Moisturizing composition containing Chinese herbal medicine compound

Abstract

The invention belongs to the technical field of cosmetics, discloses a moisturizing composition containing a Chinese herbal medicine compound, and particularly discloses a traditional Chinese medicine extract compound which comprises the following components: a radix ophiopogonis extract, a radix scrophulariae extract and a chlorella extract. The invention provides a traditional Chinese medicine extract compound prepared from radix ophiopogonis extract, radix scrophulariae extract and chlorella extract, and the traditional Chinese medicine extract compound not only can promote mitochondrial respiration, but also can promote generation of H2O in epidermal keratinocytes and dermal fibroblasts, so that generation of epidermal endogenous moisturizing factors and hyaluronic acid is promoted, and the skin moisturizing effect is improved. And the generation of hyaluronic acid in the dermis is increased, so that the comprehensive moisturizing function of the epidermis and the dermis can be maintained at the same time, and finally, the double-layer multi-moisturizing purpose of surface moisturizing and deep moisturizing is achieved.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetics, and in particular relates to a moisturizing composition containing a Chinese herbal medicine complex. Background Art

[0002] With the development of science and technology, people pay more and more attention to skin care, especially the demand for moisturizing cosmetics is increasing. Although the existing moisturizers have a certain moisturizing effect, they still have shortcomings such as being greatly affected by the environment, being allergenic, and being airtight.

[0003] The skin itself has a complex moisturizing system that can keep the skin fully hydrated under physiological conditions. The epidermis is the outermost layer of the skin and has an important barrier function. It can not only block the invasion of harmful substances from the outside, but also effectively prevent the rapid loss of internal moisture. The water content of different parts of the epidermis also varies greatly. The water content of the basal layer with vigorous metabolism can reach 70%, while the water content of the area between the granular layer and the stratum corneum is only 15% to 30%.

[0004] The epidermis is composed of the basal layer, spinous layer, granular layer and stratum corneum. Keratinocytes, as the main constituent cells of the epidermis, are the key factor in the formation of the epidermal barrier. The stem cells in the basal layer initially form undifferentiated keratinocytes in the basal layer through mitosis, and gradually differentiate into cells in the spinous layer and granular layer during the upward migration process, and finally form keratinocytes in the stratum corneum. Accounting for about 90% of epidermal cells, the dynamic balance of keratinocyte proliferation and differentiation is the premise of epidermal barrier function and a key link in skin moisturizing. The factors directly related to skin moisturizing mainly include natural moisturizing factor (NMF), aquaporin 3 (AQP3), etc. Hyaluronic acid present in the epidermis plays a certain role in the epidermal barrier function and hydration of the stratum corneum. The hyaluronic acid molecular chains are intertwined to form a continuous mesh structure. Water molecules directly interact with hyaluronic acid through polar bonds and hydrogen bonds, making hyaluronic acid like a molecular sponge, which can absorb and retain thousands of times its own weight of water.

[0005] The dermis is the next layer of the skin, which is tough and elastic. The cells in the dermis are mainly fibroblasts, which are spindle-shaped or irregular triangles. The dermis is mainly composed of collagen fibers, elastic fibers, reticular fibers and amorphous extracellular matrix, which give the skin good toughness and elasticity. Collagen is mainly produced by skin fibroblasts. 75% of the skin is composed of collagen. Collagen is rich in hydrophilic groups, and its triple helix structure can also slow down the loss of water. At the same time, collagen can also provide moisture to the epidermis. The good water retention capacity of collagen ensures that the skin contains sufficient water, so that the skin always remains moist, tender and shiny. Hyaluronic acid is one of the main matrix components of the dermis. More than 50% of the hyaluronic acid in the human body exists in the dermis, which is mainly produced by fibroblasts and can increase the water retention function of the skin.

[0006] There are many causes of dry skin, and the mechanisms are very complex. In addition to exogenous factors such as ultraviolet radiation, sudden changes in weather, and exposure to chemical agents, there are also many endogenous factors, including reduced intracellular lipids, reduced natural moisturizing factors (NMF), and abnormal mitochondrial function, which are also important causes of dry skin. Currently, topical application of emollients or moisturizers is widely used to temporarily relieve dry skin, but the strong odor of ingredients and greasy ingredients may make users feel uncomfortable. In addition, in addition to substances considered active ingredients, such as fats and moisturizers, moisturizers also contain substances that are generally considered excipients (such as emulsifiers, antioxidants, and preservatives). Recent findings indicate that the effects of actives and excipients on the skin may be more significant than previously thought, and some formulations may worsen skin conditions, such as some emulsifiers that may weaken the barrier.

[0007] The main products of the mitochondrial respiratory chain (electron transport chain) are ATP and water. Mitochondria can convert the energy generated by the oxidation of sugars, fatty acids, and amino acids into H2O and ATP through oxidative phosphorylation. The electron transport chain (aerobic respiratory chain) is located in the inner membrane of mitochondria and is composed of respiratory chain complexes I-IV and ATP synthase. In addition to participating in the decomposition of substances such as sugars, proteins, and fats, the water generated in the respiratory chain mainly maintains the water balance of cells, especially epidermal cells, continuously replenishes the lost water for the epidermis, maintains the water content of skin cells, and maintains the osmotic pressure balance of epidermal cells. It is the basis for the skin to be hydrated and shiny. More than 90% of oxygen molecules in the body are consumed in mitochondria. In the process of electron transfer in the mitochondrial respiratory chain, most of the electrons can be transferred to O2 by pigment oxidase through the respiratory chain, and oxidized to generate H2O. Blockage of the mitochondrial respiratory chain can also lead to cell Ca 2+ Increased influx of Ca into mitochondria leads to increased Ca 2+The concentration decreases. It is worth noting that when the calcium ion concentration in keratinocytes decreases, differentiation is inhibited, the expression of filaggrin in cells decreases, and the endogenous moisturizing factors are insufficiently produced. The hydration of the skin is significantly reduced, and the water is severely lost, causing dry skin. Therefore, promoting mitochondrial respiration can increase the production of intracellular H2O and endogenous moisturizing factors. As the main source of H2O in cells, mitochondria can regulate their own quality control, that is, produce healthy and to-be-cleared mitochondria through fusion and fission, remove damaged mitochondria through mitochondrial autophagy, and produce new well-functioning mitochondria through mitochondrial biogenesis. Summary of the invention

[0008] The first aspect of the present invention aims to provide a traditional Chinese medicine extract complex.

