JP2007099625A - Skin fibroblast growth promoter - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a safe skin fibroblast growth promoter that is useful for wrinkle measure regarded to be largest worries with respect to beautification, increases water holding property and keeps skin firmness and moisture, a skin care food and beverage and a skin care cosmetic containing the skin fibroblast growth promoter. <P>SOLUTION: The skin fibroblast growth promoter comprises an extract of a fermented substance, obtained by fermenting dried leaves of Alpinia speciosa with mixed bacteria of Lactobacillus plantrum, Streptococcus thermophilus and Bacillus subtilis, with hot water or 80% ethanol or the freeze-dried substance of them as an active ingredient. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a dermal fibroblast proliferation promoter containing a processed product of moon peach as an active ingredient, and a skin care food and drink and a skin care cosmetic containing the dermal fibroblast proliferation promoter.

  In the basal layer constituting the epidermis, melanocytes (pigment cells) that synthesize melanin exist, and the synthesis of melanin is synthesized in small granules surrounded by unit films called melanosomes formed in melanocytes. In this process, synthesis is initiated by a reaction in which tyrosine, which is one of amino acids, is oxidized to dopa and further to dopaquinone by an enzyme called tyrosinase. Two other tyrosinase-related proteins are known as enzymes involved in melanin synthesis. Melanin synthesized by this route is called brown to black eumelanin, and in humans, it is synthesized by skin, mucous membrane, and hair melanocytes (see, for example, Non-Patent Document 1). The number of melanocytes varies from person to person, and varies from site to site in the same person. Generally, there are few covered parts and many exposed parts. When irradiated with ultraviolet rays, melanin synthesis including an increase in tyrosinase activity increases, and the number of melanocytes also increases. There are various types of pigmentation caused by melanin deposition, with spots, freckles and senile pigmentation being typical. All of these are said to be skin care in which UV protection is effective together with a whitening agent since UV is a direct cause or an exacerbation factor (see, for example, Non-Patent Document 2).

  Conventionally, various substances capable of inhibiting the action of tyrosinase have been used in the field of cosmetics and foods in order to prevent the production of melanin by the above mechanism, and representative examples thereof include ascorbic acid, sulfurs, hydroquinone, Kojic acid, arbutin, natural plant extracts and the like. Some natural plant extracts have been tried so far. For example, a tyrosinase inhibitor containing, as an active ingredient, an extract obtained by extracting guava (Psidium guajava L.) or Erica (Erica), whitening cosmetics containing the same (for example, see Patent Document 1), water and And / or an extract of camellia seeds having an inhibitory activity on tyrosinase extracted using an organic solvent, a cosmetic or food product having a whitening function or the like containing the same, water of germinated brown rice, and water And / or a tyrosinase inhibitor containing an organic solvent extract as an active ingredient, and a functional food for whitening (see, for example, Patent Document 3), and a tyrosinase inhibitory substance selected from licorice extract, placenta extract, oxalic acid, and the like A whitening composition containing white birch extract and a reducing substance (see, for example, Patent Document 4) and a melanin inhibitor containing Hibiscus syriacus L. extract A pharmaceutical preparation, use of the inhibitor in cosmetics and foods (see, for example, Patent Document 5), a tyrosinase inhibitor containing a Yanagita extract as an active ingredient, a whitening agent and a whitening food and drink containing the same ( For example, see Patent Document 6).

  On the other hand, moon peach (Alpinia Speciosa K. Schum) belongs to the family Zingiberaceae, also known as Alpinia. It is distributed from southern Kyushu to southern China to tropical Asia. It is used as a raw material for and ropes. As for skin care using moon peaches, cautains, polyphenols (quercetin), and chlorophyll contained in moon peaches suppress adverse effects on the skin such as active oxygen and ultraviolet rays, and reduce melanin pigment deposition. Drying the leaves and / or stems of moon peaches to provide a highly safe pack that has excellent whitening effect due to its suppressive function and is resistant to spoilage without the addition of preservatives. A drying step, a pulverizing step of pulverizing the dried moon peach leaves and / or stems to obtain a moon peach powder having an average particle size of 10 to 50 μm, and the pulverized moon peach powder and a gelling agent And a gelling step of mixing into a gel to produce a pack (see, for example, Patent Document 7), rosemary, peppermint, chamomile, or ro Cosmetic soap characterized in that at least one herb made of ash, which is made of zugeronium, lavender, lemongrass, or moon peach, is blended in a soap base (see, for example, Patent Document 8) ), Leaf extract, chicken crown extract, rice extract, hematococcus algae extract, fenugreek extract, rose petal extract, and black pepper extract, For example, see Patent Document 9).

