Composition with effect of repairing skin barrier injury as well as preparation method and application of composition

Abstract

The invention provides a composition with a skin barrier injury repairing effect and a preparation method and application thereof, and belongs to the technical field of skin barrier repairing. The composition with the effect of repairing the skin barrier injury, provided by the invention, is prepared from the following raw materials in parts by weight: 16 to 24 parts of radix polygonati officinalis, 8 to 12 parts of rhizoma bletillae, 8 to 12 parts of radix sophorae flavescentis and 4 to 6 parts of liquorice root. According to the composition with the skin barrier injury repairing effect, the number of the traditional Chinese medicines is small, the four selected traditional Chinese medicines are properly and reasonably matched, and cell, animal and human body level experiments prove that the composition and the skin care product containing the composition have the obvious skin barrier repairing function and can be used for repairing the skin barrier injury. In addition, the moisturizing, anti-inflammatory and soothing effects are also realized. The composition provided by the invention has the advantages of easily available raw materials, simple preparation method, wide applicable population, safety, no irritation, long-term use and good application prospect.

Description

Technical Field

[0001] The present invention belongs to the technical field of skin barrier repair, and specifically relates to a composition having the effect of repairing skin barrier damage, and a preparation method and application thereof. Background Art

[0002] With increasing environmental pollution, enhanced ultraviolet radiation, and frequent extreme weather events, a damaged skin barrier has become a global health concern. Furthermore, modern skincare misconceptions such as over-reliance on deep cleansing, frequent exfoliation, and the misuse of acidic ingredients further accelerate the breakdown of this barrier function. The skin barrier, the body's first line of defense against external stimuli, consists of the stratum corneum and the sebum membrane. Impaired function can lead to decreased moisture retention, sensitive skin, redness, and frequent inflammation. This barrier function breakdown can contribute to the development of allergic diseases such as atopic dermatitis, food allergies, asthma, and allergic rhinitis, and can even be a major cause of recurrent disease. Clinically, epidermal barrier damage is typically treated with topical antiallergic antibiotics, which, while effective, can easily lead to drug dependency. As the pursuit of green skincare grows, Chinese herbal medicine, with its natural origins, multi-target repair mechanisms, and cultural recognition, is demonstrating unique value in contemporary skincare. However, existing traditional Chinese medicine compositions for facial skin repair suffer from a wide variety of medicinal flavors, which can lead to uncontrolled interactions between ingredients. Some have unpleasant odors (such as the fishy smell of Beauveria bassiana), resulting in a poor user experience. Alternatively, the availability of medicinal materials (such as ginseng and Ganoderma lucidum) can lead to economic and sustainability issues. Therefore, developing a natural skincare product with a minimal medicinal flavor that is effective and safe is crucial for preventing or repairing skin barrier damage. Summary of the Invention

[0003] In view of this, the purpose of the present invention is to provide a composition with a simple medicinal taste and the effect of repairing skin barrier damage, which has good barrier repair, moisturizing, anti-inflammatory and soothing effects and can significantly improve the facial skin barrier.

[0004] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0005] The invention provides a composition with the effect of repairing skin barrier damage. The composition comprises the following raw materials in parts by weight: 16-24 parts of polygonatum, 8-12 parts of bletilla striata, 8-12 parts of sophora flavescens and 4-6 parts of liquorice.

[0006] The present invention also provides a preparation method of the above-mentioned composition, comprising the following steps: mixing polygonatum, bletilla striata, sophora flavescens, liquorice and an ethanol solution, performing reflux extraction, collecting the medicinal residues and medicinal liquid 1; mixing the medicinal residues with the ethanol solution, performing reflux extraction, collecting medicinal liquid 2; mixing medicinal liquid 1 and medicinal liquid 2, and concentrating to obtain an extract; mixing the extract with water, performing water precipitation, removing the precipitate, and concentrating to obtain a composition.

[0007] Preferably, the volume fraction of the ethanol solution is 50% to 75%; the mass ratio of the total mass of Polygonatum odoratum, Bletilla striata, Sophora flavescens and Licorice to the ethanol solution is 1:6 to 10.

[0008] Preferably, the reflux extraction time is 0.5h to 2h, and the reflux extraction temperature is 80°C to 100°C.

[0009] Preferably, the mass ratio of the medicinal residue to the ethanol solution is 1:6-10.

[0010] Preferably, the mass volume ratio of the extract to water is 1 g: 2 mL to 6 mL; and the water precipitation time is 10 h to 15 h.

[0011] The present invention also provides the use of the above composition or the above preparation method in preparing a product for repairing skin barrier damage.

[0012] The present invention also provides the use of the above composition or the above preparation method in preparing products with anti-inflammatory, moisturizing and / or soothing effects.

[0013] The present invention also provides a skin care product with the effects of repairing skin barrier damage, anti-inflammatory, moisturizing and / or soothing, which is composed of the above composition and auxiliary materials, and the mass of the composition is 5% to 10% of the total mass of the skin care product.

[0014] Preferably, the types of the skin care products include cream, lotion, essence, toner or facial mask.

[0015] Beneficial effects of the present invention:

[0016] The present invention provides a composition for repairing skin barrier damage, featuring a limited number of ingredients and a well-selected, rational combination of four traditional Chinese medicines. Cell, animal, and human experiments have demonstrated that the composition and skin care products containing it significantly repair skin barrier function while also exhibiting moisturizing, anti-inflammatory, and soothing effects. The composition also boasts readily available raw materials, a simple preparation method, a broad range of applicability, and long-term safety, non-irritation, and promising application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 Effects of extracts extracted in different ways on the viability of normal HaCaT cells;

[0018] Figure 2 Effects of extracts extracted in different ways on the proliferation of HaCaT cells in the SDS-induced HaCaT injury model;

[0019] Figure 3Effects of extracts extracted by different methods on RAW264.7 cell viability;

[0020] Figure 4 The effects of extracts extracted in different ways on NO production in RAW264.7 cells;

[0021] Figure 5 This is a photo of the appearance of the facial cream of the present invention;

[0022] Figure 6 The effect of the cream of the present invention on skin lesions in a mouse model of skin mechanical barrier disorder;

[0023] Figure 7 The effect of the cream of the present invention on transepidermal water loss in a mouse model of skin mechanical barrier disorder, where * indicates p < 0.05, *** indicates p < 0.001, and NS indicates p ≥ 0.05;

[0024] Figure 8 The effects of different groups on skin pH in a mouse model of skin mechanical barrier disorder;

[0025] Figure 9 HE results of the effects of different groups on the epidermal thickness of the skin in the mouse model of skin mechanical barrier disorder (×10);

[0026] Figure 10 Statistical graph showing the effects of different groups on epidermal thickness in a mouse model of skin mechanical barrier disorder, where *** and ### indicate p < 0.001, and ns indicates p ≥ 0.05;

[0027] Figure 11 The effects of different groups on skin ceramide in the mouse model of skin mechanical barrier disorder, where * indicates p < 0.05, *** and ### indicate p < 0.001;

[0028] Figure 12 The results of the effect of the cream of the present invention on the transepidermal water loss of the test subjects;

[0029] Figure 13 The following are test images of the hemoglobin content in the skin of two subjects. DETAILED DESCRIPTION

[0030] The invention provides a composition with the effect of repairing skin barrier damage. The composition comprises the following raw materials in parts by weight: 16-24 parts of polygonatum, 8-12 parts of bletilla striata, 8-12 parts of sophora flavescens and 4-6 parts of liquorice.

