JP2012219031A - LAMININ 5 PRODUCTION PROMOTER, INTEGRIN α6β4 PRODUCTION PROMOTER, SKIN BASEMENT MEMBRANE-NORMALIZING AGENT, SKIN DAMAGE-RESTORING PROMOTER, AND AQUAPORIN 3mRNA EXPRESSION PROMOTER - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a laminin 5 production promoter, an integrin α6β4 production promoter, a skin basement membrane-normalizing agent, a skin damage-restoring promoter or an aquaporin 3mRNA expression promoter which contains, as an active ingredient, a substance which has laminin 5 production-promoting action, integrin α6β4 production-promoting action or aquaporin 3mRNA expression-promoting action, the substance being found from a natural product which is high in safety.SOLUTION: The laminin 5 production promoter, the integrin α6β4 production promoter, the skin basement membrane-normalizing agent, the skin damage-restoring promoter or the aquaporin 3mRNA expression promoter includes, as an active ingredient, an extract from Alpinia zerumbet.

Description

  The present invention relates to a laminin 5 production promoter, an integrin α6β4 production promoter, a skin basement membrane normalizing agent, a skin damage recovery promoter, and an aquaporin 3 mRNA expression promoter.

  The skin is composed of the stratum corneum, epidermis, basement membrane and dermis. The basement membrane is present at the boundary between the epidermis and the dermis and plays an important role not only in connecting the epidermis and the dermis but also in maintaining the skin function (see Non-Patent Document 1). The main skeleton of the basement membrane has a network structure composed of type IV collagen. Various glycoproteins mainly composed of laminin 5 are present at the boundary between the basement membrane and the epidermis and connect the basement membrane and the epidermis. Such laminin 5 is produced from epidermal keratinocytes present in the epidermis. Is done. In young skin, the basement membrane works to maintain the homeostatic interaction of the epidermis and dermis, ensuring moisture retention, flexibility, elasticity, and the like. Maintained.

  However, laminin 5 which is a main component of the basement membrane degrades and deteriorates when it is affected by certain external factors such as ultraviolet irradiation, drastic air drying, excessive skin washing, etc., or when aging progresses. And the basement membrane structure is destroyed (see Non-Patent Document 2). As a result, the skin's moisturizing function and elasticity are lowered, and the keratin begins to exfoliate abnormally, so the skin loses its tension and gloss, and exhibits aging symptoms such as roughness and wrinkles. As described above, changes associated with skin aging, that is, wrinkles, dullness, disappearance of texture, decrease in elasticity, and the like are associated with a decrease in basement membrane components and structural changes in basement membrane, and production of laminin 5 It is considered that aging symptoms of the skin can be prevented and improved by promoting the above.

  Laminin consists of various combinations of α chain, β chain, and γ chain, and 15 types (laminin 1 to laminin 15) are currently known. These α chain, β chain, and γ chain form a coiled triple chain structure at a site called a long arm, thereby forming a huge laminin molecule. Among them, laminin 5 (α3β3γ2) is abundantly present in the basement membrane of epithelial tissues such as skin, digestive organs, kidneys and lungs. In genetic diseases caused by congenital abnormalities of the genes encoding each chain of laminin 5 (lethal congenital epidermolysis bullosa, Herlitz junctional epidermolysis bullosa), the epidermis of the whole body may show fatal symptoms Are known. Laminin 5 is known to strongly adhere cells (high cell adhesion activity) and strongly promote cell motility (high cell motility activity) compared to other extracellular matrix molecules.

  Thus, since laminin 5 has a high cell motility activity, it is known that laminin 5 promotes cell migration in damaged skin and promotes wound healing (see Patent Document 1). That is, promoting the production of laminin 5 is important in promoting healing of skin damage that destroys the structure of the basement membrane.

  Conventionally, a skin external preparation containing laminin 5 (see Patent Document 2) has been known, and soy extract (see Patent Document 3), phenylpropanoids (Patent Document 3) have an action of promoting laminin 5 production. Literature 4), pantothenic acid (see Patent Literature 5), lysophosphatidylcholine or lysophosphatidic acid (see Patent Literature 6), coenzyme Q10 (see Patent Literature 7) and the like are known.

  On the other hand, in order for epidermal cells to adhere to extracellular matrix proteins such as type IV collagen, laminin, and fibronectin, which are constituents of the basement membrane, integrins that function as receptors are essential. Integrins are transmembrane proteins that are expressed on the cell membrane, and bind to extracellular matrix proteins such as collagen, laminin, and fibronectin, and also bind to cytoskeletal proteins such as tailin and actinin inside the cell. Involved in cell-cell binding.