  Furthermore, plant resources such as guava leaves, moon peaches, mugworts, tea leaves, etc. are finely crushed, and as a culture medium, three types of lactic acid bacteria, Streptococcus thermophilus, Lactobacillus plantarium and Bacillus subtilis, and yeast In addition, fermented foods (see, for example, Patent Document 10), which are obtained by adding molasses and water and fermenting for several days and are expected to improve blood pressure increase, have been proposed by the present applicant.

  In addition, it is known that the proliferation of dermal fibroblasts is complicatedly controlled by various cell growth factors. The main ones are platelet derived growth factor (PDGF), basic fibroblast growth factor (b-FGF), transforming growth factor-β (TGF-β), connective tissue growth factor (CTGF), etc. These growth factors have various physiological activities in addition to the control of dermal fibroblast proliferation, and are involved in wound healing and disease development. For example, PDGF and CTGF promote chemotaxis of dermal fibroblasts, b-FGF promotes angiogenesis, and TGF-β promotes matrix protein synthesis. Skin fibroblasts produce the necessary ingredients to maintain skin water retention, such as collagen and elastin. If the number of fibroblasts decreases with age, the water retention cannot be maintained and wrinkles are lost. It is said that will increase. Vitamin C is said to penetrate the dermis layer and activate fibroblasts to produce collagen, which is effective in combating wrinkles and has a skin regeneration action. In addition, female hormones that have a skin regeneration action act on fibroblasts to activate the synthesis of hyaluronic acid and collagen, and isoflavones also improve the metabolic activity of collagen in the dermis layer and give skin tension. Is expected.

JP-A-6-305978 JP 2000-159681 A JP 2001-240556 A JP 2001-335498 A JP 2003-171300 A JP 2004-83488 A Japanese Patent Laid-Open No. 2005-60260 JP 2003-55697 A JP-A-2005-47860 JP 2002-330725 A Published by Takeo Mitsui, “New Cosmetics”, pp. 14-16, March 25, 2002, published by Nanzan-do Co., Ltd. Published by Takeo Mitsui, “New Cosmetics”, pp. 26-28, March 25, 2002, published by Nanzan-do Co., Ltd.

  Control of the proliferation of skin fibroblasts is considered important in considering skin care and skin aging. By controlling the proliferation and function of dermal fibroblasts, skin aging may be prevented, and it may be possible to maintain a youthful skin forever. The problem of the present invention is useful for combating wrinkles, which is the most common cosmetic problem, and is a safe dermal fibroblast proliferation promoter that can maintain skin elasticity and moisture by increasing water retention, An object of the present invention is to provide skin care foods and beverages and skin care cosmetics containing such a dermal fibroblast proliferation promoter.

  Tsuki Peach is a ginger family perennial plant that grows in subtropics such as Okinawa and is counted as one of 164 species of aromatic plants in the world. In Okinawa, the antibacterial properties of moon peach leaves have long been used to wrap strawberries. Has been used for paper, tea, tatami mats, etc., but in recent years it has been used as a skin-friendly natural cosmetic that does not use preservatives due to its antibacterial effect. It has come to be. And this moon peach has an action to suppress the deposition of melanin pigment, and it is said that the use of cosmetics has the effect of preventing spots and freckles caused by ultraviolet rays. The present inventors have further studied about moon peach, and the processed product of moon peach using lactic acid bacteria, etc. has a stable skin fibroblast proliferation promoting action that cannot be found in unfermented moon peach. As a result, the present invention has been completed.

  That is, the present invention provides (1) a skin fiber characterized by containing as an active ingredient a fermented product obtained by fermenting moon peach with lactic acid bacteria, lactic acid bacteria and yeast, lactic acid bacteria and Bacillus subtilis, or lactic acid bacteria, yeast and Bacillus subtilis. A blast proliferation promoter, (2) the fermented product is a fermented product of moon peach leaves, (3) the dermal fibroblast proliferation promoter according to (1) above, A dermal fibroblast proliferation promoter according to (1) or (2) above, which is a fermented product by a mixed bacterium of plantalim, Streptococcus thermophilus, Bacillus subtilis, or (4) a fermented product Is an extract or a freeze-dried product thereof, and relates to the dermal fibroblast proliferation promoter according to any one of (1) to (3) above.

  The present invention also provides (5) a skin care food or drink characterized by containing the dermal fibroblast proliferation promoter according to any one of (1) to (4) above, (6) and a silymarin (Silymarin) It contains 1 or more types of components chosen from collagen peptide and hyaluronic acid, It is related with the skin care food / beverage products of the said (5) description characterized by the above-mentioned.

