CN113440456A - Application of red rice fermentation liquor - Google Patents
Application of red rice fermentation liquor Download PDFInfo
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- CN113440456A CN113440456A CN202010232010.3A CN202010232010A CN113440456A CN 113440456 A CN113440456 A CN 113440456A CN 202010232010 A CN202010232010 A CN 202010232010A CN 113440456 A CN113440456 A CN 113440456A
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Abstract
The invention belongs to the field of plant extraction and the field of daily chemicals, and particularly relates to application of red rice fermentation liquor in preparation of an external skin preparation, which is used as an anti-aging active ingredient in the external skin preparation. The red rice fermentation liquor stabilizes the skin structure, improves the skin repairing activity, delays the skin aging, improves the activity of epidermal cells and increases the thickness of the epidermis by promoting the expression of basement membrane stability related genes, extracellular matrix related genes and cell anti-aging growth factor related genes, thereby realizing the anti-aging effect, being suitable for preparing skin external agents and adding new application resources for preparing cosmetics and cosmetic additives; the preparation process of the red rice fermentation liquor is simple, the conditions are easy to control, the raw material sources are wide, and the production cost is low; the red rice fermentation liquor can produce obvious effect under lower content, the raw material is from natural food red rice, the use is safe, and the red rice fermentation liquor can be widely applied to various skin external preparations to resist skin aging.
Description
Technical Field
The invention belongs to the field of plant extraction and the field of daily chemicals, and particularly relates to application of red rice fermentation liquor.
Background
The skin is the largest organ of the human body and can protect the body from various external factors (such as ultraviolet rays, chemical substances and the like). The skin is composed of epidermis, dermis, and subcutaneous tissue, and contains skin appendages, abundant nerves, blood vessels, lymphatic vessels, and muscles. The dermis and epidermis are connected by a basement membrane structure, and the dermis is composed of fibroblasts and extracellular matrix. The up-regulation of gene expression in the skin can affect the expression change of related protein, so that the research on how to improve the expression of genes related to key parts and functions of the skin has important significance for improving the skin repair capacity, keeping the stability of the structure and delaying skin aging.
The extracellular matrix is a kind of macromolecular substance synthesized and secreted by fibroblasts, and mainly comprises collagen, elastin, glycoprotein and proteoglycan. The extracellular matrix can make our skin elastic, reduce wrinkle, and make skin plump and full of vitality. The TNC (tenascin-C) gene encodes for cytokinin C, and its primary function is to strengthen connective tissue. The cell adhesin C is taken as an important extracellular matrix protein and participates in the process of repairing tissue damage, and the increase of the expression level of the TNC can reflect the increase of the proliferation rate of the skin. The FN1(fibronectin 1) gene encodes fibronectin, which primarily mediates cell adhesion. Fibronectin can regulate the shape of cells and the organization of cytoskeleton through cell signal transduction pathways, promote cell spreading and growth, and make cell growth inseparable from skin state, and high expression of fibronectin can promote cell growth, so that the skin can be continuously renewed, and can resist aging. Vimentin, a vimentin gene, encodes vimentin, which provides elasticity not possessed by microtubules and actin and is responsible for maintaining the integrity of cytoskeleton.
A layer of thin film, called as basal membrane band, is arranged at the connecting area of the dermis and the epidermis, is an important structure for connecting the true epidermis, and also has certain selective permeability barrier function, so that the material exchange is convenient. The basement membrane contains various proteins including type IV collagen, laminin, catenin and other macromolecules, and has a very close relationship with many life phenomena such as cell morphology, cell polarity, cell metabolism, protein distribution on a plasma membrane, cell survival, cell proliferation and differentiation, cell migration and the like. The COL4A1(collagen type IV alpha 1chain) gene encodes type IV collagen, which is a major component of basement membrane. In the basement membrane, type iv collagen, laminin 5, heparin and the like form a stable and ordered grid structure, and the structure plays an important role in the construction and stabilization of the basement membrane. The up-regulation of the expression of the COL4A1 gene can promote the integrity of DEJ and delay skin aging. The ITGB1(integrin subbnit beta 1) gene encodes integrin 1, and functions as a cell adhesion, and as a transmembrane linker, a bidirectional link between the extracellular matrix and the intracellular actin backbone. Integrins also have the role of transmitting extracellular signals into the cell. The HSPG2 (heptan sulfate proteolycan 2) gene encodes Perlecan protein, a major component of basement membrane. The protein interacts with laminin, delipidated protein, type IV collagen, etc., and plays an important role in a variety of biological activities. Perlecan may also promote growth factor activity, thereby stimulating endothelial growth and regeneration. The expression up-regulation of the HSPG2 gene can promote the expression of perlecan protein, further promote the stability of a basement membrane structure and strengthen the tight connection of true epidermis. The collagen VII encoded by COL7A1(collagen type VII alpha 1chain) gene is the main collagen expressed in the connecting region of the true epidermis and plays the role of anchoring to form firm connection between the true epidermis cells. The laminin encoded by the LAMA3(laminin-3) gene is a non-collagen sugar that forms the stroma of cells, and together with collagen, forms a component of basement membrane, which is the medium by which cells adhere to the stroma and, in combination with various basement membrane components, regulates cell growth and differentiation.
