Food composition for activating nuclear receptors and method for producing same

Abstract

To provide an excellent nuclear receptor-activating food composition.SOLUTION: The inventors have found out that leaves of one or more kinds of Spinacia oleracea, Beta vulgaris, Eruca vesicaria, Lactuca sativa L. var. longifolia, or Lactuca sativa L. var. longifoli, which are often consumed as baby leaves, or processed products thereof have a nuclear receptor-activating function. That is, the leaves or processed products thereof activate one or more types among plural types of nuclear receptors, specifically, RAR, RXR, PPAR, and ER. As a result, the following effects are expected: normalization of a fasting blood sugar level, inhibition of skin inflammation, inhibition of skin cornification, inhibition of skin light aging, inhibition of skin pigmentation, inhibition of fat accumulation, inhibition of fatty liver, and increase of blood HDL cholesterol.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a food composition for activating a nuclear receptor and a method for producing the same. [Background technology]

[0002] Nuclear receptors are involved in various biological functions, and therefore factors involved in controlling their activity have attracted attention and are being actively studied. Nuclear receptors are proteins present within cells, and are known to translocate into the nucleus upon binding to ligands, where they bind to DNA and control transcription. Well-known nuclear receptors include RAR, RXR, PPAR, and ER, and are known to play various roles in the body.

[0003] It has been reported that Tamibarotene, a selective agonist of nuclear receptors RARα and RARβ, has the effect of lowering fasting blood glucose and fasting insulin concentrations (Non-Patent Document 1). In addition, it has been reported that the nuclear receptor RARα induces glucose transporter 2 (GLUT2), which is responsible for glucose transport on the pancreatic β cell membrane, and induces the expression of glucokinase (GCK), which catalyzes the phosphorylation of glucose in pancreatic β cells and plays a role in maintaining glucose homeostasis, thereby maintaining the function of insulin-producing β cells, and that an agonist of the nuclear receptor RARβ2 improves insulin sensitivity in the liver, pancreas, and kidney, and reduces serum glucose and insulin levels in both normal mice in which obesity and type 2 diabetes are induced by a high-fat diet and genetically engineered model mice of obesity and type 2 diabetes (Non-Patent Document 2). From these findings, it is believed that fasting blood glucose levels are normalized by activating nuclear receptors.

[0004] Diseases that can be treated with RAR agonists, which activate the nuclear receptor RAR, have been reported to include inflammatory diseases of the skin, keratosis, photoaging, and pigmentation. It is believed that these diseases can be suppressed by activating nuclear receptors (RAR) (Non-Patent Document 3), and therefore it is believed that activating nuclear receptors can suppress inflammatory diseases of the skin, keratosis, photoaging, and pigmentation of the skin.

[0005] It has been reported that white adipogenesis is inhibited by inhibiting the expression of zinc finger protein 423 (Zfp423), a determinant of preadipocyte differentiation, via activation of the nuclear receptor RARα (Non-Patent Document 4). Based on this, it is believed that fat accumulation is suppressed by activation of nuclear receptors.

[0006] It has been reported that all-trans retinoic acid (atRA), a derivative of vitamin A, induces the expression of FGF21, a hepatocyte-derived hormone that suppresses obesity-induced fatty liver, via the nuclear receptors RARα and RARβ, and that the nuclear receptor RARα suppresses the expression of hepatic apolipoprotein CIII (Apo-CIII), which is known to be a central factor in the development of hypertriglyceridemia (TG), thereby reducing TG levels in the liver and blood (Non-Patent Document 2), and therefore it is believed that fatty liver is suppressed by activating nuclear receptors.

[0007] It has been reported that the nuclear receptor RARγ induces the ATP-binding cassette transporter 1 (ABCA1) of macrophages (Non-Patent Document 5), and that ABCA1 exports intracellular cholesterol and phospholipids to extracellular apolipoprotein AI (apoA-I), and HDL cholesterol is formed by binding to apoA-I (Non-Patent Document 6). Based on these findings, it is believed that activation of nuclear receptors promotes the release of cholesterol from macrophage foam cells and the formation of HDL cholesterol, thereby increasing HDL cholesterol in the blood.