  Furthermore, the present invention provides (7) a skin care cosmetic comprising the dermal fibroblast proliferation promoter according to any one of (1) to (4) above, (8) and a Silymarin The skin care cosmetic according to (7) above, which contains one or more components selected from collagen peptides and hyaluronic acid.

  According to the present invention, the effect of promoting the proliferation of fibroblasts is useful for countermeasures against wrinkles, which are the most cosmetically annoying, and it is possible to increase the water retention and keep the skin firm and moisturized. Products and skin care cosmetics. Moreover, according to the present invention, in addition to maintaining water retention and keeping the skin firm and moist, it also suppresses the production of melanin, and prevents spots, buckwheat, spots, or hyperpigmenntation during pregnancy. it can.

  The dermal fibroblast growth promoter of the present invention is an active ingredient that is obtained by fermenting moon peach with lactic acid bacteria, lactic acid bacteria and yeast, lactic acid bacteria and Bacillus subtilis, or lactic acid bacteria and yeast and Bacillus subtilis. There is no particular limitation as long as the peach peach is used in the underground part such as roots and rhizomes, and the above-ground part such as stems, leaves, flowers, and fruits. The use of leaves is particularly preferred.

Examples of the lactic acid bacteria include Streptococcus, Lactobacillus, Leuconostoc, Pediococcus, Bifidobacterium, and Tetragenococcus. Bacteria belonging to any of these are preferred, and Lactobacillus is particularly preferred. The bacterium belonging to the genus Streptococcus is preferably Streptococcus thermophilus (S. thermophilus), and specific examples include Streptococcus thermophilus IFO13957 strain. In addition, as a bacterium belonging to the genus Lactobacillus, any of Lactobacillus plantarim (L. plantrum), Lactobacillus delbruckii (L. delbruckii), Lactobacillus pentosus (L. pentosus) or Lactobacillus casei (L. casei) Among these bacteria, Lactobacillus plantarim is particularly preferable. Specific examples of such Lactobacillus plantarim include IFO14712 and IFO14713 strains, Lactobacillus delbriqui as IFO13953 strain, Lactobacillus pentosus as IFO12011 strain, and Lactobacillus casei as IFO15883 strain. Moreover, as a microbe which belongs to Tetragenococcus genus, it is preferable that it is Tetrageno halophyllus (T. halophilus), and tetrageno halophyllus IFO12172 strain can be illustrated concretely. These lactic acid bacteria, dry basis 1g leaf month peach, usually ¹⁰ 3 to 10 ^7, it is preferable to use particularly ¹⁰ 6 to 10 ^7.

Moreover, as said yeast, the microbe which belongs to Candida (Candida) or Saccharomyces (Saccharomyces) is preferable. As such a bacterium belonging to the genus Candida, Candida versatilis is preferable, and IFO10038 strain can be specifically exemplified as Candida versatilis. The bacterium belonging to the genus Saccharomyces is preferably S. cerevisiae, and IFO0555 strain can be specifically exemplified as Saccharomyces cerevisiae. These yeast are added primarily to improve the aroma, in this case, dry basis 1g leaf Terminalia catappa, usually ¹⁰ 3 to 10 ^7, it is preferable to use particularly ¹⁰ 6 to 10 ^7.

Further, as the Bacillus subtilis, a B. subtilis IFO3013 strain can be specifically exemplified. These B. subtilis, dry basis 1g leaf month peach, usually ¹⁰ 3 to 10 ^7, it is preferable to use particularly ¹⁰ 6 to 10 ^7.

  As a group of microorganisms preferably used in the fermentation process of moon peach, a group of microorganisms containing lactic acid bacteria and Bacillus subtilis is preferable, and among these microorganism groups, a mixed bacteria of Lactobacillus plantarim, Streptococcus thermophilus, Bacillus subtilis It is preferable to use the same number as the number of bacteria for dry matter of moon peach leaves. By such a combination of the number of bacteria, it is possible to shorten the fermentation time and to suppress the propagation of various bacteria.

In the moon peach fermentation treatment, the dried body of moon peach leaves is pulverized to a particle size of 3 mm or less, preferably 0.5 to 1.0 mm. By setting the particle size to 3 mm or less, a sufficient contact area with the fermenting bacteria can be secured, fermentation can proceed effectively, and the particle size in the range of 0.5 to 1.0 mm. This effect can be obtained more remarkably. In order to promote the progress of fermentation, it is preferable to add 1 to 10 parts by weight, particularly about 4 to 6 parts by weight, of water to 1 part by weight of dry matter. The above-mentioned bacteria or fungal group is added to the dried pulverized product. It is preferable to mix 1 to 10% by weight (as a culture) of the fungal group after the culture of each fungus, before mixing to the medium and before adding to the medium. Fermentation is preferably performed at a temperature of 20 to 50 ° C., particularly around 40 ° C., and the fermentation time can be appropriately selected depending on the progress of fermentation under conditions such as pH and the number of bacteria, and preference. pH 4-5, if the bacteria number 10 ⁶ or more, preferably about 72 hours. At the time of fermentation treatment, aeration or deoxygenation treatment can be performed as necessary, but fermentation can be performed in static culture after the deoxygenation treatment. The fermentation format is preferably solid culture rather than liquid culture.