Growth factors are a class of polypeptides or proteins that regulate cell growth and other cellular functions by binding to high affinity, specific cell membrane receptors. Mainly secreted by various cells, act on specific target cells, and regulate cell division and matrix synthesis. CSF2(colony stimulating factor 2) gene codes granulocyte macrophage colony stimulating factor, mainly acts on myeloid progenitor cells, exerts biological effect by combining with high affinity receptors thereof, enables the granulocyte macrophage colony stimulating factor to rapidly enter cell cycle and differentiate towards granulocyte colony and macrophage colony to finally become mature cells, can prolong the life of the mature cells and enhance the functions of the mature cells, is an important cell longevity factor, and is closely related to skin aging. FGF2(fibroblast growth factor 2) gene encodes fibroblast growth factor (bFGF), a growth factor and signaling protein that regulate the proliferation and differentiation of specific cell types. In addition, FGF2 exhibits potent angiogenic effects in vivo and in vitro, stimulating smooth muscle cell growth, promoting wound healing and tissue regeneration. The egfr (epidermal growth factor receptor) gene encodes a synthetic epidermal growth factor receptor, a transmembrane glycoprotein that induces cell proliferation by binding to a ligand (epidermal growth factor). The up-regulation of the expression of the gene is beneficial to cell proliferation and realizes the effect of delaying skin aging.
The red rice is a staple food loved by people, has purple outer skin, red inner core, good rice quality, high nutritive value, slight sour taste and light taste, and can be used as rice porridge, soup and wine brewing raw materials. The red rice is rich in starch and vegetable protein, and can supplement consumed physical strength and maintain normal body temperature; the nutrient is rich in various nutrients, and iron is the most abundant, so that the nutrient has the effects of enriching blood and preventing anemia. Besides, the red rice has the functions of reducing blood pressure and blood fat, and the contained monacolin K can prevent cholesterol from being generated, so that the red rice has excellent food therapy and health care functions.
However, no relevant report about the influence of red rice fermentation liquor on the expression level of skin-related genes is found at present.
Disclosure of Invention
The invention aims to provide application of red rice fermentation liquor, in particular to application in the aspect of preparing an anti-aging skin external preparation.
The red rice fermentation liquor can stabilize skin structure, improve skin repairing activity, delay skin aging, improve epidermal cell activity, increase epidermal thickness, and realize aging resistance, and is suitable for preparing skin external preparation.
The technical scheme of the invention is as follows:
the fermented red rice liquid can be used for preparing skin external preparation as anti-aging active ingredient.
The red rice fermentation liquor is used as an active ingredient for stabilizing the skin structure, and/or an active ingredient for improving the skin repair activity, and/or an active ingredient for delaying the skin aging, so that the anti-aging effect is realized.
The red rice fermentation liquor is used as an active ingredient for improving the synthesis of protein related to basement membrane stabilization, and/or an active ingredient for improving the synthesis of protein related to extracellular matrix, and/or an active ingredient for improving the synthesis of cell anti-aging growth factors.
The red rice fermentation liquor is used as an active ingredient for promoting the expression of basement membrane stability related genes, and/or an active ingredient for promoting the expression of extracellular matrix related genes, and/or an active ingredient for promoting the expression of cell anti-aging growth factor related genes.
