KR101775122B1 - Composition for improving skin conditions and preventing or treating proliferative skin diseases containing fermentation broth of yeast lysate mixture - Google Patents
Composition for improving skin conditions and preventing or treating proliferative skin diseases containing fermentation broth of yeast lysate mixture Download PDFInfo
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- KR101775122B1 KR101775122B1 KR1020160023314A KR20160023314A KR101775122B1 KR 101775122 B1 KR101775122 B1 KR 101775122B1 KR 1020160023314 A KR1020160023314 A KR 1020160023314A KR 20160023314 A KR20160023314 A KR 20160023314A KR 101775122 B1 KR101775122 B1 KR 101775122B1
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- skin
- fermentation broth
- broth
- improvement
- yeast cell
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Images
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/14—Yeasts or derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/20—Natural extracts
- A23V2250/218—Yeast extracts
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
Abstract
Description
The present invention relates to a cosmetic composition for skin improvement and a pharmaceutical composition for the prophylaxis or treatment of proliferative skin diseases, which comprise a complex yeast cell broth fermentation broth as an active ingredient.
In addition to rapid industrialization, the generation of strong oxidizing substances such as active oxygen species due to air pollution and the increase of UV amount due to ozone layer destruction cause skin aging and pigmentation in humans. Various antioxidants such as vitamin C, tocopherol and coenzyme Q10 have been used for the production of cosmetics for the purpose of protecting the skin from these various oxidizing substances. Recently, it has been found that various physiological active substances such as whitening, anti-aging, anti-wrinkle and antioxidant exist in various plants including herbal medicines, and thus their extracts are widely used in the production of natural cosmetics or herbal cosmetics.
An abnormality in the rate of cell proliferation within keratinocytes can sometimes lead to hyperproliferation, manifesting in many proliferative skin diseases, including various forms of psoriasis, coupled with an abnormal rate of inflammation and / or apoptosis. For psoriasis, which is a typical proliferative skin disease, there is currently no superior therapy, and only inhibition therapy is present.
A yeast fermentation broth is a combination of yeast and a mixture in the incubator. When the yeast itself and yeast are fed to other plants or plants such as digestive enzymes, vitamin B group, kylate minerals, and nucleic acids excreted in the medium, Based on this, it is currently being used as a feed additive for all kinds of breeds.
Fermentation refers to the process by which microorganisms degrade organic matter using their own enzymes. It is known that raw materials after fermentation through microorganisms are at least two times to several tens of times more effective than before fermentation. Fermented metabolites contain various amino acids, organic acids and antioxidants that are good for the skin, promoting skin metabolism and making skin texture resilient and smooth. In addition, it has been reported that the fermentation process reduces the particle size, decomposes toxic substances such as heavy metals, so that it has good absorption rate and alleviates skin troubles and allergy side effects.
There have been active researches on skin condition improvement and proliferative skin disease treatment, but there is still a need for a new method for these. Accordingly, the present invention is intended to apply a new natural fermented extract to the above-mentioned necessity.
The present invention provides a cosmetic composition for skin improvement, a pharmaceutical composition for preventing or treating proliferative skin disease, a prophylactic skin disease prevention and skin-improving food additive or a health food containing the fermentation broth of a complex yeast cell broth as an active ingredient .
One embodiment of the present invention provides a cosmetic composition for skin improvement comprising a complex yeast cell broth fermentation broth as an active ingredient.
The term " compound yeast " used in the present invention refers to known yeast for fermentation generally used in the related fermentation industry, and includes, but is not limited to, lactic acid bacteria, enzymes, yeasts, fungi, actinomycetes and the like.
According to one embodiment of the present invention, the combined yeast may be selected from the group consisting of Coccidiascus , Metschnikowia , Nematospora , Schizosaccharomyces , Hanseniaspora , Nadsonia , Saccharomycodes , Wickerhamia , Lipomyces , Ambrosiozyme , arthroascus , Citeromyces , Clavispora , Cyniclomyces , Debaryomyces , Dekkera, Guilliermondella, Hansenula , Issatchenkia , Kluyveromyces , Lodderomyces , Pachysolen , Pachytichospora , Pichia , Saccharomyces , Saccharomycopsis , Schwanniomyces , Sporopachydermia , Stephanoascus , Torulaspora , Wickerhamiella , Wingea , Zygosaccharomyces , Chionosphaera , Filobasidiella , Filobasidium , Leucosporidium , Rhodosporidium , Sporidiobolus , Fibulobasidium , Sirobasidium , Holtermannia , Tremella , Aciculoconidium , Brettanomyces , Candida , Cryptococcus , Kloeckera , Malassezia , Oosporidium , Phaffia , Rhodotorula , Sarconisporon , Schizoblastosporion , Sterigmatomyces , Sympodiomyces , Trichosporon , Trigonopsis , Bullera And Sporobolomyces And preferably one or more selected from the above bacteria which are harmless to humans and commercially available, more preferably at least one selected from the group consisting of S accharomyces cerevisiae , Hansenula anomala and / or Saccharomycopsis fibuligera .
