Heat shock protein expression inducer and composition for heat shock protein expression induction

Abstract

To provide a heat shock protein expression inducer and a composition for heat shock protein expression induction by utilizing a plant-derived resorcinolic lipid.SOLUTION: A plant-derived resorcinolic lipid is used as an active ingredient of a heat shock protein expression inducer. The resorcinolic lipid is preferably alkylresorcinol. The resorcinolic lipid is preferably derived from a gramineous plant. The resorcinolic lipid is preferably derived from the hull (bran etc.) and / or whole grain of wheat and / or rye. The heat shock protein expression inducer is well suited as a material to be contained in foods and drinks, food additives, pharmaceuticals, supplements, cosmetics, animal feed, etc.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a functional material using a plant-derived component, and more particularly to a heat shock protein expression inducer using a plant-derived resorcinol lipid. [Background technology]

[0002] Resorcinol lipids are lipophilic substances having alkyl groups at the 2-, 4-, 5-, and / or 6-positions of the aromatic ring of resorcinol (1,3-dihydroxybenzene). Resorcinol lipids are naturally found in large amounts in the seeds of grasses, particularly in their outer skins (bran), ginkgo leaves, cashew nut shell oil, etc. In recent years, there have been reports of various functionalities of resorcinol lipids for humans, such as oral antibacterial properties, immunosuppression, antidiabetic properties, anti-inflammatory properties, intestinal flora regulation, anticholesterol properties, anti-obesity properties, sleep improvement, anti-aging properties, circadian rhythm regulation, and liver function improvement (e.g., Patent Documents 1 to 11).

[0003] On the other hand, heat shock proteins are a group of proteins whose expression level increases when cells are exposed to stressful conditions such as heat, thereby protecting the cells. They are generally known to have a function of assisting protein folding, or to be involved in specific transcriptional control through their function. It is believed that the effect of inducing the expression of heat shock proteins can confer resistance to stresses experienced by the living body to humans and animals (Non-Patent Document 1). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Patent No. 2613474 [Patent Document 2] JP 2013-091612 A [Patent Document 3] Patent No. 2014-139166 [Patent Document 4] Patent No. 2015-020993 [Patent Document 5] Patent No. 2015-218130 [Patent Document 6] Patent No. 2015-231986 [Patent Document 7] Patent No. 2016-132641 [Patent Document 8] Patent No. 2016-153387 [Patent Document 9] Patent No. 5926616 [Patent Document 10] Patent No. 5951448 [Patent Document 11] International Publication No. 2020 / 235546 Brochure [Non-patent literature]

[0005] [Non-Patent Document 1] "The charm of heat shock proteins (HSP70)" by Yoko Ito, Journal of Oncology, Vol. 77, No. 3, May 2014, pp. 222-226 Summary of the Invention [Problem to be solved by the invention]

[0006] However, there have been no reports to date on the effect of resorcinol lipids on inducing the expression of heat shock proteins.

[0007] An object of the present invention is to provide a heat shock protein expression inducer and a composition for inducing heat shock protein expression, which utilizes a plant-derived resorcinol lipid. [Means for solving the problem]

[0008] To achieve the above object, in a first aspect, the present invention provides a heat shock protein expression inducer comprising a plant-derived resorcinol lipid as an active ingredient.

[0009] In the heat shock protein expression inducer provided by the present invention, the resorcinol lipid is preferably an alkylresorcinol.

[0010] In the above heat shock protein expression inducer, the resorcinol lipid is preferably derived from a grass family plant.

[0011] In the heat shock protein expression inducer, the resorcinol lipid is preferably derived from the husk and / or whole grain of wheat and / or rye.

[0012] In the above-mentioned heat shock protein expression inducer, the resorcinol lipid is preferably a solvent extract of a plant containing resorcinol lipid.

[0013] In order to achieve the above-mentioned object, in a second aspect, the present invention provides a composition for inducing heat shock protein expression, which contains the above-mentioned heat shock protein expression inducer and is provided as a food or beverage, a food additive, a pharmaceutical, a supplement, a cosmetic, or an animal feed.