[0031] The present invention has no particular limitation on the specific sources of the above raw materials; conventional commercially available raw materials in the art can be used. In the present invention, the amount of Polygonatum odoratum is preferably 18-22 parts, more preferably 19-21 parts; the amount of Bletilla striata is preferably 9-11 parts, more preferably 10 parts; the amount of Sophora flavescens is preferably 9-11 parts, more preferably 10 parts; and the amount of Licorice is preferably 5 parts.

[0032] In the composition described in the present invention, Polygonatum odoratum has the effect of improving the skin's moisturizing ability and promoting the proliferation and differentiation of keratinocytes. Its main active ingredient, polysaccharides (such as steroidal saponins and flavonoid glycosides), can increase the expression of filaggrin and strengthen the skin's water-locking barrier; activate aquaporins and improve the dry state. Bletilla striata strengthens the skin's physical barrier structure and promotes damage repair. Its main active ingredients, bibenzyl and phenanthrene components, can promote collagen synthesis and accelerate epidermal wound healing; the polysaccharide components form a moisturizing film and reduce transepidermal water loss. Sophora flavescens can inhibit skin inflammatory reactions and the proliferation of pathogenic microorganisms. Its active ingredients, matrine and oxymatrine, reduce the release of inflammatory factors (TNF-α, IL-6) by inhibiting the NF-κB pathway; flavonoid components can inhibit the growth of pathogenic bacteria such as Staphylococcus aureus and Propionibacterium acnes. Licorice can synergistically fight inflammation and antioxidants and balance the overall activity of the composition. Its active ingredients, glycyrrhizic acid and glycyrrhetinic acid, inhibit histamine release and COX-2 enzyme activity, reducing redness, swelling and itching; flavonoid components scavenge free radicals and reduce oxidative stress damage.

[0033] The composition of the present invention achieves skin barrier repair through the following synergistic pathways: Polygonatum odoratum, as the main drug, has the effect of nourishing yin and moistening dryness, can nourish the skin, repair the skin barrier, and improve the dry skin condition. Bletilla striata, as the ministerial drug, has the effects of astringing and stopping bleeding, reducing swelling and promoting tissue regeneration, assisting Polygonatum odoratum in nourishing the skin and helping to alleviate skin inflammation. Sophora flavescens, as the adjuvant drug, has the effects of clearing away heat and detoxifying, drying dampness and relieving itching, and helps to alleviate skin inflammation and skin itching. Licorice, as the guiding drug, has the effects of harmonizing medicinal properties and detoxifying, and plays a role in harmonizing various drugs, making the prescription more balanced. At the same time, its detoxifying effect helps to relieve skin inflammation and promote the repair of the skin barrier.

[0034] The present invention also provides a preparation method of the above-mentioned composition, comprising the following steps: mixing polygonatum, bletilla striata, sophora flavescens, liquorice and an ethanol solution, performing reflux extraction, collecting the medicinal residues and medicinal liquid 1; mixing the medicinal residues with the ethanol solution, performing reflux extraction, collecting medicinal liquid 2; mixing medicinal liquid 1 and medicinal liquid 2, and concentrating to obtain an extract; mixing the extract with water, performing water precipitation, removing the precipitate, and concentrating to obtain a composition.

[0035] The present invention uses alcohol extraction and water precipitation to achieve better barrier repair and anti-inflammatory effects, and is highly safe. In the present invention, the low-concentration ethanol solution (<50%) has a high solvent polarity, close to water, and is suitable for extracting water-soluble components in the composition, such as polysaccharides, saponins (partially water-soluble saponins), and proteins and amino acids. The main skin care functions of the extracted components are moisturizing and barrier repair, but the anti-inflammatory ability is insufficient. The medium-concentration ethanol solution (50% to 75%) has a moderate ratio of ethanol to water, taking into account both polarity and fat solubility, and is suitable for extracting medium-polarity components in the composition, such as flavonoids, alkaloids, saponins, etc. The main skin care functions of the extracted components are moisturizing and barrier repair, and can also take into account anti-inflammatory and antibacterial effects. Among them, glycyrrhizic acid, the core anti-inflammatory component in licorice, is best extracted in medium-concentration ethanol. High-concentration ethanol (>75%) has a high ethanol concentration and low polarity, and is suitable for extracting fat-soluble components, such as volatile oils, sterols, fat-soluble pigments, etc. The main skin care functions of the extracted ingredients are antibacterial and oil control, but the moisturizing and barrier repair ingredients are relatively small. In order to ensure the efficacy of the composition of the present invention, the volume fraction of the ethanol solution is preferably 50% to 75%, more preferably 65% ​​to 72%.

[0036] In the present invention, the mass ratio of the total mass of Polygonatum odoratum, Bletilla striata, Sophora flavescens, and Glycyrrhiza uralensis to the ethanol solution is preferably 1:6-10, more preferably 1:7-9. This mass ratio of the total mass of the Chinese medicinal raw materials to the ethanol solution as defined in the present invention ensures a high extraction effect while preventing solvent waste due to excessive addition of ethanol solution. In the present invention, if the reflux extraction temperature is too low, the extraction of the active ingredients will be poor, while if it is too high, the activity of the active ingredients will be weakened or even lost. The reflux extraction temperature is preferably 80°C-100°C, more preferably 85°C-95°C. To minimize the manpower and material resources required for extraction while ensuring efficacy, the reflux extraction is preferably performed twice, and the reflux extraction time is preferably 0.5h-2h, more preferably 1h-1.5h. In the present invention, the mass ratio of the medicinal residue to the ethanol solution is preferably 1:6-10, more preferably 1:7-9. The present invention does not specifically limit the specific method of concentration; conventional concentration methods in the art can be used. The extract preferably contains 0.8g / mL-1.2g / mL of the medicinal material. In the present invention, the mass volume ratio of the extract to water is preferably 1g:2mL~6mL, more preferably 1g:3mL~4mL, and the water precipitation time is preferably 10h~15h, more preferably 11h~14h. In the present invention, when the precipitate is concentrated, it is preferably concentrated to a concentration of the medicinal material in the composition of 0.8g / mL~1.2g / mL.

