the other in a ratio of 90 to 10 wt%.

[0048] Another specific embodiment of the composition of the present invention is a composition containing any three specific components in a total amount of 10 to 100 wt% relative to the entire composition, including a composition containing 10 to 50 wt% of the first component, 30 to 40 wt% of the second component, and 60 to 10 wt% of the third component among the three components.

[0049] Below, formulation examples according to specific embodiments of the composition of the present invention will be shown, but the present invention is not limited to these formulation examples, and the present invention can take various forms as long as the object of the present invention can be solved.

[0050] (Example 1: Lotion) The composition of the present invention in the form of a lotion was prepared by mixing 0.01 part by mass of trehalose, 0.01 part by mass of ginger extract, 0.01 part by mass of Cassia japonica, 10 parts by mass of glycerin, 3 parts by mass of diglycerin, 12 parts by mass of 1,3-butylene glycol, 3 parts by mass of pentylene glycol, 0.1 part by mass of sodium hyaluronate, 0.01 part by mass of citric acid, 0.02 part by mass of sodium citrate, 0.1 part by mass of xanthan gum, 0.15 part by mass of methylparaben, 0.2 part by mass of carbomer, 0.03 part by mass of sodium hydroxide, and the remainder of the mixture being 100 parts by mass.

[0051] (Example 2: Shampoo) The composition of the present invention in the form of a shampoo was prepared by mixing 0.02 parts by weight of hop extract, 0.01 parts by weight of caffeine, 7.5 parts by weight of sodium laureth sulfate, 4.2 parts by weight of cocamidopropyl betaine, 3 parts by weight of cocamide DEA, 0.1 parts by weight of 1,3-butylene glycol, 0.225 parts by weight of polyquaternium-10, 0.15 parts by weight of citric acid, 0.05 parts by weight of sodium citrate, 0.9 parts by weight of phenoxyethanol, and the remainder water, making the total 100 parts by weight.

[0052] (Example 3: Soap) The composition of the present invention in the form of a soap was prepared by mixing and solidifying 0.5 parts by weight of aspartame, 0.2 parts by weight of melilot extract, 0.1 parts by weight of elastin, 2 parts by weight of glycerin, 1 part by weight of olive oil, 0.1 parts by weight of tetrasodium EDTA, 0.2 parts by weight of tetrasodium etidronate, and the remainder of the soap base, making a total of 100 parts by weight.

[0053] (Formulation example 4: Emulsion) The composition of the present invention was prepared in the form of an emulsion by mixing 0.1 part by mass of ceramide, 0.1 part by mass of alanine, 0.1 part by mass of arginine, 3 parts by mass of sucrose fatty acid ester, 12 parts by mass of glycerin, 6 parts by mass of squalane, 24 parts by mass of dimethyl silicone oil, 1 part by mass of polypropylene glycol, 0.06 part by mass of thickener, 0.2 part by mass of phenoxyethanol, 5 parts by mass of ethanol, 0.01 part by mass of sodium hydroxide, and the remainder purified water, making the total 100 parts by mass.

[0054] (Formulation Example 5: Cosmetic cream) Based on a total of 100 parts by mass, the following ingredients are added: alanine 0.1 part by mass, fructooligosaccharide 0.1 part by mass, squalane 15.0 parts by mass, octyldodecyl myristate 4.0 parts by mass, hydrogenated soybean phospholipid 0.2 parts by mass, butyl alcohol 2.4 parts by mass, hardened oil 1.5 parts by mass, stearic acid 1.5 parts by mass, lipophilic glycerin monostearate 1.5 parts by mass, polyglyceryl monostearate 0.5 parts by mass, behenyl alcohol 0.8 parts by mass, polyglyceryl monomyristate 0.7 parts by mass, white beeswax 0.3 parts by mass, d-δ-tocopherol 0.1 parts by mass, methylparaben 0.3 parts by mass, C10-30 alkyl-modified carboxyvinyl polymer 0.2 parts by mass, carboxyvinyl polymer 0.1 parts by mass, 1,3-butanediol 18.0 parts by mass, sodium hydroxide 0.1 parts by mass of the above-mentioned powder and the remainder of the powder were mixed together to prepare a composition of the present invention in the form of a cosmetic cream.