[0008] In this way, activation of nuclear receptors is thought to lead to the normalization of fasting blood glucose levels, inhibition of inflammatory skin diseases, inhibition of skin keratosis, inhibition of skin photoaging, inhibition of skin pigmentation, inhibition of fat accumulation, inhibition of fatty liver, and an increase in blood HDL cholesterol. [Prior art documents] [Non-patent literature]

[0009] [Non-Patent Document 1] Hiroyuki Tsuchitani. "The usefulness of retinoids as a treatment for non-alcoholic fatty liver disease." YAKUGAKU ZASSHI 132.8 (2012): 903-909.-1 [Non-Patent Document 2] Saeed, Ali, et al. "Disturbed vitamin A metabolism in non-alcoholic fatty liver disease (NAFLD)." Nutrients 10.1 (2018): 29. [Non-Patent Document 3] Beckenbach, Lisa, et al. "Retinoid treatment of skin diseases." European Journal of Dermatology 25.5 (2015): 384-391. [Non-Patent Document 4] Wang, Bo, et al. "Retinoic acid inhibits white adipogenesis by disrupting GADD45A-mediated Zfp423 DNA demethylation." Journal of Molecular Cell Biology 9.4 (2017): 338-349. [Non-Patent Document 5] Costet, Philippe, et al. "Retinoic acid receptor-mediated induction of ABCA1 in macrophages." Molecular and Cellular Biology 23.21 (2003): 7756-7766. [Non-Patent Document 6] Yokoyama, Shinji. "Assembly of high-density lipoprotein." Arteriosclerosis, Thrombosis, and Vascular Biology 26.1 (2006): 20-27. Summary of the Invention [Problem to be solved by the invention]

[0010] The problem to be solved by the present invention is to provide a novel food composition for activating a nuclear receptor. [Means for solving the problem]

[0011] Based on the above, the inventors of the present application have conducted extensive research and found that the leaves of one or more of spinach, beet, arugula, red romaine, and green romaine, which are often eaten as baby leaves, or their processed products, have a nuclear receptor activating effect. That is, the leaves or their processed products activate multiple types of nuclear receptors, specifically, one or more of RAR, RXR, PPAR, and ER. As a result, it is expected to normalize fasting blood glucose levels, suppress inflammatory skin diseases, suppress skin keratosis, suppress photoaging of the skin, suppress skin pigmentation, suppress fat accumulation, suppress fatty liver, and increase HDL cholesterol in the blood. From this perspective, the present invention is defined as follows.

[0012] The food composition for activating nuclear receptors according to the present invention contains any one or more of leaves of spinach, beet, arugula, red romaine, and green romaine, or a processed product thereof. In other words, the raw material contributing to nuclear receptor activation is any one or more of leaves of spinach, beet, arugula, red romaine, and green romaine, or a processed product thereof. The component contributing to nuclear receptor activation is a component contained in spinach, beet, arugula, red romaine, and green romaine. The aforementioned leaves are preferably within 30 days after germination. The nuclear receptor is preferably any one or more of RAR, RXR, PPAR, and ER.

[0013] The food composition for activating a nuclear receptor according to the present invention contains baby leaves or processed products thereof. The baby leaves are young leaves of one or more of spinach, beet, arugula, red romaine, and green romaine or processed products thereof. The nuclear receptor is preferably one or more of RAR, RXR, PPAR, or ER.

[0014] The food composition for normalizing fasting blood glucose level according to the present invention contains a component that contributes to nuclear receptor activation, and originates from the leaves of one or more of spinach, beet, arugula, red romaine, and green romaine. The component that contributes to nuclear receptor activation is preferably a component that contributes to RAR activation, and the leaves are preferably from within 30 days after germination.

[0015] The food composition for suppressing inflammatory skin diseases according to the present invention contains a component that contributes to nuclear receptor activation, and originates from the leaves of one or more of spinach, beet, arugula, red romaine, and green romaine. The component that contributes to nuclear receptor activation is preferably a component that contributes to RAR activation, and the leaves are preferably from within 30 days after germination.