The term " culture medium " used in the present invention includes culture media for fermentation which are commonly used in the related fermentation industry, preferably cultivation media prepared using only ingredients registered in international cosmetic raw materials, Preferably, it may be a medium prepared using only glucose, hydrolyzed corn protein and yeast extract, which are known to be usable as raw materials in the cosmetics manufacturing industry. Specifically, the medium may be 1 to 5% by weight of glucose, 1 to 5% by weight of corn protein and 0.5 to 3% by weight of yeast extract, preferably 2% by weight of glucose, 2% by weight of hydrolyzed corn protein and 1% by weight of yeast extract.
As used herein, the term " crushing " means that the fermentation broth of the complex yeast is treated with lysozyme and reacted at 37 ° C for 3 to 9 hours, preferably 6 hours, followed by sonication or homogenization .
According to one embodiment of the present invention, the fermentation broth may be contained in an amount of 2 to 10% by weight, preferably 5% by weight, based on the total weight of the composition.
According to one embodiment of the present invention, the skin improvement is characterized in that the skin is selected from the group consisting of antioxidant, dermal cell growth promotion, collagen increase, collagenase reduction, immune cell activity inhibition, inflammation inhibition, skin hydration control, Improvement of skin barrier, improvement of skin texture, reduction of skin erythema and reduction of pigmentation.
Another embodiment of the present invention provides a cosmetic comprising the cosmetic composition for skin improvement.
The cosmetic may have formulations of ointments, solutions, suspensions, emulsions, pastes, gels, creams, lotions, serums, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays , Preferably a formulation of a skin toner, a water cream or a cream, but is not limited thereto.
An embodiment of the present invention provides a pharmaceutical composition for preventing or treating a proliferative skin disease, which comprises a fermentation broth for multiple yeast cell culture as an active ingredient.
According to one embodiment of the present invention, the combined yeast is selected from the group consisting of Coccidiascus , Metschnikowia , Nematospora , Schizosaccharomyces , Hanseniaspora , Nadsonia , Saccharomycodes , Wickerhamia , Lipomyces , Ambrosiozyme , arthroascus , Citeromyces , Clavispora , Cyniclomyces , Debaryomyces , Dekkera, Guilliermondella, Hansenula , Issatchenkia , Kluyveromyces , Lodderomyces , Pachysolen , Pachytichospora , Pichia , Saccharomyces , Saccharomycopsis , Schwanniomyces , Sporopachydermia , Stephanoascus , Torulaspora , Wickerhamiella , Wingea , Zygosaccharomyces , Chionosphaera , Filobasidiella , Filobasidium , Leucosporidium , Rhodosporidium , Sporidiobolus , Fibulobasidium , Sirobasidium , Holtermannia , Tremella , Aciculoconidium , Brettanomyces , Candida , Cryptococcus , Kloeckera , Malassezia , Oosporidium , Phaffia , Rhodotorula , Sarconisporon , Schizoblastosporion , Sterigmatomyces , Sympodiomyces , Trichosporon , Trigonopsis , Bullera And Sporobolomyces And preferably one or more selected from bacterial strains which are harmless to the human body and are commercially available.
According to one embodiment of the present invention, the fermentation extract may be contained in an amount of 2 to 10% by weight, preferably 5% by weight based on the total weight of the composition.
According to one embodiment of the present invention, the proliferative dermatosis is selected from the group consisting of actinic keratosis, basal cell carcinoma, squamous cell carcinoma, fibrous histiocytoma, elevated dermatofibrosis, angioma, flammei, A malignant melanoma, metastatic carcinoma, and psoriasis, in a mammal, including a human, a human,
The pharmaceutical composition of the present invention may be formulated into various oral or parenteral dosage forms, preferably parenteral dosage forms, more preferably ointments.
Examples of the formulations for oral administration include, but are not limited to, tablets, pills, hard, soft capsules, liquids, suspensions, emulsions, syrups, granules and the like. These formulations may contain, in addition to the active ingredient, a diluent such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine, a lubricant such as silica, talc, stearic acid and its magnesium or calcium salt and / Or polyethylene glycol). The tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine, optionally mixed with starch, agar, alginic acid Or a disintegrating or boiling mixture such as its sodium salt and / or an absorbent, a colorant, a flavoring agent and a sweetening agent. The formulations may be prepared by conventional mixing, granulating or coating methods.
Formulations for parenteral administration include injections, ointments, suppositories, and the like, but are not limited thereto. Depending on the characteristics of each formulation, appropriate additives such as solvents can be selected according to common knowledge of those skilled in the art.
Another embodiment of the present invention provides a pharmaceutical preparation containing the pharmaceutical composition for preventing or treating the proliferative skin disease.
Another embodiment of the present invention provides a food additive for prevention of proliferative skin diseases and a composition for improving skin comprising a fermentation broth for multiple yeast cell culture as an active ingredient.
Another embodiment of the present invention provides a health functional food for prevention of proliferative skin disease and skin improvement comprising a fermentation broth for multiple yeast cell culture as an active ingredient.
