JP2003171225A - COMPOSITION FOR INTEGRINalpha6beta4 PRODUCTION PROMOTION - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a composition for integrinα6β4 production promotion for preventing, checking and ameliorating skin aging. <P>SOLUTION: This composition for integrinα6β4 production promotion comprises a plant of Hedera helix L., Echinacea purpurea, Cucurbita pepo L., Taraxacum officinale and Angelica archangelica L., or its extract. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to laminin-5, which is present in the cell membrane of epidermal basal cells and is a constituent of epidermal basal membrane.
The present invention relates to a composition containing a plant or an extract thereof, which promotes the production of integrin α6β4, which is a receptor for Escherichia coli, and its use.

[0002]

2. Description of the Related Art Conventionally, aging, sun exposure, environmental stress, and mental stress have been involved in changes caused by skin aging, such as wrinkles, dullness, loss of texture, and loss of elasticity. It is known. If we consider these as microscopic changes inside the skin, elastic fibers such as type I collagen, type III collagen and elastin in the dermis are reduced or denatured, and type IV collagen or laminin, which is a component of the basement membrane, is reduced or denatured. Damage to the basement membrane and abnormal turnover of epidermal cells have occurred.

The basement membrane is a dense membrane composed of extracellular matrix proteins such as type IV collagen, laminin and fibronectin, and is present at the junction between the epidermis and the dermis. The basement membrane is responsible for the connection between the epidermis and the dermis, is important for maintaining the structure of the skin, and is involved in controlling the turnover of the epidermis and transmitting information between the epidermis and the dermis. On the other hand, since the basement membrane is overlapped and flattened in aged skin, it is considered that the structural change of the basement membrane causes skin aging (Lavker, R. et al .: J. In.
vest. Dermatol. , 73: 59-389,
1979, Lavker, R .; et al. : Derm
atol. Clin. , 4: 379-389, 198.
6). From the above, a composition containing a component that promotes the production of laminin-5, which is a constituent of the skin basement membrane, has been developed in recent years for the purpose of preventing, preventing, and improving damage to the skin basement membrane due to aging. (Japanese Patent Laid-Open No. 11-3432)
26, JP 2000-226308 A, Japanese Patent Application 2000-3
31318, Japanese Patent Application No. 2001-151485).

In order for epidermal cells to adhere to extracellular matrix proteins such as type IV collagen, laminin and fibronectin which are constituents of basement membrane, integrins acting as their receptors are indispensable. Integrin is a heterodimer in which α and β chains associate non-covalently, is expressed on the cell membrane and binds to extracellular ligands such as collagen, laminin, fibronectin, and intracellularly, such as tailin and α-actinin. It binds to the cytoskeletal proteins of and is involved in the binding between cells and extracellular matrix or between cells. Currently, 16 types of α chains and 9 types of β chains have been reported.
By combining β chains, various molecules having different ligand specificities are formed. Integrin, through cell adhesion, cell proliferation, cell differentiation, cell motility, apoptosis,
It is responsible for a wide range of biological functions such as morphogenesis, immune response, wound healing, blood coagulation, and bone resorption (Hyne).
S.R. O. : Cell, 69: 11-25, 199.
2, Rouslati, E .; et al. : Cell,
77: 477-478, 1994).

Up to now, integrin α6β4 and integrin α3β1 have been known as receptors for laminin-5, which is one of the laminin family, and α6β4 is mainly involved in cell adhesion, and α3β1 is mainly involved in cell motility. (Rousselle, P. et al.
et al. : J. Cell Biol. , 114: 5
67-576, 1991, Verrndo, P .; et
al. : Lab. Invest. , 71: 567-57.
4, 1994). Integrin α6β4 has a molecular weight of 15
It is a dimer of 0kD α6 chain and 205kD β4 chain.
Only α6 has been reported as an α chain forming a dimer with 4 (Suzuki, S .: EMBO J., 9:
757-763, 1990). Integrin α6β4
Is expressed in epithelial cells, Schwann cells, nerve cells, some endothelial cells, etc., and is present on the basal side of epidermal basal cells in the skin and binds to laminin 5 present in the basement membrane to form epidermis and basal cells. Responsible for binding between membranes (Roussell
e, P. et al. : J. Cell Biol. , 11
4: 567-576, 1991).