[0009] The purpose of the second aspect of the present invention is to provide an application of the traditional Chinese medicine extract complex of the first aspect of the present invention.

[0010] The third aspect of the present invention is to provide a cosmetic.

[0011] The fourth aspect of the present invention aims to provide a skin softener.

[0012] The fifth aspect of the present invention aims to provide a method for preparing the skin softening lotion according to the fourth aspect of the present invention.

[0013] In order to achieve the above object, the technical solution adopted by the present invention is:

[0014] The first aspect of the present invention provides a traditional Chinese medicine extract complex, comprising the following components: an Ophiopogon japonicus extract, a Scrophularia ningpoensis extract and a Chlorella vulgaris extract.

[0015] In some embodiments of the present invention, the Chinese medicine extract complex comprises the following components in parts by weight: 1 to 10 parts of Ophiopogon japonicus extract, 1 to 10 parts of Scrophularia ningpoensis extract, and 1 to 5 parts of Chlorella vulgaris extract.

[0016] In some preferred embodiments of the present invention, the Chinese medicine extract complex comprises the following components in parts by weight: 1 to 6 parts of Ophiopogon japonicus extract, 1 to 6 parts of Scrophularia ningpoensis extract and 1 to 3 parts of Chlorella vulgaris extract.

[0017] In some embodiments of the present invention, the Ophiopogon japonicus extract is an ethanol extract of Ophiopogon japonicus.

[0018] In some embodiments of the present invention, the Ophiopogon japonicus extract is prepared by the following preparation method: Ophiopogon japonicus is mixed with alcohol and subjected to reflux extraction to obtain the Ophiopogon japonicus extract.

[0019] In some preferred embodiments of the present invention, the mass volume ratio of the Ophiopogon japonicus to the alcohol is 1:(5-12); further 1:(8-12).

[0020] In some preferred embodiments of the present invention, the reflux extraction time is 1 to 3 hours; further 1 to 2 hours.

[0021] In some preferred embodiments of the present invention, multiple reflux extractions are performed during the preparation process.

[0022] In some preferred embodiments of the present invention, solid-liquid separation and concentration are performed after the reflux extraction to obtain the Ophiopogon japonicus extract.

[0023] In some embodiments of the present invention, the Scrophularia ningpoensis extract is an aqueous extract of Ophiopogon japonicus.

[0024] In some embodiments of the present invention, the Scrophulariaceae extract is prepared by the following preparation method: mixing Scrophulariaceae with water, heating for extraction, condensing and refluxing for 1 to 2 hours, and concentrating to obtain the Scrophulariaceae extract.

[0025] In some preferred embodiments of the present invention, the mass volume ratio of the ginseng to water is 1:(5-12); further 1:(8-10).

[0026] In some embodiments of the present invention, the Chlorella extract is a Chlorella alkali extract.

[0027] In some embodiments of the present invention, the Chlorella extract is prepared by the following preparation method: mixing Chlorella with sodium hydroxide solution, ultrasonic extraction, solid-liquid separation, collecting liquid; and alcohol precipitation of the liquid to obtain the alcohol precipitate, which is the Chlorella extract.

[0028] In some preferred embodiments of the present invention, the mass volume ratio of Chlorella to sodium hydroxide solution (0.6 mol / mL) is 1:20-30.

[0029] The second aspect of the present invention provides the use of the Chinese medicine extract complex of the first aspect of the present invention in any one of (1) to (8):

[0030] (1) Preparation of cosmetics;

[0031] (2) preparing products that inhibit skin moisture loss;

[0032] (3) Increase the content of hyaluronic acid in cells;

[0033] (4) preparing a product containing hyaluronic acid in cells;

[0034] (5) Increase the expression of genes related to moisture retention in cells;

[0035] (6) preparing products that increase the expression of genes related to moisture retention in cells;

[0036] (7) Promote mitochondrial respiration of cells;

[0037] (8) Prepare products that promote mitochondrial respiration in cells.

[0038] In some embodiments of the present invention, the cells described in (3) to (8) include epidermal cells and dermal cells.

[0039] In some embodiments of the present invention, the moisturizing-related genes in (5) include but are not limited to AQP3 and / or Filaggrin.

[0040] Mitochondrial respiration:

[0041] The main products of the mitochondrial respiratory chain (electron transport chain) are ATP and water. Mitochondria can convert the energy generated by the oxidation of sugars, fatty acids, and amino acids into H2O and ATP through oxidative phosphorylation. The electron transport chain (aerobic respiratory chain) is located in the inner membrane of mitochondria and is composed of respiratory chain complexes I-IV and ATP synthase. Complex I is mainly composed of flavin mononucleotide (FMN), NAD dehydrogenase, and three Fe-S centers. It accepts electrons from NADH and transfers electrons in NADH to ubiquinone (UQ). Complex II: Succinate-ubiquinone oxidoreductase, composed of succinate dehydrogenase, flavin adenine dinucleotide (FAD), and iron-sulfur clusters, its main function is to transfer electrons from FADH2 to UQ. Complex III: Ubiquinone-cytochrome c oxidoreductase, composed of two cytochrome b (Cytb560 and Cytb566), one cytochrome c (Cytc) and one iron-sulfur protein (Fe-S), whose function is to transfer electrons from UQH2 to Cytc. Complex IV: Cytochrome oxidase, mainly composed of cytochrome oxidase complex, cytochrome a and a3 (Cyta, Cyta3), can transfer electrons from cytochrome C to O2, and catalyze and activate the combination of O2 and H+ to generate H2O. ATP synthase catalyzes the phosphorylation of ADP to produce ATP. ATP and water are two important substances in organisms. ATP is the energy source required for all life activities of organisms, and participates in many important physiological metabolism and immune regulation processes. It plays an important role in skin whitening, anti-wrinkle, freckle removal, UV protection, and regulation of skin disease signal pathways. The water produced in the respiratory chain is the most basic component in organisms. In addition to participating in the decomposition of sugars, proteins, fats and other substances, the water generated in the respiratory chain mainly maintains the water balance of cells, especially epidermal cells. It continuously replenishes the lost water for the epidermis, maintains the water content of skin cells, and maintains the osmotic pressure balance of epidermal cells. It is the basis for skin hydration and luster.

[0042] More than 90% of oxygen molecules in the body are consumed in the mitochondria. In the electron transfer process of the mitochondrial respiratory chain, most electrons can be transferred to O2 by the respiratory chain pigment oxidase, and oxidized to generate H2O. However, 1% to 2% of single electrons leak from the respiratory chain and are directly transferred to O2 to generate O2·ˉ. O2·ˉ is quickly decomposed into H2O2 by the superoxide dismutase (SOD) of the mitochondria. The once generated, uncharged H2O2 moves across the mitochondrial membrane into the cytoplasm. Hydrogen peroxide can be removed by glutathione peroxidase (GPX) in the mitochondrial matrix or catalase in the cytoplasm. When cells are stimulated by external factors (such as ultraviolet rays, pollution, etc.), the number of electrons increases, and the number of electrons transferred by complex III is insufficient to catalyze all the electrons, resulting in electron accumulation, which causes negatively charged electrons to directly combine with O2 to generate O2·ˉ, which is free at the end of the electron transport chain in the form of oxygen free radicals O2·ˉ, resulting in the generation of a large amount of ROS, which leads to a decrease in the generation of ATP and water, and further damages the mitochondrial inner membrane, reduces membrane potential, and causes mitochondrial function disorders. At the same time, obstruction of the mitochondrial respiratory chain can also lead to cell Ca 2+ Increased influx of Ca into mitochondria leads to increased Ca 2+ Concentration decreases. It is worth noting that when the calcium ion concentration in keratinocytes decreases, differentiation is inhibited, the expression of Faliggrin in cells decreases, and the endogenous moisturizing factor is insufficiently produced, the hydration of the skin is significantly reduced, and the water is severely lost, causing dry skin. Therefore, promoting mitochondrial respiration can increase the production of intracellular H2O and endogenous moisturizing factors.

[0043] The third aspect of the present invention provides a cosmetic comprising the traditional Chinese medicine extract complex according to the first aspect of the present invention.

[0044] In some embodiments of the present invention, the mass percentage of the Ophiopogon japonicus extract in the cosmetic is 0.01% to 0.08%.

[0045] In some preferred embodiments of the present invention, the mass percentage of the Ophiopogon japonicus extract in the cosmetic is 0.02% to 0.07%.

[0046] In some more preferred embodiments of the present invention, the mass percentage of the Ophiopogon japonicus extract in the cosmetic is 0.025% to 0.05%.

[0047] In some embodiments of the present invention, the mass percentage of the Scrophularia ningpoensis extract in the cosmetic is 0.01% to 0.05%.

[0048] In some preferred embodiments of the present invention, the mass percentage of the Scrophularia ningpoensis extract in the cosmetic is 0.02% to 0.04%.

[0049] In some more preferred embodiments of the present invention, the mass percentage of the Scrophularia ningpoensis extract in the cosmetic is 0.025% to 0.03%.

[0050] In some embodiments of the present invention, the mass percentage of the Chlorella extract in the cosmetic is 0.01% to 0.08%.

[0051] In some preferred embodiments of the present invention, the mass percentage of the Chlorella extract in the cosmetic is 0.02% to 0.07%.

[0052] In some more preferred embodiments of the present invention, the mass percentage of the Chlorella extract in the cosmetic is 0.02% to 0.05%.

[0053] In some embodiments of the present invention, the cosmetics include water-based cosmetics, gel-based cosmetics, emulsion-based cosmetics or cream-based cosmetics.

[0054] The fourth aspect of the present invention provides a skin softening lotion, comprising the Chinese medicine extract complex of the first aspect of the present invention and cosmetically acceptable excipients.

[0055] In some embodiments of the present invention, the cosmetically acceptable excipients include one or more of a chelating agent, a humectant, a thickener, an emulsifier, a pH adjuster, an antioxidant and a preservative.

[0056] In some embodiments of the present invention, the chelating agent includes but is not limited to disodium EDTA and / or tetrasodium EDTA.

[0057] In some embodiments of the present invention, the moisturizing agent includes, but is not limited to, one or more of glycerin, propylene glycol, sodium hyaluronate, 2-hexanediol, butylene glycol and saccharide isomers.

[0058] In some embodiments of the present invention, the emulsifier includes anionic, cationic, amphoteric and nonionic emulsifiers. The anionic emulsifiers are, for example, phosphates such as potassium lauryl phosphate and C20-22 alcohol phosphate. The nonionic emulsifier can be glycerol esters such as glyceryl stearate, polyglycerol esters such as polyglycerol-10 stearate, polyglycerol-6 oleate, polyglycerol-3 caprylate, polyglycerol-4 caprylate, etc., polysorbates such as polysorbate-20, polysorbate-60, polysorbate-80, etc.; polyethers such as ceteareth-25, ceteareth-6, etc.