  An integrin is a heterodimer in which an α chain and a β chain are associated with each other in a non-covalent manner, and various molecules having different ligand specificities are formed by a combination of the α chain and the β chain. Among these, integrin α6β4 and integrin α3β1 are known as laminin 5 receptors. Integrin α6β4 is expressed in epithelial cells, Schwann cells, nerve cells, some endothelial cells, etc., and is present on the basement membrane side of epidermal cells in the skin, and binds to laminin 5 present in the basement membrane, It plays a role in maintaining normal skin structure. It is known that the congenital abnormality of the gene encoding each chain of integrin α6β4 causes lethal congenital epidermolysis bullosa like laminin 5.

  The binding of integrin α6β4 and laminin 5 is known not only to stabilize cells by adhesion but also to transmit signals into epidermal cells via integrin α6β4 to initiate normal growth of epidermal cells. . Integrin α6β4 is known to decrease expression in epidermal cells with aging, which is considered to contribute to skin aging. Therefore, by promoting the expression of integrin α6β4, normal proliferation of epidermal cells is promoted, which leads to repair of basement membrane damage and maintenance of normal skin structure, and can prevent and improve skin aging symptoms. .

  Based on such an idea, for example, an extract of litchi and oren extract (see Patent Document 8), an extract of Japanese kizuta (see Patent Document 9), and the like are known as having integrin α6β4 expression promoting action.

  In skin cells, it is known that aquaporins, known as water channels, are expressed on the cell membrane and play a role of taking in low-molecular substances such as interstitial water into cells.

  In humans, the presence of 13 types of aquaporins (AQP0 to AQP12) is known. In epidermal cells, aquaporin 3 (AQP3) is mainly present, and it is considered to play a role of taking in low-molecular compounds such as glycerol and urea involved in water retention action in addition to water.

  However, since aquaporin 3 decreases with aging, and this has been suggested to contribute to a decrease in the water retention function, by promoting the expression of aquaporin 3, the water retention capacity and barrier due to aging are increased. It is considered that functions and the like can be controlled (see Non-Patent Document 3). Based on such an idea, for example, tocopheryl retinoate (see Patent Document 10), an extract obtained from a Nozenhalen plant (see Patent Document 11), and the like are known as having aquaporin 3 expression promoting action. .

JP 2006-63033 A Japanese Patent Laid-Open No. 10-147515 JP 2004-217618 A JP 2007-77169 A JP 2005-179243 A JP 2000-226308 A JP 2007-63160 A JP 2006-045087 A JP 2003-171225 A JP 2006-290873 A JP 2004-168732 A

Marinkovich MP et al., “J. Cell. Biol.”, 1992, 199, p. 695-703 Lavker et al., “J. Invest. Dermatol.”, 1979, Vol. 73, p. 59-66 “Fragrance Journal”, 2006, Vol. 34, No. 10, p. 19-23

  The present invention finds a laminin 5 production promoting action, an integrin α6β4 production promoting action or an aquaporin 3 mRNA expression promoting action among highly safe natural products, and a laminin 5 production promoting agent comprising the same as an active ingredient, integrin α6β4. An object is to provide a production promoter, a skin basement membrane normalizing agent, a skin damage recovery promoter, or an aquaporin 3 mRNA expression promoter.

  In order to solve the above problems, the laminin 5 production promoter, integrin α6β4 production promoter, skin basement membrane normalizing agent, skin damage recovery promoter or aquaporin 3 mRNA expression promoter of the present invention is obtained by using an extract from moon peach. It is contained as an active ingredient.

  According to the present invention, laminin 5 production promoter having excellent laminin 5 production promoting action and high safety, integrin α6β4 production promoting agent having excellent integrin α6β4 production promoting action and high safety, Skin laminin 5 production promoting action and / or integrin α6β4 production promoting action and highly safe skin basement membrane normalizing agent, excellent laminin 5 production promoting action and / or integrin α6β4 production promoting action and safety It is possible to provide a high skin damage recovery promoter and an aquaporin 3 mRNA expression promoter that has an excellent aquaporin 3 mRNA expression promoting action and is highly safe.

Embodiments of the present invention will be described below.
The laminin 5 production promoter, integrin α6β4 production promoter, skin basement membrane normalizing agent, skin damage recovery promoter, or aquaporin 3 mRNA expression promoter of the present embodiment contains an extract from moon peach as an active ingredient. is there.