[0037] The present invention also provides the use of the above composition or the above preparation method in preparing a product for repairing skin barrier damage or in preparing a product with anti-inflammatory, moisturizing and / or soothing effects. In the present invention, the type of the product preferably includes skin care products.

[0038] The present invention also provides a skin care product with the effects of repairing skin barrier damage, anti-inflammatory, moisturizing and / or soothing, which is composed of the above composition and auxiliary materials, and the mass of the composition is 5% to 10% of the total mass of the skin care product.

[0039] In the skin care product of the present invention, the excipients preferably include those acceptable in the cosmetics field, and the skin care product is administered topically. Preferred types of skin care products include face creams, lotions, serums, toners, or facial masks. When the skin care product is a face cream, the excipients preferably include one or more of an emollient, an emulsifier, a humectant, a thickener, a preservative, a penetration enhancer, and an antioxidant. In the present invention, the emollient is preferably one or more of isononyl isononanoate, C9-12 alkanes, polydimethylsiloxane, and coconut oil caprylate / caprate. The emulsifier is preferably one or more of polyglyceryl-3 cetyl ether, sesame seed oil, malic acid, polyglyceryl-6 distearate, jojoba esters, cetyl alcohol, and polyglyceryl-3 beeswax. The humectant is preferably one or more of butylene glycol, glycerin, 1,2-hexanediol, hydrolyzed sodium hyaluronate, and sodium hyaluronate. The thickener is preferably one or more of ammonium acryloyldimethyltaurate / VP copolymer, xanthan gum, and behenyl alcohol. The preservative is preferably one or more of phenoxyethanol, ethylhexylglycerin, and p-hydroxyacetophenone. The penetration enhancer is preferably one or more of phosphatidylcholine, hydrogenated lecithin, and ethoxydiglycol. The antioxidant is preferably one or more of ascorbyl palmitate and tocopherol (vitamin E). In the present invention, to further enhance the repairing effect of the topical cream, active ingredients such as Centella asiatica extract, allantoin, niacinamide, and ceramide NS / NG / NP / AP may be added.

[0040] In the present invention, when the skin care product is a facial cream, the dosage of the facial cream is preferably 0.1g to 10g per application, more preferably 0.5g to 5g per application, and most preferably 1g to 3g per application. Alternatively, depending on the area of ​​application, the application amount of the facial cream is preferably 0.01g to 0.5g per square centimeter of skin, more preferably 0.05g to 0.2g per square centimeter of skin. The facial cream can preferably be applied 1 to 4 times daily, more preferably 1 to 2 times daily. The specific dosage and frequency of application can be appropriately adjusted based on the user's age, weight, specific application area, and individual response.

[0041] In vitro experiments have shown that the composition provided by the present invention can significantly promote the proliferation of normal HaCaT cells and effectively reduce SDS-induced HaCaT cell damage, demonstrating its potential for skin repair. The composition can also promote the proliferation of RAW264.7 macrophages and significantly inhibit LPS-induced nitric oxide production in these cells, demonstrating its anti-inflammatory activity. Furthermore, the composition of the present invention exhibits an inhibitory effect on hyaluronidase in in vitro experiments, suggesting its moisturizing effect.

[0042] In vivo experiments (mouse model) showed that a cream containing the composition of the present invention significantly reduced transepidermal water loss (TEWL) in a mouse model of skin barrier damage, effectively lowered skin surface pH, and reduced the compensatory increase in ceramide synthesis triggered by barrier damage. These results suggest that a cream containing the composition of the present invention has the effect of repairing epidermal barrier function.

[0043] Human efficacy evaluation (volunteer testing) showed that after 28 days of continuous topical application of a cream containing the composition of the present invention, transepidermal water loss (TEWL) and hemoglobin levels were significantly lower than initial values ​​(indicating a reduction in erythema). These results demonstrate that creams containing the composition of the present invention effectively strengthen the skin barrier function, improve the condition of damaged skin, and exhibit significant skin repair efficacy.

[0044] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0045] In the following examples, unless otherwise specified, all methods are conventional.

[0046] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0047] Example 1

[0048] A composition with the effect of repairing skin barrier damage is prepared from 20 parts of polygonatum, 10 parts of bletilla striata, 10 parts of sophora flavescens and 5 parts of liquorice.

[0049] The preparation method is:

[0050] Mixing polygonatum, bletilla striata, sophora flavescens, and liquorice with a volume fraction of 70% ethanol solution, wherein the mass ratio of the total mass of polygonatum, bletilla striata, sophora flavescens, and liquorice to the ethanol solution is 1:8, performing reflux extraction at 90°C for 1 hour, and collecting the medicinal residues and medicinal liquid 1; mixing the medicinal residues with a volume fraction of 70% ethanol solution that is 8 times the mass of the medicinal residues, performing reflux extraction at 90°C for 1 hour, and collecting medicinal liquid 2; mixing medicinal liquid 1 and medicinal liquid 2, and concentrating to 90 g to obtain an extract; mixing the extract with water at a mass-to-volume ratio of 1 g:4 mL, performing water precipitation, and water precipitation at 4°C for 12 hours, removing the precipitate, and concentrating by rotary evaporation until the extract has a volume of 1 g / mL to obtain a composition.

[0051] Example 2

[0052] A composition with the effect of repairing skin barrier damage is prepared from 16 parts of polygonatum, 8 parts of bletilla striata, 8 parts of sophora flavescens and 4 parts of liquorice.

[0053] The preparation method is:

[0054] Polygonatum odoratum, Bletilla striata, Sophora flavescens, and Glycyrrhiza uralensis are mixed with a 50% ethanol solution by volume, wherein the mass ratio of the total mass of the Polygonatum odoratum, Bletilla striata, Sophora flavescens, and Glycyrrhiza uralensis to the ethanol solution is 1:6, and the mixture is refluxed at 80°C for 2 hours, and the medicinal residue and medicinal liquid 1 are collected; the medicinal residue is mixed with a 50% ethanol solution by volume that is 6 times the mass of the medicinal residue, and the mixture is refluxed at 80°C for 2 hours, and medicinal liquid 2 is collected; the medicinal liquid 1 and the medicinal liquid 2 are mixed and concentrated to 80 g to obtain an extract; the extract is mixed with water at a mass-to-volume ratio of 1 g:2 mL, and the mixture is precipitated in water at 4°C for 10 hours, the precipitate is removed, and the mixture is concentrated by rotary evaporation to an extract of 0.8 g / mL to obtain a composition.