  In such a fermentation treatment, carbohydrates and proteins can be added as an agent for fermenting bacteria. The carbohydrate as the assimilating agent is preferably a commercially available sugar such as glucose, sucrose, molasses, etc., and the amount of these added is preferably 1 to 10% by weight per medium, particularly around 3% by weight. The protein as an assimilating agent is preferably rice bran, bran or the like, and the addition amount thereof is preferably 1 to 5% by weight per medium. These assimilating agents may be used alone or in combination of two or more.

  As the fermented product as an active ingredient of the dermal fibroblast growth promoter of the present invention, the fermented product or the fermented product is subjected to one or more treatments such as drying treatment, sterilization treatment, extraction treatment, enzyme treatment and the like. Examples of the treated product are listed. For example, after completion of fermentation, it can be dried by a drying process so that the moisture value becomes 10% by weight or less. The drying method may be heat drying or freeze drying. In the case of heat drying, it is preferable that the product temperature is 100 ° C. or lower because the deactivation of the physiologically active component can be prevented. Further, after drying, if necessary, sterilization can be performed by a known method such as heating. As the fermented processed product as an active ingredient of the dermal fibroblast growth promoter of the present invention, an extract obtained by extracting and extracting moon peach after drying can be particularly preferably exemplified. Examples of the solvent used for the extraction treatment include lower monohydric alcohols such as water (hot water), methyl alcohol, and ethyl alcohol, liquid polyhydric alcohols such as glycerin, propylene glycol, and 1,3-butylene glycol, and hexane. One type or two or more types of nonpolar solvents such as As a preferable extraction method, for example, a hot water extraction with water at 70 to 90 ° C. for 10 to 60 minutes, preferably a hot water extraction with water at 80 ° C. for 20 minutes and a filtration, or a concentration of 80% by volume And a method of performing extraction for 24 hours at room temperature and then filtering. Furthermore, a freeze-dried product obtained by freeze-drying a hot water extract or a freeze-dried product obtained by removing a solvent from a solvent extraction product such as alcohol and then freeze-drying can be advantageously used. It is also possible to use a mixture obtained by mixing these extracts with a solvent.

  The above extract, an extract obtained by distilling off the solvent from the extract, or a spray-dried product or freeze-dried product of these extracts can be used as a dermal fibroblast proliferation promoter as they are. You may perform the refinement | purification process for deodorizing, decoloring, etc. in the range which does not affect an effect | action. Furthermore, it can be used as an active skin fibroblast proliferation promoter by performing separation and purification treatment such as liquid-liquid partition extraction, various chromatography, ion exchange, membrane separation and the like.

The dermal fibroblast growth promoter comprising as an active ingredient the fermentation product of moon peach obtained as described above is itself or a pharmaceutically acceptable binder, stabilizer, excipient, dilution. Can be used as liquids, powders, granules, tablets, capsules, etc. by mixing with agents, pH buffers, disintegrants, solubilizers, solubilizers, etc. It can be used as the skin care food / beverage product of the invention, or added to and blended with a cosmetic / cosmetic material to obtain the skin care cosmetic of the present invention. The skin care food and drink and skin care cosmetics of the present invention can be used in combination with known skin care ingredients. Examples of such skin care ingredients include silymarin, collagen peptide, hyaluronic acid, coenzyme Q ₁₀ , glucosamine, vitamin C and the like, and these can be used alone or in combination of two or more.

The above silymarin is a generic name for flavonolignans extracted from Asteraceae Maria thistle, and the main components that have been identified include silybin, silydianin, silycristin, isosiribin, etc., extracted from the seeds of the Asteraceae maria thistle And it is marketed as an extract raw material obtained as a dry powder by spray drying. As the silymarin used in the present invention, commercially available silymarin can be used as it is, and the collagen peptide, hyaluronic acid, coenzyme Q ₁₀ , glucosamine, vitamin C and the like are all commercially available. Can do. For example, in the case of a wrinkle-preventing composition comprising fermented moon peach extract (lyophilized product), silymarin, collagen peptide, hyaluronic acid, fermented moon peach extract (lyophilized product) 15 to 25% by weight, silymarin 0.1 Preferred examples include -1.0 wt%, collagen peptide 70-80 wt%, and hyaluronic acid 0.1-0.2 wt%.