  Here, in the present embodiment, the “extract from moon peach” includes an extract obtained by using moon peach as an extraction raw material, a diluted or concentrated solution of the extract, and a dried product obtained by drying the extract. Or any of these crudely purified products or purified products.

The extraction raw material used in the present embodiment is Tsukitomo (scientific name: Alpinia speciosa (Wendl.) K. Schum.).
Moon peach (Alpinia speciosa (Wendl.) K. Schum.) Is a perennial evergreen herb belonging to the genus Glyceraceae and is distributed from southern Kyushu to India and is easily available from these regions . Tsuki-Peach is called Sannin in Okinawa and is used in Muchy, a traditional confectionery since the Ryukyu dynasty. Examples of the part that can be used as an extraction raw material include a leaf part, a stem part, an above-ground part, a flower part, a fruit part, a seed part, a root part, a whole plant, or a mixture of these parts. is there.

  An extract from moon peach can be obtained by drying the raw material for extraction and then pulverizing the raw material as it is or using a crusher and subjecting it to extraction with an extraction solvent. Drying may be performed in the sun or using a commonly used dryer. Moreover, after performing pretreatment, such as degreasing, with a nonpolar solvent such as hexane, it may be used as an extraction raw material. By performing pretreatment such as degreasing, extraction with a polar solvent of moon peach can be performed efficiently.

  As the extraction solvent, it is preferable to use a polar solvent, and examples thereof include water and hydrophilic organic solvents. These are used alone or in combination of two or more at room temperature or a temperature below the boiling point of the solvent. It is preferable.

  Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, and those subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, buffering, and the like. Therefore, the water that can be used as the extraction solvent in this embodiment includes purified water, hot water, ion exchange water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.

  Examples of hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene. Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycol, propylene glycol and glycerin.

  When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when using a mixed solution of water and a lower aliphatic alcohol, it is preferable to mix 1 to 90 parts by volume of a lower aliphatic alcohol with respect to 10 parts by volume of water. When using a mixed solution, it is preferable to mix 1 to 40 parts by volume of a lower aliphatic ketone with 10 parts by volume of water, and when using a mixed solution of water and a polyhydric alcohol, water 10 It is preferable to mix 10 to 90 parts by volume of a polyhydric alcohol with respect to the volume part.

  The extraction treatment is not particularly limited as long as the soluble component contained in the extraction raw material can be eluted in the extraction solvent, and can be performed according to a conventional method. For example, the extraction raw material is immersed in an extraction solvent 5 to 15 times (mass ratio) of the extraction raw material, the soluble components are extracted at room temperature or under reflux, and then filtered to remove the extraction residue. A liquid can be obtained. When the solvent is distilled off from the obtained extract, a paste-like concentrate is obtained, and when this concentrate is further dried, a dried product is obtained.

  The extract obtained as described above can be used as an active ingredient of laminin 5 production promoter, integrin α6β4 production promoter, skin basement membrane normalizing agent, skin damage recovery promoter or aquaporin 3 mRNA expression promoter. However, a concentrated solution or a dried product is easier to use.

  In addition, since the extract from moon peach has a peculiar odor, it can be purified for the purpose of decolorization, deodorization, etc. within a range that does not cause a decrease in its physiological activity. Since it is not used in a large amount when it is blended, etc., there is no practical problem even if it is not purified.

  Since the extract from moon peach obtained as described above has an excellent laminin 5 production promoting action, integrin α6β4 production promoting action and aquaporin 3 mRNA expression promoting action, laminin 5 production promoting agent, integrin α6β4 production It can be used as an active ingredient of a promoter, a skin basement membrane normalizing agent, a skin damage recovery promoter, or an aquaporin 3 mRNA expression promoter.

  The laminin 5 production promoter, integrin α6β4 production promoter, skin basement membrane normalizing agent, skin damage recovery promoter or aquaporin 3 mRNA expression promoter of this embodiment may consist of only an extract from moon peach, An extract from moon peach may be formulated.

  The laminin 5 production promoter, integrin α6β4 production promoter, skin basement membrane normalizing agent, skin damage recovery promoter or aquaporin 3 mRNA expression promoter of this embodiment is a pharmaceutically acceptable carrier such as dextrin, cyclodextrin, etc. It can be formulated into an arbitrary dosage form such as powder, granule, tablet, and liquid according to a conventional method using an arbitrary auxiliary agent. In this case, as an auxiliary agent, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring / flavoring agent, and the like can be used. Laminin 5 production promoter, integrin α6β4 production promoter, skin basement membrane normalizing agent, skin damage recovery promoter or aquaporin 3 mRNA expression promoter may be used in other compositions (eg, skin external preparations, cosmetic foods and beverages). In addition to being used in combination, it can also be used as an ointment, a solution for external use, a patch and the like.