[0055] Example 3

[0056] A composition with the effect of repairing skin barrier damage is prepared from 24 parts of polygonatum, 12 parts of bletilla striata, 12 parts of sophora flavescens and 6 parts of liquorice.

[0057] The preparation method is:

[0058] Mixing polygonatum, bletilla striata, sophora flavescens, and liquorice with a volume fraction of 75% ethanol solution, wherein the mass ratio of the total mass of polygonatum, bletilla striata, sophora flavescens, and liquorice to the ethanol solution is 1:10, performing reflux extraction at 100°C for 0.5h, and collecting the medicinal residues and medicinal liquid 1; mixing the medicinal residues with a volume fraction of 75% ethanol solution that is 10 times the mass of the medicinal residues, performing reflux extraction at 100°C for 0.5h, and collecting medicinal liquid 2; mixing medicinal liquid 1 and medicinal liquid 2, and concentrating to 100g to obtain an extract; mixing the extract with water at a mass-to-volume ratio of 1g:6mL, performing water precipitation, and performing water precipitation at 4°C for 15h, removing the precipitate, and concentrating by rotary evaporation until the extract has a volume of 1.2g / mL to obtain a composition.

[0059] Comparative Example 1

[0060] The difference from Example 1 is the preparation method. The rest is the same as Example 1. The preparation method is:

[0061] Mix polygonatum, bletilla striata, sophora flavescens, licorice and water, with the mass ratio of the total mass of polygonatum, bletilla striata, sophora flavescens and licorice to water being 1:8. Decoction and extract for 1 hour, then filter. Add 8 times the mass of water to the residue again, extract for 1 hour, filter, combine the two filtrates, and concentrate to an extract of 1 g / mL to obtain a water extract for later use.

[0062] Comparative Example 2

[0063] The difference from Example 1 is that the preparation method is different, and the preparation method is:

[0064] Mix Polygonatum odoratum, Bletilla striata, Sophora flavescens, and Glycyrrhiza uralensis with water in a mass ratio of 1:8. Decoction and extract for 1 hour, then filter. Add 8 times the weight of the residue to water and decoct and extract for another hour. Filter. Combine the two filtrates, concentrate to 90 mL, add 360 mL of 70% ethanol, and incubate at 4°C for 12 hours. Remove the precipitate by suction, and concentrate the extract by rotary evaporation to 1 g / mL. This extract is the aqueous-alcoholic precipitate and set aside.

[0065] Comparative Example 3

[0066] The difference from Example 1 is the preparation method. The rest is the same as Example 1. The preparation method is:

[0067] Mix polygonatum, bletilla striata, sophora flavescens, and licorice with a 70% volume fraction ethanol solution, with the mass ratio of the total mass of polygonatum, bletilla striata, sophora flavescens, and licorice to the 70% volume fraction ethanol solution being 1:8, perform reflux extraction at 90°C for 1 hour, and collect the medicinal residues and medicinal solution 1; mix the medicinal residues with a 70% volume fraction ethanol solution 8 times the mass of the medicinal residues, perform reflux extraction at 90°C for 1 hour, and collect medicinal solution 2; mix medicinal solutions 1 and 2, and concentrate the extract to 1 g / mL to obtain an alcohol extract for later use.

[0068] Test Example 1

[0069] In vitro experiments on the extracts obtained from Example 1 and Comparative Examples 1 to 3

[0070] (1) Human immortalized epidermal cells (HaCaT) cell viability assay

[0071] HaCaT cells were cultured at 6×10 4The cells were inoculated in a 96-well plate at a density of 100 μL / well and placed in an incubator for 24 hours. The extracts of Example 1 (alcohol extraction and water precipitation), Comparative Example 1 (water extraction), Comparative Example 2 (water extraction and alcohol precipitation) and Comparative Example 3 (alcohol extraction) were added and incubated for 24 hours. Six replicates were set up in each group. The culture medium in the 96-well plate was then removed, and 100 μL of serum-free culture medium containing 10% CCK8 solution was added to each well. After incubation in the incubator for 1 hour, the cells were taken out and the OD value of each well was detected at 450 nm using a microplate reader to calculate the proliferation of HaCaT cells. Cell viability (%) = (OD value of the experimental group - OD value of the blank control group) / (OD value of the negative group - OD value of the blank control group) × 100%.

[0072] The results are as follows Figure 1 As shown, the extracts (extracts) prepared by the alcohol extraction and alcohol extraction and water precipitation methods showed a good effect on promoting cell viability, and showed a clear concentration dependence. The extract (extract) prepared by the water extraction method had almost no cytotoxicity to HaCaT cells, but had no significant pro-proliferation effect. The water extraction and alcohol precipitation method showed some cytotoxicity at concentrations above 25 μg / mL.

[0073] (2) Cell viability detection in the SDS-induced HaCaT injury model

[0074] 10,000 HaCaT cells per well were seeded in a 96-well plate and incubated overnight in a cell culture incubator. The supernatant was then removed by aspiration, 100 μL of 80 μg / mL SDS solution was added, and the cells were incubated for another 24 h. The supernatant was discarded, and extract solutions of different concentrations from Example 1 (alcohol extraction and water precipitation), Comparative Example 1 (water extraction), Comparative Example 2 (water extraction and alcohol precipitation), and Comparative Example 3 (alcohol extraction) were added, incubated for 24 h, and cell viability was detected using the CCK-8 method. The control group was a group without SDS and extract, and the model group was a group with SDS but no extract.

[0075] The results are as follows Figure 2 As shown in the results, after SDS treatment, the survival rate of HaCaT cells decreased significantly, while after administration of the extract (extract) of the present invention, the cell survival rate was significantly improved. Among them, the Chinese herbal extracts (extracts) prepared by alcohol extraction and alcohol extraction water precipitation methods with a crude drug concentration of 100 μg / mL or above have good epidermal barrier repair effects.

[0076] (3) Viability detection of mouse monocyte macrophage RAW264.7 cells

[0077] Take macrophages in the logarithmic growth phase and scrape them with a disposable spatula. Centrifuge at 1000 rpm for 4 minutes and remove the supernatant. Count the corresponding number of cells and add the corresponding culture medium to 1×10 5The density of cells / mL was inoculated into 96-well plates, 100 μL per well. The experiment set up an untreated control group, each extract embodiment 1 (alcohol extraction and water precipitation), comparative example 1 (water extraction), comparative example 2 (water extraction and alcohol precipitation) and comparative example 3 (alcohol extraction) different dosage groups, 6 replicates per group, and the total incubation time was 24h. Afterwards, the 96-well plate culture medium was removed, the culture solution containing 10% CCK8 was prepared, and the cells were incubated in a cell culture incubator for 1h. The OD450 value was measured using a microplate reader to calculate the cell survival rate. Cell survival rate (%) = (extract group OD value-blank OD value) / (untreated control group OD value-blank OD value) × 100%, the blank group refers to the PBS well, and the untreated control group refers to the macrophage + culture medium culture, and the effect of different concentrations of extract on macrophage activity was evaluated.