  Examples of the types and forms of the skin care food and drink of the present invention include, for example, yogurt, drink yogurt, juice, milk, soy milk, liquor, coffee, tea, sencha, oolong tea, sports drinks, pudding, cookies, bread Baked confectionery such as cake, rice cracker, Japanese confectionery such as sheep crab, bread and confectionery such as frozen confectionery, jelly and chewing gum, noodles such as udon and soba, fish paste products such as kamaboko, ham and fish sausage, miso and soy sauce , Seasonings such as dressing, mayonnaise, sweeteners, dairy products such as cheese and butter, various prepared dishes such as tofu, konjac, other boiled fish, dumplings, croquettes, salads, honey, royal jelly, dietary supplements, Sick and infant food, emergency food, diet food, etc. can be mentioned, usually in packaging and instructions etc. It is useful for combating wrinkles, which is considered the most cosmetic problem, and is sold with the indication that it can maintain the elasticity and moisture of the skin by increasing water retention.

  Examples of the type and form of the skin care cosmetics of the present invention include, for example, lotions, emulsions, creams, foundations, base materials, white powder, blushers, sunscreens, etc. For example, it is useful for combating wrinkles, which are considered the most troublesome cosmetics, and it is displayed and sold to show that it can maintain water elasticity and moisturize the skin.

  EXAMPLES Hereinafter, although an Example demonstrates this invention more concretely, the technical scope of this invention is not limited to these illustrations.

(Preparation of hot water extract of fermented moon peach)
30 g of dried moon peach leaves were pulverized to a particle size of 0.1 to 3 mm and placed in a container. 30 g of water and 0.9 g of molasses were added to a container containing crushed moon peach. Separately, after each culture of Lactobacillus plantarim, Streptococcus thermophilus, and Bacillus subtilis was mixed at a ratio of 1: 1: 1, 10% by weight with respect to the weight of moon peach The mixture was added, the container was sealed, and fermentation was performed by static culture. The fermentation temperature was 40 ° C. and the fermentation time was 72 hours. Then, after drying at 60 degreeC until the moisture value became 10 weight% or less with a dryer, the sterilization process (130 degreeC steam, 5-15 seconds) was performed, and 30 g of fermented moon peach was obtained (henceforth "fermented moon peach"). Sometimes called "tea"). Then, add water so that it becomes a ratio of 10 mL with respect to 1 g of fermented moon peach tea, extract with hot water at 80 ° C. for 20 minutes, pool the supernatant, and again so that the ratio becomes 10 mL with respect to 1 g of precipitate. Water was added, hot water extraction was performed at 80 ° C. for 20 minutes, and a hot water extract was obtained together with the supernatant. Moreover, fermented moon peach freeze-dried product (hot water extract) (yield 9.12%) obtained by freeze-drying this hot-water extract was prepared.

(Preparation of fermented moon peach alcohol extract)
80% ethanol was added so that it might become a ratio of 10 mL with respect to 1 g of fermented moon peach tea prepared similarly to Example 1, and it extracted at room temperature for 24 hours, this extract liquid was concentrated and dried, and concentration 20 mg / The extract was prepared by adding DMSO to make up to ml and re-dissolving.

(Skin fibroblast proliferation test: Use of hot water extract)
Skin fibroblasts (derived from a human newborn, purchased from Dainippon Pharmaceutical Co., Ltd., 1 × 10 ⁶ ) were cultured for 4 days in a 25 cm ² flask using an α-MEM medium containing 10% FBS. Thereafter, the cells were collected by trypsin treatment, transferred to a 24-well plate at 7 × 10 ³ cells / mL, and cultured for 12 hours in an α-MEM medium containing 5% FBS. Next, 5 μL of a sample of the hot water extract of fermented moon peach as a sample or a 5-fold dilution was added to the dermal fibroblasts and cultured at 37 ° C. and 5% CO _{2 for} 3 days. As a control, no extract was added, and a non-fermented dried product of moon peach leaves was also cultured under the same conditions. The cell viability is measured by the MTT method, which is one of the colorimetric methods, and the cell growth rate is expressed as cell growth rate (%) = [(AC) − (BD) / (BD)] × 100. In the formula, A: MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide) added, hot water extract added, B: MTT added, no hot water extract added, C : No MTT added, hot water extract added, D: no MTT added, no hot water extract added. The results are shown in FIG. As shown in FIG. 1, the cell growth rate of the product obtained by adding the hot water extract of the processed product of the moon peach fermentation of the present invention is about 12 to 17% higher than that of the control, and about 9 compared with that not fermenting the moon peach. It was found to be ˜11% higher. Although the 5-fold dilution has better cell growth rate than the stock solution, the reason is unknown.