Integrin β4 gene and laminin 5
The gene mutation is congenital junctional epidermolysis bullosa (Herlitz's junctional epidermis), which is a serious genetic disease characterized by peeling between the epidermis and the dermis and blistering.
dermolysis bullosa), it has been revealed that the adhesion between integrin α6β4 and laminin 5 is essential for maintaining normal skin structure (Aberdam, D. et al .:
Nature Genet. , 6: 299-304, 1
994, Pulkkinen, L .; et al. : Na
pure Genet. , 6: 293-298, 199.
4, Vidal, F.F. et al. : Nature Ge
net. , 10: 229-234, 1995). Also,
Integrin β4 gene knockout mice show epidermal detachment similar to congenital junctional epidermolysis bullosa, and basal cells undergo apoptosis. Therefore, laminin-5-integrin α6β4 binding is important for basal cell survival. Is known (Dawling, J.e.
t al. : J. Cell Biol. , 134: 559
-572, 1996).

Furthermore, laminin 5 and integrin α6β
4 binding is involved in cell proliferation signals Ras-MA
It is known to be involved in the proliferation of epidermal cells via the P kinase pathway (Mainiero, F. et a.
l. : EMBO J. , 16: 2365-2375, 1
997, Mainiero, F .; et al. : EMB
OJ. , 17: 3940-3951, 1998). Thus, integrin α6β4 is a receptor for laminin 5, which plays a role in binding between epidermis and dermis, is important for maintaining normal skin structure, and plays an important role for survival and proliferation of epidermal basal cells.

On the other hand, since the expression of integrin β4 in epidermal keratinocytes decreases with aging, it has been suggested that there is a causal relationship between aging of the skin and decreased expression of integrin α6β4 (Le Varlet B. et. a
l. : J. Invest. Dermatol. Symp
osium Proceedings, 3: 172-1
79, 1998). Therefore, it is expected that skin aging can be prevented, prevented or improved by promoting the production of integrin α6β4 in epidermal cells and preventing, preventing or improving the decrease in the bond between the skin basement membrane and basal cells due to aging. It was

[0009]

However, by promoting the production of integrin α6β4 in epidermal cells and promoting the binding between epidermal basal cells and laminin-5, it is possible to prevent, prevent, and improve skin aging. Compositions intended to do so have not been developed. Therefore, an object of the present invention is to provide a composition for promoting integrin α6β4 production for preventing, preventing and improving skin aging.

[0010]

[Means for Solving the Problems] The present inventors examined the activity of promoting the production of integrin α6β4 in epidermal keratinocytes of various plants. As a result, they have found that plants of Izodaceae, Echinacea, pumpkin, dandelion, and Angelica significantly promote the production of integrin α6β4, and completed the present invention.

That is, the present invention relates to a composition for promoting the production of integrin α6β4, which contains one or more plants selected from the group consisting of Ivy, Echinacea, squash, dandelion, and Angelica, or an extract thereof.

The present invention also relates to the use of a composition containing these plants or an extract thereof as a composition for suppressing skin aging or an external preparation for skin.

[0013]

BEST MODE FOR CARRYING OUT THE INVENTION Hedera helix L. used in the present invention is a plant belonging to the genus Hedera of the family Araliaceae. Native to Europe and the Caucasus. Used as an anti-convulsant, anti-pertussis agent, analgesic, dissipative.
Ingredients include saponins, flavonoids, organic acids and the like. It has cleansing, anti-inflammatory and slimming effects and is widely used in cosmetics. It is especially used in facial cleansers, creams and shampoos for acne and oily skin.

Echinacea (Echinace) used in the present invention
a purpurea, E. angustifolia, E. pallida) is a plant belonging to the genus Echinacea of the Asteraceae family. Native to North America, as a panacea among Native Americans, toothache, sore throat, colds,
It has been used for treatment of infectious diseases for about 400 years.
At the end of the 19th century, it was introduced to Europe and cultivation started. Ingredients include polysaccharides, caffeic acid derivatives, alkylamides and the like. It is said to enhance immunity, and it is effective in treating and preventing colds, infectious diseases, and skin diseases, and inflammation (“Functional research of medicinal herbs”, Health Industry Newspaper).