Further, the basement membrane firmness related gene is at least one of COL4A1 gene, LAMA3 gene, ITGB1 gene, HSPG2 gene and COL7A1 gene;
the extracellular matrix protein related gene is at least one of VIM gene, FN1 gene and TNC gene;
the cell anti-aging growth factor related gene is at least one of a CSF2 gene, an FGF2 gene and an EGFR gene.
The red rice fermentation liquor is used as an active ingredient for improving the synthesis of type IV collagen, and/or an active ingredient for improving the synthesis of laminin, and/or an active ingredient for improving the synthesis of integrin 1, and/or an active ingredient for improving the synthesis of perlecan protein, and/or an active ingredient for improving the synthesis of type VII collagen, so that the basement membrane is stabilized, the skin structure is stabilized, and the anti-aging effect is exerted;
the red rice fermentation liquor is used as an active ingredient for improving the synthesis of vimentin, and/or an active ingredient for improving the synthesis of fibronectin and/or an active ingredient for improving the synthesis of cell adhesin C protein, so that the skin structure is stabilized, the skin repairing activity is improved, and the anti-aging is realized;
the red rice fermentation liquor is used as an active component for improving the synthesis of colony stimulating factors, and/or an active component for improving the synthesis of fibroblast growth factors, and/or an active component for improving the synthesis of epidermal growth factor receptors, so that the skin repairing activity is improved, the skin aging is delayed, and the anti-aging is realized.
In another embodiment of the present invention, the red rice fermented liquid is used as an active ingredient for increasing the activity of epidermal cells and/or an active ingredient for increasing the thickness of epidermis to achieve anti-aging.
The red rice fermentation liquor is stock solution of red rice liquid fermentation product filtrate, or dilution, concentrated solution, extract or dried product prepared from the stock solution of the red rice liquid fermentation product filtrate.
The red rice fermentation liquor is a product obtained by metabolizing Lactobacillus plantarum (L.plantarum) in a culture medium containing red rice. Fermenting red rice with Lactobacillus plantarum liquid, centrifuging the obtained fermentation liquid, collecting supernatant, filtering, and collecting filtrate to obtain red rice fermentation liquid. Or diluting, concentrating, or making into extract or dry powder.
The preparation method of the red rice fermentation liquor comprises the following steps: (1) crushing red rice, mixing the crushed red rice with water to prepare a culture medium, inoculating lactobacillus plantarum, and carrying out constant-temperature culture for 16-48 hours at the temperature of 30-40 ℃ and at the speed of 150-250 rpm for liquid fermentation; (2) centrifuging the fermentation liquor for 10-60 min at the temperature of 4-10 ℃ and the rpm of 1000-8000, and taking supernatant; (3) adjusting the pH of the supernatant to 5.0-7.0, heating at 80-90 ℃ for 30-60 min, centrifuging at 1000-8000 rpm for 10-30 min, and taking the supernatant; (4) filtering the supernatant with a filter membrane with the pore diameter of 0.05-0.22 mu m under the pressure of 1-5 bar, and taking the fermentation filtrate. The preferable fermentation temperature is 35-40 ℃.
In the culture medium, the content of the red rice is 1 wt% -5 wt%, and the balance is water. Preferably, the red rice is crushed into particles of 20-200 meshes. Preferably, the red rice is produced in the region of pan-himalayas.
A skin external preparation having effects of stabilizing skin structure, and/or improving skin repair activity, and/or delaying skin aging, and/or improving epidermal cell activity, and/or increasing epidermal thickness comprises red rice fermentation broth, wherein the content of red rice fermentation broth is 0.001-99.9 wt%, preferably 0.01-60 wt%, more preferably 0.05-25 wt%, based on the stock solution of red rice liquid fermentation product filtrate.
In a preferred embodiment of the present invention, the content of the red rice fermented liquid is 0.5 wt% to 10 wt% based on the stock solution of the red rice liquid fermented product filtrate.
The skin external agent is a product which is spread on any part of the surface of a human body by smearing, spraying or other similar methods so as to clean, maintain, beautify, modify, change the appearance or correct the smell of the human body and keep a good state. The skin external preparation can be basic cosmetics, facial cosmetics, head care products, and body care products for cleaning, caring, beautifying or caring skin.
The formulation of the skin external preparation is not particularly limited, and may be determined according to the use condition, such as cleansing cream, essence, lotion, cream, lotion (such as smoothing toner, toner), mask, makeup remover, gel, aerosol or spray.
