JP2019210245A - CXCL12 expression promoter - Google Patents

Abstract

To find a substance that can control, from outside the body, CXCL12 composing the niche of pigment stem cells and involved in the suppression of pigment stem cell differentiation, and to provide this as an agent for fundamentally and efficiently preventing and/or improving gray hair.SOLUTION: Provided is a CXCL12 (stromal cell-derived factor-1) expression promoter, and the suppressing agent of pigment stem cell differentiation, characterized by containing as active ingredients truffle extract and one or two extracts selected from the extract of Panax ginseng that has been heat-dried after steam heat treatment.SELECTED DRAWING: None

Description

  The present invention relates to a CXCL12 expression promoter and a pigment stem cell differentiation inhibitor.

  The color of skin and hair is greatly influenced by melanin produced by melanocytes. It is known that the origin of melanocytes in vivo is pigment stem cells present in the bulge region in the hair follicle. Melanoblasts differentiated from pigment stem cells migrate to the epidermis and hair bulbs and acquire expression of a series of melanin synthesis-related enzymes such as Tyrosinase (TYR), TYR-related protein-1 (TYRP1), and Dopachrome tautomerase (DCT) However, it differentiates into mature melanocytes with melanin synthesis ability, and is involved in pigment production of skin and hair, respectively (Non-patent Document 1). Therefore, abnormalities in the maintenance and differentiation of pigment stem cells cause gray hair. For example, as one of the mechanisms of graying, it has become clear that pigment stem cells are ectopically differentiated into melanocytes in the bulge region, leading to depletion of pigment stem cells and loss of the source of melanocytes. (Non-Patent Document 2).

  Conventionally, the main method for improving gray hair is to activate melanocytes present in the hair follicle and promote the production of melanin and the proliferation of melanocytes. However, such a method does not control the pigment stem cells that are the origin of melanocytes, is limited or coping therapy, and cannot improve the fundamental gray hair.

  In general, stem cells are controlled by a microenvironment surrounding a stem cell called a niche (Non-patent Document 3). In the hair follicle bulge region, hair follicle stem cells exist so as to surround the periphery of the pigment stem cells and function as niche cells of the pigment stem cells. Therefore, if attention is paid to the properties of hair follicle stem cells and the niche of pigment stem cells can be appropriately controlled, it can be said that it is extremely effective as a method for fundamentally improving gray hair, as compared with conventional methods of melanocyte activation and proliferation promotion.

  On the other hand, in the previous researches by the present inventors, CXCL12 (also known as stromal cell-derived factor-1: SDF-1), which is a kind of chemokine, is specifically expressed in hair follicle stem cells in the bulge region in the hair follicle. The expression level is lower in the hair root of gray hair than in the hair root of black hair, and CXCL12 produced by hair follicle stem cells suppresses ectopic differentiation of pigment stem cells and suppresses expression of melanocyte differentiation-related genes. (Patent Document 1). Therefore, if CXCL12 that constitutes the pigment stem cell niche that is the origin of this melanocyte can be controlled, a fundamental and effective white hair prevention and / or improvement effect can be achieved compared to conventional approaches that target only melanocytes. I can expect. In addition, since it is difficult to control factors such as CXCL12 in vivo, development of drugs that can be controlled from the outside is awaited.

JP2018-027920

Nishimura EK, Jordan SA, Oshima H, Yoshida H, Osawa M, Moriyama M, Jackson IJ, Barrandon Y, Miyachi Y, Nishikawa S, Dominant role of the niche in melanocyte stem-cell fate determination, Nature. 2002 Apr 25; 416 (6883): 854-60. Nishimura EK, Granter SR, Fisher DE, Mechanisms of hair graying: incomplete melanocyte stem cell maintenance in the niche, Science. 2005 Feb 4; 307 (5710): 720-4. Epub 2004 Dec 23. Mesa KR, Rompolas P, Greco V., The Dynamic Duo: Niche / Stem Cell Interdependency, Stem Cell Reports. 2015 Jun 9; 4 (6): 961-6.