[0014] To achieve the above object, in a third aspect, the present invention provides use of a plant-derived resorcinol lipid for preparing a composition for inducing expression of a heat shock protein.

[0015] In the use of plant-derived resorcinolic lipids in the present invention, it is preferred that said resorcinolic lipids are alkylresorcinols.

[0016] In the above-mentioned use, the resorcinol lipid is preferably derived from a grass family plant.

[0017] Also, in said uses, it is preferred that said resorcinol lipids are derived from the husk and / or whole kernels of wheat and / or rye.

[0018] In the above uses, it is also preferred that the resorcinol lipid comprises a solvent extract of a plant containing resorcinol lipid.

[0019] In addition, in the above-mentioned use, the composition for inducing expression of a heat shock protein is preferably provided as a food or drink, a food additive, a pharmaceutical, a supplement, a cosmetic, or an animal feed. Effect of the Invention

[0020] According to the present invention, a heat shock protein expression inducer and a composition for inducing heat shock protein expression can be provided by utilizing a plant-derived resorcinol lipid. [Brief description of the drawings]

[0021] [Figure 1] 1 is a graph showing the results of investigating the gene expression level of the heat shock protein HSP90 in the liver in the animal test of Test Example 2. The results are shown as the average value and its standard deviation in each group (n=2). [Diagram 2] 1 is a graph showing the results of investigating the gene expression level of heat shock protein HSP90 in soleus muscle in the animal test of Test Example 2. [Diagram 3] 1 is a graph showing the results of examining the gene expression level of the heat shock protein HSP90 in brown adipose tissue (BAT) in the animal test of Test Example 2. The results are shown as the average value and its standard deviation in each group (n=2). [Figure 4] 1 is a graph showing the results of examining the gene expression level of heat shock protein HSP70 in the cell test of Test Example 3. The results are shown as the average value and its standard deviation in each group (n=3). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022] In the present invention, a plant-derived resorcinol lipid is used as an active ingredient of the heat shock protein expression inducer.

[0023] As described above, resorcinol lipids are lipophilic substances having alkyl groups at the 2nd, 4th, 5th, and / or 6th positions of the aromatic ring of resorcinol (1,3-dihydroxybenzene). In nature, they are often found in plants belonging to the Anacardiaceae, Ginkgoaceae, Proteaceae, Ardisiaceae, Primulaceae, Myristicae, Iridaceae, Araceae, Fabaceae, Poaceae, etc., and in mugwort of the Asteraceae family. Among these plants, Poaceae plants such as wheat and rye contain relatively abundant resorcinol lipids in their edible parts (seeds), and therefore can be suitably used as a raw material for the resorcinol lipids used in the present invention. For example, the outer skin (bran, etc.) and whole grains of the edible parts (seeds) of wheat, rye, etc. contain about 0.015 to 0.3 mass% of an alkylresorcinol compound represented by the following formula (I). Fermented products such as miso, which is made from wheat or rye, can also be exemplified as preferred raw materials for the resorcinol lipids used in the present invention.

[0024] The resorcinol lipid used in the present invention typically includes an alkylresorcinol compound represented by the following formula (I).

[0025] [ka] In formula (I), R1 represents a saturated or unsaturated linear or branched alkyl group. The number of carbon atoms of the alkyl group represented by R1 is not limited, but is preferably 1 to 27, more preferably 3 to 27, and further preferably 5 to 27.

[0026] In formula (I), R1 is preferably a saturated or unsaturated linear alkyl group, more preferably a saturated linear alkyl group. Examples of the saturated linear alkyl group represented by R1 include methyl, n-propyl, n-pentyl, n-heptyl, n-nonyl, n-undecyl, n-tridecyl, n-pentadecyl, n-heptadecyl, n-nonadecyl, n-henicosyl, and n-tricosyl.

[0027] In formula (I), the position and number of unsaturated bonds in the unsaturated linear alkyl group represented by R1 are not particularly limited. Examples of the unsaturated linear alkyl group include the above-mentioned saturated linear alkyl group having an unsaturated bond at any position on the carbon chain.