[0059] In some embodiments of the present invention, the thickener can be selected from carbomers such as carbomer 934, carbomer 940, carbomer 980, polyacrylamide such as Seppic's 305 (polyacrylamide / C13-14 isoparaffin / laureth-7), etc., and can also be selected from one or more of xanthan gum, stearic acid, cellulose, disodium hydrogen phosphate, cetostearyl alcohol, magnesium aluminum silicate, hydroxyethyl cellulose and xanthan gum.

[0060] In some embodiments of the present invention, the pH adjuster includes but is not limited to one or more of sodium hydroxide, potassium hydroxide, triethanolamine and arginine.

[0061] In some embodiments of the present invention, the anti-allergic agent includes, but is not limited to, one or more of dipotassium glycyrrhizinate, gentiana root extract, ecotolein, allantoin and panthenol.

[0062] In some embodiments of the present invention, the antioxidant includes but is not limited to one or more of p-hydroxyacetophenone, pentaerythritol tetrakis (di-tert-butyl hydroxyhydrocinnamate), polydatin, phloretin, resveratrol and ferulic acid.

[0063] In some embodiments of the present invention, the skin conditioning agent includes but is not limited to one or more of silica, talc, bisabolol, tocopheryl acetate, tetrahydromethylpyrimidine carboxylic acid, collagen, xanthan gum, allantoin, ceramide, cholesterol, phytosteryl isostearate, trehalose and glycerin.

[0064] In some embodiments of the present invention, the emollient includes, but is not limited to, one or more of isododecane, isohexadecane, neopentyl glycol diheptanoate, polydimethylsiloxane, isononyl isononanoate, and caprylic triglyceride.

[0065] In some embodiments of the present invention, the preservatives include preservatives conventionally used in cosmetic hygiene regulations, such as imidazolidinyl urea, parabens such as methylparaben and propylparaben, phenoxyethanol, disodium ethylenediaminetetraacetic acid, etc., and an appropriate amount of phenoxyethanol is preferred in the present invention.

[0066] In some embodiments of the present invention, the mass percentage of the Ophiopogon japonicus extract in the skin softener is 0.01% to 0.08%.

[0067] In some preferred embodiments of the present invention, the mass percentage of the Ophiopogon japonicus extract in the skin softener is 0.02% to 0.07%.

[0068] In some more preferred embodiments of the present invention, the mass percentage of the Ophiopogon japonicus extract in the skin softener is 0.025% to 0.05%.

[0069] In some embodiments of the present invention, the mass percentage of the Scrophularia ningpoensis extract in the skin softening water is 0.01% to 0.05%.

[0070] In some preferred embodiments of the present invention, the mass percentage of the Scrophularia ningpoensis extract in the skin softening water is 0.02% to 0.04%.

[0071] In some more preferred embodiments of the present invention, the mass percentage of the Scrophularia ningpoensis extract in the skin softening water is 0.025% to 0.03%.

[0072] In some embodiments of the present invention, the mass percentage of the Chlorella extract in the skin softener is 0.01% to 0.08%.

[0073] In some preferred embodiments of the present invention, the mass percentage of the Chlorella extract in the skin softener is 0.02% to 0.07%.

[0074] In some more preferred embodiments of the present invention, the mass percentage of the Chlorella extract in the skin softener is 0.02% to 0.05%.

[0075] The fifth aspect of the present invention provides a method for preparing the skin softener of the fourth aspect of the present invention, comprising the following steps: mixing cosmetically acceptable excipients with water, dissolving at 80-90°C, cooling to 40-45°C, and mixing with a traditional Chinese medicine extract complex to obtain the skin softener.

[0076] The beneficial effects of the present invention are:

[0077] The invention provides a traditional Chinese medicine extraction compound composed of ophiopogon japonicus extract, scrophularia ningpoensis extract and chlorella vulgaris extract. The traditional Chinese medicine extraction compound can not only promote mitochondrial respiration, but also promote the production of H2O in epidermal keratinocytes and dermal fibroblasts, thereby promoting the production of endogenous moisturizing factors and hyaluronic acid in the epidermis, and increasing the production of hyaluronic acid in the dermis, so as to simultaneously maintain the comprehensive moisturizing function of the epidermis and dermis, and finally achieve the double-layer multiple moisturizing purpose of surface moisturizing and deep moisturizing.

[0078] Specifically, the present invention establishes a human keratinocyte (HaCaT) and skin fibroblast (HFF-1) culture model, and constructs a cell stress model using tBHP or ultraviolet irradiation. On this basis, it is verified whether the Chinese medicine extraction complex has the effect of promoting mitochondrial respiration, and whether it has a promoting effect on the expression of related moisturizing factors in cells. The results show that the Chinese medicine extraction complex can significantly promote the mitochondrial respiration of epidermal cells, promote the expression of aquaporin AQP3 and natural moisturizing factor Filaggrin in HaCaT cells, and increase the generation of intracellular hyaluronic acid content. The Chinese medicine extraction complex can significantly promote the mitochondrial respiration of dermal cells and increase the generation of intracellular hyaluronic acid content. BRIEF DESCRIPTION OF THE DRAWINGS

[0079] Figure 1 The effects of different proportions of Chinese herbal extracts on the mitochondrial respiratory chain of epidermal cells; wherein A is the effect of different proportions of Chinese herbal extracts on mitochondrial respiratory chain complex I of epidermal cells, B is the effect of different proportions of Chinese herbal extracts on mitochondrial respiratory chain complex II of epidermal cells, C is the effect of different proportions of Chinese herbal extracts on mitochondrial respiratory chain complex III of epidermal cells, and D is the effect of different proportions of Chinese herbal extracts on mitochondrial respiratory chain complex IV of epidermal cells; in the figure, c is the control group, S1~S4 represent embodiments 1~4 respectively, and D1~D5 represent comparative examples 1~5 respectively.