  Therefore, the present invention has found a substance that constitutes a pigment stem cell niche and can control CXCL12 involved in suppression of pigment stem cell differentiation from the outside of the living body, and can prevent and / or prevent gray hair fundamentally and efficiently. It is an object to provide as a drug for improvement.

  As a result of intensive studies to solve the above problems, the inventors of the present invention significantly increased the expression of CXCL12 in hair follicle stem cells by extracting truffle, extract of ginseng that had been subjected to predetermined pretreatment, and a mixture thereof. And the present invention has been completed.

That is, the present invention includes the following inventions.
(1) CXCL12 (stromal cell) characterized by containing as an active ingredient one or two extracts selected from truffle extract and steamed ginseng extract after steam heat treatment -expression factor of derived factor-1).
(2) Differentiation of pigment stem cells, characterized by containing as an active ingredient one or two extracts selected from truffle extract and steamed ginseng extract after steam heat treatment Inhibitor.
(3) The agent according to (1) or (2), wherein the ginseng is dried or raw ginseng.

  Since the CXCL12 expression promoter of the present invention can significantly increase the expression of CXCL12 involved in the suppression of pigment stem cell differentiation, it is effective for fundamental prevention and / or improvement of gray hair.

  The CXCL12 expression promoter according to the present invention contains, as an active ingredient, one or two extracts selected from a truffle extract and an extract of ginseng that has been heat-dried after steam heat treatment. To do. In the present invention, “promoting CXCL12 expression” refers to promoting CXCL12 expression at a biological level or a culture level.

  CXCL12 (also known as stromal cell-derived factor-1) is a ligand for the chemokine receptor CXCR4,7, which is a G protein-coupled receptor (GPCR). As described above, CXCL12 is specifically expressed in hair follicle stem cells in the hair follicle bulge region, which is a pigment stem cell niche (the ecological position of stem cells), and CXCL12 produced by hair follicle stem cells is expressed in hair follicle bulges. It is known that it has the action of suppressing the differentiation of pigment stem cells in the region and suppressing the expression of melanocyte differentiation-related genes (Japanese Patent Laid-Open No. 2018-027920). As shown in the examples described later, the extract of truffles, the extract of ginseng that has been heat-dried after steam heat treatment (hereinafter sometimes referred to as “Suji ginseng”), and mixtures thereof, Has CXCL12 expression promoting effect. Accordingly, one or two extracts selected from the truffle extract and the repaired ginseng extract promote the expression of CXCL12 to differentiate the pigment stem cells in the bulge region in the hair follicle, which is the niche of the pigment stem cells. Since suppression becomes possible, it can be used as an active ingredient of a pigment stem cell differentiation inhibitor. In the present invention, “inhibition of differentiation of pigment stem cells” means suppression of ectopic differentiation of pigment stem cells in the hair follicle bulge region (niche) that is originally maintained in an undifferentiated state, and in vitro of isolated pigment stem cells. Inhibition of differentiation at the same time, and maintenance of undifferentiation at the biological level or culture level. Therefore, the pigment stem cell differentiation inhibitor of the present invention can also be referred to as a pigment stem cell undifferentiated state maintenance agent or function maintenance agent.

  Moreover, CXCL12 can prevent the decrease or depletion of pigment stem cells that are the source of melanocytes by suppressing the ectopic differentiation of pigment stem cells in the bulge region in the hair follicle. Therefore, from the extract of truffles and the extract of repaired ginseng The selected one or two kinds of extracts can be used as an active ingredient for preventing and / or improving white hair. In the present invention, “prevention and / or improvement of white hair” includes prevention of white hair generation, improvement of the degree (number and range) of white hair, reduction in the progress of whitening, change from white hair to black hair, accompanied by white hair. Examples include suppression of hair loss, suppression of decrease in gloss and elasticity of hair associated with white hair. Moreover, the prevention and / or improvement effect of gray hair includes both the case of showing a direct action mechanism on the hair and the case of showing a percutaneous action mechanism on the head.