(Skin fibroblast proliferation test: use of 80% ethanol extract)
The 80% ethanol extract prepared in Example 2 was dissolved in DMSO and used at a final concentration of 10 μg / mL or 4 μg / mL, and the cell growth rate was determined in the same manner as in Example 3. The results are shown in FIG. From FIG. 2, it was found that the cell proliferation rate when the ethanol extract was added was about 5% at 10 μg / mL and about 18% at 4 μg / mL compared to the control. It was also found that 10 μg / mL was about 61% higher and 4 μg / mL was about 18% higher than the one that did not ferment moon peach. In this experiment, the cell growth rate is better at 4 μg / mL than at 10 μg / mL, but the reason is unknown.

(Monitor test of fermented moon peach tea)
A questionnaire survey of fermented moon peach tea was conducted for 20 women in their 20s to 50s. 2 g of fermented moon peach tea obtained in Example 1 was placed in 1 liter of water and ignited, and boiled for 5 minutes after boiling. This boiled liquid was ingested daily by 500 mL or more, and after 8 weeks, a questionnaire survey was conducted on “skin tension”, “skin moisture” and “skin gloss”. The questionnaire was conducted with a five-level evaluation (improvement, slightly improved, unchanged, slightly worsened, worsened). Table 1 shows the results of the questionnaire regarding skin tension, Table 2 shows the results of questionnaire about moisture, and Table 3 shows the results of questionnaire regarding skin gloss.

  According to the questionnaire results (Table 1) about skin tension, 12 out of 21 (57%) answered that they improved or slightly improved, and according to the questionnaire results (Table 2) about skin moisture , 10 out of 21 (47%) replied that it was improved or slightly improved, and according to the questionnaire results (Table 3) on skin gloss, 9 out of 21 replied that it improved or slightly improved As is clear from the above, it has been found that the skin fibroblast proliferation promoter of the present invention has an effect on “skin elasticity”, “skin moisture” and “skin gloss”.

(Monitor test of fermented moon peach extract grains)
A 55-day monitoring test was conducted on 6 women in their 20s and 40s. The skin state before monitoring was measured for skin (cheek) erythema (redness) and elasticity with a skin viscoelasticity measuring device CUTERTERMER 580. Fermented moon peach extract granules (granule of freeze-dried fermented moon peach extract in Example 1; 200 mg × 4 grains) were taken every day during the monitoring period before going to bed (20: 00-22: 00), On the 55th day after the start of monitoring, the skin erythema and elasticity were similarly measured. As for the measurement method, for the erythema, the probe is directly applied to each part and data processing is performed on the program.For the elasticity, the probe is applied to the measurement part and suctioned, the pressure is released, the displacement with respect to time is measured, and the elasticity is calculated from the waveform. Calculated and evaluated by calculation. The measurement environment was a temperature of 25 ± 1 ° C. and a humidity of 53 to 63%. The results are shown in FIG. 3 for individual data for erythema redness (cheek), and the result of statistical processing (t-test) taking the average is shown in FIG. 3 and 4, all subjects tended to decrease, and the results of statistical processing showed a significant decrease after ingestion compared to before intake.

The results of the elasticity measurement test are shown in FIG. 5 for the waveform model, FIG. 6 for the individual data for elasticity, and FIG. 7 for the average value. As shown in the waveform model of FIG. 5, a value obtained by subtracting the minimum value at the time of releasing pressure from the maximum value at the time of suction and dividing by the maximum value at the time of suction is evaluated as elasticity. Except for one subject, all subjects tended to increase their elasticity. The vertical axis of the graph is the value of R2 shown in the waveform model, but the closer it is to 1, the closer it is to a perfect elastic body, and it is evaluated that there is elasticity. As shown in FIGS. 6 and 7, statistical processing was performed by taking an average. As a result, although there was no significant difference, there was an increase in elasticity after ingestion compared with before ingestion. As described above, it was found that the effects of fermented moon peach extract grains on the skin physical properties include an increase in skin elasticity and an improvement in erythema (redness).
[Reference Example 1]