[0080] Figure 2 The effects of different proportions of Chinese herbal extracts on the mitochondrial respiratory chain of dermal cells; wherein A is the effect of different proportions of Chinese herbal extracts on mitochondrial respiratory chain complex I of dermal cells, B is the effect of different proportions of Chinese herbal extracts on mitochondrial respiratory chain complex II of dermal cells, C is the effect of different proportions of Chinese herbal extracts on mitochondrial respiratory chain complex III of dermal cells, and D is the effect of different proportions of Chinese herbal extracts on mitochondrial respiratory chain complex IV of dermal cells; in the figure, c is the control group, S1~S4 represent embodiments 1~4 respectively, and D1~D5 represent comparative examples 1~5 respectively.

[0081] Figure 3 The figure shows the effects of different proportions of Chinese herbal extracts on the mRNA expression levels of AQP3 (A) and Filaggrin (B). In the figure, c is the control group, S1-S4 represent Examples 1-4 respectively, and D1-D5 represent Comparative Examples 1-5 respectively.

[0082] Figure 4The figures show the effects of different proportions of Chinese herbal extracts on the hyaluronic acid content in the epidermis. In the figure, C is the control group, S1 to S4 represent Examples 1 to 4 respectively, and D1 to D5 represent Comparative Examples 1 to 5 respectively.

[0083] Figure 5 The figures show the effects of different proportions of Chinese herbal extracts on the hyaluronic acid content in the dermis. In the figure, C is the control group, S1-S4 represent Examples 1-4 respectively, and D1-D5 represent Comparative Examples 1-5 respectively. DETAILED DESCRIPTION

[0084] The present invention is further described in detail below through specific examples.

[0085] It should be understood that these embodiments are only used to illustrate the present invention and are not used to limit the scope of the present invention.

[0086] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0087] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0088] Preparation of the Chinese herbal extracts involved in the examples:

[0089] Preparation of Ophiopogon japonicus extract: Weigh 100 g of the raw material, crush it through a 60-mesh sieve, add 65% ethanol at a solid-liquid ratio of 1:10 (g / mL) for slight boiling reflux extraction for 2 hours, filter and take the supernatant; add 65% ethanol at a solid-liquid ratio of 1:8 (g / mL) to the filtered residue for a second slight boiling reflux extraction, the extraction time is 2 hours, filter and discard the residue, centrifuge at 5000r / min for 15 minutes, combine the filtrate, vacuum concentrate to 0.05L at (60±3)℃, and store at 4℃ in the dark for later use.

[0090] Preparation of Scrophulariaceae extract: After the raw drug of Scrophulariaceae is crushed in an appropriate amount, it is extracted with 10 times and 8 times the amount of distilled water respectively, heated at a constant temperature of 40-45°C. Both decoctions are condensed and refluxed for 1.5 hours. The drug liquids are combined, filtered, and concentrated under reduced pressure to obtain a brown-colored decoction with a sweet and aromatic smell and a slightly bitter taste. It is stored at 4°C away from light for later use.

[0091] Preparation of Chlorella extract: Weigh an appropriate amount of dry Chlorella powder, mix it with 0.6mol / mL NaOH solution at a solid-liquid ratio of 1:25g / mL, ultrasonically treat the mixed solution (ultrasonic power is 200W, ultrasonic time is 20min), react in a water bath at 80℃ for a period of time, centrifuge at 8000r / min for 10min, collect the supernatant, add 4 times the volume of anhydrous ethanol, and let it stand at 4℃ overnight. The next day, discard the supernatant after centrifugation, dissolve the lower precipitate with distilled water, which is the Chlorella solution, and store it at 4℃ away from light for later use

[0092] Example 1

[0093] A traditional Chinese medicine extract complex comprises, by weight, 1 part of ophiopogon japonicus extract, 1 part of scrophularia ningpoensis extract and 2 parts of chlorella vulgaris extract.

[0094] The Chinese herbal medicine extract complex can be obtained by mixing the three extracts.

[0095] Example 2

[0096] A traditional Chinese medicine extract complex comprises, by weight, 2 parts of ophiopogon japonicus extract, 2 parts of scrophularia ningpoensis extract and 1 part of chlorella vulgaris extract.

[0097] The Chinese herbal medicine extract complex can be obtained by mixing the three extracts.

[0098] Example 3

[0099] A traditional Chinese medicine extract complex comprises, by weight, 3 parts of ophiopogon japonicus extract, 4 parts of scrophularia ningpoensis extract and 3 parts of chlorella vulgaris extract.

[0100] The Chinese herbal medicine extract complex can be obtained by mixing the three extracts.

[0101] Example 4

[0102] A traditional Chinese medicine extract complex comprises, by weight, 5 parts of ophiopogon japonicus extract, 3 parts of scrophularia ningpoensis extract and 2 parts of chlorella vulgaris extract.

[0103] The Chinese herbal medicine extract complex can be obtained by mixing the three extracts.

[0104] Comparative Example 1

[0105] A traditional Chinese medicine extract comprises 2 parts of ophiopogon japonicus extract by weight.

[0106] Comparative Example 2

[0107] A traditional Chinese medicine extract comprises 2 parts of Scrophularia ningpoensis extract by weight.

[0108] Comparative Example 3

[0109] A traditional Chinese medicine extract comprises 2 parts of chlorella extract by weight.

[0110] Comparative Example 4

[0111] A traditional Chinese medicine extract complex comprises, by weight, 1 part of ophiopogon japonicus extract and 1 part of scrophularia ningpoensis extract.

[0112] Comparative Example 5

[0113] A traditional Chinese medicine extract compound comprises, by weight, 1 part of Scrophularia ningpoensis extract and 1 part of Chlorella vulgaris extract.