  The truffle used in the present invention (scientific name: Tuber spp.) Is an ascomycete belonging to the order of Tuberaceae, Tuberaceae, and is agglomerated in the ground and the seed layer is open to the outside. Absent. Many fruiting bodies have a strong scent, and animals such as squirrels and rabbits are dug out for food. Truffle is one of the world's three delicacies and is used in fine French cuisine. As a kind of truffle used for this invention, a white truffle (Tuber magnatum Pico) and a black truffle (Tuber melanosporum Vitt) are preferable.

  In the present invention, truffle extract refers to fruit body or mycelium extract, but fruit body extract is preferred. In addition, fruit bodies or mycelia may be used as they are for extraction, or may be subjected to treatments such as drying, pulverization and shredding.

  The extraction method is not particularly limited, and can be performed by stirring or column extraction using water or hot water, or a mixed solvent of water and an organic solvent. Examples of organic solvents include lower alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohols (1,3-butylene glycol, propylene glycol, glycerin, etc.) , Ketones (acetone, methyl ethyl ketone, etc.), acetonitrile, esters (ethyl acetate, butyl acetate, etc.), hydrocarbons (hexane, heptane, liquid paraffin, etc.), ethers (ethyl ether, tetrahydrofuran, propyl ether, etc.) It is done. Among these, polar solvents such as lower alcohols and liquid polyhydric alcohols are preferable, water-soluble organic solvents such as ethanol, 1,3-butylene glycol, and propylene glycol are more preferable, and one or more of these may be used. . Particularly preferred extraction solvents include water or water-ethanol mixed polar solvents. The amount of the solvent used is not particularly limited. For example, it may be 10 times or more, preferably 20 times or more with respect to the truffle (dry weight). For convenience of operation, it is preferably 100 times or less. Moreover, although extraction temperature and time depend on the kind of solvent to be used, for example, it is 10-100 degreeC, Preferably it is 30-90 degreeC, 30 minutes-24 hours, Preferably it can illustrate 1 to 10 hours. In addition, the extract may be used as it is in the extracted solution, but if necessary, in a range that does not affect the effect, concentration (concentration by organic solvent, vacuum concentration, membrane concentration, etc.), dilution, filtration, You may use, after performing processing, such as decoloring by activated carbon, deodorizing, ethanol precipitation. Further, the extracted solution may be subjected to a treatment such as concentration to dryness, spray drying, freeze drying, etc., and used as a dried product.

  Panax ginseng CAMey (scientific name: Panax ginseng CAMey, also known as ginseng, ginseng, ginseng) used in the present invention is a perennial plant belonging to the genus Araliaceae genus Panax, and is a herbal medicine "carrot" (Japanese name: The basic plant of carrot, scientific name: Ginseng Radix). The dried ginseng root is used as a herbal medicine, but due to the difference in the manufacturing method, the root skin was peeled off and dried `` white ginseng '' and steamed and dried with the skin `` It is broadly divided into “red ginseng”.

  In the present invention, as for ginseng, any of a part of the plant body such as leaves, stems, fruits, fruit skins, flowers, flower buds, seeds, whole plants, roots, rhizomes or the like, or a mixture thereof can be used. However, roots (particularly lateral roots), leaves and stems are preferred.

  In the present invention, as the ginseng, either dried ginseng or raw ginseng can be used, but dried ginseng is preferred. In the case of dried ginseng, it is preferably dried until the water content is 20% or less, preferably 10% or less. The moisture content can be measured using a method such as loss on drying of the Japanese Pharmacopoeia. The drying method is not particularly limited as long as it is a method usually used as a method for drying a plant and the water content falls within the above range. For example, natural drying (air drying), sun drying, ventilation drying, hot air drying, etc. , Spray drying, vacuum drying, vacuum drying and the like.