  When the laminin 5 production promoter, integrin α6β4 production promoter, skin basement membrane normalizing agent, skin damage recovery promoter or aquaporin 3 mRNA expression promoter of this embodiment is formulated, the content of extract from moon peach is However, it is not particularly limited, and can be appropriately set according to the purpose.

  Note that the laminin 5 production promoter, integrin α6β4 production promoter, skin basement membrane normalizing agent, skin damage recovery promoter or aquaporin 3 mRNA expression promoter of the present embodiment is optionally laminin 5 production promoter, integrin Other natural extracts having an α6β4 production promoting action or an aquaporin 3 mRNA expression promoting action can be blended with an extract from moon peach and used as an active ingredient.

  As a method for administering the laminin 5 production promoter, integrin α6β4 production promoter, skin basement membrane normalizing agent, skin damage recovery promoter or aquaporin 3 mRNA expression promoter of this embodiment to a patient, transdermal administration, oral administration, and the like are possible. A suitable method for prevention / treatment or the like may be selected as appropriate depending on the type of disease.

  In addition, the dose of the laminin 5 production promoter, integrin α6β4 production promoter, skin basement membrane normalizing agent, skin damage recovery promoter or aquaporin 3 mRNA expression promoter of this embodiment is also determined according to the disease type, severity, patient's It may be appropriately increased or decreased depending on individual differences, administration methods, administration periods, and the like.

  The laminin 5 production promoter of this embodiment can promote the production of laminin 5 through the laminin 5 production promoting action of the extract from moon peach. Thereby, reconstruction of the basement membrane structure can be induced, and skin aging symptoms such as wrinkles, dullness, disappearance of texture, and decrease in elasticity can be prevented or improved. However, the laminin 5 production promoter of the present embodiment can be used for all purposes that are meaningful for exerting the laminin 5 production promoting action in addition to these uses. For example, laminin 5 deficiency (deficiency) It can be used as a prophylactic or therapeutic agent for diseases caused by (such as epidermolysis bullosa).

  The integrin α6β4 production promoter of this embodiment can promote the production of integrin α6β4 through the integrin α6β4 production promoting action of the extract from moon peach. Thereby, reconstruction of the basement membrane structure can be induced, and skin aging symptoms such as wrinkles, dullness, disappearance of texture, and decrease in elasticity can be prevented or improved. However, the integrin α6β4 production promoter of the present embodiment can be used for all purposes that are meaningful for exerting the integrin α6β4 production promoting action in addition to these uses. For example, deficiency (deficiency) of integrin α6β4 It can be used as a prophylactic or therapeutic agent for diseases caused by (such as epidermolysis bullosa).

  The skin basement membrane normalizing agent of this embodiment can promote the production of laminin 5 and / or integrin α6β4 through the laminin 5 production promoting action and / or the integrin alpha 6β4 production promoting action of the extract from moon peach. As a result, restructuring of the basement membrane structure can be induced, and skin aging symptoms such as wrinkles, dullness, loss of texture, and decreased elasticity can be prevented or improved. However, the skin basement membrane normalizing agent of the present embodiment can be used for all purposes meaningful for exerting laminin 5 production promoting action and / or integrin α6β4 production promoting action in addition to these uses.

  The skin damage recovery promoter of this embodiment can promote the recovery of skin damage through the laminin 5 production promoting action and / or the integrin α6β4 production promoting action of the extract from moon peach. In addition to preventing or improving skin aging symptoms such as dullness, loss of texture, and loss of elasticity, it is possible to treat and improve wounds in the skin. However, the skin damage recovery promoting agent of the present embodiment can be used for all uses that are meaningful for exerting laminin 5 production promoting action and / or integrin α6β4 production promoting action in addition to these uses. It can be used for applications such as skin wound treatment.

  The aquaporin 3 mRNA expression promoting agent of the present embodiment can promote the expression of aquaporin 3 through the aquaporin 3 mRNA expression promoting action of the extract from moon peach, thus improving the water retention ability and barrier function of the skin. It is possible to prevent or improve skin aging symptoms such as skin dryness, wrinkle formation, and reduced elasticity. However, the aquaporin 3 mRNA expression promoting agent of the present embodiment can be used for all uses other than these uses that are meaningful for exerting the aquaporin 3 mRNA expression promoting action.