[0028] In formula (I), the position or number of branches of the saturated or unsaturated branched alkyl group represented by R1, or the position or number of unsaturated bonds, is not particularly limited.

[0029] Preferred examples of the compound of formula (I) above include the following.

[0030] 5-Pentylresorcinol [Olivetol, or 1,3-dihydroxy-5-n-pentylbenzene (C5:0)] 5-heptylresorcinol [or 1,3-dihydroxy-5-n-heptylbenzene (C7:0)] 5-Nonylresorcinol [or 1,3-dihydroxy-5-n-nonylbenzene (C9:0)] 5-Undecylresorcinol [or 1,3-dihydroxy-5-n-undecylbenzene (C11:0)] 5-Tridecylresorcinol [or 1,3-dihydroxy-5-n-tridecylbenzene (C13:0)] 5-Pentadecylresorcinol [or 1,3-dihydroxy-5-n-pentadecylbenzene (C15:0)] 5-heptadecylresorcinol [or 1,3-dihydroxy-5-n-heptadecylbenzene (C17:0)] 5-Nonadecylresorcinol [or 1,3-dihydroxy-5-n-nonadecylbenzene (C19:0)] 5-Henicosylresorcinol [or 1,3-dihydroxy-5-n-henicosylbenzene (C21:0)] 5-tricosylresorcinol [or 1,3-dihydroxy-5-n-tricosylbenzene (C23:0)] 5-Pentacosylresorcinol [or 1,3-dihydroxy-5-n-pentacosylbenzene (C25:0)] 5-heptacosylresorcinol [or 1,3-dihydroxy-5-n-heptacosylbenzene (C27:0)]

[0031] The plant-derived resorcinol lipid used in the present invention may be, for example, a commercially available product, or may be extracted from a plant by a conventional method. For example, the commercially available product of the compound of formula (I) above can be purchased from ReseaChem GmbH, SIGMA-ALDRICH, etc.

[0032] Examples of the extraction method from plants include a method in which the raw material is immersed in an extraction solvent at room temperature or heated under normal or increased pressure while stirring as necessary, a reflux extraction method, etc. Before being added to the extraction solvent, the raw material may be cut, crushed, squeezed, dried, or a combination thereof as necessary.

[0033] Examples of the extraction solvent include alcohols that are liquid at room temperature, such as lower alcohols such as methanol, ethanol, n-propanol, isopropanol, and n-butanol, or polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, and glycerin; ethers such as diethyl ether and propyl ether; esters such as butyl acetate and ethyl acetate; ketones such as acetone and ethyl methyl ketone; hexane; and organic solvents such as chloroform. These solvents may be used alone or in combination of two or more. Among the above solvents, from the viewpoints of operability and environmental impact, it is preferable to use alcohols that are liquid at room temperature, such as lower alcohols having 1 to 4 carbon atoms, and it is more preferable to use ethanol from the viewpoint of safety due to residual solvents.

[0034] The extraction solvent may further include a water-containing organic solvent in which an aqueous component is contained in the organic solvent. From the viewpoint of maintaining high extraction efficiency, the content of the aqueous component in the water-containing organic solvent is usually 50% by volume or less, preferably 30% by volume or less, more preferably 20% by volume or less. The water-containing organic solvent may preferably be a water-containing alcohol in which an aqueous component is contained in the alcohol that is liquid at room temperature as described above, more preferably a water-containing ethanol.