According to embodiments of the present invention, the health functional food may be, for example, but not limited to beverage, gum, tea, vitamin complex, and other health supplement foods. The health functional food may be in the form of a capsule, tablet, powder, granule, liquid, ring, flake, paste, syrup, gel, beverage, jelly or varnish.
According to an embodiment of the present invention, the composite yeast cell broth fermentation broth may contain 0.05 to 3% by weight, based on the total weight of the health functional food.
According to an embodiment of the present invention, the health functional food may include a food supplementary additive suitable in view of common knowledge of those skilled in the art, for example, various nutrients, vitamins, electrolytes, flavors, colorants, , Alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks and the like.
The present invention provides a novel use of a composition containing a fermentation broth of a complex yeast cell broth as an active ingredient for skin improvement or prevention and treatment of proliferative skin diseases. The fermented extract of the present invention is useful as an antioxidant, dermal cell growth promoting, collagen increasing, collagenase reduction, immune cell activity inhibition, inflammation inhibition, skin hydration control, skin wrinkle improvement, dermis intimacy increase, skin elasticity improvement, Improvement of skin texture, reduction of skin erythema and reduction of pigment deposition. In addition, the fermented extract of the present invention can be used for the treatment and / or prophylaxis of diseases such as actinic keratosis, basal cell carcinoma, squamous cell carcinoma, fibrous histiocytoma, hypertrophic dermatofibroma, hemangioma, flamma, hematoma, Kaposi sarcoma, mastocytosis, And proliferative skin diseases such as dysplasia, nevus cellulata, malignant melanoma, malignant melanoma, metastatic carcinoma and psoriasis.
Fig. 1 shows the effect of the present invention on the decomposition of active oxygen of the mixed yeast cell broth fermentation broth, alpha -tocopherol and BHT.
FIG. 2 shows the cell activity of human dermal fibroblasts (nHDF) according to the concentration of the yeast cell broth fermentation broth of the present invention.
FIGS. 3 and 4 show changes in expression of COL1A1 and collagenase MMP1 in human dermal fibroblasts, respectively.
FIG. 5 shows the cell activity of mouse macrophages (RAW 264.7) according to the concentration of the yeast cell broth fermentation broth of the present invention.
Figure 6 shows the activity of luciferase in immune cells according to the concentration of the fermentation broth of the complex yeast cell of the present invention.
FIG. 7 and FIG. 8 show changes in IL-1β and TNF-α gene expression according to the concentration of the fermentation broth of the complex yeast cell of the present invention, respectively.
FIGS. 9 to 17 are graphs showing changes in skin moisture content, skin oil content, skin wrinkle degree, dermal density, skin elasticity, transdermal water content, and skin permeability of a subject's cream and toner containing the fermentation broth of the complex yeast cell of the present invention, Loss amount, improvement degree of skin texture, degree of skin erythema, and change of pigmented area.
Hereinafter, the present invention will be described in more detail with reference to one or more embodiments. However, these embodiments are illustrative of one or more embodiments, and the scope of the present invention is not limited to these embodiments.
1. Preparation of sample
1.1. Preparation of a culture broth of multiple yeast cell crushing fermentation
Saccharomyces cerevisiae , Hansenula anomala And Saccharomycopsis fibuligera was inoculated on a yeast culture medium (containing 2% by weight of glucose, 2% by weight of hydrolyzed corn protein and 1% by weight of yeast extract), and fermented until a cell count of 1 x 10 10 CFU / ml or more was confirmed. The fermentation broth was treated with 0.001% by weight of lysozyme, reacted at 37 ° C. for 6 hours, disrupted using an ultrasonic grinder, and filtered through 500 mesh and 0.2 μm filter paper to obtain a liquid yeast cell lysate . The above-mentioned disruption liquid was obtained by using a rotary vacuum concentrator and a freeze drier to obtain a dried cell disruption product from the above-mentioned complex yeast cell disruption liquid.
100 g of the thus-obtained complex yeast cell lysate was mixed with purified water in an amount of 10% by weight, and fermented by inoculation with the previously known microorganism Bifidobacterium longum for fermentation. The fermented culture was filtered through 500 mesh and 0.2 mu m filter paper to obtain a fermentation broth of the liquid yeast cell lysate. As a result of carrying out a related test example of the composition according to the present invention thus formed, it was confirmed that there is no abnormality in the skin stability.
1.2. Manufacture of skin toner
The composition of the ingredients for making a skin toner containing the composition of the present invention is shown in the following Table 1 and is prepared by the following procedure. The A phase (water phase) was completely dissolved at room temperature and the B phase (alcohol phase) was heated to 50 DEG C using a heater. The phase B was mixed with the phase A and solubilized at 1000 rpm for 10 minutes using a disperser. The mixture was cooled, filtered, and maintained at a pH of 5.2 to complete a skin toner. The combined yeast cell broth fermentation broth was prepared by adding 5 wt% of the fermentation broth.