The skin external preparation contains red rice fermentation broth, and also contains other active ingredients including one or more of moisturizing active ingredient, whitening active ingredient, moisturizing active ingredient, antioxidant active ingredient, wrinkle resisting active ingredient, speckle removing active ingredient, acne removing active ingredient, sunscreen active ingredient, acne removing/acne active ingredient, dandruff removing active ingredient, antiallergic active ingredient or sebaceous gland inhibiting active ingredient.
The skin external preparation further comprises one or more of thickener, solubilizer, cosolvent, antiseptic, surfactant, stabilizer, softener, aerosol solvent, aromatic, toner, penetration enhancer, controlled release agent, sustained release agent, humectant, emulsifier, plasticizer, or pearling agent.
Compared with the prior art, the invention has the beneficial effects that:
the red rice fermentation liquor stabilizes the skin structure, improves the skin repairing activity, delays the skin aging, improves the activity of epidermal cells and increases the thickness of the epidermis by promoting the expression of basement membrane stability related genes, extracellular matrix related genes and cell anti-aging growth factor related genes, thereby realizing the anti-aging effect, being suitable for preparing skin external agents and adding new application resources for preparing cosmetics and cosmetic additives; the preparation process of the red rice fermentation liquor is simple, the conditions are easy to control, the raw material sources are wide, and the production cost is low; the red rice fermentation liquor can produce obvious effect under lower content, the raw material is from natural food red rice, the use is safe, and the red rice fermentation liquor can be widely applied to various skin external preparations to resist skin aging.
Drawings
FIG. 1 shows the results of gene expression related to the stability of the basement membrane of the blank control group (NT) and the fermentation broth of red rice.
FIG. 2 shows the results of extracellular matrix-related gene expression in a blank control (NT) and in a red rice-added fermentation broth.
FIG. 3 shows the results of the expression of the anti-senescence growth factor-associated genes in the control blank (NT) and the cells supplemented with red rice fermentation broth.
FIG. 4 shows the results of staining of the histological sections of the epidermis model.
Detailed Description
EXAMPLE 1 preparation of Red Rice fermentation broth
The substrate raw material of the fermentation product is red rice, and is produced in the Tibetan Linzhi city in the area of pan-Hi-Ma-La-Yao. And cleaning red rice seeds with deionized water, drying in the air, pulverizing with a pulverizer, and sieving with a 60-mesh sieve to obtain the red rice powder. Mixing red rice powder with deionized water to obtain fermentation culture medium, and sterilizing at high temperature under high pressure. Wherein the red rice flour contains 1-5 wt% of red rice flour, and the balance of water, and the percentages are mass percentages.
Lactobacillus plantarum (Lactobacillus plantarum) is inoculated into the fermentation medium, and the Lactobacillus plantarum is cultured at the constant temperature of 35-40 ℃ and 150-250 rpm for 36 hours.
Preparing red rice fermentation filtrate:
(1) centrifuging the fermentation product for 10-30 min at 4-10 ℃ and 6000rpm, and taking supernatant;
(2) adjusting the pH value to 5.0-7.0, carrying out water bath at 80-90 ℃ for 30-60 min, centrifuging at 1000-4000 rpm for 10-30 min, and taking supernatant;
(3) selecting a filter membrane with the aperture of 0.05, 0.10 or 0.22 mu m, and pre-soaking the filter membrane in 75% (v/v) ethanol; the filter and the equipment are cleaned by 75% (v/v) ethanol and dried by a 70 ℃ oven for use;
(5) the membrane filtration device is firstly circulated and soaked in 0.5% NaOH aqueous solution for 5-10 min, then is circulated or soaked in 0.5% NaHSO3 aqueous solution for 5-10 min, and then is cleaned by deionized water. After the membrane is arranged in equipment, sterilizing water is added for cleaning, and then the water is emptied;
(6) and (3) inoculating the supernatant obtained in the step (1) for filtration, controlling the filtration speed (1.0-2.0L/h) by adjusting the pressure (less than or equal to 5bar), and storing the filtered fermentation filtrate in a sterile container.
The obtained red rice fermentation filtrate had a dry matter content of 1.5 wt%.