[0035] The conditions for extraction, such as temperature and time, can be appropriately set depending on the type of extraction solvent used and the extraction conditions, but the extraction temperature is preferably 2 to 100°C, and the extraction time is preferably about 30 minutes to 60 hours. The amount of the extraction solvent may be preferably about 200 to 2000 parts by mass per 100 parts by mass of the raw material. After the extraction process, the mixture containing the extract and the residue may be subjected to filtration or centrifugation, etc., as necessary, to remove the solid components that are the residue. Furthermore, the removed solid components may be subjected to an extraction operation using the extraction solvent again, and this operation may be repeated several times. The obtained extract may be subjected to liquid chromatography or the like to prepare an extract containing further purified resorcinol lipids. When purified in this way, the degree of purification may be, for example, typically in the range of 1% by mass or more, more typically in the range of 5% by mass or more, and even more typically in the range of 10% by mass or more, as the content of the resorcinol lipids derived from the above-mentioned plant in any form. For example, the content may typically be in the range of 100% by mass or less, more typically in the range of 90% by mass or less, and even more typically in the range of 80% by mass or less. The obtained extract may be used in the present invention as it is, or after further treatment such as concentration, freeze-drying, hot air drying, pulverization, powderization, classification, dilution, mixing with water, etc., as necessary.

[0036] As shown in the examples below, resorcinol lipids derived from plants have the effect of inducing the expression of heat shock proteins. The present invention provides a heat shock protein expression inducer based on the attributes of resorcinol lipids. In general, "heat shock proteins" are a group of proteins whose expression increases when cells are exposed to a stressful environment such as heat to protect the cells. Generally, they are known to have the function of helping protein folding or to be involved in specific transcriptional control. Typical molecular species in humans include Hsp70, Hsp90, Hsp104, Hsp60, Hsp47, Hsp32, Hsp40, Hsp27, Hsp10, and Hsp100. According to the present invention, the effect of inducing the expression of heat shock proteins can confer resistance to stresses that the living body is subjected to, such as heat, ultraviolet rays, pathogens, viruses, wounds, inflammation, nutritional deficiencies, hypoxia, free radicals, arteriosclerosis, hyperlipidemia, toxic substances, and ischemia.

[0037] The dosage of the heat shock protein expression inducer provided by the present invention may be appropriately changed depending on the species, symptoms, age, sex, etc. of the individual to which it is applied. The dosage for humans may usually be 0.01 to 10 g of the above-mentioned plant-derived resorcinol lipid per adult per day. The daily dosage may be administered once or in several divided doses. It is more preferable to administer it orally. The subject of application is not limited to humans, and it may also be applied to animals such as dogs and cats.

[0038] The form of the heat shock protein expression inducer provided by the present invention is not particularly limited as long as it is a form in which the above-mentioned plant-derived resorcinol lipid can be ingested. For example, the form may be solid, semi-solid or liquid, or may be various forms such as tablets, chewable tablets, powders, capsules, granules, drinks, gels, syrups, liquid foods for tube-enteral nutrition, etc. In addition, it may be in the form of, for example, health foods, functional foods, foods for specified health uses, foods for sick people, food additives therefor, medicines, supplements, cosmetics, animal feed for livestock, racehorses, ornamental animals, pets, etc. By containing the above-mentioned plant-derived resorcinol lipid as a part of each form, it can be made into a form that is easy to ingest. That is, the heat shock protein expression inducer provided by the present invention may be in the form of a composition containing it, which can be said to provide a composition for inducing heat shock protein expression. In this case, the composition for inducing heat shock protein expression may be substantially composed of the above-mentioned plant-derived resorcinol lipid. Therefore, for example, in any form, the content of the resorcinol lipid derived from the above plant can be, for example, typically in the range of 0.1% by mass or more, more typically in the range of 0.5% by mass or more, and even more typically in the range of 1.0% by mass or more. Also, for example, it can be, for example, typically in the range of 100% by mass or less, more typically in the range of 90% by mass or less, and even more typically in the range of 80% by mass or less.

[0039] Typical pharmaceutical forms include, for example, oral preparations such as tablets, capsules, granules, powders, syrups, dry syrups, liquids, and suspensions, inhalants, transdermal preparations, enteral preparations such as suppositories, and parenteral preparations such as drops and injections. The liquid preparations such as the above liquids and suspensions may be dissolved or suspended in water or other suitable medium immediately before administration, and the above tablets and granules may be surface-coated by a known method. The above injections may be solutions or suspensions of the above plant-derived resorcinol lipids in sterile distilled water or sterile saline, optionally containing a solubilizing agent.