[0078] The results are as follows Figure 3 As shown, the extract (extract) of the present invention prepared by the alcohol extraction and water precipitation method showed a good effect in promoting the viability of RAW264.7 cells, and showed a clear concentration-dependent effect. The extracts (extracts) prepared by the alcohol extraction and water extraction and alcohol precipitation methods had a second-best effect in promoting the proliferation of RAW264.7 cells. The extract (extract) prepared by the water extraction method showed some cytotoxicity.

[0079] (4) Detection of NO content in RAW264.7 cell inflammation model

[0080] 1×10 5 Cells were seeded in a 12-well plate at a density of 1 mL / well and cultured for 12 h (24 h). After adherence, different concentrations of extracts from Example 1 (ethanol extraction and water precipitation), Comparative Example 2 (water extraction and alcohol precipitation), and Comparative Example 3 (ethanol extraction) were added and incubated for 24 h. LPS (final concentration of 1.0 μg / mL) and IFN-γ (final concentration of 4 ng / mL) were then added to the treatment group and the model control group. Only culture medium was added to the blank control group and cultured for another 12 h (24 h). The supernatant was collected. The specific steps were performed according to the instructions of the NO test kit. OD values ​​were measured at 540 nm, and the NO content and inhibition rate were calculated. The NO content was substituted into the standard curve for calculation. NO inhibition rate (%) = (average NO content in the model control group - average NO content in the treatment group) / average NO content in the model control group × 100%, where the model control group refers to the group that only added LPS to stimulate macrophages.

[0081] The results are as follows Figure 4 As shown, the extract (extract) prepared by the alcohol extraction and water precipitation method in Example 1 inhibited the NO level most significantly, indicating that it had an anti-inflammatory effect and the best effect.

[0082] (5) In vitro hyaluronidase inhibition study

[0083] First, a standard curve for the enzyme activity of hyaluronidase reacting with the substrate HA was constructed using the DNS method. The reaction solution was prepared according to Table 1, mixed by vortexing, and reacted at 37°C for 30 minutes, followed by a boiling water bath for 5 minutes. 200 μL of the reaction solution was added to 400 μL of the DNS solution, mixed by vortexing, and then heated in a boiling water bath for 5 minutes. 150 μL of sample was added to each well of a 96-well plate, with three replicates per well. The A540 value was measured using a microplate reader. A standard curve was constructed by plotting the OD value at A540 against the enzyme concentration. The results showed good linearity within the hyaluronidase addition range of 200 to 500 μL (i.e., hyaluronidase solution concentrations of 400 to 1000 U / mL). The linear equation was: y = 0.0004x + 0.2338, r = 0.9940. The raw data are shown in Table 1.

[0084] Table 1 Reaction solution

[0085]

[0086]

[0087] Preparation contains the extract that the different concentrations embodiment 1 (alcohol extraction and water precipitation), comparative example 1 (water extraction), comparative example 2 (water extraction and alcohol precipitation) and comparative example 3 (alcohol extraction) of 600U / mL hyaluronidase make, the concentration range of wherein contained extract is 0~0.05% (the enzymatic activity of hyaluronidase solid powder is 400~1000U / mg, calculates according to 500U / mg), standby.Get each group of solution 500 μ L, add 0.5%HA500 μ L respectively, 37 ℃ of water-baths 30min, boiling water bath 5min makes enzyme inactivation afterwards, get 200 μ L reaction solutions, add DNS solution 400 μ L, boiling water bath 5min, get 150 μ L and add 96 orifice plates, every sample establishes 3 replicate holes, microplate reader measures A540 value, according to the linear regression equation calculation solution residual enzyme activity, the results are shown in Table 2.

[0088] Table 2 Enzyme activity

[0089]

[0090] As can be seen from Table 2, the extracts from different processes all inhibited hyaluronidase, and the higher the concentration, the better the inhibition effect. Among them, the water extraction and water extraction and alcohol precipitation processes had the best enzyme inhibition rates, and the alcohol extraction and water precipitation process was the second most effective.

[0091] The above results indicate that all four extracts significantly promoted the proliferation of SDS-damaged HaCaT cells. At extract concentrations of 50-150 μg / mL, the proliferation effect ranked as follows: alcohol extraction and water precipitation > water extraction and alcohol precipitation > water extraction > alcohol extraction. In the LPS-induced inflammation model, alcohol extraction and water precipitation > water extraction and alcohol precipitation > alcohol extraction were the most effective methods. Water extraction was excluded due to significant cytotoxicity (cell viability <80%). Hyaluronidase inhibition (moisturizing) revealed that the inhibitory activity of each process ranked as follows: water extraction > water extraction and alcohol precipitation > alcohol extraction and water precipitation > alcohol extraction.

[0092] Based on a comprehensive assessment of efficacy balance and safety: Core efficacy weight distribution: Barrier repair must prioritize proliferation promotion (weight 40%) and anti-inflammation (weight 40%), with moisturizing as an auxiliary function (weight 20%). In the present invention, the alcohol extraction and water precipitation method is determined to be the best process because it performs dual optimality in core efficacy (proliferation promotion and anti-inflammation). Although the moisturizing activity is not the highest, it is still within the effective range and has a certain moisturizing activity (inhibition rate>30%); and the alcohol extraction and water precipitation method can completely avoid the risk of cytotoxicity.

[0093] Example 4

[0094] A facial cream having the effects of repairing skin barrier damage, anti-inflammatory, moisturizing and / or soothing, the preparation method of which is as follows:

[0095] Weigh 71 g of water, 2.6 g of butylene glycol, 7 g of the composition obtained in Example 1, 0.86 g of 1,2-hexanediol, 0.5 g of ammonium acryloyldimethyltaurate / VP copolymer, 0.2 g of allantoin, 0.07 g of hydrolyzed sodium hyaluronate, 0.05 g of xanthan gum, and 0.02 g of sodium hyaluronate and heat to 85°C (referred to as phase A);

[0096] In another container, 3 g of isononyl isononanoate, 2.8 g of polyglyceryl-3 cetyl ether, 0.525 g of sesame seed oil, 0.175 g of malic acid, 2.7 g of C9-12 alkane, 2 g of behenyl alcohol, 0.64 g of polyglyceryl-6 distearate, 0.19 g of jojoba esters (purchased from Shanghai Puen Biochemical Technology Co., Ltd.), 0.085 g of cetyl alcohol, 0.085 g of polyglyceryl-3 beeswax ester, 1 g of polydimethylsiloxane, 0.3 g of coconut oil caprylate / caprate, and 0.1 g of HDI / trimethylol hexyl lactone crosspolymer were added, and the raw materials were heated to 85° C., recorded as phase B;