  In the present invention, before extracting the above-mentioned dried ginseng or raw ginseng, steam heat treatment and heat drying treatment corresponding to the repair treatment (processing treatment of steaming and drying) performed during the processing of the herbal medicine are performed. The steam heating process is a process of heating an object in a high humidity atmosphere, for example, an atmosphere having a humidity of 80% or more, using hot water, saturated steam, superheated steam, reduced pressure (vacuum) steam or the like as a heat medium. Steam heating may be performed by bringing the heat medium into direct contact with the object, or the object may be heated indirectly through a heat exchanger. Heating may be performed under normal pressure or under pressure. As conditions for the steam heat treatment, the temperature is preferably 70 to 180 ° C, more preferably 100 to 150 ° C, and still more preferably 100 to 120 ° C. Although time changes with temperature, 1 to 15 hours are preferable, 2 to 10 hours are more preferable, and 4 to 8 hours are further more preferable. These temperature and time conditions are merely examples, and can be appropriately changed depending on the relationship between temperature and time. Moreover, what is necessary is just to perform the steam heat processing in this invention using a continuous type or batch type steamer (steamer), an autoclave, etc. FIG.

  Since the ginseng that has been steam-heated contains moisture, it is heat-dried. As temperature of heat drying, 40-85 degreeC is preferable and 50-70 degreeC is more preferable. As a drying method, ventilation drying, hot air drying, microwave drying, or the like can be used. The drying time (heating time) varies depending on the heating temperature, the water content of ginseng after the steam heat treatment, and the total amount to be dried, and is not specified, but is in the range of about 6 to 24 hours.

  After the steam heat treatment as described above, the ginseng (Shuji ginseng) that has been heat-dried is extracted. The extraction method is not particularly limited, and can be performed by stirring or column extraction using water or hot water, or a mixed solvent of water and an organic solvent. Examples of organic solvents include lower alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohols (1,3-butylene glycol, propylene glycol, glycerin, etc.) , Ketones (acetone, methyl ethyl ketone, etc.), acetonitrile, esters (ethyl acetate, butyl acetate, etc.), hydrocarbons (hexane, heptane, liquid paraffin, etc.), ethers (ethyl ether, tetrahydrofuran, propyl ether, etc.) It is done. Among these, polar solvents such as lower alcohols and liquid polyhydric alcohols are preferable, water-soluble organic solvents such as ethanol, 1,3-butylene glycol, and propylene glycol are more preferable, and one or more of these may be used. . Particularly preferable extraction solvents include water, water-ethanol mixed polar solvents, and 1,3-butylene glycol. The amount of the solvent used is not particularly limited. For example, it may be 10 times or more, preferably 20 times or more of the above ginseng (dry weight). However, in the case of concentration or isolation after extraction. For convenience of operation, it is preferably 100 times or less. Moreover, although extraction temperature and time depend on the kind of solvent to be used, for example, it is 10-100 degreeC, Preferably it is 30-90 degreeC, 30 minutes-24 hours, Preferably it can illustrate 1 to 10 hours. In addition, the extract may be used as it is in the extracted solution, but if necessary, in a range that does not affect the effect, concentration (concentration by organic solvent, vacuum concentration, membrane concentration, etc.), dilution, filtration, You may use, after performing processing, such as decoloring by activated carbon, deodorizing, ethanol precipitation. Further, the extracted solution may be subjected to a treatment such as concentration to dryness, spray drying, freeze drying, etc., and used as a dried product.

  In the present invention, any one of the truffle extract and the repaired ginseng extract may be used, but using both together is preferable because the CXCL12 expression promoting effect is enhanced. In the case where the extract of truffle and the extract of repair ginseng are used in combination, the mixing ratio is not limited, but is preferably 1:10 to 10: 1, more preferably 1: 5 to 5: 1, and still more preferably. 1: 2 to 2: 1, most preferably 1: 1.