[0040] The above pharmaceutical forms may contain various pharma- ceutically acceptable carriers and base materials, such as excipients, stabilizers, pH adjusters, and other additives, as necessary, and may further contain other medicinal ingredients, such as various vitamins, minerals, and herbal medicines.

[0041] Typical cosmetic forms may include, for example, creams, lotions, milks, ointments, poultices, and the like.

[0042] The above-mentioned cosmetic forms may contain various carriers and base materials acceptable for use as cosmetics, such as dispersants, suspending agents, emulsifiers, stabilizers, pH adjusters, antioxidants, and other additives, as necessary, and may further contain other medicinal ingredients, such as various vitamins, minerals, and herbal medicines.

[0043] Typical forms of food and beverages include tea beverages such as green tea, oolong tea, and black tea, coffee beverages, soft drinks, jelly drinks, sports drinks, milk beverages, carbonated drinks, fruit juice drinks, lactic acid bacteria beverages, fermented milk beverages, powdered beverages, cocoa beverages, alcoholic beverages, mineral water, and other beverages; spreads such as butter, jam, furikake, and margarine; mayonnaise, shortening, cream, dressings, bread, cooked rice, noodles, pasta, miso soup, tofu, milk, yogurt, soups or sauces, and sweets (e.g., biscuits, cookies, chocolate, candy, cake, ice cream, chewing gum, and tablets).

[0044] In the form of the above-mentioned food and beverage products, they may contain, as necessary, other food and beverage ingredients acceptable for food, various nutrients, various vitamins, minerals, amino acids, various oils and fats, various additives (for example, taste components, sweeteners, acidulants such as organic acids, surfactants, pH adjusters, stabilizers, antioxidants, colorants, flavors), etc., or they may further contain other medicinal ingredients such as various vitamins, minerals, herbal medicines, etc.

[0045] Typical food additives may have a composition and form that allows them to be incorporated into foods and beverages, as exemplified above.

[0046] Typical animal feeds can have compositions and forms that are substantially similar to those of foods and beverages, as exemplified above, except that they are for animals.

[0047] Typical forms of the supplement include oral compositions among those mentioned above, such as tablets, chewable tablets, powders, capsules, granules, drinks, gels, syrups, etc. EXAMPLES

[0048] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples.

[0049] <Preparation Example 1> ~Preparation of alkylresorcinol~ Rye bran (100 kg) was mixed with 5 times the amount of 99% ethanol and extracted with stirring under heating (60°C, 1 hour). After that, solid-liquid separation was performed to obtain filtrate A, and the same amount of 99% ethanol was added to the residue, which was then rinsed and further separated into solid and liquid to obtain filtrate B. The obtained filtrate A and filtrate B were combined and concentrated under reduced pressure to obtain 3.9 kg of rye bran ethanol extract.

[0050] 500 g of the above extract was dissolved in 2 L of hexane and purified by silica normal phase chromatography. The column used was Yamazen Corporation's Hi-Flash Column Silica 5 L Catalog No. W007 (column inner diameter x column length: 60 x 180 mm, packing amount: 250 g). The chromatography conditions were a flow rate of 70 mL / min, detection wavelength of 280 nm, and mobile phase (hexane / ethyl acetate: 90 / 10 (v / v) for 9 minutes → 80 / 20 (v / v) for 15 minutes → 60 / 40 (v / v) step gradient for 16 minutes), and the eluate was collected between 30 and 40 minutes of elution. The collected material was dried under reduced pressure to obtain 106 g of dried material.

[0051] <Test Example 1> -Analysis by high performance liquid chromatography- The amount of alkylresorcinol in the dried product obtained in Preparation Example 1 was quantified by high performance liquid chromatography. Hereinafter, alkylresorcinol may be referred to as "ARs" or "AR".

[0052] Specifically, the analytical column used was Inertsil ODS4 (GL Science), and the analytical conditions were a flow rate of 1 mL / min, detection wavelength of 275 nm, column temperature of 40°C, and a mobile phase gradient of 89 v / v% methanol → (5 min) → 92 v / v% methanol → (25 min) → 100% methanol. Commercially available Olivetol (AR with a linear alkyl chain of 5 carbon atoms: Sigma-Aldrich) was used as the alkylresorcinol standard. The elution positions of ARs with different alkyl chain lengths were specified in advance, and the measured peak areas were converted by multiplying the molecular weight of each AR divided by the molecular weight of Olivetol as a coefficient.