Fermentation broth
1.3. Manufacture of moisture cream
The composition of the composition for making the water cream containing the composition of the present invention is shown in Table 2 below. Concretely, the A phase was heated to 80 DEG C and stirred, and the B phase was heated to 80 DEG C to dissolve, and then the mixture was put into the phase A and emulsified at 2000 rpm for 10 minutes. The C phase was evenly mixed, and the mixture was put on the A + B phase at 80 ° C and neutralized at 2000 rpm for 5 minutes. The solution was cooled to 30 DEG C while stirring slowly, and adjusted to pH 6.15. The combined yeast cell broth fermentation broth was prepared by adding 5 wt% of the fermentation broth.
1.4. Manufacture of cream
The constituents for preparing the cream containing the composition of the present invention are shown in Table 3 below. Concretely, the A phase was heated to 80 DEG C and stirred, and the B phase was heated to 80 DEG C to dissolve, and then the mixture was put into the phase A and emulsified at 2000 rpm for 10 minutes. The C phase was evenly mixed, and the mixture was put on the A + B phase at 80 ° C and neutralized at 2000 rpm for 5 minutes. Subsequently, the mixture was gradually cooled to 30 DEG C while being mixed and stirred, and adjusted to pH 6.23. The combined yeast cell broth fermentation broth was prepared by adding 5 wt% of the fermentation broth.
2. Experimental methods and results
Example 1. Confirmation of the effect of decomposition of active oxygen by the fermentation broth of a complex yeast cell broth
In order to confirm the effect of improving the beauty of skin by the fermentation broth of multiple yeast cells, we investigated whether active oxygen species, which plays a major role in skin aging, are removed by the yeast cell culture broth.
In order to confirm the effect of decomposition of active oxygen in the fermentation broth of multiple yeast cells, free radical scavenging with 1,1-diphenyl-2-picryl hydrazyl (DPPH), which is a free radical, Activity measurement method was used. 4 mL of methanol was added to the test tube, and the mixed yeast cell broth fermentation broth was added by concentration. Then, 1 mL of 0.15 mM DPPH solution was added and reacted at room temperature for 30 minutes, and the absorbance was measured at 520 nm. Initial (Ai) and blank (Ab) without substrate and DPPH were measured, and positive control group compared α-tocopherol and BHT.
As a result of the analysis, the complex yeast cell broth fermentation broth according to the present invention exhibited a higher active oxygen decomposition ability at a lower concentration than the positive control groups, -tocopherol and BHT (see Table 4 and FIG. 1). Therefore, it was confirmed that the degradation of active oxygen was promoted by the fermentation broth of the yeast cell culture broth, which was effective in improving the skin.
Example 2 Confirmation of Growth Promoting Effect of Human Dermal Fibroblast by Combined Yeast Cell Lysate Fermentation Medium
In order to confirm the improvement of skin cosmetic effect by the mixed yeast cell broth fermentation broth, it was experimentally examined whether the fibroblast growth to synthesize collagen, which is a constituent of the dermis, was promoted by the composite yeast cell broth fermentation broth.
Human dermal fibroblast (nHDF) was purchased from Lonza. The cell culture medium was supplemented with 10% fetal bovine serum (FBS; Gibco), 10% penicillin (100 unit / ml) and streptomycin (100 g / ml) in Dulbecco's modified Eagle's sediment (DMEM; Gibco) / mL) was added and used. The cultivation was carried out under the conditions of 37 ° C, 95% humidity, 5% CO 2 incubator, and cells between 3 and 10 generations were used for the experiment. In order to determine the degree of cytotoxicity of human dendritic fibroblast nHDF cells treated with the mixed yeast cell broth fermentation broth, cells were cultured for 24 hours after treatment with each concentration of the mixed yeast cell broth fermentation broth , And then the viability of the cells was measured using MTT of the cells.
As a result of the analysis, the cell growth was increased in a concentration-dependent manner after the treatment of the mixed yeast cell culture broth fermentation broth (see Table 5 and FIG. 2, below), compared with the control (untreated) group.
Example 3 Confirmation of Increased Expression of Col1A1 and Decreased Expression of MMP1 by Fragmented Fermentation Medium of Complex Yeast Cells
In order to confirm the skin cosmetic improvement effect of the mixed yeast cell broth fermentation broth, the expression of COL1A1, one of typical collagen expressed by human dermal fibroblasts, and MMP1, a collagenase, was measured. In order to confirm the degree of collagen expression change by the mixed yeast cell broth fermentation broth, human dermal fibroblasts were treated with various concentrations of the mixed yeast cell broth fermentation broth and cultured for 24 hours.
Herein, quantitative real-time PCR (qRT-PCR), which is a method of continuously detecting a fluorescent substance by attaching a fluorescent substance to a DNA product amplified during a polymerase chain reaction (PCR) PCR). SYBR Green I (Invitrogen) was used to quantitatively determine the expression of COL1A1 and MMP1 gene expression in the mixed yeast cell broth fermentation broth by using the fluorescence value emitted by the PCR product. The reaction solution was prepared by mixing 0.2 μM primer, 50 mM KCl, 20 mM Tris / HCl pH 8.4, 0.8 mM dNTP, 0.5 U Extaq DNA polymerase, 3 mM MgCl 2 and 1 × SYBR green. The denaturation, annealing and polymerization were carried out at 94 ° C for 30 seconds, 58 ° C for 30 seconds, and 72 ° C for 30 seconds, respectively, after primary denaturation at 94 ° C for 3 minutes using Linegene K (BioER) 40 cycles and fluorescence intensity was measured after each cycle. PCR results were verified by melting curve for each result. The threshold cycle (Ct) value of each gene was normalized to the Ct value of β- actin, and the amount of change in the Ct value was compared and analyzed. The Ct value was the number of cycles when the amount of fluorescence generated by the PCR product (the number of amplified PCR products) reached a constant reference value above the basal value, and the amount of gene expression was confirmed through the Ct value. The respective genes used in the experiment are shown in Table 6 below.