  When one or two extracts selected from truffle extract and extract of repair ginseng are used as an agent for preventing and / or improving gray hair, they can be used as they are. Can be mixed with an appropriate additive or the like as long as it does not impair hair, and can be in the form of a composition such as cosmetics, quasi-drugs, and pharmaceuticals for preventing and / or improving gray hair. Especially, the composition for hair formulated into the formulation form suitable for using for a scalp and hair is preferable.

  The composition for hair can be produced according to a method known in the art by selecting various ingredients, additives, bases, and the like that are usually used in an external composition for skin according to the type and mixing them appropriately. it can. Examples of ingredients, additives, and bases to be blended include diluents (purified water, ethanol, etc.), fats and oils (olive oil, coconut oil, evening primrose oil, jojoba oil, castor oil, hardened castor oil, etc.), waxes ( Lanolin, beeswax, carnauba wax, etc.), hydrocarbons (liquid paraffin, squalene, squalane, petrolatum, etc.), fatty acids (lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, etc.), higher alcohols (myristyl alcohol, Cetanol, cetostearyl alcohol, stearyl alcohol, behenyl alcohol, etc.), esters (isopropyl myristate, isopropyl palmitate, cetyl octanoate, glyceryl trioctanoate, octyldodecyl myristate, octyl stearate, stearyl stearate, etc.), organic acids Citric acid, lactic acid, α-hydroxyacetic acid, pyrrolidone carboxylic acid, etc.), saccharides (maltitol, sorbitol, xylobiose, N-acetyl-D-glucosamine, etc.), surfactant, silicone oil, humectant, thickener, UV Examples include absorbents, sequestering agents, cooling agents, antioxidants, stabilizers, preservatives, flame retardants, bactericides, fragrances, and coloring agents.

Moreover, as long as the said composition for hair does not have a bad influence on the effect of this invention, you may include the component conventionally known as an effective component for hair growth and hair growth. For example, plant extract such as assembly extract, citrus extract, vitamin B ₆ , vitamin E and derivatives thereof, vitamins such as biotin, pantothenic acid and derivatives thereof, glycyrrhizic acid and derivatives thereof, nicotinic acid ester, serine, methionine, etc. Examples thereof include amino acids, ceforanthin, carpronium chloride, minoxidil, nicorandil, acetylcholine derivatives, cyclosporine, and female hormone agents such as estradiol, and mixtures thereof.

  In the present invention, the hair composition widely refers to those used for the scalp and hair, and any composition can be used as long as it is applicable to the scalp and hair, and the dosage form is not particularly limited. For example, any of liquid, emulsion, cream, gel, paste, spray and the like may be used. Specific product forms include cream, lotion, emulsion, ointment, gel, hair shampoo, hair rinse, hair treatment, hair conditioner, scalp treatment, hair spray, hair pack, hair essence, hair tonic, hair liquid, hair mousse, etc. Can be mentioned.

  Although the content of one or two extracts selected from the extract of truffle and the extract of repair ginseng in the hair composition taking these product forms differs depending on the form, it cannot be specified. In general, it is 0.0001 to 20% by weight (w / w), preferably 0.001 to 10% by weight (w / w) based on the total weight of the composition. About the addition method of the 1 type or 2 types of extract chosen from the extract of truffle and the extract of repair ginseng, it may add in advance or may be added in the middle of manufacture, and it considers workability suitably. Just choose.

  One or two extracts selected from a truffle extract and an extract of repaired ginseng are also used to maintain the undifferentiated state of the cells when the pigment stem cells are cultured for use in regenerative medicine. It can also be used as a medium additive. Pigment stem cells cultured in a medium to which the medium additive is added can survive for a certain period while maintaining undifferentiation. The undifferentiated state maintenance period of the cells can be appropriately changed according to the purpose of culture, the type of basal medium, the culture temperature, and the like.

  Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to these.