[0053] As a result, it was confirmed that the dried product obtained in Preparation Example 1 contained 23.1 g of ARs per 100 g. Hereinafter, the dried product obtained in Preparation Example 1 may be referred to as "ARs-containing dried product."

[0054] <Test Example 2> ~Animal Test~ The effects of feeding alkylresorcinols to animals were investigated.

[0055] Specifically, non-alcoholic steatohepatitis (NASH) model mice (Charles River Japan, C57BL / 6 NASH male mice) were used, and were fed a choline-deficient high-fat diet (A06071302: Research diets) from the age of 4 weeks. After being brought in at the age of 7 weeks, the mice were housed 5 per cage after a 2-day acclimation period. The mice were housed in a temperature-controlled, humidity-controlled room maintained on a 12-hour light-dark cycle (light period: 8:00-20:00), and after the acclimation period, the mice were allowed to freely consume the diet appropriate for each test group.

[0056] Control group: Choline and methionine deficient high fat sample (A06071302) Alkylresorcinol intake group: A choline- and methionine-deficient high-fat sample (A06071302) containing 0.3% by mass of the ARs-containing dried solid obtained in Preparation Example 1 (hereinafter, this intake group may be referred to as the "ARs 0.3% group")

[0057] The test diet was administered for 28 days, and on the final day, the animals were sacrificed under isoflurane anesthesia, after which the liver, soleus muscle, and brown adipose tissue (hereinafter sometimes referred to as "BAT") were immediately extracted and frozen on dry ice, then cut into 1.5 mL tubes and stored at -80°C. Brown fat was extracted from the dorsal scapula, and soleus muscle was extracted from the triceps surae muscle.

[0058] Total RNA was prepared from each tissue using the "NucleoSpin (registered trademark) RNA II Kit (Takara Bio Inc.)" according to the product instructions.

[0059] cDNA synthesis from total RNA was carried out using "Protoscript (registered trademark) II First strand cDNA Synthesis Kit (New England Biolabs)" according to the product instructions.

[0060] Quantitative RT-PCR using the synthesized cDNA was performed using a "MyGo Pro&Mini Single tube individual bag (Funakoshi Co., Ltd.)" according to the product's instructions. Specifically, the mRNA expression level of HSP90, a heat shock protein, was measured using the PCR primers shown in Table 1 and the temperature cycle program shown in Table 2. The PCR measurement values ​​were normalized using the expression level of β-actin as a housekeeping gene. The results were compared relative to the control group, which was set at 100%.

[0061] [Table 1]

[0062] Table 2 shows the temperature program for the PCR reaction.

[0063] [Table 2]

[0064] As a result, as shown in FIG. 1, the expression level of mouse HSP90 mRNA in the liver of the ARs 0.3% group was significantly increased compared to the control group.

[0065] Furthermore, as shown in FIG. 2, the mRNA expression level of mouse HSP90 in the soleus muscle of the ARs 0.3% group was significantly increased compared to the control group.

[0066] Furthermore, as shown in FIG. 3, the expression level of mouse HSP90 mRNA in BAT in the ARs 0.3% group was significantly increased compared to the control group.

[0067] <Test Example 3> ~Cell test~ We investigated the effects of stimulating cultured cells with alkylresorcinols. Specifically, we investigated the effects of stimulating human hepatoma-derived cells (HepG2) by adding alkylresorcinols to the culture medium.

[0068] The medium was prepared by mixing the following medium ingredients in the appropriate volumes: E-MEM (FUJIFILM Wako Pure Chemical Industries, Ltd.): 450 mL - Inactivated Fetal Bovine Serum (FBS): 50mL Penicillin-streptomycin solution (FUJIFILM Wako Pure Chemical Industries, Ltd.): 5 mL

[0069] A frozen stock of human hepatoma-derived cells (HepG2) was thawed in a water bath at 37° C. to obtain 1 mL of cell solution, which was then added to 5 mL of medium, centrifuged at 1000 rpm for 3 minutes, and the supernatant was removed.