  In addition, the laminin 5 production promoter, integrin α6β4 production promoter, skin basement membrane normalizing agent, skin damage recovery promoter or aquaporin 3 mRNA expression promoter of the present embodiment is excellent in laminin 5 production promotion and integrin α6β4 production promotion. Since it has an action or an aquaporin 3 mRNA expression promoting action, it is suitable, for example, for blending into an external preparation for skin or food and drink. In this case, an extract from moon peach may be added as it is, or a laminin 5 production promoter, an integrin α6β4 production promoter, a skin basement membrane normalizing agent, a skin damage formulated from an extract from moon peach A recovery promoter or an aquaporin 3 mRNA expression promoter may be added.

  Here, the topical skin preparation is not limited in its category, and includes a wide range of skin cosmetics, quasi-drugs, pharmaceuticals, and the like used transdermally. Creams, milky lotions, beauty essences, lotions, packs, foundations, lip balms, bath salts, hair nicks, hair lotions, soaps, body shampoos and the like.

  The food and drink is not limited in its category, and includes a wide variety of foods such as general foods, health foods, health functional foods, quasi drugs, and pharmaceuticals that are taken orally.

  In addition, the laminin 5 production promoter, integrin α6β4 production promoter, skin basement membrane normalizing agent, skin damage recovery promoter or aquaporin 3 mRNA expression promoter of the present embodiment is excellent in laminin 5 production promotion and integrin α6β4 production promotion. Since it has an action or aquaporin 3 mRNA expression promoting action, it can be suitably used as a reagent for research related to the production mechanism of laminin 5, the production mechanism of integrin α6β4, or the expression mechanism of aquaporin 3 mRNA.

  In addition, the laminin 5 production promoter, integrin α6β4 production promoter, skin basement membrane normalizing agent, skin damage recovery promoter or aquaporin 3 mRNA expression promoter of the present embodiment is suitably applied to humans. However, it can be applied to animals other than humans (for example, mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as the respective effects are exhibited.

  Hereinafter, although a test example is shown and this invention is demonstrated concretely, this invention is not restrict | limited to each following example at all. In this test example, a freeze-dried product of moon peach leaf extract BG (manufactured by Maruzen Pharmaceutical Co., Ltd., sample 1) was used as an extract from moon peach.

[Test Example 1] Laminin 5 production promoting action test Laminin 5 production promoting action was tested as follows for the moon peach extract (sample 1).

Normal human epidermal keratinocyte (NHEK) in an 80 cm ² flask using medium for human normal epidermal keratinocytes (KGM) at 37 ° C. and 5% CO ₂ -95% air. The cells were collected by pre-culture and trypsin treatment. The collected cells were diluted with a medium (KGM-BPE) obtained by removing BPE from KGM so that the cell density was 1.0 × 10 ⁵ cells / mL, and then seeded at 500 μL per well in a 24-well plate. ° _C., and cultured for two days under the conditions of 5% CO 2 -95% air.

After completion of the culture, the medium was removed, and 500 μL of KGM-BPE medium added with the test sample (sample 1, sample concentration see Table 1 below) or KGM-BPE medium without sample added was added to each well. ° _C., and cultured for 48 hours under the conditions of 5% CO 2 -95% air. After completion of the culture, 100 μL of the supernatant was transferred to an ELISA plate and adsorbed on the plate at 37 ° C. for 2 hours, and then the amount of adsorbed laminin 5 was used for monoclonal anti-human laminin 5 antibody (mouse IgG, manufactured by Chemicon). It was measured by the ELISA method. From the obtained measurement results, the laminin 5 production promotion rate (%) was calculated by the following formula.

Laminin 5 production promotion rate (%) = A / B × 100
In the formula, A represents “absorbance at a wavelength of 405 nm when a test sample is added”, and B represents “absorbance at a wavelength of 405 nm when no sample is added”.
The results are shown in Table 1.

  As shown in Table 1, it was confirmed that the moon peach extract (sample 1) has an excellent laminin 5 production promoting action.

[Test Example 2] Integrin α6β4 production promoting effect test The moon peach extract (sample 1) was tested for the integrin α6β4 production promoting effect as follows.

Human normal neonatal epidermal keratinocytes (NHEK) were pre-cultured in an 80 cm ² flask using human normal epidermal keratinocyte medium (KGM) at 37 ° C. and 5% CO ₂ -95% air. Cells were collected by treatment. The collected cells are diluted with KGM medium so that the cell density becomes 1.0 × 10 ⁵ cells / mL, and then seeded at 100 μL per well in a 96-well plate, 37 ° C., 5% CO ₂ -95% air. For 2 days.