[Example 1] Production example of truffle and ginseng extract A truffle and ginseng extract was produced as follows.

(Production Example 1) Preparation of hot water extract of truffle 1 L of purified water was added to 100 g of crushed truffle, extracted at 90-100 ° C. for 2 hours, filtered, the filtrate was concentrated, freeze-dried and truffle 5.1 g of a hot water extract was obtained.

(Production Example 2) Preparation of a hot water extract of raw ginseng root (repair treatment / steam heating: 105 ° C., heat drying: 50 ° C.) Harvested raw ginseng (water content 80%) at 105 ° C. for 8 hours After steaming, it was dried at 50 ° C. (compatible with “Japanese red ginseng” of Japanese Pharmacopoeia) After adding 800 mL of purified water to 40 g of this dried ginseng and extracting at 95-100 ° C. for 2 hours, filtering, concentrating the filtrate to dryness, and steaming heat treatment (105 ° C.) of raw ginseng root 17.5 g of hot water extract was obtained.

(Production Example 3) Preparation of a hot water extract of dried ginseng root (salvage treatment / steam heating: 105 ° C., heat drying: 50 ° C.) Steamed dried ginseng root (water content 9%) at 105 ° C. for 8 hours. And then dried at 50 ° C. After adding 800 mL of purified water to 40 g of this dried ginseng and extracting at 95-100 ° C. for 2 hours, filtering, concentrating the filtrate, freeze-drying, and steam-heated (105 ° C.) dried ginseng root 18.7 g of a hot water extract was obtained.

(Production Example 4) Preparation of 50% ethanol extract of dried ginseng root (salvage treatment / steam heating: 105 ° C., heat drying: 70 ° C.) Dried ginseng root (water content 9%) at 105 ° C. for 8 hours After steaming, it was dried at 70 ° C. 500 g of purified water and 500 mL of ethanol were added to 100 g of this dried ginseng, extracted at room temperature for 7 days, filtered, and the filtrate was concentrated to dryness and dried by steam heat treatment (105 ° C.). 43 g of 50% ethanol extract was obtained.

(Production Example 5) Preparation of 1,3-butylene glycol extract of dried ginseng leaves and stems (repair treatment / steam heating: 100 ° C., heat drying: 60 ° C.) Dried ginseng leaves and stems (water content 10% ) Was steamed at 100 ° C. for 4 hours and then dried at 60 ° C. 200 g of 1,3-butylene glycol was added to 20 g of this dried ginseng, extracted at room temperature for 7 days, filtered and steam-heated (100 ° C.) 1,3-butylene glycol in the roots and stems of dried ginseng 141 g of extract was obtained.

(Comparative Production Example 1) Preparation of hot water extract of raw ginseng root (uncured / no steam heat treatment) Harvested raw ginseng root after drying at 65 ° C. (white ginseng) 40 g of purified water After adding 800 mL and extracting at 95-100 degreeC for 2 hours, it filtered, the filtrate was concentrated and lyophilized | freeze-dried, and 12.5g of hot water extract of the raw ginseng root was obtained.

[Example 2] Truffle and Panax ginseng CXCL12 expression promoting effect Truffle extract and Panax ginseng extract produced with each production agent of Example 1 constitutes a pigment stem cell niche and is responsible for maintenance of hair follicle stem cells In order to analyze how CXCL12 is affected, these extracts were allowed to act on hair follicle stem cells, and the gene expression level of CXCL12 at that time was analyzed.

(1) Culture of human hair follicle stem cells Human hair was collected by hair removal, and a tissue containing a bulge region of the hair follicle tissue was collected using a scalpel or the like. After washing with PBS (−), trypsin (BD Biosciences) treatment was performed. Thereafter, the cells were isolated and recovered using a cell strainer (manufactured by FALCON). The collected cells were seeded on a culture plate and maintained using KG2 medium (manufactured by KURABO) until confluent. Confluent cells were collected, seeded again on a culture plate, then engrafted, and proliferating cells were used as hair follicle stem cells in the following tests.