[0114] Effect Example

[0115] 1. Detection of mitochondrial respiratory chain complex I to IV activity

[0116] The specific detection process is as follows:

[0117] (1) HaCaT (HFF-1) cells were cultured at 1.5×10 5 / well were inoculated in 6-well plates, and a blank control group was set up. When the cells grew to a single cell layer confluence density of about 40%, the calcium chloride solution prepared in advance was added to make Ca 2+ The concentration reached 10 mM, and the cells were cultured under this condition for 24 h. Then, 42.5 μg of the Chinese medicine extract complex of Examples 1 to 4 and the Chinese medicine extract or Chinese medicine extract complex of Comparative Examples 1 to 5 were added and cultured for 4 h. Then, the cells were placed in an ultraviolet radiation instrument and the UVB radiation dose was set to 200 mJ / cm 2 After the ultraviolet radiation, the cells were cultured for 6 h, the culture medium was discarded, the cells were washed 3 times with pre-cooled PBS, 1 mL of the extract was added, and the cells were homogenized on ice with a homogenizer.

[0118] (2) Centrifuge at 4°C, 600 g for 10 min. Transfer the supernatant to another centrifuge tube, discard the precipitate, and centrifuge at 4°C, 11,000 g for 15 min to obtain the supernatant and precipitate.

[0119] (3) The supernatant was discarded, and 400 mL of the extract was added to the precipitate, and ultrasonic disruption was performed (power 200 W, ultrasonic 5 s, interval 10 s, repeated 15 times) for the determination of complex I-IV enzyme activity and protein content.

[0120] (4) Preheat the microplate reader for more than 30 minutes, adjust the wavelength to 340 nm, and zero the spectrophotometer with distilled water. Add the working solution to the 96-well plate, mix it immediately, perform the test, record the absorbance value, and calculate it. The larger the absorbance value, the stronger the activity of mitochondrial respiratory chain complex I-IV.

[0121] The extract and working solution were purchased from Solebao mitochondrial respiratory chain I (catalog number: BC0515), II (catalog number: BC3230), III (catalog number: BC3240), and IV (catalog number: BC0945) detection kits.

[0122] Table 1 Effects of different proportions of Chinese herbal extracts on the mitochondrial respiratory chain of epidermal cells

[0123]

[0124] Table 2 Effects of different proportions of Chinese herbal extracts on the mitochondrial respiratory chain of dermal cells

[0125]

[0126] Note: In Table 1 and Table 2, * indicates significant difference between each group and blank control group, where * represents P < 0.05, ** represents P < 0.01, *** represents P < 0.001, and **** represents P < 0.0001.

[0127] In Tables 1 and 2, △ represents the significant difference between each embodiment and the best comparison example (mitochondrial respiratory chain complex I in epidermal cells is comparison example 1, mitochondrial respiratory chain complex II is comparison example 5, mitochondrial respiratory chain complex III is comparison example 4, and mitochondrial respiratory chain complex IV is comparison example 4; mitochondrial respiratory chain complex I in dermal cells is comparison example 5, mitochondrial respiratory chain complex II is comparison example 5, mitochondrial respiratory chain complex III is comparison example 5, and mitochondrial respiratory chain complex IV is comparison example 5), wherein △ represents P < 0.05, △△ represents P < 0.01, △Δ△ represents P < 0.001, and △ΔΔ△ represents P < 0.0001.

[0128] The effects of the Chinese herbal extract complexes of Examples 1 to 4 and Comparative Examples 1 to 5 on the mitochondrial respiratory chain of epidermal cells are shown in Tables 1 and Figure 1 The effects on the mitochondrial respiratory chain of dermal cells are shown in Table 2 and Figure 2 As shown. It can be seen that the activities of mitochondrial respiratory chain complexes I-IV of all the Chinese herbal extract complexes in Examples 1 to 4 are significantly higher than those in the comparative examples, indicating that the Chinese herbal extract complexes in Examples 1 to 4 can promote the mitochondrial respiration of epidermal cells and dermal cells, and the promotion effect is significantly higher than that of comparative examples 1 to 5. Among them, the effect of the Chinese herbal extract complex in Example 1 on promoting the mitochondrial respiration of epidermal cells and dermal cells is more significant.

[0129] Moreover, by comparing Examples 1 to 4 with the best comparative example, there are significant differences, indicating that the effect is obtained based on the complementary enhancement effect of the three. The combination of Ophiopogon japonicus, Scrophularia ningpoensis and Chlorella vulgaris is significantly better than the effect of using them alone or in combination of two to promote mitochondrial respiration.

[0130] 2. qPCR detection of mRNA levels of moisturizing-related genes in HaCaT cells

[0131] HaCaT cells were grown at 1.5×10 5 / well were inoculated in 6-well plates, and a blank control group was set up. When the cells grew to a single cell layer confluence density of about 40%, the calcium chloride solution prepared in advance was added to make Ca 2+ The concentration reached 10 mM, and the cells were cultured for 24 h under this condition. Then, the Chinese medicine extract complexes of Examples 1 to 4 and Comparative Examples 1 to 5 were added and cultured for 4 h. Then, the cells were placed in an ultraviolet radiation instrument and the UVB radiation dose was set to 200 mJ / cm 2 , and irradiate the cells. After the ultraviolet radiation was completed, the cells were cultured for 6 hours, and the total RNA was extracted by cell lysis. qPCR was used to detect the mRNA expression levels of AQP3 and Filaggrin as auxiliary detection indicators of moisturizing (the higher the mRNA expression levels of AQP3 and Filaggrin, the more significant the moisturizing effect). The test results of AQP3 and Filaggrin are shown in Tables 3 and Figure 3 shown.