(Measurement of melanin production)
The amount of melanin and the amount of DNA were measured, and the amount of melanin produced per DNA was measured. Further, as a test sample, a freeze-dried product of fermented moon peach hot water extract in Example 1 and a solution of L (+) sodium ascorbate (manufactured by Wako) dissolved in PBS (−) solution as vitamin C. In addition, a solution sterilized by filtration with a 0.22 μm filter was used. The sample solution was prepared to be 1% or less with respect to the medium so as not to affect cell growth.
(1) Preparation of measurement sample: B16 melanoma cells (purchased from Riken Cell Bank, B16melanoma 4A5: Car No RCB0557) are added as a cell in 10% FBS-MEM (pH 7.4) medium and a sample solution of a predetermined concentration is added. The cells were cultured at 37 ° C. and 5% CO _{2 for} 3 days. The medium was removed, the cell surface was washed twice with PBS (−) solution, 1 mL of 50 mM Tris-HCl buffer (pH 7.5) was then added, and the cells were collected with a cell scraper (manufactured by Nunc), and the sonicator (Yamato powersonic It was crushed with Model 150). This was used as a cell lysate.
(2) Measurement of melanin amount: 0.5 mL of 2N NaOH was added to 0.5 mL of the cell disruption solution of (1) and heated at 80 ° C. to dissolve melanin. After cooling, the absorbance at 400 nm was measured. A calibration curve was prepared using a melanin standard solution, and the amount of melanin was calculated.
(3) Measurement of DNA amount: DAPI (4 ′, 6-Diamidino-2-phenylindole (DAPI) [SIGMA] 1 mg / 20 mL Tris-HCI buffer (pH 7.5)) in 80 μm of the cell disruption solution of (1) 4 mL of Tris-HCI buffer (pH 7.5) was diluted at the time of use and mixed, and the mixture was placed in the dark for 30 minutes. The excitation wavelength (Ex) was 350 nm, and the fluorescence intensity was measured (Shimadzu spectofluorophotometer RF-540). A calibration curve was prepared using a DNA standard solution, and the amount of DNA was calculated.

Concentration of non-fermented moon peach and fermented moon peach (moon peach tea) was 0 (control), 25, 50, 100, 150 (μg / mL medium), and vitamin C (comparative example) The effect (effect) exerted on the amount of melanin produced by B16 melanoma cells when 50 (μg / mL medium) is used is determined by adopting the methods for measuring melanin and measuring DNA in (2) and (3) above. It was. The results are shown in FIG. From FIG. 8, compared with the control, both the non-fermented moon peach and the fermented moon peach (moon peach tea) have a high degree of inhibition of melanin production, and the concentration is in the range of 25 to 100 (μg / mL medium). The higher the value, the higher the suppression force, but 150 (μg / mL medium) is somewhat inferior to 100 (μg / mL medium). It was found that the amount of melanin produced was suppressed from vitamin C at any concentration. However, there was almost no difference between the unfermented moon peach and the fermented moon peach (moon peach tea).
[Reference Example 2]

(Measurement of intracellular tyrosinase activity)
The tyrosinase activity and protein amount in the cells were measured, and the enzyme activity per protein was determined.
(1) Preparation of measurement sample: A sample of a predetermined concentration was added to the medium by the same method as in Reference Example 1, and B16 melanoma cells were cultured. The medium was removed, and the cell surface was washed twice with PBS (−) solution. Then, 1 mL of 0.5% DOC (deoxycorticosterone) was added, and the cells were collected with a cell scraper and disrupted with a sonicator. This was centrifuged (11000 rpm, 20 min, 4 ° C.), and the supernatant was used as an enzyme solution.
(2) Tyrosinase activity measurement: 0.5 mL of the enzyme solution was added to 1 mL of 0.05% L-DOPA (L-DOPA) /0.1 M phosphate buffer solution preincubated for 10 minutes at 37 ° C. Absorbance at 475 nm was measured over time (1 min) for 10 minutes with incubation.
In order to measure auto-oxidation of L-DOPA (L-DOPA), the absorbance of 0.5 mL DOC added to 1 mL of 0.05% L-DOPA / 0.1 M phosphate buffer solution was also measured over time. Measured to be a baseline. Reaction time (min) on the horizontal axis and O.D. from the start of measurement per protein on the vertical axis. D. The slope after the start of the reaction was determined from the graph with the amount of change at 475 nm, and was defined as tyrosinase activity. Comparison was made at each concentration with the tyrosinase activity in the control as 100%.
(3) Protein amount quantification (Lowry method)
Reagent _{A: 2% Na 2 CO 3} -in0.1N NaOH
Reagent B: 0.5% CuSO ₄ .5H ₂ O-1% potassium or sodium tartrate Reagent C: Reagent A / Reagent B = 50/1 mixed. Every time new reagent D: phenol reagent (diluted with distilled water twice when using a commercial product)
Protein standard solution: Albumin preparation about 5 mg / 10 mL. Weigh accurately.
[Operation] A sample solution containing 5 to 100 μg of protein in 0.2 mL is taken into a test tube, and 1 mL of reagent C is added and mixed. After standing at room temperature for 10 minutes or more, add 0.2 mL of reagent D and mix quickly. Colorimetry is performed at an absorbance of 750 nm within 20 minutes to 2 hours. A calibration curve was prepared by diluting a standard albumin solution and using 30 to 200 μg / mL.