(2) CXCL12 expression promoting effect in hair follicle stem cells Each extract obtained in the above production example was added to the hair follicle stem cells so that the final concentration was 100 μg / mL. However, for the mixed extract of the truffle extract and the ginseng extract, two kinds of extracts were added at 50 μg / mL so that the final concentration of the mixed extract was 100 μg / mL. The extract was added, the cells after 48 hours were collected, and the gene expression level of CXCL12 in hair follicle stem cells was analyzed.

  CXCL12 gene expression analysis was performed as follows. The hair follicle stem cells after the addition of the extract were washed twice with PBS (−), and then RNA was extracted from the cells with Trizol Reagent (Invitrogen). 2-STEP real-time PCR kit (Applied Biosystems) was used to reverse-transcribe the extracted RNA into cDNA, then ABI7300 (Applied Biosystems) was used to perform real-time PCR (95 ° C .: 15 seconds) , 60 ° C .: 30 seconds, 40 cycles), and the expression of CXCL12 was confirmed. Other operations were carried out in accordance with established methods.

Primer set for CXCL12:
5′-CATGCCCGATTCTTCGAAAGC-3 ′ (SEQ ID NO: 1)
5′-CGAGTGGGTCTAGCGGAAAG-3 ′ (SEQ ID NO: 2)
Primer set for 18srRNA (internal standard):
5′-CCGAGCCCCCTGGATAC-3 ′ (SEQ ID NO: 3)
5′-CAGTTCCGAAAACCAACAAAAATAGA-3 ′ (SEQ ID NO: 4)

  As for the CXCL12 expression promoting effect, CXCL12 mRNA expression level in hair follicle stem cells (control) cultured without adding an extract was calculated as a ratio with respect to the expression level of 18s ribosomal RNA (18srRNA) as an internal standard. The value of the gene relative expression level (CXCL12 gene expression level / 18srRNA gene expression level) is set to 1, and the value of the CXCL12 gene relative expression level in hair follicle stem cells cultured with the extract added is calculated and evaluated. did. The results are shown in Table 1 below.

  As shown in Table 1, hairs cultured in a medium supplemented with truffle extract (Production Example 1), steam heat treatment followed by heat drying treatment (Shuji) Panax ginseng extract (Production Examples 2-5) It was shown that the expression level of the CXCL12 gene was increased and the CXCL12 gene expression was promoted in the stem cells, compared to the extract of uncured ginseng (white ginseng) (Comparative Production Example 1). Moreover, the extract of the ginseng (manufacture examples 3-5) dried before the steam heat treatment was more effective than the extract of the ginseng (red ginseng) (manufacture example 2). Furthermore, when the extract of truffle (manufacture example 1) and the extract of ginseng (manufacture examples 2-5) were used together, it was confirmed that the effect was remarkably enhanced. Therefore, the extract of truffles, the extract of repaired ginseng, and the mixture thereof have an undifferentiated state by suppressing the ectopic differentiation of pigment stem cells in the bulge region in the hair follicle based on the CXCL12 gene expression promoting action. While maintaining, it can suppress the expression of melanocyte differentiation-related genes, and thus can be said to be able to fundamentally suppress and prevent graying.

  INDUSTRIAL APPLICABILITY The present invention can be used in the field of manufacturing cosmetics and quasi-drugs aimed at preventing and / or improving gray hair.

Claims (3)

  CXCL12 (stromal cell-derived factor) characterized by containing as an active ingredient one or two extracts selected from truffle extract and steamed ginseng extract after steam heat treatment -1) expression promoter.   An agent for inhibiting differentiation of pigment stem cells, comprising as an active ingredient one or two extracts selected from truffle extract and extract of ginseng that has been heat-dried after steam heat treatment.   The agent according to claim 1 or 2, wherein the ginseng is dried or raw ginseng.