Dandelion (Ta) used in the present invention
raxacum officinale) is a plant of the dandelion genus of the family Asteraceae. Native to Eastern Europe, leaves are used as raw salads, roasted roots are used as a substitute for coffee, and flowers are used as ingredients for wine. Ingredients include triterpenes, sterols,
Contains sugars, vitamins and potassium. The root is effective for indigestion and constipation, stimulates the liver and gallbladder, increases bile secretion, and is effective for liver damage and jaundice. The leaves have a strong diuretic effect, and also have a cleaning effect on blood and tissues,
It is also effective as a remedy for skin diseases and rheumatism (“Herbal Taizen” Shogakukan).

Angelica used in the present invention
ca archangelica L.) is a plant belonging to the genus Shishido of the Umbelliferae family. Ingredients include angelica lactone, angelicin, angelic acid, bergapten and folic acid, vitamin B1
Including B vitamins such as 2. It has the effect of improving blood circulation and regulating the secretion of female hormones, and is widely used for anemia, menstrual cramps, menstrual irregularities, coldness, etc. (“Bioactivity of functional herbs” Tokiwa Plant Chemistry Research Institute).

The pumpkin (Cucurbita) used in the present invention
pepo L.) is a plant of the genus Pumpkin of the family Cucurbitaceae. Native to North America, it is well known as a vegetable and is cultivated all over the world. Seeds are nutritious and are edible. The components include fatty acids, lignans, tocopherols, selenium which is a trace element, and the like. In Europe, oily extracts of seeds are used for the treatment of urinary system diseases such as hypersensitivity bladder and early stage prostatic hypertrophy (“Bioactivity of functional herbs” Tokiwa Botanical Institute).

The plants in the present invention include leaves, stems, buds, flowers,
All parts such as above-ground parts such as woody part and bark part (bark), underground part such as roots and tubers, seeds and resin can be used. The plant in the present invention is a dried product obtained by drying them themselves, a crushed product thereof, squeezed juice obtained by press-extracting themselves, water itself or an organic solvent such as alcohol, ether or acetone of the plant itself or its dried product. The crude extract and the crude extract are distributed, and include all of the extract fractions obtained by stepwise purification by various chromatography such as column chromatography, but the organic solvent extract of the plant itself is preferable. Alcohol extracts are particularly preferred. These may be used alone or in combination of two or more.

The effective compounding amount of the plant composition of the present invention is appropriately selected and determined according to the plant preparation method, formulation form and the like, and is not particularly limited. For example, from the dry pulverized product of the plant of the present invention When an extract extracted with an organic solvent such as alcohol, ether, or acetone is used as a preparation, its dry weight is 0.001 to 30.
Weight percent is suitable.

Ivy, Echinacea, pumpkin,
A composition of the present invention containing a plant of dandelion, angelica or an extract thereof is useful for promoting the production of integrin α6β4, activating basal cells, and promoting regeneration.
Enhances the function of basal cells that have been reduced by UV irradiation,
Since it strengthens the binding force between the basement membrane and basal cells, it not only suppresses photoaging of the skin and the like, but also has a regenerating action of photoaging.

The composition of the present invention includes oils and fats such as vegetable oils, higher fatty acids, higher alcohols, silicones, anionic surfactants, cationic surfactants, amphoteric surfactants,
Nonionic surfactants, preservatives, sugars, sequestering agents, polymers such as water-soluble polymers, thickeners, powder components,
An ultraviolet absorber, an ultraviolet blocker, a moisturizing agent such as hyaluronic acid, a fragrance, a pH adjusting agent, a desiccant and the like can be contained. Vitamins, skin stimulants, blood circulation promoters, indigenous bacteria control agents, active oxygen scavengers, anti-inflammatory agents, anti-cancer agents,
Other medicinal and physiologically active ingredients such as a whitening agent and a bactericidal agent may be contained.

Examples of the oils and fats are camellia oil, evening primrose oil, macadamia nut oil, olive oil, rapeseed oil, corn oil, sesame oil, jojoba oil, germ oil, wheat germ oil,
Liquid oils and fats such as glycerin trioctanoate, cocoa butter,
Solid oils such as coconut oil, hardened coconut oil, palm oil, palm kernel oil, sorghum, sorghum kernel oil, hardened oil, and hardened castor oil, beeswax, candelilla wax, cotton wax, nuka wax,
Waxes such as lanolin, lanolin acetate, liquid lanolin, and sugar cane wax are mentioned.

Examples of hydrocarbons include liquid paraffin, squalene, squalane, and microcrystalline wax.

As the higher fatty acid, for example, lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, docosahexaenoic acid (D
HA), eicosapentaenoic acid (EPA) and the like.