[0016] The food composition for inhibiting skin keratosis according to the present invention contains a component that contributes to nuclear receptor activation, and originates from the leaves of one or more of spinach, beet, arugula, red romaine, and green romaine. The component that contributes to nuclear receptor activation is preferably a component that contributes to RAR activation, and the leaves are preferably from within 30 days after germination.

[0017] The food composition for suppressing skin photoaging according to the present invention contains a component that contributes to nuclear receptor activation, and originates from the leaves of one or more of spinach, beet, arugula, red romaine, and green romaine. The component that contributes to nuclear receptor activation is preferably a component that contributes to RAR activation, and the leaves are preferably from within 30 days after germination.

[0018] The food composition for inhibiting skin pigmentation according to the present invention contains a component that contributes to nuclear receptor activation, and originates from the leaves of one or more of spinach, beet, arugula, red romaine, and green romaine. The component that contributes to nuclear receptor activation is preferably a component that contributes to RAR activation, and the leaves are preferably from within 30 days after germination.

[0019] The food composition for inhibiting fat accumulation according to the present invention contains a component that contributes to nuclear receptor activation, and originates from the leaves of one or more of spinach, beet, arugula, red romaine, and green romaine. The component that contributes to nuclear receptor activation is preferably a component that contributes to RAR activation, and the leaves are preferably from within 30 days after germination.

[0020] The food composition for suppressing fatty liver according to the present invention contains a component that contributes to nuclear receptor activation, and originates from the leaves of one or more of spinach, beet, arugula, red romaine, and green romaine. The component that contributes to nuclear receptor activation is preferably a component that contributes to RAR activation. Preferably, the leaves are within 30 days of germination.

[0021] The food composition for increasing blood HDL cholesterol according to the present invention contains a component that contributes to nuclear receptor activation, which is preferably derived from the leaves of one or more of spinach, beet, arugula, red romaine, and green romaine, and the nuclear receptor activation contributing component is preferably a component that contributes to RAR activation, and the leaves are preferably from within 30 days after germination.

[0022] The method for producing a food composition for activating a nuclear receptor according to the present invention includes at least an extraction step. Here, the component that contributes to nuclear receptor activation is extracted from the leaves of one or more of spinach, beet, arugula, red romaine, and green romaine. The extraction is preferably performed with water, aqueous ethanol, or an organic solvent. The leaves are preferably from within 30 days after germination. Effect of the Invention

[0023] The present invention makes it possible to provide a novel food composition for activating nuclear receptors. That is, by providing a component that contributes to the activation of nuclear receptors, a food composition is provided that contributes to normalizing fasting blood glucose levels, suppressing inflammatory skin diseases, suppressing skin keratosis, suppressing photoaging of the skin, suppressing skin pigmentation, suppressing fat accumulation, suppressing fatty liver, and increasing HDL cholesterol in the blood. [Brief description of the drawings]

[0024] [Figure 1] Figure showing nuclear receptor activation ability in young leaves [Diagram 2] Diagram showing nuclear receptor activation ability in adult leaves DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] <Food composition for activating a nuclear receptor according to the present embodiment> The contributing component of the food composition for activating a nuclear receptor according to the present embodiment (hereinafter referred to as "the composition") is one or more kinds of leaves or processed products thereof from spinach, beet, arugula, red romaine, and green romaine. Details of the leaves of the items and the processed products thereof will be described later.

[0026] <Leaves according to the present embodiment> In the present embodiment, the leaves used may be within 90 days after germination, preferably within 50 days after germination. Generally, baby leaves refer to leafy vegetables that are about 10 to 30 days after germination (hereinafter referred to as "young leaves"), and can be used more preferably than leafy vegetables that are more than 30 days after germination (hereinafter referred to as "adult leaves"). However, it is particularly preferable to use young leaves that are 14 to 22 days after germination. The leaves may include the petioles. The type of baby leaves is not particularly limited, but examples include vegetables of the Brassicaceae, Amaranthaceae, Asteraceae, Umbelliferae, Liliaceae, and Chenopodiaceae families. Specific examples include spinach, beet, arugula, red romaine, green romaine, red oak, mizuna, spinach, mustard, sunny lettuce, green curl, lettuce, green mustard, red mustard greens, romaine, tatsoi, small red lettuce, rolo rossa, red Asian mustard, pino green, white kale, red kale, Detroit, pak choi, etc. Among the above, it is preferable to use one or more of spinach (Spinacia oleracea), beet (Beta vulgaris), arugula (Eruca vesicaria), red romaine (Lactuca sativa L. var. longifolia), and green romaine (Lactuca sativa L. var. longifolia) as those exhibiting nuclear receptor activation activity.