As a result of analysis, the expression of Col1A1 mRNA was increased in a concentration-dependent manner after the treatment of the yeast cell broth fermentation broth compared with the control group (see Table 7 and FIG. 3). In addition, the expression of MMP1 mRNA decreased in a concentration-dependent manner after the treatment of the yeast cell broth fermentation broth compared to the control (see Table 8 and FIG. 4, below).
Expression (%)
Example 4. Confirmation of growth inhibition effect of immune cells by a fermentation broth of a complex yeast cell broth
In order to confirm the skin cosmetic improvement effect of the mixed yeast cell broth fermentation broth, it was experimentally examined whether the growth of mouse macrophage (RAW 264.7), which acts on inflammation and erythema of the skin, is inhibited by the mixed yeast cell broth fermentation broth. Mouse immune cells were purchased from Korean Cell Line Bank (KCTC). Cell culture medium was prepared by adding 10% FBS (Gibco), 10% penicillin (100 units / mL) and streptomycin (100 g / mL) to DMEM (Gibco). The cultivation was carried out at 37 ° C, 95% humidity, 5% CO 2 incubator conditions, and cells between 3 and 10 generations were used for the experiment. In order to determine the degree of cytotoxicity of RAW 264.7 cells treated with the mixed yeast cell broth fermentation broth, the cells were cultured for 24 hours in the respective yeast cell broth fermentation broth And then the activity of the cells was measured.
As a result of the analysis, the growth of the cells was decreased in a concentration-dependent manner after the treatment of the yeast cell broth fermentation broth compared to the control (untreated group) (see Table 9 and FIG.
Example 5 Confirmation of Inhibitory Effect of NF-κB Transcription by Fermentation Culture Medium of Combined Yeast Cells
In order to confirm the skin cosmetic improvement effect of the mixed yeast cell broth fermentation broth, the transfection ability of NF-κB, which is a transcription factor of cytokine which plays an important role in the immune response, was inhibited by the culture of the yeast cell broth fermentation broth.
NF-κB reporter assay was performed to determine the degree of inhibition of NF-κB transcription by the yeast cell culture. The promoter region of the gene known to be transcribed by NF-κB was cloned into a vector containing the luciferase gene and inserted into the immune cells. After NF-κB transcription was activated by treatment with LPS inducing NF-κB transcription, the culture broth of the complex yeast cell culture broth was treated at each concentration and luciferase activity was measured. The luciferase activity is determined by the upstream region of the IgG kappa NF- kappa B region (SEQ ID NO: 5-GGGGACTTTCC-3) oligonucleotide, the NF-kB binding site of the IgG kappa chain gene, with the minimal IL-8 promoter (position -67 to +44) The gene was inserted into the immune cells according to the reagent manual using lipofectamine plus (Gibco-BRL), which was constructed using a luciferase reporter gene construct constructed to be positioned upstream of the gene. The transfection ability of the NF-κB was activated by treating LPS with the gene-inserted immune cells, and the culture broth of the complex yeast cell broth was treated at a concentration of 0, 1, 10, 100 mg / L and cultured for 24 hours . The cultured immune cells were directly added with 0.1 mL dissolution buffer (0.1 M HEPES, pH 7.6, 1% Triton-X, 1 mM DTT, and 2 mM EDTA) at the time of collection to collect cell lysates, The cells were centrifuged at rpm to obtain cell proteins. Protein concentration was measured using a Bradford assay (Bio-Rad Laboratories), and then 20 μg of protein was added to a luciferase assay mixture (25 mM glycylglycine, 15 mM MgSO 4 , 1 mg / mL BSA, ATP and 1 mM D-luciferin (Analytical Luminescence Laboratory), and the luminescence was measured three times repeatedly for 20 seconds using Monolight 2010 (Analytical Luminescence Laboratory) to obtain an average value.
As a result of the analysis, the transactivity of NF-κB increased by LPS after the treatment of the yeast cell broth fermentation broth was decreased in a concentration-dependent manner (see Table 10 and FIG. 6).
(Drainage)
Example 6. Confirmation of Inflammatory Cytokine Synthesis Inhibitory Effect by Crushed Fermentation Medium of Complex Yeast Cells
In order to confirm the improvement effect of skin cosmetic effect on the mixed yeast cell broth fermentation broth, it was confirmed that the synthesis of the inflammatory cytokine inducing the immune response was inhibited by the culture broth of the mixed yeast cell culture broth.