Examples of higher alcohols include straight chain alcohols such as lauryl alcohol, stearyl alcohol, cetyl alcohol and cetostearyl alcohol, and branched chain alcohols such as monostearyl glycerin ether, lanolin alcohol, cholesterol, phytosterol and octyldodecanol. Can be mentioned.

Examples of silicones include chain polysiloxanes such as dimethylpolysiloxane and methylphenylpolysiloxane, and cyclic polysiloxanes such as decamethylcyclopentasiloxane.

As the anionic surfactant, for example, fatty acid salt such as sodium laurate, higher alkyl sulfate ester salt such as sodium lauryl sulfate, alkyl ether sulfate ester salt such as POE lauryl sulfate triethanolamine, N-acyl sarcosinic acid, Sulfosuccinate,
Examples thereof include N-acyl amino acid salts.

Examples of the cationic surfactant include alkyl trimethyl ammonium salts such as stearyl trimethyl ammonium chloride, benzalkonium chloride and benzethonium chloride.

Examples of the amphoteric surfactant include betaine-based surfactants such as alkyl betaine and amidobetaine.

Examples of nonionic surfactants include sorbitan fatty acid esters such as sorbitan monooleate and hydrogenated castor oil derivatives.

Examples of preservatives include methylparaben and ethylparaben. Examples of the sequestering agent include edetate salts such as disodium ethylenediaminetetraacetate, edetic acid and sodium edetate.

Examples of the polymer include gum arabic, tragacanth gum, galactan, guar gum, carrageenan, pectin, agar, quince seed, dextran,
Pullulan carboxymethyl starch, collagen, casein, gelatin methyl cellulose, methyl hydroxypropyl cellulose, hydroxyethyl cellulose, sodium carboxymethyl cellulose (CMC), sodium alginate carboxy vinyl polymer (CARB0
POL) and other vinyl polymers.

Examples of the thickener include carrageenan, tragacanth gum, quince seed, casein, dextrin, gelatin, CMC, hydroxyethyl cellulose, hydroxypropyl cellulose carboxyvinyl polymer, guar gum, xanthan gum and bentonite.

As the powder component, talc, kaolin, mica, silica, zeolite, polyethylene powder, polystyrene powder, cellulose powder, inorganic white pigment, inorganic red pigment, titanium oxide coated mica, titanium oxide coated talc, colored titanium oxide coated. Examples thereof include pearl pigments such as mica, and organic pigments such as Red No. 201 and Red No. 202.

Examples of the ultraviolet absorber include paraaminobenzoic acid, phenyl salicylate, isopropyl paramethoxycinnamate, octyl paramethoxycinnamate, and 2,4-dihydroxybenzophenone. Examples of ultraviolet blocking agents include titanium oxide, talc, carmine, bentonite, kaolin, zinc oxide and the like.

As a moisturizer, polyethylene glycol,
Propylene glycol, dipropylene glycol, 1,
3-butylene glycol, 1,2-pentanediol,
Glycerin, diglycerin, polyglycerin, xylitol, maltitol, maltose, sorbitol, glucose, fructose, sodium chondroitin sulfate, sodium hyaluronate, sodium lactate, pyrrolidonecarboxylic acid, cyclodextrin and the like can be mentioned.

As the medicinal component, vitamin A oil, vitamin A such as retinol, vitamin B ₂ such as riboflavin, etc.
S, B ₆ such as pyridoxine hydrochloride, L- ascorbic acid, L- ascorbic acid phosphoric acid ester, L- ascorbic acid monopalmitate, L- ascorbic acid dipalmitate ester, L- ascorbic acid 2-glucoside , Vitamin C such as calcium pantothenate, vitamin D ₂ such as vitamin D ₂ and cholecalciferol; vitamin E such as α-tocopherol, tocopherol acetate, DL-α-tocopherol nicotinate Can be mentioned. Whitening agents such as placenta extract, glutathione, Yukinoshita extract, skin activating agents such as royal jelly, beech tree extract, blood circulation promoters such as capsaicin, zingerone, cantharisin tincture, ictamol, caffeine, tannic acid, γ-oryzanol, glycyrrhizic acid. Examples thereof include derivatives, glycyrrhetinic acid derivatives, anti-inflammatory agents such as azulene, amino acids such as arginine, serine, leucine and tryptophan, maltose sucrose condensates of resident bacterial control agents, lysozyme chloride and the like.