[0027] <Processed products of leaves> The function of the processed products of leaves is to activate nuclear receptors. The form of the processed products of leaves is not particularly limited, but it is preferably an extract, which can be obtained by a normal extraction method used for extracting plants. The extraction method can be set appropriately. For preparation of the processed products, the leaves can be used as they are or after drying and crushing. Components contained in the leaves and the processed products thereof contribute to the activation of nuclear receptors, and examples of components that are assumed to contribute include lutein, α-carotene, β-carotene, retinoic acid, β-cryptoxanthin, resveratrol, lycopene, lycopene metabolites, etc.

[0028] The extraction solution used in preparing the processed leaf product is preferably water or aqueous ethanol from the viewpoint of edible use, but is not limited thereto and may be appropriately selected from solvents commonly used in this type of extraction. Examples include alcohols such as methanol, ethanol, propanol, and butanol; polyhydric alcohols such as ethylene glycol, propylene glycol, 1,2-butylene glycol, 1,3-butylene glycol, 1,4-butylene glycol, and 2,3-butylene glycol; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; linear or cyclic ethers such as diethyl ether and tetrahydrofuran; polyethers such as polyethylene glycol; dichloromethane, dichloroethane, chloroform, etc. Examples of suitable fatty acids include halogenated hydrocarbons such as hexane and carbon tetrachloride; hydrocarbons such as pentane, hexane, cyclohexane, and petroleum ether; aromatic hydrocarbons such as benzene and toluene; pyridines; supercritical carbon dioxide; edible oils such as rapeseed oil and soybean oil; fatty acid esters of glycerin such as monoacylglycerol, diacylglycerol (DAG), and triacylglycerol (TAG); medium-chain fatty acids (MCTs) with 5 to 12 carbon atoms, such as caprylic acid with 8 carbon atoms and capric acid with 10 carbon atoms; fats and oils such as squalane and squalene, waxes, and other oils.

[0029] The extract may be used as it is, or a highly active fraction may be separated by suitable separation means, such as gel filtration, chromatography, precision distillation, etc. The extract obtained may be diluted, concentrated, or freeze-dried, and then used in the form of a powder or paste. The extract obtained by the above-mentioned method may be dissolved in a solvent different from the extraction solvent.

[0030] <Nuclear receptors> Nuclear receptors are ligand-dependent transcription factors whose ligands are mainly lipid-soluble low molecular weight compounds in the body. When the ligand binds, they migrate into the nucleus and bind to DNA to control transcription. They regulate various biological phenomena such as development, cell proliferation, differentiation, morphogenesis, and metabolism, and are involved in the onset of various diseases. Representative nuclear receptors include retinoic acid receptors (RAR), retinoid X receptors (RXR), peroxisome proliferator-activated receptors (PPAR), and estrogen receptors (ER).

[0031] RAR is known to play an important role mainly in development, homeostasis, and immunity. There are three subtypes of RAR: RARα, RARβ, and RARγ. The present composition activates one or more of the above three subtypes.

[0032] RXR is known to play a role in promoting the function of RXR by forming a heterodimer with other nuclear receptors. There are three subtypes of RXR: RXRα, RXRβ, and RXRγ. The present composition activates one or more of the above three subtypes.

[0033] PPAR is known to play an important role mainly in lipid and sugar metabolism. There are three subtypes of PPAR: PPARα, PPARβ / δ, and PPARγ. The present composition activates any one or more of the above three subtypes.

[0034] ER is known to be mainly involved in female reproduction, bone formation, and alleviation of menopausal symptoms. There are two subtypes of ER, ERα and ERβ. Of the two subtypes, this composition activates ERβ.