[0070] The precipitated cells were suspended in 10 mL of medium and collected at 75 cm 2 / Tissue Culture Flask (AGC Technoglass Co., Ltd.) plus 5% CO 2 The cells were cultured in an incubator at 37°C for 3 days.

[0071] A 24-well plate was used, with a cell concentration of 1.0 x 10 cells per well. 6 The cell solution was added so that the cell concentration became 10 cells / mL, and the ARs-containing dried product obtained in Preparation Example 1 was added thereto so that the final concentration in terms of ARs became 10 μg / mL, and the mixture was incubated at 37° C. for 18 hours. After that, the medium was removed, the cells were washed three times with 1000 μL of PBS(-), washed once with 300 μL of 0.5 mg / mL trypsin solution (containing 0.2 mg / mL EDTA·4Na), and then 300 μL of the same trypsin·EDTA solution was added and incubated at 37°C for 5 minutes. 600 μL of medium was added to completely detach the cells, and the entire amount of the obtained cell suspension was transferred to a 1.5 mL sterile tube and centrifuged at 3000 rpm for 5 minutes, the supernatant was removed, and the cells were collected. The collected cells were stored at -80°C until the preparation of total RNA.

[0072] Total RNA was prepared from the cells collected after the above culture in a conventional manner, and the mRNA expression level of HSP70, a heat shock protein, was measured by quantitative RT-PCR in the same manner as in Test Example 2. The results were compared relative to the control group, which was set at 100%.

[0073] [Table 3]

[0074] As a result, as shown in FIG. 4, the addition of alkylresorcinols induced the expression level of human HSP70 mRNA.

[0075] (Sequence Number) SEQ ID NO:1: Forward primer for PCR amplification of mouse HSP90 gene SEQ ID NO:2: Reverse primer for PCR amplification of mouse HSP90 gene SEQ ID NO: 3: Forward primer for PCR amplification of human HSP70 gene SEQ ID NO: 4: Reverse primer for PCR amplification of the human HSP70 gene SEQ ID NO: 5: Forward primer for PCR amplification of mouse and human β-actin genes SEQ ID NO:6: Reverse primer for PCR amplification of mouse and human β-actin genes

Claims

1. A heat shock protein expression inducer that contains plant-derived resorcinol lipids as its active ingredient.

2. The heat shock protein expression inducer according to claim 1 , wherein the resorcinol lipid is an alkylresorcinol.

3. The heat shock protein expression inducer according to claim 1 , wherein the resorcinol lipid is derived from a grass family plant.

4. 2. The heat shock protein expression inducer of claim 1, wherein the resorcinol lipid is derived from the husk and / or whole grain of wheat and / or rye.

5. 2. The heat shock protein expression inducer according to claim 1, wherein the resorcinol lipid comprises a solvent extract of a plant containing resorcinol lipid.

6. A composition for inducing heat shock protein expression, comprising the heat shock protein expression inducer according to any one of claims 1 to 5, and being provided as a food or drink, a food additive, a pharmaceutical, a supplement, a cosmetic, or an animal feed.

7. Use of a plant-derived resorcinol lipid for the preparation of a composition for inducing expression of a heat shock protein.

8. 8. The use according to claim 7, wherein the resorcinol lipid is an alkylresorcinol.

9. The use according to claim 7, wherein the resorcinol lipid is derived from a grass plant.

10. 8. The use according to claim 7, wherein the resorcinol lipids are derived from the husk and / or whole grain of wheat and / or rye.

11. 8. The use according to claim 7, wherein the resorcinol lipids comprise a solvent extract of a plant containing resorcinol lipids.

12. The use according to any one of claims 7 to 11, wherein the composition for inducing expression of a heat shock protein is provided as a food or drink, a food additive, a medicine, a supplement, a cosmetic, or an animal feed.

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