[0097] Add the raw materials of phase B into phase A, homogenize, and maintain the temperature at 85℃ for emulsification; cool to 75℃, add 1g of isononyl isononanoate and homogenize; cool to 60℃, add 2g of 1,2-pentanediol, 0.09g of ethylhexylglycerin, and 0.00075g of p-hydroxyacetophenone; continue to cool to 40℃, add 0.5g of Centella asiatica extract, 0.2g of niacinamide, 0.06g of p-hydroxyacetophenone, 0.00002g of glycerol, 0.00001g of phosphatidylcholine, 0.0000003g of ceramide NP, and 0.0000003g of ceramide AP. 0.0000002g and hydrogenated lecithin, ethoxydiglycol, ascorbyl palmitate, 1,2-hexanediol, DZ052 composite ceramide liposome (purchased from Shanghai Zhina Biotechnology Co., Ltd., raw material number 0106326) 0.0000005g, tocopherol (vitamin E) 0.00009g, finally add water to the total amount of 100g, stir evenly, and obtain a face cream. Among them, the appearance of the face cream is shown in the figure Figure 5 .

[0098] Test Example 2

[0099] In vitro experiment of the cream obtained in Example 4

[0100] 1. Experimental Materials

[0101] 1.1 Experimental Reagents

[0102] Sodium hydroxide (AR), concentrated hydrochloric acid (GR), glacial acetic acid (AR), p-dimethylaminobenzaldehyde (AR), anhydrous sodium carbonate (AR), anhydrous sodium acetate (AR), anhydrous calcium chloride (AR), acetylacetone (AR), sodium hyaluronate (USP), hyaluronidase (BR); dipotassium glycyrrhizate, purity ≥98%.

[0103] 1.2 Test substances

[0104] Sample: the cream prepared in Example 4 diluted into a 10% aqueous solution;

[0105] Negative / solvent control: buffer;

[0106] Positive control: 1% dipotassium glycyrrhizate solution.

[0107] 2. Experimental Methods

[0108] Set up four groups, A, B, C, and D, with three replicates per group. Add calcium chloride, buffer solution, sample solution, sodium hydroxide, and acetylacetone solution to test tubes, mix thoroughly, and then place in boiling water, ice water, and room temperature for reaction. Add P-DAB colorimetric reagent to all four tubes. After the reaction is complete, measure the absorbance of each group at 530 nm.

[0109] Hyaluronidase inhibition rate (%) = [(CD) - (AB)] / (CD) × 100%

[0110] Wherein: A is the absorbance of the reaction solution containing sample and enzyme; B is the absorbance of the reaction solution containing sample and no enzyme; C is the absorbance of the reaction solution containing enzyme but no sample; D is the absorbance of the reaction solution without sample and enzyme.

[0111] Under the condition of ensuring the effectiveness of the test system (hyaluronidase inhibition rate of the positive control > 50%), if the hyaluronidase inhibition rate of the test sample is higher than that of the negative control, and there is a significant difference between the two (P < 0.05), it can be determined that the test sample has a soothing effect.

[0112] 3. Experimental Results

[0113] According to the laboratory method (XYJC-SOP-CE-021 hyaluronidase inhibition rate method), the soothing effect of the facial cream prepared in Sample Example 4 was tested. The test results are shown in Table 3, which shows that when the experimental system is effective (hyaluronidase inhibition rate of the positive control is >50%), when the sample concentration is 10%, the hyaluronidase inhibition rate is 43.888%, which is significantly different from that of the negative control (P<0.05), indicating that the sample has a soothing effect.

[0114] Table 3 Test results

[0115] name Test results unit P-value Example 4 obtained facial cream 43.888±5.735 ％ <0.05 Negative control group 0.802±1.388 ％ / Positive control group 85.972±4.349 ％ <0.05

[0116] Test Example 3

[0117] In vivo experiment of the cream prepared in Example 4

[0118] (1) Establishment of a mouse model of skin mechanical barrier disorder

[0119] Twenty male BALB / c mice were randomly divided into a blank group, a model group, a matrix control group and a drug administration group, with 5 mice in each group. Except for the blank control group, a skin mechanical barrier damage model was established by the tape method. The specific method is as follows: stick the medical tape tightly to the shaved mouse back skin and then quickly tear it off. Each mouse was stimulated twice a day for modeling, and the "tape sticking-tearing off" process was repeated 5 times each time for each modeling. The interval between the two modeling stimulations was 8 hours, and the modeling process lasted for 5 consecutive days. The modeling was successful when the mouse skin was red but without bleeding. After that, the mice in the matrix group and the drug administration group were given a single cream (the difference between the single cream here and Example 4 is that it does not contain the composition of Example 1, and the rest are the same as Example 4) and a face cream intervention, 5 mg / mouse, for 5 consecutive days. At the same time, the blank group and the model group did not receive any intervention. The specific administration method is as follows: use a syringe to draw 5 mg of cream or face cream and apply it to the damaged part of the mouse skin mechanical barrier. After 24 hours of each administration, the condition of the damaged skin tissue was observed.

[0120]

[0121] The results are as follows Figure 6 As shown in the figure, after modeling, the modeling sites on the backs of mice in each group showed symptoms such as redness, sloughing, and thickening, indicating that the modeling was successful. After the last administration of each group, the skin of mice in the matrix group was still red and swollen, while the redness and swelling of the skin of mice in the administration group (cream prepared in Example 4) subsided or showed a significant trend of subsiding. After the last administration, the skin barrier recovery rate of mice in the administration group was statistically significant compared with that of the model group and the matrix group, and was very significant. There was no significant difference between the model group and the matrix group. This shows that the cream obtained in Example 4 has a good skin barrier repair function.

[0122] (2) Determination of transepidermal water loss

[0123] 24 hours after the last administration, the transepidermal water loss (TEWL) of the mouse back skin was measured using a skin detector (Tewametermobile). Transepidermal water loss (TEWL), also known as transdermal water loss, indicates the loss of water from the deep dermis through evaporation of the epidermis. It is an important parameter to describe the skin barrier and is closely related to the water content of the skin's stratum corneum. The higher the TEWL value, the more water is lost through the skin and the poorer the barrier function of the stratum corneum. Testing environment: room temperature 20-25°C, relative humidity 50%-60%. During the test, the instrument is in full contact with the skin of the test area and kept as vertical as possible. Repeat the measurement multiple times on the same test area and take the average value.