[0035] <Nuclear receptor activators> Nuclear receptor activators contribute to the normalization of fasting blood glucose levels, the suppression of inflammatory skin diseases, the suppression of skin keratosis, the suppression of photoaging of the skin, the suppression of skin pigmentation, the suppression of fat accumulation, the suppression of fatty liver, and the increase in blood HDL cholesterol.

[0036] <Normalization of fasting blood glucose levels> Fasting blood glucose levels are blood glucose levels obtained from venous blood drawn after fasting for 10 hours or more. According to the Japan Diabetes Society's "Scientifically Based Diabetes Diagnosis Guidelines 2013," fasting blood glucose levels below 110 mg / dL are considered to be in the normal range, so normalization of fasting blood glucose levels in humans means that fasting blood glucose levels of 110 mg / dL or higher are in the normal range. Furthermore, since levels between 100 and 109 mg / dL are considered to be high normal, normalization also includes levels below 100 mg / dL.

[0037] <Suppression of inflammatory skin diseases> Suppression of inflammatory skin diseases refers to suppressing the occurrence of diseases accompanied by inflammation of the skin, such as acne and rosacea, or alleviating the symptoms.

[0038] <Suppression of skin keratosis> Suppression of skin keratosis refers to suppressing the occurrence of hereditary keratosis such as palmoplantar keratosis, ichthyosis, and Darier's disease, and acquired keratosis such as psoriasis, pityriasis rubra pilaris, and lichen, or alleviating the symptoms.

[0039] <Skin photoaging inhibition> Suppression of skin photoaging refers to the inhibition of photoaging phenomena such as wrinkles, sagging, and dryness caused by ultraviolet rays and the like.

[0040] <Skin Pigmentation Inhibition> Suppression of skin pigmentation refers to the inhibition of pigmentation on the skin, such as spots that occur after skin inflammation or spots caused by ultraviolet rays.

[0041] <Suppression of fat accumulation> Since obesity due to visceral fat accumulation leads to lifestyle-related diseases such as diabetes and dyslipidemia, suppression of fat accumulation refers to the suppression of an increase in visceral fat mass and the reduction in lipid droplets determined histologically in visceral fat.

[0042] <Suppression of fatty liver> Fatty liver is a condition in which neutral fats accumulate in 30% or more of liver cells, and suppression of fatty liver refers to suppressing the accumulation of neutral fats in liver cells.

[0043] <Increase in Blood HDL Cholesterol> Blood HDL cholesterol plays a role in transporting excess cholesterol in the blood to the liver, and is called "good cholesterol" because the higher the blood concentration, the less likely one is to develop arteriosclerosis. In humans, the normal range of blood HDL cholesterol is 40 mg / dL or higher, so an increase in blood HDL cholesterol in humans refers to a value approaching 40 mg / dl from less than 40 mg / dl, or reaching 40 mg / dl or higher.

[0044] <Outline of the method for producing the present composition> The process for producing the present composition (hereinafter referred to as "the present method") is as follows. The present method mainly comprises extraction.

[0045] <Extraction> The purpose of preparing an extract from the leaves by extraction is to concentrate the contributing components. The extraction method may be a known method, for example, shaking extraction using water or an organic solvent.

[0046] The components contained in the processed product of the extracted leaves contribute to the activation of nuclear receptors, and examples of components that are thought to contribute include lutein, α-carotene, β-carotene, retinoic acid, β-cryptoxanthin, resveratrol, lycopene, and lycopene metabolites.