In order to confirm the degree of expression of inflammatory cytokine by the yeast cell culture broth, LPS-treated immune cells were treated with various concentrations of the mixed yeast cell culture broth and cultured for 24 hours. The gene expression changes were then measured by qRT-PCR. SYBR Green I (Invitrogen) was used to quantitatively determine changes in the expression of IL-1β and TNF-α gene in the yeast cell culture broth of multiple yeast cells by using the fluorescence value emitted by the PCR product . The reaction solution was prepared by mixing 0.2 μM primer, 50 mM KCl, 20 mM Tris / HCl pH 8.4, 0.8 mM dNTP, 0.5 U Extaq DNA polymerase, 3 mM MgCl 2 and 1 × SYBR green. The denaturation, annealing and polymerization were carried out at 94 ° C for 30 seconds, 60 ° C for 30 seconds, and 72 ° C for 30 seconds, respectively, after primary denaturation at 94 ° C for 3 minutes using Linegene K (BioER) 40 cycles and fluorescence intensity was measured after each cycle. PCR results were verified by melting curve for each result. The Ct value of each gene was normalized to the Ct value of β - actin, and the change of the Ct value was compared and analyzed. The Ct value was the number of cycles when the amount of fluorescence generated by the PCR product (the number of amplified PCR products) reached a constant reference value above the basal value, and the amount of gene expression was confirmed through the Ct value. Primers for each gene used in the experiment are shown in Table 11 below. L-NG-monomethyl arginine and acetate salt (L-NG-Monomethylarginine, Acetate Salt; L-NMMA) were used as the positive control.
As a result of the analysis, the expression of IL-1β mRNA increased by LPS in a concentration-dependent manner after the treatment of the mixed yeast cell culture fermentation broth was decreased as compared with the LPS-treated group (see Table 12 and FIG. In addition, the expression of TNF-α mRNA increased by LPS in a concentration-dependent manner after the treatment of the yeast cell culture broth of composite yeast cells was decreased as compared with the LPS treatment group (see Table 13 and FIG. 8).
(Drainage)
(Drainage)
Example 7. Confirmation of changes in the skin water content index by the pulverized fermentation broth of complex yeast cells
In order to confirm the improvement effect of skin cosmetics by the fermentation broth of complex yeast cells, skin moisture was analyzed using a corneometer and a sebumeter.
First, in order to confirm the effect of the mixed yeast cell broth fermentation broth on the improvement of skin oiliness, 30 adult women over 30 years old were selected and cream and toner were used. Cream and toner were used for the control, the positive control and the experimental group for 2 weeks and 4 weeks, respectively, and skin moisture index was measured according to the instrument manual using a moisture analyzer (MPA5, Courage-Khazaka Electronic GmbH). Using a moisture analyzer, the same test person measured the right ball portion of the subject to be measured three times in succession. Skin oil index was measured according to the instrument manual using a sebum analyzer (MPA5, Courage-Khazaka Electronic GmbH). Using the sebum analyzer, the same person in charge of the test measured the area of the forehead which is the measurement site of the subject.
As a result of the analysis, the skin moisture was increased in a time-dependent manner after treatment of the mixed yeast cell broth fermentation broth (see Table 14 and FIG. 9), as compared with the control (no treatment) group. Also, in the case of oil, the skin moisture was increased in a concentration-dependent manner after the treatment of the mixed yeast cell crushing fermentation broth (see Tables 15 and 10 below), as compared with the control (untreated group).
Example 8. Confirmation of skin wrinkle-improving effect by the crushed fermentation broth of complex yeast cells
In order to confirm the effect of improving the wrinkles of the skin by the fermentation broth of the mixed yeast cells, skin wrinkles were analyzed using a skin analyzer.
First, in order to confirm the effect of the mixed yeast cell broth fermentation broth on the improvement of skin wrinkles, 30 adult women aged 30 years or older were selected to use the cream and toner prepared above. Cream and toner were used for 2 weeks and 4 weeks for the control, the positive control and the experimental group, respectively. The degree of facial wrinkling was measured using PRIMOS Lite (field of
As a result, compared with the control group (untreated group), the wrinkles of the tail of the tail were decreased in a time-dependent manner after treatment of the mixed yeast cell crushing fermentation broth (see Table 16 and FIG.
Example 9. Confirmation of Skin Thickening Effect by Crushed Fermentation Medium of Complex Yeast Cells
Increased proliferation of dermal fibroblast and increased expression of collagen were observed by the fermentation broth of multiple yeast cells. Thus, it was analyzed whether the thickness of the dermal layer was increased by the collagen produced, and the effect of improving the skin thickness by the fermentation broth of the composite yeast cell broth was confirmed.
First, in order to examine the influence of the mixed yeast cell broth fermentation broth on skin thickness, 30 adult women over 30 years old were selected and cream and toner were used. The control, positive control and experimental groups were allowed to use cream and toner for 2 and 4 weeks, respectively, and then measured according to the instrument manual using a DUB-skin scanner (tpm taberna pro medicum). A DUB-skin scanner was applied with a gel for ultrasound examination and the probe was placed at right angles to the skin, and then the same test person was measured with the same pressure on the right side of the subject's right eye.