[0055] (Example 6: Pack) The composition of the present invention was prepared in the form of a pack by mixing 0.1 part by mass of placenta, 0.01 part by mass of gallic acid, 20.0 parts by mass of polyvinyl alcohol, 5.0 parts by mass of glycerin, 20.0 parts by mass of ethanol, 6.0 parts by mass of kaolin, 0.2 parts by mass of preservative, 0.1 part by mass of fragrance, and the remainder being purified water, making the total 100 parts by mass.

[0056] (Formulation Example 7: Tablets) The composition of the present invention in the form of tablets was prepared by mixing and tableting 10 parts by weight of barley leaf powder, 8 parts by weight of ginger extract, 5 parts by weight of caffeine, 5 parts by weight of melilot, 20 parts by weight of crystalline cellulose, 50 parts by weight of lactose, 4 parts by weight of magnesium stearate, and the remainder of corn starch (total 100 parts by weight).

[0057] (Formulation Example 8: Tablets) The composition of the present invention in the form of a tablet was prepared by mixing and tableting 0.5 parts by mass of alanine, 8 parts by mass of Cassia japonica powder, 5 parts by mass of melilot, 0.5 parts by mass of fructooligosaccharides, 20 parts by mass of crystalline cellulose, 50 parts by mass of lactose, 4 parts by mass of magnesium stearate, and the remainder of corn starch, totaling 100 parts by mass.

[0058] (Formulation Example 9: Granules) The composition of the present invention was prepared in the form of granules by mixing and granulating 15 parts by mass of hop powder, 5 parts by mass of Cassia japonica extract, 10 parts by mass of lactose, 1 part by mass of calcium stearate, and the remainder of the crystalline cellulose, totalling 100 parts by mass.

[0059] (Formulation Example 10: Capsules) The composition of the present invention was prepared in the form of a capsule by mixing 10 parts by weight of mint extract, 20 parts by weight of ginger extract, 5 parts by weight of elastin, 8 parts by weight of lecithin, and the remainder of olive oil (total 100 parts by weight). The mixture was used as the content liquid and encapsulated in a capsule shell to prepare the composition of the present invention in the form of a capsule.

[0060] (Example 11: Liquid) The composition of the present invention was prepared in the form of a liquid by mixing 0.84 parts by mass of young barley leaf extract, 1 part by mass of arginine, 1 part by mass of alanine, 10 parts by mass of fructose glucose liquid sugar, 1 part by mass of citric acid, 0.02 parts by mass of sodium benzoate, 2 parts by mass of a flavor preparation, 0.05 parts by mass of sucralose, 0.03 parts by mass of acesulfame potassium, and the remainder being purified water, making a total of 100 parts by mass.

[0061] The present invention will be described in more detail below with reference to examples. However, the present invention is not limited to these examples, and the present invention can take various forms as long as the object of the present invention can be achieved. EXAMPLES

[0062] It has been demonstrated, as described below, that the specific ingredients trehalose, aspartame, barley, ceramide, placenta, alanine, mint and hops have a particularly significant UCP1 expression promoting effect.

[0063] (1) Test substance Of the specific components, trehalose, aspartame, ceramide, placenta, alanine, mint and hops were commercially available.

[0064] The barley used was dried powder of barley stalks and leaves prepared as follows. That is, barley stalks and leaves harvested at a height of about 30 cm were washed with water, and pre-treated by removing adhering mud and the like, and cutting into pieces of about 5 to 10 cm in size. The pre-treated stalks and leaves were blanched once with hot water at 90 to 100°C for 90 to 120 seconds, and then cooled with cold water. The obtained stalks and leaves were then dried in a dryer with hot air at 80°C to 130°C for 20 to 180 minutes until the moisture content was 5% by mass or less. The dried stalks and leaves were coarsely pulverized to a size of about 1 mm using a mixer. The obtained barley stalks and leaves were finely pulverized using a pulverizer so that 90% or more passed through a 200 mesh division, and dried powder of barley stalks and leaves was obtained.

[0065] (2) Preparation of collagenase solution A collagenase solution was prepared by shaking a DMEM medium containing 0.2% (W / V) collagenase and 1% (W / V) BSA at room temperature for 1 hour.