[0047] <Food composition for activating nuclear receptors> The present composition can be added to or mixed with food, beverage, feed, or pet food. Alternatively, the present composition can be added to or mixed with food and beverage products that indicate that a disease or condition can be prevented or improved by activating a nuclear receptor, i.e., health foods, functional foods, foods for the sick, and foods for specified health uses. Specifically, the present composition can be used in various formulations such as fine granules, tablets, granules, powders, capsules, syrups, liquids, and liquid diets. Foods in formulations can contain additives such as excipients, binders, disintegrants, lubricants, antioxidants, colorants, and flavorings, as necessary. In addition, the present composition can be produced by mixing the contributing component with a carrier acceptable for use as a food, such as a suitable excipient, and then using conventional means. Furthermore, it is also possible to use the product as it is as a food, and the type and form of the food are not particularly limited. Examples of the food include liquid food compositions such as soups, juices, vegetable drinks, fruit drinks, vegetable and fruit drinks, milk, dairy drinks, whey drinks, lactic acid bacteria drinks, tea drinks, alcoholic drinks, coffee drinks, carbonated drinks, soft drinks, water drinks, cocoa drinks, jelly drinks, sports drinks, and diet drinks, puddings, yogurt, etc. It can be added to or mixed with any semi-solid food composition, breads, noodles, confectioneries, spreads, seasonings, etc. EXAMPLES

[0048] <Example 1> The leaf stalks of spinach (variety: Green Spinach) grown in soil 16-20 days after germination (young leaves) and 36-40 days after germination (adult leaves), beet (variety: Detroit) grown hydroponically 16-20 days after germination (young leaves) and 36-40 days after germination (adult leaves), arugula (variety: Arugula (Rocket)) grown in soil 14-18 days after germination (young leaves) and 36-40 days after germination (adult leaves), red romaine (variety: Red Romaine) grown hydroponically 18-22 days after germination (young leaves) and 38-42 days after germination (adult leaves), and green romaine (variety: Green Romaine) grown hydroponically 18-22 days after germination (young leaves) and 38-42 days after germination (adult leaves) were cut with scissors and stored at -80°C. The leaves were pre-frozen in a 1000-mbar ultra-low temperature freezer (As One Corporation). They were then quickly transferred to a vacuum freeze dryer (FZ-6BT, Labconco Corporation) and vacuum dried for 5 to 7 days. Drying was completed when the valves of the vacuum chamber and the cold trap were closed and it was confirmed that there was no change in the set vacuum of 0.040 mbar for 20 to 30 minutes. The leaves were then pulverized in a grinder (Tokyo Unicom, High Speed ​​Mill T-351) to obtain freeze-dried samples. The powdered samples were then subjected to extraction. The amount of powdered sample (g) per 1 ml of final sample is shown in Table 1 for young leaves and Table 2 for adult leaves. The specific extraction method will be described later.

[0049] [Table 1]

[0050] [Table 2]

[0051] For the extracted samples, the nuclear receptors PPARα, PPARδ, PPARγ, RXRα, RXRβ, RXRγ, vitamin D receptor (VDR), androgen receptor (AR), glucocorticoid receptor (GR), progesterone receptor (PR), and thyroid hormone receptor α (TRα) were evaluated by the reporter assay described below using CV-1 cells (purchased from the JCRB Cell Bank, National Institutes of Biomedical Innovation, Health and Nutrition). The nuclear receptors RARα, RARβ, RARγ, ERα, ERβ, ERRα, ERRβ, and ERRγ were evaluated by the reporter assay described below using COS-7 cells (purchased from ATCC (American Type Culture Collection)). The nuclear receptors pregnane receptor (PXR), farnesoid X receptor (FXR), liver X receptor α (LXRα), liver X receptor β (LXRβ), hepatocyte nuclear factor 4α (HNF-4α), RAR orphan receptor α (RORα), and transcription factor (Nrf2) belonging to the CNC family were evaluated by the reporter assay described below using Hep G2 cells (purchased from the RIKEN BioResource Research Center (RIKEN BRC)). The evaluation results are shown in Figure 1 for young leaves and Figure 2 for adult leaves. Detailed measurement methods for nuclear receptor and Nrf2 activity and explanations of the figures will be described later.

[0052] <Water extraction> Sterile water was added to the powder samples of spinach and beet leaves to make the concentration 50 mg / ml, and sterile water was added to the powder samples of arugula, red romaine, and green romaine leaves to make the concentration 100 mg / ml. After shaking for 24 hours at 4°C and centrifuging (220×g, 10 to 30 minutes), the supernatant was sterilized by filtration using a membrane filter with a pore size of 0.2 μm.