[0132] Table 3 Effects of different proportions of Chinese herbal extracts on the mRNA expression levels of AQP3 and Filaggrin

[0133] combination AQP3(Relative mRNA levels(fold)) Filaggrin(Relative mRNA levels(fold)) Blank Group 1 1 Example 1 <![CDATA[2.655±0.03536**** △ΔΔ△ ]]> <![CDATA[3.17±0.08485**** △ΔΔ△ ]]> Example 2 <![CDATA[2.115±0.02121**** △ΔΔ△ ]]> <![CDATA[2.425±0.0495**** △ΔΔ△ ]]> Example 3 <![CDATA[2.285±0.1344**** △ΔΔ△ ]]> <![CDATA[2.125±0.0495**** △ΔΔ△ ]]> Example 4 <![CDATA[2.095±0.07778**** △ΔΔ△ ]]> <![CDATA[2.685±0.0495**** △ΔΔ△ ]]> Comparative Example 1 0.475±0.1061**** 0.835±0.03536 Comparative Example 2 0.365±0.02121**** 0.735±0.02121** Comparative Example 3 0.78±0.01414* 0.74±0.02828** Comparative Example 4 1.08±0.02828 1.09±0.09899 Comparative Example 5 1.275±0.02121** 1.21±0.01414*

[0134] Note: In Table 3, * indicates significant difference between each group and blank control group, where * represents P < 0.05, ** represents P < 0.01, *** represents P < 0.001, **** represents P < 0.0001. △ represents significant difference between each example in improving mRNA expression level of AQP3 and Filaggrin and comparative example 5 (the best effect of improving mRNA expression level of AQP3 and Filaggrin is comparative example 5), where △ represents P < 0.05, △△ represents P < 0.01, △Δ△ represents P < 0.001, △ΔΔ△ represents P < 0.0001.

[0135] From Table 3 and Figure 3It can be seen that the effects of increasing the mRNA expression levels of AQP3 and Filaggrin in all the composition groups in the examples are significantly higher than those in the comparative examples, indicating that the Chinese medicine extract complexes in Examples 1 to 4 can increase the amount of aquaporin AQP3 and filaggrin, significantly increase the expression of moisturizing-related genes in epidermal cells, thereby improving the moisturizing effect, and the improvement effect is significantly higher than that in comparative examples 1 to 5. Among them, the effect of the Chinese medicine extract complex in Example 1 on improving the moisturizing effect is more significant.

[0136] Moreover, by comparing Examples 1 to 4 with the best comparative example, there are significant differences, indicating that the effect is obtained based on the complementary enhancement effect of the three. The combination of Ophiopogon japonicus, Scrophularia ningpoensis and Chlorella vulgaris is significantly better than the effect of using them alone or in combination of two to enhance the moisturizing effect.

[0137] 3. ELISA detection of hyaluronic acid content in HaCaT (HFF-1) cells

[0138] The cells were 1.5×10 5 / well were inoculated in 6-well plates, and a blank control group was set up. When the cells grew to a single cell layer confluence density of about 40%, the calcium chloride solution prepared in advance was added to make Ca 2+ The concentration reached 10 mM. The cells were cultured for 24 h under this condition. Then different proportions of Chinese herbal extracts were added and cultured for 4 h. Then, the cells were placed in an ultraviolet radiation instrument and the UVB radiation dose was set to 200 mJ / cm 2 , and irradiate the cells. After the ultraviolet radiation is completed, continue to culture for 6 hours, and pre-wash the cells with PBS to remove extracellular proteins. Add protein extraction buffer containing phosphatase inhibitors and protease inhibitors (both at a concentration of 100nM) and mix thoroughly. Place the mixture on ice for a certain period of time to break the cell membrane and release intracellular proteins. Centrifuge at 4°C, 12000×g to remove residual substances such as cell debris and nucleic acids, and collect the supernatant. Quantify by protein determination, and dilute the sample to the required concentration in 1× cell extraction buffer PTR. Add working solution and samples to a 96-well plate. After incubation, record the OD at 450nm and calculate (the larger the OD value, the higher the hyaluronic acid content in the sample. Calculation method: Substitute the OD value into the standard curve to get the result). The test results of the hyaluronic acid content in the epidermis are shown in Tables 4 and Figure 4 As shown in Table 5 and Figure 5 shown.

[0139] Table 4 Effects of different proportions of Chinese herbal extracts on the content of hyaluronic acid in the epidermis

[0140]

[0141]

[0142] Table 5 Effects of different proportions of Chinese herbal extracts on hyaluronic acid content in the dermis

[0143] Experimental Grouping Hyaluronic acid (ng / mL) Blank Group 1 Example 1 <![CDATA[2.81±0.05657**** △ΔΔ△ ]]> Example 2 <![CDATA[2.155±0.07778**** △ΔΔ△ ]]> Example 3 <![CDATA[2.285±0.09192**** △ΔΔ△ ]]> Example 4 <![CDATA[2.315±0.1202**** △ΔΔ△ ]]> Comparative Example 1 0.72±0.02828* Comparative Example 2 0.445±0.0495*** Comparative Example 3 0.935±0.03536 Comparative Example 4 1.07±0.04243 Comparative Example 5 1.175±0.1626

[0144] Note: In Tables 4 and 5, * indicates significant difference between each group and the blank control group, where * represents P < 0.05, ** represents P < 0.01, *** represents P < 0.001, and **** represents P < 0.0001. △ represents significant difference between each example in increasing the content of hyaluronic acid in the epidermis and dermis and Comparative Example 5 (Comparative Example 5 has the best effect on the epidermis and dermis), where △ represents P < 0.05, △△ represents P < 0.01, △Δ△ represents P < 0.001, and △ΔΔ△ represents P < 0.0001.