Example 2 Red Rice fermentation broth for Gene expression Activity test
1. Preparation of skin model
Human primary fibroblasts (NHDF) and human primary keratinocytes (NHEK) were extracted from skin tissue, respectively. Reviving human primary fibroblasts, mixing the human primary fibroblasts with collagen after culture and digestion, performing dermal stage culture for 3 days, then digesting and inoculating human primary keratinocytes to the surface of a dermal stage cultured skin model, sequentially performing epidermal immersion culture and gas-liquid phase culture, and culturing for 26 days to obtain the 3D skin model for detection.
On the 4 th day of submerged culture after the inoculation of human primary keratinocytes on the skin model surface at the dermal stage, the cells were grouped into a blank control (NT) group, a red rice fermentation broth (1%, v/v) group and a red rice fermentation broth (5%, v/v) group, depending on the medium used.
Diluting red rice fermentation liquor into a culture medium for culture according to the same proportion on the 1 st day and the 4 th day of gas-liquid phase culture, and collecting skin for subsequent RNA extraction on the 8 th day of gas-liquid phase culture.
Extraction of RNA
Clipping 3D skin model tissue about 2 x 3mm, and placing into 2mL EP tube; adding 1mL of Trizol, adding 2 steel balls, homogenizing by using a tissue disruptor, and carrying out 50Hz ultrasonic treatment for 5 min; centrifuging at 9000rpm and 4 deg.C for 1min, and transferring the supernatant to a new 1.5mL EP tube; adding 200 mu L of chloroform, slightly reversing for several times, uniformly mixing, and standing for 5 minutes at room temperature; centrifuging at 9000rpm and 4 deg.C for 15 min; transferring the upper water phase (about 400 μ L) into a new 1.5mL EP tube, adding isopropanol with the same volume, mixing, and standing at room temperature for 10 min; centrifuging at 9000rpm and 4 deg.C for 10 min; discarding the supernatant, washing the precipitate with pre-cooled 70% anhydrous ethanol for 1 time, and centrifuging at 8000rpm and 4 deg.C for 5 min; discarding the supernatant, drying the precipitate in air for 5-10 minutes, and dissolving in 30 μ L of DEPC (diethyl cokenate) water; the RNA concentration was determined spectrophotometrically.
3. Analysis of Gene expression
RNA purification and gene quantitative analysis were performed according to QuantiGene R2.0 Plex Assay kit, gene expression was analyzed using luminex platform, and expression levels of the relevant genes were detected, with the results shown in FIGS. 1-3.
(1) Expression of red rice fermentation liquor for improving stability-related genes of basement membrane
As shown in fig. 1, when compared to the blank control group (NT), the genes related to the basal layer, such as COL7a1, LAMA3, ITGB1, HSPG2, and COL4a1, were up-regulated to different degrees after the addition of the red rice fermentation broth, and the degree of up-regulation of the basal layer-related proteins by 5% of the red rice fermentation broth was greater than 1% of the red rice fermentation broth. For example, 1% red rice broth up-regulated the expression of COL4A1 gene by 1.99 fold and 5% red rice broth up-regulated the expression of COL4A1 gene by 2.59 fold compared to NT group. Collagen IV coded by COL4A1 gene and many molecules such as laminin 5 and heparin form a stable and ordered grid structure, thereby playing a role of stabilizing a basement membrane, and the up-regulation of COL4A1 gene can also promote the integrity of DEJ and delay skin aging. The ITGB1 gene encodes synthetic integrin 1 as a transmembrane linker that functions both in cell adhesion and signaling between the extracellular matrix and the intracellular actin scaffold. The basement membrane glycan protein (Perlecan) coded by the HSPG2 gene can promote the activity of growth factors and the firmness of a basement membrane structure, and strengthen the tight connection of true epidermis. The collagen VII encoded by COL7A1 gene is the main collagen expressed in the connecting area of true epidermis and plays the role of anchoring to form firm connection between true epidermis cells. Laminin encoded by the LAMA3 gene is a mediator of cell adhesion to substrates and binds to various basement membrane components, regulating cell growth and differentiation.
As can be seen from the figure, the red rice fermentation liquor can promote the up-regulation of the expression of the genes related to the basal layer to play an anti-aging role from multiple aspects.