  Concentrations of non-fermented moon peach and fermented moon peach (moon peach tea) were 0 (control), 25, 50, 100, 150 (μg / mL medium) and vitamin C (comparative example) The effect (effect) exerted on the amount of melanin produced by B16 melanoma when 50 (μg / mL medium) was used was determined by employing the methods for measuring tyrosinase activity and protein quantification described in (2) and (3) above. The results are shown in FIG. FIG. 9 shows that the tyrosinase activity of both the non-fermented moon peach and the fermented moon peach (moon peach tea) is suppressed in a concentration-dependent manner compared to the control, and tyrosinase compared to vitamin C. It was found that the activity was suppressed. However, there was almost no difference between the non-fermented moon peach and the fermented moon peach (moon peach tea). At 100,150 (μg / mL medium), the non-fermented moon peach is more fermented. It was found that the tyrosinase inhibitory effect is superior to moon peach.

(Preparation of dermal fibroblast growth-promoting tablet)
Lyophilized 20.47% by weight of fermented moon peach hot water extract in Example 1, 75.0% by weight of collagen peptide (manufactured by Zelize Co., Ltd.), 0.15% by weight of hyaluronic acid (Horus Co., Ltd.) Manufactured) and silymarin (manufactured by Tokiwa Plant Chemical Research Co., Ltd.) at a ratio of 0.5% by weight to obtain granular tablets.

(Preparation of skin care food and drink containing skin fibroblast growth promoter)
100 mg of the freeze-dried fermented moon peach extract in Example 1 is added to 50 mL of water, and citric acid, flavor, vitamin C, glucose, sugar, isomerized sugar, etc., are added as appropriate, and mixed and stirred. And got the juice.

(Preparation of skin care cosmetics containing skin fibroblast growth promoter)
A hot water extract of fermented moon peach (fermented moon peach extract), purified water, glycerin, and urea in Example 1 were mixed in the following composition to obtain a skin care cosmetic.
Purified water 93%
Glycerin 3%
Urea 3%
Fermented moon peach extract 1%

It is a figure which shows the skin fibroblast proliferation measurement result using the hot-water extract of this invention (fermented moon peach) with control (no addition) and moon peach (no fermentation). It is a figure which shows the dermal fibroblast proliferation measurement result using the 80% ethanol extraction freeze-dried material of this invention (fermented moon peach) with control (no addition) and moon peach (no fermentation). It is a figure which shows the individual data of the erythema (redness) (cheek) taking this invention (fermented moon peach grain). It is a figure which shows the average value of the erythema (redness) taking this invention (fermented moon peach grain). It is a figure which shows the waveform model of the elasticity (cheek) of taking this invention (fermented moon peach grain). It is a figure which shows the data according to elasticity individual for taking this invention (fermented moon peach grain). It is a figure which shows the average value about the elasticity of taking this invention (fermented moon peach grain). It is a figure which shows the melanin production inhibitory effect of this invention (non-fermented moon peach, fermented moon peach) with control (no addition) and vitamin C. It is a figure which shows the inhibitory effect of the tyrosinase activity of this invention (no fermented moon peach, fermented moon peach) with a control (no addition) and vitamin C.

Claims (8)

A skin fibroblast growth promoter characterized by comprising as an active ingredient a fermented product obtained by fermenting moon peach with lactic acid bacteria, lactic acid bacteria and yeast, lactic acid bacteria and Bacillus subtilis, or lactic acid bacteria and yeast and Bacillus subtilis. The dermal fibroblast growth promoter according to claim 1, wherein the fermented product is a fermented product of moon peach leaves. The dermal fibroblast proliferation promoter according to claim 1 or 2, which is a fermented product of a mixed bacterium of Lactobacillus plantarim, Streptococcus thermophilus, and Bacillus subtilis. The dermal fibroblast growth promoter according to any one of claims 1 to 3, wherein the fermented product is an extract or a lyophilized product thereof. A skin care food or drink comprising the dermal fibroblast proliferation promoter according to any one of claims 1 to 4. Furthermore, the skin care food-drinks of Claim 5 containing the 1 or more types of component chosen from silymarin (Silymarin), a collagen peptide, and hyaluronic acid. A skin care cosmetic comprising the dermal fibroblast growth promoter according to any one of claims 1 to 4. The skin care cosmetic according to claim 7, further comprising at least one component selected from silymarin, collagen peptide, and hyaluronic acid.