Furthermore, chamomile extract, parsley extract,
Beech tree extract, wine yeast extract, grapefruit extract, honeysuckle extract, rice extract, grape extract, hop extract, rice bran extract, loquat extract, pearl oyster extract, quinine extract, senburi extract, merilito extract, birch extract, licorice extract, peony extract, soapless extract, Loofah extract, capsicum extract, lemon extract, gentian extract, perilla extract, aloe extract, rosemary extract, sage extract, thyme extract, tea extract, seaweed extract, cucumber extract, clove extract, carrot extract, horse chestnut extract, hamamelis extract, mulberry extract, etc. Can be mentioned various extracts.

The composition of the present invention is, for example, an aqueous solution, an oil solution,
Liquid agents such as emulsions and suspensions, semi-solid agents such as gels and creams,
It can be applied in the form of solid agents such as powder, granules, capsules, microcapsules, and solids. Prepared into these forms by conventionally known methods, lotions, emulsions, gels, creams, ointments, plasters, poultices, aerosols, suppositories, injections, powders, granules, tablets, pills. , Various forms such as syrup, troche, etc. These can be applied to the body by applying, sticking, spraying, drinking and the like. Especially among these dosage forms, lotion,
Emulsions, creams, ointments, plasters, poultices, aerosols and the like are suitable as external skin preparations. As cosmetics,
Skin lotion, milky lotion, cream, skin cosmetics such as packs, makeup base lotion, makeup cream, milky liquid or creamy or ointment type foundation, lipstick, eye color, cheek color makeup cosmetics, hand cream, Body creams such as leg creams and body lotions, bath salts, oral cosmetics,
It can be a hair cosmetic.

[0040]

EXAMPLES Next, the present invention will be further described with reference to examples and comparative examples, but the present invention is not limited thereto. [Preparation of plant extract] Ivy (Hedera helix)
L.) leaves, Echinacea purpurea
L. Moench) root, medicinal pumpkin (Cucubitae pepo L.)
Seeds, dandelion (Taraxacum officinale)
G.) root, Angelica archangelica L.
Roots, stinging nettle (Urtica dioica L.) roots, and Gentiana lutea L. roots were finely chopped, and 3 kg of 99.5% ethanol was added to each 1 kg of the roots, and the pieces were immersed overnight at room temperature. This was filtered to obtain each extract. Here, extracts of nettle nettle (Urtica dioica L.) and gentiana (Gentiana lutea L.) were prepared for comparison. When each extract was concentrated, the evaporation residues were 65 g, 23 g, 135 g, 45 g and 78 g, respectively.
It was g, 56 g, and 43 g.

[Culturing of normal human epidermal keratinocytes] Fetus-derived normal human epidermal keratinocytes; NHEK (Asahi Techno Glass) was used in a medium for epidermal keratinized cells; KGM (Asahi Techno Glass) 37
The cells were cultured in a -5 ° C-5% CO ₂ incubator. KGM
Is human epidermal growth factor (0.1 ng / ml) and insulin (5.0 μg / m) in epidermal keratinocyte basal medium.
l), hydrocortisone (0.5 μg / ml), gentamicin (50 μg / ml), amphotericin B
(50 μg / ml) and bovine pituitary extract (2 ml) were added. In this experiment, cells at passage number 3 to 5 were used.

[RT-PCR (Reverse tra
nscriptase-polymerase cha
of integrin α6β4 production promoting activity of each plant extract by in reaction] NHEK to KGM
Suspension to 1.25 × 10 ⁵ / ml, and add 4 ml
/ Well was seeded on a 6-well plate and cultured for 24 hours.
Each plant extract prepared by the above method and KGM supplemented with fetal bovine serum (FBS) were treated with 4 ml / well and further cultured for 24 hours. After culturing, TRIZOL
Total RNA was recovered using reagents (Lifetech Oriental) according to the protocol. Fetal bovine serum (FB
S) is a substance known as a factor that promotes the expression of integrin α6 mRNA and integrin β4 mRNA, and was used as a control here.