As a result of the analysis, the thickness of the skin increased in a time-dependent manner after treatment of the mixed yeast cell broth fermentation broth (see Table 17 and Fig. 12 below), as compared with the control (no treatment) group.
Example 10. Confirmation of skin elasticity improvement effect by crushed fermentation broth of complex yeast cell
The effect of improving the skin elasticity was confirmed in order to confirm the improvement effect of the skin cosmetic by the fermentation broth of the complex yeast cell broth. In order to confirm the effects of the fermentation broth of complex yeast cells on skin thickness, 30 adult women over 30 years old were selected and cream and toner were used. After using cream and toner for 2 weeks and 4 weeks for control, positive control and experimental group respectively, the degree of change of skin elasticity was measured with DermaLab USB elasticity probe. The DermaLab USB elastic probe shows the skin change and resilience with time of inhalation and inhalation time. The same test person measured the probe three times after fixation to the left ball with tape.
As a result of analysis, skin elasticity was increased in a time-dependent manner after treatment of the mixed yeast cell broth fermentation broth (see Table 18 and FIG. 13), as compared to the control (no treatment) group.
Example 11 Confirmation of Improvement of Skin Barrier Effect by Fragmented Fermentation Medium of Complex Yeast Cells
The effect of improving the skin barrier was confirmed in order to confirm the skin cosmetic improvement effect of the mixed yeast cell broth fermentation broth.
First, 30 adult women aged 30 years or older were selected to use the prepared cream and toner in order to confirm the effect of the fermentation broth of the complex yeast cell broth on skin barrier improvement. Cream and toner were used for 2 weeks and 4 weeks for the control, the positive control and the experimental group, respectively. Then, the TEWL (TransEpidermal Water Loss) was measured using a DermaLab USB TEWL probe (Cortex Technology, Inc.) It was measured according to the manual of the apparatus.
As a result of analysis, the amount of transdermal water loss was decreased in a time-dependent manner after treatment of the mixed yeast cell broth fermentation broth (see Table 19 and FIG. 14), as compared to the control (no treatment) group.
Example 12. Confirmation of effect of improving skin texture by fermentation broth of complex yeast cells
In order to confirm the skin cosmetic improvement effect of the mixed yeast cell broth fermentation broth, skin texture improving effect was confirmed.
First, in order to confirm the effect of the mixed yeast cell broth fermentation broth on skin texture improvement, 30 adult women over 30 years old were selected and cream and toner were used. After 2 and 4 weeks of cream and toner were applied to the control, positive control, and experimental groups respectively, the skin texture was measured using PRIMOS light (field of view 45 × 30-simple, flexible 3D measuring, GFMesstechnik GmbH) Respectively. Using the PRIMOS light, the same test person measured the area of the forehead of the subject three times in succession and measured the same area.
As a result of analysis, the depth of wrinkles formed on the skin to determine skin texture was decreased in a time-dependent manner after treatment of the mixed yeast cell broth fermentation broth (see Table 20 and FIG. 15), as compared to the control (untreated group).
Example 13. Compound Yeast Confirming the effect of improving the erythema of the cell broth fermentation broth
Through the above Examples 5, 6 and 7, the inhibition of the proliferation of the immune cells and the decrease of the expression of inflammatory cytokines by the fermentation broth of the complex yeast cell broth were confirmed. Thus, the effect of reducing the erythema by the reduced cytokine was examined and the effect of improving the erythema of the yeast cell broth fermentation broth was confirmed.
First, in order to confirm the effect of the mixed yeast cell broth fermentation broth on erythema improvement, 30 adult women over 30 years old were selected and the cream and toner were used. The control, positive control, and experimental groups were allowed to use cream and toner for two and four weeks, respectively, followed by spectrophotometer (Spectrophotometer CR-2600D, Konica Minolta) according to the instrument manual. The same test person measured the right side of the subject's right side of the ball three times in succession, and the mean value was calculated and used for the analysis. The values measured in the spectrophotometer are L *, a *, and b *, and the red value indicates a lot of red as the value of a * increases to a value indicating erythema.
As a result of analysis, erythema decreased in a time-dependent manner after treatment of the mixed yeast cell broth fermentation broth (see Table 21 and FIG. 16 below), as compared with the control (untreated group).
Example 14 Confirmation of Improvement of Pigmentation Deposition by Crushed Fermentation Medium of Complex Yeast Cells
In order to confirm the skin cosmetic improvement effect of the mixed yeast cell broth fermentation broth, the effect of improving pigment deposition was confirmed.
First, in order to confirm the effect of the mixed yeast cell broth fermentation broth on the improvement of pigment deposition, 30 adult women over 30 years old were selected and cream and toner were used. The control, positive control and experimental groups were allowed to use cream and toner for 2 and 4 weeks, respectively, and then measured according to the instrument manual using ANTERA 3D (Miravex). The same examiner used the left ball area three times in a row for analysis.