(2) Method for improving expression of extracellular matrix related genes by red rice fermentation extracting solution
As can be seen from fig. 2, the extracellular matrix-related genes were up-regulated to different degrees after adding the red rice fermentation broth compared to the blank control (NT), and the degree of extracellular matrix-related proteins in 5% red rice fermentation broth was greater than that in 1% red rice fermentation broth. For example, compared with NT group, 1% red rice fermentation liquor can up-regulate VIM gene expression by 1.39 times, and 5% red rice fermentation liquor can up-regulate VIM gene expression by 1.96 times. VIM gene encodes vimentin, which provides elasticity not possessed by microtubules and actin and is responsible for maintaining the integrity of cytoskeleton. FN1 gene encodes synthetic fibronectin, which regulates the shape of cells and the tissue of cytoskeleton through cell signal transduction pathway, promotes the cell spreading and growth, and ensures the cell growth to be inseparable from the skin state, so that the skin can be continuously renewed, and the skin can be resisted from aging. The cytoadhesin C encoded by the TNC gene is involved in the tissue damage repair process and cell proliferation. As can be seen from the figure, the red rice fermentation liquor can promote the up-regulation of the expression of the genes related to the cell matrix, increase the skin elasticity, reduce the generation of wrinkles and slow down the aging process.
(3) Method for improving cell anti-aging growth factor related gene expression by red rice fermentation liquor
As shown in fig. 3, after the anti-senescence cell growth factor related gene was added to the blank control group (NT), the anti-senescence cell growth factor related gene expression was up-regulated to a different extent, and the degree of up-regulation of the anti-senescence cell growth factor by 5% of the red rice fermentation broth was greater than that by 1% of the red rice fermentation broth. For example, 1% red rice broth up-regulated the CSF2 gene expression by 7.69-fold and 5% red rice broth up-regulated the CSF2 gene expression by 14.76-fold compared to the NT group. The CSF2 gene coded colony stimulating factor is an important cell longevity factor, and can prolong the life of mature cell and strengthen the function of mature cell. FGF2 gene encodes fibroblast growth factor that regulates the proliferation and differentiation of specific cell types, stimulates the growth of smooth muscle cells, promotes wound healing and tissue regeneration. The EGFR gene encodes a synthetic epidermal growth factor receptor that binds to a ligand (epidermal growth factor) to induce cell proliferation. The up-regulation of the expression of the gene is beneficial to cell proliferation and realizes the effect of delaying skin aging. Therefore, the red rice fermentation liquid can promote the up-regulation of the expression of the cell anti-aging growth factor related gene and slow down the aging process.
From the above gene expression test results, it can be seen that, compared to the blank control group (NT), the expressions of the basal layer protein-related gene, the extracellular matrix protein-related gene, and the anti-aging cytokine gene are increased to different degrees after the red rice fermentation broth is added. Therefore, the red rice fermentation liquor can improve the skin repair activity, stabilize the skin structure and delay the skin aging.
Example 3 efficacy testing of red rice fermentation broth on 3D recombinant human epidermal model
The in vitro human recombinant epidermal model can simulate the growth cycle of normal human epidermis, and the safety and the efficacy of a test object are evaluated through the proliferation and the differentiation of epidermal keratinocytes and the structure and the components of the epidermis.
The test method comprises the following steps:
3D human recombinant epidermal models were constructed using suspended cell culture chambers Transwell (Corning) matched to 12-well plates. And when the human keratinocytes in the culture flask are expanded to 70-80% confluence, digesting and harvesting the cells, re-suspending the cells by using a 3D epidermal model culture solution, and inoculating the cells into a Transwell. After 3D epidermis model culture inoculation, the submerged phase culture is maintained in the initial stage, and the gas-liquid phase culture is started from the 4 th day until the end. The experiment was divided into a blank control group and a 5% red rice broth treatment group, each group was set with 4 replicate wells, 3 of which were used for MTT cell activity test and 1 for histochemical staining and analysis.
And (3) testing results:
according to the results of the MTT test (table 1), the 5% red rice fermentation broth showed no cytotoxic effect and had a very significant enhancing effect on epidermal cell activity.