RT-PCR using the recovered total RNA
Integrin α6mRN by high (Toyobo)
A, integrin β4 mRNA and G3PDHmRN
The expression promoting activity of A was measured. The G3PDH gene is known as a housekeeping gene, and since it is constitutively expressed in all cells, it is widely used as a control gene. As a PCR primer, integrin α6 was 5'-GAA CTG TGT GAA CA.
T CAG A-3 ′ and 5′-ATC CTTACA G
Using CA TGG TAT CG-3 ', integrin β
4 is 5'-CGG ATG CTG CTT ATT GAG
AAC-3 'and 5'-GAG GGT GGA GGA
Using TGT GCT TAG-3 ', G3PDH is 5'
-ACC ACA GTC CAT GCC ATC AC-
3'and 5'-TCC ACCACC CTG TTG C
TG TA-3 'was used. The reaction conditions for PCR were that integrin α6 and integrin β4 were treated at 94 ° C. for 7 minutes, then 94 ° C., 1 minute, 56 ° C., 1 minute, 74 ° C., 1 minute for 2 minutes.
After repeating 5 times, it was treated at 74 ° C for 10 minutes and stored at 4 ° C. On the other hand, G3PDH was treated at 94 ° C for 7 minutes,
C., 1 minute, 58.degree. C., 1 minute, 74.degree. C., 1 minute was repeated 20 times, then 74.degree. C., 10 minutes, and stored at 4.degree. RT
-PCR products were electrophoresed, stained with ethidium bromide and photographed.

The results of treating each plant extract with 0.1% are shown in FIG. Treating the plant extract with 0.1% means that the solid content concentration is 0.1% by weight.
This means that a plant extract solution adjusted to 100 mg / ml with ethanol is added by 1% by weight of the medium amount for treatment. Expression of integrin α6 mRNA and integrin β4 mRNA was significantly promoted when treated with extracts of Echinacea vulgaris leaves, Echinacea purpurea roots, medicinal pumpkin seeds, dandelion roots, and Angelica root extracts compared to untreated controls. It was On the other hand, the root of the nettle,
When it was treated with the extract of gentian root, no promoting activity on the expression of integrin α6 mRNA and integrin β4 mRNA was observed. The expression of G3PDH was similar regardless of whether or not the plant extract was treated. From these results, it was found that the Ivy, Echinacea, pumpkin, dandelion, and Angelica of the present invention act selectively to promote the production of integrin α6β4.

FIG. 2 shows the results obtained when the extracts of leaves of A. auricularis were treated at different concentrations. The concentration in FIG. 2 is also the same as above, and represents the solid content concentration with respect to the amount of medium. Treatment with the extract of leaves of P. mellifera caused integrin α6 mRNA and integrin β4 in a concentration-dependent manner.
Expression of mRNA was promoted. Moreover, when treated with 0.1% and 1.0% of the leaf extract of Ivy,
Similar activity was observed when BS was treated with 0.1% and 1.0%. The expression of G3PDH was similar regardless of whether or not the plant extract was treated.

[Measurement of photodamage suppression effect of each plant extract]
NHEK was suspended in KGM at 2 × 10 ⁵ / ml and treated with each plant extract and fetal bovine serum (FBS), which is a factor that promotes the expression of integrin α6 mRNA and integrin β4 mRNA. 1 of this suspension
A 24-well plate was seeded at ml / well and cultured for 24 hours. Then, the culture solution was discarded, the cells were washed with PBS (-), PBS (-) was added again, and ultraviolet rays were irradiated at 70 mJ / cm ² without covering the plate. As a control, cells of a group not irradiated with ultraviolet rays were also prepared. After further culturing for 24 hours, the cells were washed with PBS (-) to remove floating cells, and then 5% glutaraldehyde aqueous solution was added at room temperature for 1 hour.
The cells were fixed by treating for 5 minutes. After washing with tap water and drying with a drier, fluorescent staining solution {5 μg / ml Hoc
hst33342 (Sigma-Aldrich C
o. ) And 0.001% polyoxyethylene (1
0) Aqueous solution containing octyl phenyl ether (Wako Pure Chemical Industries)} was treated at room temperature in the dark for 1.5 hours. After washing with tap water and drying with a dryer, the fluorescence intensity was measured with a fluorescence plate reader to calculate the cell viability.

The results of treating each plant extract with 0.1% are shown in Table 1. The cell viability in this experiment was expressed as 100% when the plant extract was untreated and the ultraviolet ray was not irradiated. In the case of non-irradiation with ultraviolet rays, the cell survival rate was similar regardless of whether or not each plant extract was treated. On the other hand, when the cells were irradiated with ultraviolet rays at 70 mJ / cm ² , the cell viability in the non-treated state was about 55%, whereas the plant extracts of Izodaceae, Echinacea, squash, dandelion and Angelica were used. When treated, the cell viability increased significantly. On the other hand, the root of the nettle,
No increase in cell viability was observed when each gentian root extract was treated.