As a result of the analysis, the pigmented area decreased in a time-dependent manner after the treatment of the yeast cell broth fermentation broth compared with the control (no treatment group) (see Table 22 and Fig. 17 below).
The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than the foregoing description, and all changes or modifications derived from the meaning and scope of the claims and equivalents thereof are included in the scope of the present invention. .
Claims (12)
The complex yeast includes Saccharomyces cerevisiae , Hansenula anomala and Saccharomycopsis fibuligera ,
The skin improvement can be used to improve the skin, such as antioxidant, dermal cell growth promotion, collagen increase, collagenase reduction, immune cell activity inhibition, inflammation suppression, skin irritation control, skin wrinkle improvement, dermis intimacy increase, skin elasticity improvement, , Reduction of skin erythema and reduction of pigmentation. ≪ RTI ID = 0.0 > 21. < / RTI >
Wherein the fermentation broth is contained in an amount of 2 to 10% by weight based on the total weight of the composition.
The complex yeast includes Saccharomyces cerevisiae , Hansenula anomala and Saccharomycopsis fibuligera ,
The proliferative dermatological disease may be selected from the group consisting of actinic keratosis, basal cell carcinoma, squamous cell carcinoma, fibrous histiocytoma, hypertrophic dermatosarcoma, angioma, flamma, hematoma, Kaposi sarcoma, mastocytosis, A malignant melanoma, a metastatic carcinoma, and psoriasis, wherein the composition is at least one selected from the group consisting of benign cell nevus, malignant melanoma, malignant melanoma, metastatic carcinoma and psoriasis.
Wherein the fermentation broth is contained in an amount of 2 to 10% by weight based on the total weight of the composition.
The complex yeast includes Saccharomyces cerevisiae , Hansenula anomala and Saccharomycopsis fibuligera ,
The proliferative dermatologic disease may be selected from the group consisting of actinic keratosis, basal cell carcinoma, squamous cell carcinoma, fibrous histiocytoma, hypertrophic dermatosarcoma, hemangioma, flamglobulin, xanthoma, Kaposi sarcoma, mastocytosis, , Malignant melanoma, malignant melanoma, metastatic carcinoma, and psoriasis.
The skin improvement can be used to improve the skin, such as antioxidant, dermal cell growth promotion, collagen increase, collagenase reduction, immune cell activity inhibition, inflammation suppression, skin irritation control, skin wrinkle improvement, dermis intimacy increase, skin elasticity improvement, , Reduction of skin erythema, and decrease of pigmentation. The food additive for proliferative skin disease prevention and skin improvement.
The complex yeast includes Saccharomyces cerevisiae , Hansenula anomala and Saccharomycopsis fibuligera ,
The proliferative dermatologic disease may be selected from the group consisting of actinic keratosis, basal cell carcinoma, squamous cell carcinoma, fibrous histiocytoma, hypertrophic dermatosarcoma, hemangioma, flamglobulin, xanthoma, Kaposi sarcoma, mastocytosis, , Malignant melanoma, malignant melanoma, metastatic carcinoma, and psoriasis.
The skin improvement can be used to improve the skin, such as antioxidant, dermal cell growth promotion, collagen increase, collagenase reduction, immune cell activity inhibition, inflammation suppression, skin irritation control, skin wrinkle improvement, dermis intimacy increase, skin elasticity improvement, , Reduction of skin erythema, and reduction of pigment deposition. The health functional food for prophylactic skin disease prevention and skin improvement.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20220053786A (en) | 2020-10-23 | 2022-05-02 | 주식회사 로렌츄컴퍼니 | Functional mulled wine for skin improvement and prevention of female diseases |
| CN114933979A (en) * | 2021-11-26 | 2022-08-23 | 北京工商大学 | Saccharomycopsis fibuligera strain SF-1, extracellular protein prepared by using same and preparation method thereof |
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| US20090068160A1 (en) | 2007-09-04 | 2009-03-12 | L'oreal | Use of a lysate of bifidobacterium species for treating sensitive skin |
| US20150093462A1 (en) * | 2013-09-30 | 2015-04-02 | Elc Management Llc | Watery Lotion Skin Care Compositions And Methods |
| KR101583600B1 (en) | 2015-06-10 | 2016-01-21 | 장영웅 | Novel Strain Bacillus megaterium, And Uses Thereof |
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| US20090068160A1 (en) | 2007-09-04 | 2009-03-12 | L'oreal | Use of a lysate of bifidobacterium species for treating sensitive skin |
| US20150093462A1 (en) * | 2013-09-30 | 2015-04-02 | Elc Management Llc | Watery Lotion Skin Care Compositions And Methods |
| KR101583600B1 (en) | 2015-06-10 | 2016-01-21 | 장영웅 | Novel Strain Bacillus megaterium, And Uses Thereof |
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| KR20220053786A (en) | 2020-10-23 | 2022-05-02 | 주식회사 로렌츄컴퍼니 | Functional mulled wine for skin improvement and prevention of female diseases |
| CN114933979A (en) * | 2021-11-26 | 2022-08-23 | 北京工商大学 | Saccharomycopsis fibuligera strain SF-1, extracellular protein prepared by using same and preparation method thereof |
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