According to the results of staining and quantitative analysis of the sections of the epidermis model (table 1 and fig. 4), the morphology of the integral structure, stratum corneum and granular layer was more regular in the epidermis to which 5% of red rice fermentation broth was added, compared to the blank control group. The total thickness of the epidermis layer is one of the most important indexes for measuring the state of the epidermis, and the epidermis microscopic pictures of all samples are quantitatively analyzed by Image Pro-Plus software, the average thickness of the epidermis in each group of pictures is counted (Table 2), and the result shows that the epidermis treated by the 5% red rice fermentation liquor can obviously increase the thickness of the epidermis (P < 0.01).
From the test results, compared with the blank control group, the activity of the epidermal cells is obviously improved and the thickness of the epidermis is obviously increased after the red rice fermentation liquor is added. Therefore, the red rice fermentation liquor can improve the activity of epidermal cells and increase the thickness of epidermis, thereby realizing anti-aging.
TABLE 1 MTT method for cell viability assay
| Cell viability (%) | |
| Blank control group | 100 |
| 5% Red Rice fermentation broth | 114.7 |
TABLE 23D skin epidermal thickness
| Skin thickness (pixel) | |
| Blank control group | 102.6 |
| 5% Red Rice fermentation broth | 119.9 |
Claims (10)
1. The application of the red rice fermentation liquor in the preparation of the skin external agent is characterized in that the red rice fermentation liquor is used as an anti-aging active ingredient.
2. Use according to claim 1, characterized in that the red rice fermentation broth is used as an active ingredient for stabilizing the skin structure, and/or for improving the skin repair activity, and/or for delaying skin aging, achieving anti-aging.
3. The use according to claim 2, wherein the red rice fermentation broth is used as an active ingredient for enhancing the synthesis of proteins related to basement membrane firmness, and/or for enhancing the synthesis of proteins related to extracellular matrix, and/or for enhancing the synthesis of anti-senescence growth factors of cells.
4. The use according to claim 2, wherein the red rice fermentation broth is used as an active ingredient for promoting the expression of basement membrane firmness-related genes, and/or an active ingredient for promoting the expression of extracellular matrix-related genes, and/or an active ingredient for promoting the expression of anti-aging growth factor-related genes.
5. The use according to claim 4, wherein the basement membrane firmness associated gene is at least one of COL4A1 gene, LAMA3 gene, ITGB1 gene, HSPG2 gene and COL7A1 gene;
the extracellular matrix protein related gene is at least one of VIM gene, FN1 gene and TNC gene;
the cell anti-aging growth factor related gene is at least one of a CSF2 gene, an FGF2 gene and an EGFR gene.
6. The use according to claim 2, wherein the red rice fermentation broth as an active ingredient for increasing collagen type IV synthesis, and/or an active ingredient for increasing laminin synthesis, and/or an active ingredient for increasing integrin 1 synthesis, and/or an active ingredient for increasing perlecan protein synthesis, and/or an active ingredient for increasing collagen type VII synthesis stabilizes the basement membrane, achieves a firm skin structure, and exerts an anti-aging effect;
the red rice fermentation liquor is used as an active ingredient for improving the synthesis of vimentin, and/or an active ingredient for improving the synthesis of fibronectin and/or an active ingredient for improving the synthesis of cell adhesin C protein, so that the skin structure is stabilized, the skin repairing activity is improved, and the anti-aging is realized;
the red rice fermentation liquor is used as an active component for improving the synthesis of colony stimulating factors, and/or an active component for improving the synthesis of fibroblast growth factors, and/or an active component for improving the synthesis of epidermal growth factor receptors, so that the skin repairing activity is improved, the skin aging is delayed, and the anti-aging is realized.
7. The use according to claim 1, the red rice fermentation broth is used as an active ingredient for increasing the activity of epidermal cells and/or increasing the thickness of epidermis to achieve anti-aging.
8. The use according to any one of claims 1 to 7, wherein the red rice fermentation broth is a stock solution of red rice liquid fermentation product filtrate, or a dilution, concentrate, extract or dried product prepared from the stock solution of red rice liquid fermentation product filtrate.
9. A skin external preparation having effects of stabilizing skin structure, and/or improving skin repair activity, and/or delaying skin aging, and/or increasing epidermal cell activity, and/or increasing epidermal thickness, characterized by comprising red rice fermentation broth.
10. The external preparation for skin as claimed in claim 9, wherein the content of the red rice fermentation broth is, based on the stock solution of the red rice liquid fermentation product filtrate: 0.001 wt% -99.9 wt%.
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