[0053] <Ethanol extraction> The powder samples were suspended in 70% ethanol at 100 mg / ml (w / v) and shaken on a shaker at room temperature for 3 hours. After filtering through a membrane filter with a pore size of 0.2 μm, the filtrate was dried under reduced pressure at room temperature. The resulting dried product was dissolved in dimethyl sulfoxide (DMSO) at a concentration of 200 mg / ml or 400 mg / ml.

[0054] <Mixed Solvent Extraction> The powder sample was added to a mixed solution of hexane:ethanol:acetone:toluene (hereinafter referred to as the mixed solvent) at 100 mg / ml and shaken on a shaker at room temperature for 3 hours. It was then centrifuged (890×g, 10-30 minutes) and the supernatant was collected. After sterilization by filtration using a membrane filter with a pore size of 0.2 μm, the filtrate was dried under reduced pressure at room temperature. The obtained dried product was dissolved in DMSO at a concentration of 200 mg / ml or 400 mg / ml.

[0055] <Method for measuring nuclear receptor activity and Nrf2: reporter assay> African green monkey kidney-derived cell lines CV-1, COS-7, and human hepatoma cell line Hep G2 were each incubated at 2x10 5 The cells were seeded in a 6-well plate at 1000 cells / well and cultured in DMEM (10% serum) for 1 day. A chimeric protein expression plasmid of Gal4 DNA binding domain (Gal4-DBD) and nuclear receptor ligand binding domain (NR-LBD), a reporter plasmid (pGal4-Luc) containing Gal4 DNA response element and firefly luciferase gene, and an internal standard plasmid (pGL4.75hRluc-CMV or pGL4.73hRluc-SV40) with a gene constitutive expression promoter (CMV, SV40) linked upstream of Renilla luciferase gene as an internal standard were mixed in a weight ratio of 1:0.9:0.1 and dissolved in Opti-MEM at a concentration of 10 μg / ml (total DNA amount).

[0056] For the transcription factor Nrf2, a reporter plasmid (GSTA2-Luc) containing the promoter region of glutathione S-transferase A2 (GSTA2), one of the target genes of Nrf2, and the firefly luciferase gene, and an internal standard plasmid (pGL4.73hRluc-SV40) in which a gene constitutive expression promoter (SV40) was linked upstream of the Renilla luciferase gene as an internal standard, were mixed in a weight ratio of 1.9:0.1 and dissolved in Opti-MEM at a concentration of 10 μg / ml (total DNA amount).

[0057] At the same time, a gene transfer reagent (X-tremeGENE HP; Roche) was added at 1 / 43 volume, and after leaving it for 15 minutes, 200 μl of this mixture was added to each well and cultured for 6 hours to transfer the gene. The cell lines and plasmids used in the assay of each receptor are shown in Table 3.

[0058] The transfected cells were dispersed with trypsin and placed in a 96-well plate at 1.6 x 10 4 cells / well (CV-1), 2.0x10 4 The wells were seeded again at 100 cells / well (COS-7, Hep G2). At this time, the culture medium was replaced with DMEM medium (10% activated charcoal-treated FBS) without phenol red. After 1 hour, the same DMEM medium as above containing each concentration of the test substance was added to each well. As negative controls (solvent controls), 0.5% DMSO (70% ethanol extraction, mixed solvent extraction) and 10% ultrapure water (water extraction) were used.

[0059] The test concentration for each sample was set based on the maximum concentration that showed no cytotoxicity in a previous cytotoxicity test on CV-1 cells, COS-7 cells, and Hep G2 cells.

[0060] After culturing for 48 hours in a CO2 incubator, the cells were washed with phosphate-buffered saline (PBS) and lysed using a dual luciferase assay system (Promega). A substrate solution containing luciferin was then added, and firefly and Renilla luciferase activities were measured using a plate reader (Luminescencer, AB-2350EX, ATTO). The above operations were performed using three wells per sample (including positive and negative controls), and the average value of the three wells was used as data. The positive controls used for quality control in this experiment are shown in Table 4. DMSO (final concentration 0.5%) was used as a solvent for adding the substances in Table 4.

[0061] [Table 3]

[0062] [Table 4]

[0063] <Data Analysis> The transcription activity (internal standard correction value of luciferase activity) of a gene dependent on a nuclear receptor or a transcription factor was defined as follows.