[0124] The results are as follows Figure 7As shown, the TEWL values ​​of mice increased after modeling. Compared with the blank group, the TEWL values ​​of mice in the model and matrix groups increased, and there was a statistically significant difference. The TEWL values ​​of mice in the treatment group showed significant improvement compared with the model group, and there was no statistical difference compared with the blank group, indicating that the cream can promote the repair of the skin's stratum corneum barrier.

[0125] (3) Skin pH measurement

[0126] 24 hours after the last dose, measure the pH of the mouse back skin using a pH meter. Testing environment: room temperature 20-25°C, relative humidity 50%-60%. During the test, the instrument should be in full contact with the skin of the test site and kept as vertical as possible. Repeat the measurement three times at the same test site, and calculate the average value.

[0127] See the results Figure 8 The skin pH value increased after the skin barrier was damaged. Compared with the blank group, the pH value of the skin of mice in the matrix group and the model group increased. The pH value of the skin of mice in the drug group decreased significantly and was close to that of the blank group, indicating that the cream has a good skin barrier repair function.

[0128] (5) HE staining and ceramide determination

[0129] Twenty-four hours after the last dose, the hair around the dorsal area of ​​the rat's ear was shaved and the rat was anesthetized with isoflurane. After anesthesia, a 1.25 cm × 1.25 cm section of skin from the mouse's back where the model was created was obtained. A portion was fixed in 4% paraformaldehyde solution for subsequent tissue embedding, sectioning, and staining. The remaining portion was frozen in liquid nitrogen until further use, and ceramide content in the skin was subsequently determined by ELISA.

[0130] The results are as follows Figures 9 to 11 The results of the effect of the cream prepared in Example 4 on the epidermal thickness of barrier disorder mice are shown in Figure 9 and Figure 10 The epidermal tissue structure of mice in the blank group was intact, while the epidermis of mice in the model group was significantly thickened, with partial stratum corneum shedding, acanthosis and downward protrusions, accompanied by inflammatory cell infiltration. The cream prepared in Example 4 showed significant improvement after application, and compared with the matrix group, the cream group was able to significantly reduce epidermal thickness.

[0131] The results of the effect of the cream prepared in Example 4 on the ceramide in the skin of barrier disorder mice are shown in Figure 11 Compared with the blank group, the ceramide content in the model group was significantly increased. After applying the blank matrix and the cream prepared in Example 4, the ceramide content decreased, and the difference in the cream group was significant compared with the blank matrix group (P<0.05).

[0132] Test Example 4

[0133] Human body efficacy experiment of the facial cream prepared in Example 4

[0134] 1. Test sample information: facial cream prepared in Example 4.

[0135] 2. Test basis: T / CAB 0152-2022 "Test Methods for Cosmetics for Anti-wrinkle, Firming, Moisturizing, Oil Control, Repairing, Nourishing, and Soothing Efficacy".

[0136] 3. Test purpose: To evaluate the effectiveness and safety of skin improvement after subjects use repair cosmetics.

[0137] 4. Test design:

[0138] Planned number of recruits: 32, number of screened: 31, number of enrolled: 30.

[0139] Test period: 2025.01.08~2025.02.05.

[0140] 5. Test environment: The test should be carried out in a constant environment with a temperature of 21±1°C and a relative humidity of 50±10%RH.

[0141] 6. Test instruments: VISIA-7 skin image analyzer, HQ-TEWL transepidermal moisture tester, skin melanin and hemoglobin tester ( MX18).

[0142] 7. Test method:

[0143] 7.1 Volunteer requirements:

[0144] Inclusion criteria: healthy men or women aged between 18 and 60; those who judged whether they had sensitive skin based on subjective symptoms, excluding subjects with non-sensitive skin; those who felt that their skin had symptoms such as dryness, scaling, and redness; those who could understand the testing process, voluntarily participated in the trial and signed the informed consent form; and those who could use the product according to the required dosage.

[0145] Exclusion criteria: pregnant or breastfeeding women, or those who are planning to get pregnant in the near future; those who have used any anti-inflammatory drugs on the test area in the past two months; those who have clinically unhealed inflammatory skin diseases; those who have had an allergic disease attack and treatment or used hormones or other anti-inflammatory drugs in the past month; those whose test results are affected by scars, pigmentation, atrophy, port-wine stains or other blemishes on the skin to be tested; those who have participated in related clinical trials in the past three months; and those who are deemed unsuitable for participation in the trial by other clinical assessments.

[0146] Subject Restrictions: During the study, subjects are prohibited from using or taking any reagents that could affect the test. Cleaning products and usage of the test area should be consistent with daily routine. Subjects should primarily remain indoors and avoid prolonged exposure to light. If a subject experiences an adverse event or violates the study protocol during the study, or if the researcher deems the subject unsuitable for continuing the study, the subject must withdraw from the clinical study and state the reason for withdrawal.

[0147] 7.2 Safety Assessment: Adverse reactions are judged according to the following three-level criteria:

[0148] Mild: easily tolerated, causing minimal discomfort and not affecting normal life.

[0149] Moderate: Sufficient to interfere with normal life.

[0150] Severe: interferes with normal life.

[0151] Relationship of adverse reactions to the study:

[0152] Unrelated: obviously related to other factors; Impossible: very likely related to other factors; Possible: occurs after use, but may also be caused by other factors; Likely: occurs after use, but cannot be explained by other factors; Very likely: occurs after use, but cannot be explained by other factors, and reappears after use.

[0153] 7.3 Subjects

[0154] A total of 30 subjects were included, with 0 dropouts. The number of valid cases was 30, including 8 males and 22 females. The youngest age was 21 years old, the oldest was 39 years old, and the average age was 29.0 years old.

[0155] 7.4 Test steps:

[0156] 7.4.1 Screening:

[0157] Recruit subjects as required and have them sign informed consent forms; ask subjects relevant questions based on the inclusion and exclusion criteria, and record them together with the test screening results.

[0158] 7.4.2 Cleaning:

[0159] Before the test begins, the research center must provide the subjects with a facial cleansing product that has no therapeutic effect. The test area should be cleaned and then dried with a lint-free paper towel. The cleaning product and water temperature should remain consistent for each follow-up visit.

[0160] 7.4.3 Testing:

[0161] Eligible subjects underwent baseline skin assessment and testing prior to product use, including instrument testing and standard image acquisition and data recording. The same testing was repeated 28 days after product use, for a total of two testing time points (D0 and D28).

[0162] a) Epidermal water loss test:

[0163] At each visit point, the transepidermal water loss (TEWL) of the skin was tested using an HQ-TEWL transepidermal moisture tester. The test was performed three times and the average value was taken.

[0164] b) Hemoglobin content test:

[0165] At each visit time point, skin melanin and hemoglobin tester ( MX18) Test the hemoglobin content of the test site three times and take the average value.