Table 2 shows the results obtained by treating the extracts of leaves of E. chinensis with different concentrations. The extract of leaves of P. aequinata increased the cell viability in a concentration-dependent manner.

[0049]

[Table 1]

[0050]

[Table 2]

The formulation examples of the present invention are shown below. [Formulation Example 1] Cream A cream was produced according to the following formulation (unit: mass%). (1) Stearyl alcohol 6.0% (2) Stearic acid 2.0% (3) Hydrogenated lanolin 4.0% (4) Squalane 9.0% (5) Octyldodecanol 10.0% (6) POE (25) Cetyl alcohol ether 3.0% (7) Glycerin monostearate 2.0% (8) Ivy extract 0.1% (9) Preservative Amount (10) Perfume Amount (11) 1,3 Butylene Glycol 6.0% (12) PEG 1500 4.0% (13) Purified water Residual

[Production Method] 80 parts of the above components (1) to (10)
Dissolve by heating at ℃ to obtain an oil phase. The components (11) to (13) are heated and dissolved at 70 ° C. to obtain an aqueous phase. The water phase was gradually added to the oil phase to emulsify, and the mixture was cooled to 40 ° C with stirring and further
The mixture was stirred and cooled to ℃ to obtain a cream.

[Formulation Example 2] Tablet A tablet was produced according to the following formulation (unit is mass%). (1) Chrysanthemum radix extract 20.0% (2) Lactose 65.0% (3) Corn starch 14.0% (4) Guar gum 1.0%

[0054]

INDUSTRIAL APPLICABILITY As described above, the integrin α6β4 of the present invention containing a specific plant or its extract
The production promoter promotes the production of integrin α6β4,
By promoting adhesion between laminin 5 and epidermal basal cells, it promotes structure maintenance and function improvement of epidermis and basement membrane. Therefore, by preventing, preventing and improving structural abnormalities and functional decline of the epidermis and basement membrane for aged skin, especially skin damaged by ultraviolet rays, youthful without wrinkles, spots, dullness and sagging. The skin condition can be maintained.

[0055]

[Sequence list]

SEQUENCE LISTING <110> FANCL Inc. <120> Composition for promoting integrin α6β4 production <130> PC22-033 <160> 6

[0056] <210> 1 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> Synthetic RNA <400> 1 gaactgtgtg aacatcaga 19

[0057] <210> 2 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> Synthetic RNA <400> 2 atccttacag catggtatcg 20

[0058] <210> 3 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> Synthetic RNA <400> 3 cggatgctgc ttattgagaa c 21

[0059] <210> 4 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> Synthetic RNA <400> 4 gagggtggag gatgtgctta g 21

[0060] <210> 5 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> Synthetic RNA <400> 5 accacagtcc atgccatcac 20

[0061] <210> 6 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> Synthetic RNA <400> 6 tccaccaccc tgttgctgta 20

[Brief description of drawings]

FIG. 1 shows the results of RT-PCR when normal human epidermal keratinocytes were treated with each plant extract.

FIG. 2 is a diagram showing the results of RT-PCR when normal human epidermal keratinocytes were treated with the extract of leaves of Eriocarpus chinensis in a concentration-dependent manner.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61K 35/78 A61K 35/78 T A61P 17/00 A61P 17/00 43/00 107 43/00 107 111 111 111 F Term (reference) 4C083 AA111 AA112 AC022 AC072 AC122 AC182 AC242 AC422 AD042 AD512 CC02 CC05 DD31 EE12 4C088 AB16 AB19 AB26 AB40 AC03 AC04 AC05 AC06 AC08 AC11 AC13 BA07 BA09 BA10 CA03 CA05 CA14 MA07 MA63 NA14 ZA89 ZB22 ZC02

Claims (3)

[Claims] 1. A composition for promoting the production of integrin α6β4, which comprises one or more kinds of plants selected from Izodaceae, Echinacea, pumpkin, dandelion, and Angelica, or an extract thereof. 2. A composition for suppressing photoaging, which comprises one or more plants selected from the group consisting of Ikizuki, Echinacea, squash, dandelion, and Angelica, or an extract thereof. 3. A skin external preparation comprising the composition according to claim 1 or 2.