[0145] From Table 4, Figure 4 , Table 5 and Figure 5 It can be seen that the effects of increasing the content of hyaluronic acid in the epidermis and dermis by using the Chinese medicine extract complexes in all the examples are significantly higher than those in the comparative examples, indicating that the Chinese medicine extract complexes in Examples 1 to 4 can increase the content of hyaluronic acid, thereby improving the moisturizing effect, and the moisturizing effect is significantly higher than that in Comparative Examples 1 to 5. Among them, the effect of the Chinese medicine extract complex in Example 1 on improving the moisturizing effect is more significant.

[0146] Moreover, by comparing Examples 1 to 4 with the best comparative example, there are significant differences, indicating that the effect is obtained based on the complementary enhancement effect of the three. The combination of Ophiopogon japonicus, Scrophularia ningpoensis and Chlorella vulgaris is significantly better than the effect of using them alone or in combination of two to enhance the moisturizing effect.

[0147] Application Examples

[0148] The skin softening lotion containing the extracts of Ophiopogon japonicus, Scrophularia ningpoensis and Chlorella vulgaris (ie Example 2) is used for skin moisturizing.

[0149] A skin softener, the formula of which is shown in Table 6.

[0150] Table 6 Toner formula

[0151]

[0152] The preparation method of the above-mentioned skin softening water includes the following steps: putting component A into a water phase pot, stirring at 80-85°C until completely dissolved; then cooling to 40-45°C, adding phase B, stirring evenly, discharging after passing the inspection, and the preparation is completed.

[0153] The moisturizing effect of the above-mentioned softening water was tested as follows:

[0154] Experimental design: 30 valid subjects were selected to participate in the test. The moisture content of the inner arm was tested on humans. The arm was divided into three areas, namely the blank control area (no sample was applied), the matrix area (the same as the preparation method of the skin softener in the application example, except that it does not contain the Chinese medicine extract complex of Example 2) and the sample area (skin softener group). Before the test, the staff cleaned the inner forearms of both hands of the subjects; the group number measurement area on the inner forearm of the subject was marked, and the initial value (0h) of the moisture of the skin stratum corneum was tested; the inner forearm of the subject was measured at (2.0±0.1) mg / cm 2 The dosage was evenly applied once to the blank control group, matrix group and sample group test area, and the blank control area (no sample was applied), matrix area, and sample test area were randomly distributed in the inner area of ​​the left or right forearm of the subject, and the skin moisture content test probe was used to test the moisture content of the stratum corneum of the skin 2 hours after application. The results are shown in Table 7.

[0155] Table 7 2h skin stratum corneum moisture content

[0156] Grouping Before use 2h after use Moisture content change Moisture content change rate blank 27.80 28.90 1.1 3.96％ Matrix Group 26.89 32.03 5.14 19.10％ Sample set 27.86 46.97 19.11 68.60％

[0157] As can be seen from Table 7, the use of the skin softener containing Ophiopogon japonicus, Scrophularia ningpoensis and Chlorella vulgaris extract (Example 2) can significantly increase the moisture content in the skin. This shows that the Chinese medicine extract complex composed of Ophiopogon japonicus, Scrophularia ningpoensis and Chlorella vulgaris extract can significantly increase the moisture content in the skin and has a good moisturizing effect.

[0158] The embodiments of the present invention are described in detail above in conjunction with the accompanying drawings, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present invention. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. A Chinese medicine extract complex comprising the following components: Ophiopogon japonicus extract, Scrophularia ningpoensis extract and Chlorella vulgaris extract.

2. The Chinese medicine extract complex according to claim 1, characterized in that: The traditional Chinese medicine extract compound comprises the following components in parts by weight: 1 to 10 parts of ophiopogon japonicus extract, 1 to 10 parts of scrophularia ningpoensis extract and 1 to 5 parts of chlorella vulgaris extract.

3. The Chinese medicine extract complex according to claim 1 or 2, characterized in that: The Ophiopogon japonicus extract is an ethanol extract of Ophiopogon japonicus; and / or, the Scrophularia ningpoensis extract is a water extract of Scrophularia ningpoensis; and / or, the Chlorella vulgaris extract is an alkali extract of Chlorella vulgaris.

4. Use of the Chinese medicine extract compound according to any one of claims 1 to 3 in any one of (1) to (8): (1) Preparation of cosmetics; (2) preparing products that inhibit skin moisture loss; (3) Increase the content of hyaluronic acid in cells; (4) preparing a product containing hyaluronic acid in cells; (5) Increase the expression of genes related to moisture retention in cells; (6) preparing products that increase the expression of genes related to moisture retention in cells; (7) Promote mitochondrial respiration of cells; (8) Prepare products that promote mitochondrial respiration in cells.

5. The use according to claim 4, characterized in that: The cells described in (3) to (8) include epidermal cells and dermal cells.

6. A cosmetic comprising the Chinese medicine extract complex according to any one of claims 1 to 3.

7. A skin softening lotion comprising the Chinese medicine extract complex according to any one of claims 1 to 3 and cosmetically acceptable excipients.

8. The skin softener according to claim 7, characterized in that: The cosmetically acceptable excipients include at least one of a chelating agent, a moisturizing agent, a thickening agent, an emulsifier, a pH adjuster, an antioxidant, a skin conditioning agent, an emollient and a preservative.

9. The skin softener according to claim 7 or 8, characterized in that: The mass percentage of the Ophiopogon japonicus extract in the skin softening water is 0.01% to 0.08%; and / or the mass percentage of the Scrophularia ningpoensis extract in the skin softening water is 0.01% to 0.05%; and / or the mass percentage of the Chlorella vulgaris extract in the skin softening water is 0.01% to 0.08%.

10. The method for preparing the skin softener according to any one of claims 7 to 9, comprising the following steps: mixing cosmetically acceptable excipients with water, dissolving at 80-90°C, cooling to 40-45°C, and mixing with a traditional Chinese medicine extract complex to obtain the skin softener.

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