[0066] (3) Preparation of brown adipocyte fluid ICR mice (Kyudo) were euthanized with diethyl ether, and brown adipose tissue was extracted from the back. Adipose tissue, blood vessels, muscles, etc. attached around the brown adipose tissue were removed from the obtained brown adipose tissue using tweezers, and the brown adipose tissue was washed three times in DMEM. The brown adipose tissue was then immersed in a collagenase solution and subjected to a shredding process in which the brown adipose tissue was finely chopped in the solution with scissors. A cell dispersion was obtained by incubating the shredded brown adipose tissue in the collagenase solution at 37°C for 1 hour. The obtained cell dispersion was filtered through a cell strainer with a pore size of 100 μm, and then subjected to centrifugation (800 rpm, 5 min, 20°C). The precipitated cells were then suspended in DMEM to obtain a cell suspension. The obtained cell suspension was then filtered using a 25 μm filter sterilized by autoclaving. The filtered supernatant was centrifuged (800 rpm, 5 min, 20° C.), and the precipitated cells were suspended in DMEM containing 10 vol% FBS to prepare a brown adipocyte solution.

[0067] (4) Preparation of medium As a growth medium, DMEM containing 10 vol% FBS was used.

[0068] A differentiation-inducing medium was prepared by adding 2.5 μM dexamethasone, 10 μg / ml insulin and 0.5 mM IBMX to the proliferation medium.

[0069] As a maintenance medium, a growth medium containing 10 μg / ml insulin was prepared using the insulin solution described above.

[0070] The medium containing the test substance was prepared as follows: each test substance was dissolved in DMSO, then diluted with the maintenance medium to adjust the final DMSO concentration to 0.5 vol%, and sterilized by filter. After sterilization, the test substances were diluted to the prescribed concentration, i.e., hops, trehalose, aspartame, barley, ceramide, placenta, alanine, and mint, to 20 μg / ml using the maintenance medium containing 0.5 vol% DMSO.

[0071] (5)Cell culture 75cm2 5 ml of the collagen coating solution was added to the flask, and the flask was allowed to stand at room temperature for about 1 hour. The collagen coating solution was then removed by suction, and the flask was washed twice with 5 ml of PBS to obtain a collagen-coated flask.

[0072] The brown adipocyte liquid was added to the collagen-coated flask, and the brown adipocytes were cultured for 24 hours in a 5% CO2 incubator at 37°C. The cultured brown adipocytes were then washed once with growth medium, which was then replaced with fresh growth medium, and cultured until they became subconfluent.

[0073] After the culture, the brown adipocytes were subjected to trypsinization to cause them to float. The resulting floating cells were seeded into each well of a collagen-coated 24-well plate (collagen coating solution 350 μL / well, PBS washing 350 μL / well × 2 times).

[0074] Brown adipocytes were cultured in a growth medium until they became confluent. After removing the medium from each well, 500 μl of differentiation-inducing medium was added to each well and cultured for 48 hours.

[0075] After removing the differentiation-inducing medium, 500 μl of medium containing 20 μg / ml of the test substance was added and cultured for 6 days. The medium was changed every other day. As a control, 500 μl of medium not containing the test substance was added and cultured for 6 days. The medium was changed every other day.

[0076] After 8 days of culture, the supernatant was collected, RNA was extracted using an RNeasy Mini Kit (Qiagen), and cDNA was synthesized using a QuantiTect Reverse Transcription Kit (Qiagen).

[0077] Using the obtained cDNA as a template and a primer for the UCP1 gene (Qiagen), quantitative real-time PCR was performed with the Rotor-Gene SYBR Green PCR Kit (Qiagen) to measure the mRNA expression level of UCP1. As an endogenous control, the mRNA expression level of GAPDH was measured using a GAPDH primer (Qiagen).

[0078] (6) Evaluation The results of measuring the mRNA expression level of UCP1 when each of the test substances, namely hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint, was administered (relative expression level when the mRNA expression level of the control was set at 1.00) are shown in Table 1. [Table 1]

[0079] As shown in Table 1, when hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint were used as test substances, the amount of UCP1 expression was high. [Industrial Applicability]

[0080] The compositions of the present invention contain specific components that can be administered either parenterally or orally, and are particularly useful for subjects suffering from cellulite, obesity and associated diseases, and contribute to the health and well-being of such subjects.

Claims

1. A composition for promoting energy consumption, comprising crushed barley stalks and leaves as an active ingredient (excluding herbal medicines containing aspergillus oryzae, germ and young barley leaves).

2. A composition for promoting thermogenesis, comprising crushed barley stalks and leaves as an active ingredient (excluding herbal medicines containing aspergillus oryzae, germ and young barley leaves).

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