After completion of the culture, 100 μL of a KGM medium to which a test sample (sample 1, sample concentration is shown in Table 2 below) or a sample-free KGM medium was added was added to each well at 37 ° C., 5% CO ₂ -95%. The cells were cultured for 72 hours under air conditions. After completion of the culture, the medium was removed, the cells were fixed to the plate, and the amount of integrin α6β4 expressed on the cell surface was measured by ELISA using a monoclonal anti-human integrin β4 antibody (mouse IgG, manufactured by Chemicon). From the obtained measurement results, the integrin α6β4 production promotion rate (%) was calculated by the following formula.

Integrin α6β4 production promotion rate (%) = A / B × 100
In the formula, A represents “absorbance at a wavelength of 405 nm when a test sample is added”, and B represents “absorbance at a wavelength of 405 nm when no sample is added”.
The results are shown in Table 2.

  As shown in Table 2, it was confirmed that the moon peach extract (sample 1) has an excellent integrin α6β4 production promoting action.

[Test Example 3] Aquaporin 3 (AQP3) mRNA expression promoting action test The moon peach extract (sample 1) was tested for AQP3 mRNA expression promoting action as follows.

Human normal neonatal epidermal keratinocytes (NHEK) were pre-cultured in an 80 cm ² flask using medium for normal human epidermal keratinocytes (KGM) under conditions of 37 ° C., 5% CO ₂ -95% air, and trypsin. Cells were collected by treatment. The collected cells are diluted with KGM medium so as to have a cell density of 20 × 10 ⁴ cells / mL, and then seeded at 2 mL each in a 35 mm petri dish (manufactured by FALCON) (40 × 10 ⁴ cells / Petri dish) at 37 ° C. They were cultured for 24 hours under the conditions of _5% CO 2 _-95% air.

After culturing, the medium was removed, and 2 mL of KGM medium added with the test sample (sample 1, sample concentration as shown in Table 3 below) or KGM medium without sample was added to each dish at 37 ° C., 5% CO _2. The cells were cultured for 24 hours under the condition of -95% air. After culturing, the medium is removed, and total RNA is extracted with ISOGEN (Nippon Gene, Cat. No. 311-02501), and the amount of each RNA is measured with a spectrophotometer so that it becomes 200 ng / μL. Total RNA was prepared.

  Using this total RNA as a template, the expression levels of AQP3 and the internal standard GAPDH mRNA were measured. Detection was performed by real-time 2-step RT-PCR reaction using a real-time PCR device Smart Cycler (Cepheid) and TaKaRa SYBR Prime Script RT-PCR kit (Perfect Real Time) (Takara Bio, code No. RR063A). . The expression level of AQP3 mRNA was calculated based on the total RNA preparation prepared from the cells cultured in “addition of test sample” and “no sample”, and a correction value was calculated from the value of GAPDH mRNA. When the correction value was set to 100, a correction value for “addition of test sample” was calculated. From the obtained results, the AQP3 mRNA expression promotion rate (%) was calculated by the following formula.

AQP3 mRNA expression promotion rate (%) = A / B × 100
In the above formula, A represents “correction value when test sample is added” and B represents “correction value when sample is not added”.
The results are shown in Table 3.

  As shown in Table 3, it was confirmed that the moon peach extract (sample 1) has an excellent AQP3 mRNA expression promoting action.

  The laminin 5 production promoter, integrin α6β4 production promoter, skin basement membrane normalizing agent, and skin damage recovery promoter of the present invention are used to prevent or prevent skin aging symptoms such as wrinkles, dullness, disappearance of texture, and decrease in elasticity. Useful for improvement. In particular, the skin damage recovery promoter of the present invention is useful for treating or improving skin wounds. Moreover, the aquaporin 3 mRNA expression promoter of the present invention is useful for improving the moisture retention ability and barrier function of the skin.

Claims (5)

  A laminin 5 production promoter comprising an extract from moon peach as an active ingredient.   An integrin α6β4 production promoter comprising an extract from moon peach as an active ingredient.   A skin basement membrane normalizing agent comprising an extract from moon peach as an active ingredient.   A skin damage recovery accelerator comprising an extract from moon peach as an active ingredient.   An aquaporin 3 mRNA expression promoter characterized by containing an extract from moon peach as an active ingredient.