[0064] (Internal standard correction value) = (firefly luciferase activity by Gal4-Luc or GSTA2-Luc) / (Renilla luciferase activity by hRluc-CMV or hRluc-SV40) Activity was evaluated as a ratio to the negative control (activity value in sample / activity value in negative control), and significant activity was defined as a value (negative control ratio) of 2.0 or more without significant damage to the cells (no significant decrease in Renilla luciferase activity value). However, when no activity was observed based on the value of only the firefly luciferase activity by Gal4-Luc, it was considered to have no activity even if the internal standard correction value was 2.0 or more.

[0065] Figures 1 and 2 show the results of comparing the nuclear receptor activation ability of young and adult leaves, respectively. Compared to the negative control, 3+ was 101 times or more (underlined values ​​are 500 times or more), 2+ was 51 to 100 times, and 1+ was 2 to 50 times (underlined values ​​are 10 times or more). Classification was performed using the maximum value obtained in the assay. [Industrial Applicability]

[0066] The field in which the present invention is useful is the production of health foods and foods with functional claims.

Claims

1. A food composition for activating a nuclear receptor, comprising: It contains arugula leaves or processed products thereof, The leaves are within 30 days of germination; The nuclear receptor is any one of RAR, RXR, PPAR, and ER, or Two or more.

2. The food composition for activating a nuclear receptor according to claim 1, The RAR is any one or two of RARα, RARβ, and RARγ. or more, wherein the RXR is any one of RXRα, RXRβ, and RXRγ. One or more of the PPARs are selected from the group consisting of PPARα, PPARγ, and PPA Rδ is any one or more, The ER is ERβ.

3. The food composition for activating a nuclear receptor according to claim 2, The nuclear receptor is a RAR.

4. A food composition for normalizing fasting blood glucose level, comprising: It contains a nuclear receptor activation contributing component, Its origin is the arugula leaf, The leaves are within 30 days of germination; The nuclear receptor activation contributing component is an RAR activation contributing component.

5. A food composition for suppressing inflammatory skin diseases, comprising: It contains a nuclear receptor activation contributing component, Its origin is the arugula leaf, The leaves are within 30 days of germination; The nuclear receptor activation contributing component is a RAR activation contributing component.

6. A food composition for inhibiting skin keratosis, comprising: It contains a nuclear receptor activation contributing component, Its origin is the arugula leaf, The leaves are within 30 days of germination; The nuclear receptor activation contributing component is an RAR activation contributing component.

7. A food composition for suppressing photoaging of the skin, comprising: It contains a nuclear receptor activation contributing component, Its origin is the arugula leaf, The leaves are within 30 days of germination; The nuclear receptor activation contributing component is an RAR activation contributing component.

8. A food composition for inhibiting skin pigmentation, comprising: It contains a nuclear receptor activation contributing component, Its origin is the arugula leaf, The leaves are within 30 days of germination; The nuclear receptor activation contributing component is an RAR activation contributing component.

9. A food composition for inhibiting fat accumulation, comprising: It contains a nuclear receptor activation contributing component, Its origin is the arugula leaf, The leaves are within 30 days of germination; The nuclear receptor activation contributing component is an RAR activation contributing component.

10. A food composition for suppressing fatty liver, comprising: It contains a nuclear receptor activation contributing component, Its origin is the arugula leaf, The leaves are within 30 days of germination; The nuclear receptor activation contributing component is an RAR activation contributing component.

11. A food composition for increasing blood HDL cholesterol, comprising: It contains a nuclear receptor activation contributing component, Its origin is the arugula leaf, The leaves are within 30 days of germination; The nuclear receptor activation contributing component is an RAR activation contributing component.

12. A method for producing a food composition for activating a nuclear receptor, comprising at least the following components: Extraction: The substance extracted here is a nuclear receptor activation contributing component, which is extracted from arugula leaves, The leaves are within 30 days of germination; The nuclear receptor activation contributing component is an RAR activation contributing component.

13. 13. The method of claim 12, The component contributing to nuclear receptor activation is extracted with water, aqueous ethanol, or an organic solvent.

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