[0166] 7.4.4 Sample issuance:

[0167] The test product will be distributed to the subjects, along with instructions for use that clearly define how to use the product (including the intended area of ​​application, method of use, dosage per application, frequency of use, and precautions). This ensures that the subjects use the product correctly and continuously until the end of the testing period. At the final follow-up visit, the subjects will bring the test product home, and the researcher will weigh and record the amount of test product used.

[0168] 7.4.5 Adverse reaction records:

[0169] If a subject experiences an adverse skin reaction during product use, the test should be terminated immediately and the subject should receive appropriate medical treatment. Adverse reactions should be recorded.

[0170] 7.5 Test site: face.

[0171] 7.6 Instructions for use: Take 0.3-0.5g of repair cream in the palm of your hand, apply evenly on the face, and massage gently until absorbed.

[0172] 8 Statistical analysis methods:

[0173] 8.1 Data Analysis Methods:

[0174] 8.1.1 Descriptive Statistics: Statistical analysis software shall be used to perform descriptive statistics on the test values ​​of each tested part, including quantity, mean, standard deviation, minimum value, median value and maximum value.

[0175] 8.1.2 Analysis of Differences: Calculate the difference between the initial value of the test site and the values ​​measured at other time points. If the instrumental analysis data conform to a normal distribution, perform statistical analysis using the paired t-test. If the test data do not conform to a normal distribution, perform statistical analysis using the Wilcoxon signed-rank test. All statistical methods use a two-tailed test, with a significance level of α = 0.05. Significance symbols: "ns" indicates no statistical difference, P ≥ 0.050; P < 0.050 indicates a significant difference; 0.010 ≤ P < 0.050, denoted by "*"; 0.001 ≤ P < 0.010, denoted by "**"; P < 0.001, denoted by "***".

[0176] 8.1.3 Subject self-assessment: Calculate the percentage of satisfaction with the questionnaire.

[0177] 8.2 Result determination:

[0178] After using the product during the test period, if the measured values ​​of the skin's transepidermal water loss and hemoglobin content are significantly different from the initial values ​​(p<0.05), and the mean of the measured values ​​is better than the initial value, then the test product is deemed to have a repair effect during the test period.

[0179] 9. Analysis of test results:

[0180] 9.1 Test results of epidermal water loss at the test site, see Figure 12 .

[0181] On D0, TEWL values ​​were measured in 30 subjects, with a mean of 27.52, a median of 27.09, a standard deviation of 2.97, a minimum of 21.89, and a maximum of 32.55. On D28, TEWL values ​​were measured in 30 subjects, with a mean of 18.89, a median of 18.29, a standard deviation of 2.45, a minimum of 15.65, and a maximum of 23.36. The calculated rate of change in TEWL values ​​on D28 was -31.36%, which was statistically significant using a paired T-test. Rate of change = (D28 mean - D0 mean) / D0 mean * 100%.

[0182] 9.1.1 Descriptive Statistics of Transepidermal Water Loss

[0183] After continuous use of the cream obtained in Example 4 for 28 days, the transepidermal water loss value of the subjects decreased by 31.36% compared with the initial value. The D28 measurement value was significantly different from the D0 initial value (p < 0.05), and the measurement values ​​were better than the initial value.

[0184] 9.2 The test results of skin hemoglobin content in the test site are shown in Table 4, including the test pictures of two volunteers. Figure 13 shown.

[0185] After using the sample continuously for 28 days, the subjects' skin hemoglobin content decreased by 27.52% compared with the initial value. The D28 measurement values ​​were significantly different from the D0 initial values ​​(p < 0.05), and the measurement values ​​were better than the initial values.

[0186] Table 4 Test results of skin hemoglobin content in the tested parts

[0187] Initial value D0 average value Initial value D28 average value D28 change rate 262.72±4.51 190.43±4.65 -27.52％

[0188] Note: The data in the table are mean ± standard error.

[0189] 9.3 Subject Questionnaire Results

[0190] The results of the subjects’ self-assessment satisfaction survey are shown in Table 5.

[0191] Table 5 Results of the self-assessment satisfaction survey of the subjects

[0192]

[0193]

[0194] Note: 1 point means strongly disagree, 2 points means disagree, 3 points means neutral / neutral, 4 points means agree, and 5 points means strongly agree.

[0195] After using the test product for 28 consecutive days, the results of the questionnaire survey on the subjects' satisfaction with the product showed:

[0196] Q1: 96.67% of the subjects agreed that the product had a pleasant smell;

[0197] Q2: 93.33% of the subjects agreed that the product made the skin feel refreshed and non-sticky when used, and was easy to apply;

[0198] Q3: 96.67% of the subjects agreed that their skin felt delicate and smooth after using the product;

[0199] Q4: 96.67% of the subjects agreed that the product is easily permeable and absorbed;

[0200] Q5: 100.00% of the subjects agreed that the product was very hydrating after use;

[0201] Q6: 90.00% of the subjects agreed that the product had an effect on improving skin irritation such as redness, itching, and stinging after use;

[0202] Q7: 96.67% of the subjects agreed that this product can effectively strengthen the skin barrier after use;

[0203] Q8: 100.00% of the subjects agreed that they were generally satisfied with the product after using it.

[0204] 10. Conclusion

[0205] This test refers to the standard T / CAB 0152-2022 "Test Methods for Seven Efficacy of Cosmetics: Anti-wrinkle, Firming, Moisturizing, Oil Control, Repairing, Nourishing, and Soothing" to conduct a human efficacy evaluation test on the repair efficacy of the test samples. A total of 30 subjects participated in this test, of which 0 withdrew, and there were 30 valid subjects.

[0206] (1) Test results: After 30 subjects used the cream obtained in Example 4 continuously for 28 days, the transepidermal water loss of the skin decreased by 31.36%, and the skin hemoglobin content decreased by 27.52%. The D28 measured values ​​were significantly different from the initial values ​​on D0 (P<0.05), and the measured values ​​were better than the initial values. The results show that the cream obtained in Example 4 has a repairing effect on facial skin.

[0207] (2) Self-evaluation results of the subjects: The number of subjects with satisfaction scores of ≥4 points for each question was no less than 27; among them, 90.00% of the subjects agreed that the product had an effect on improving skin irritation such as redness, itching, and stinging after use; and 96.67% of the subjects agreed that the product could effectively strengthen the skin barrier after use.

[0208] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A composition having the effect of repairing skin barrier damage, characterized in that: The invention comprises the following raw materials in parts by weight: 16-24 parts of polygonatum, 8-12 parts of bletilla striata, 8-12 parts of sophora flavescens and 4-6 parts of liquorice.

2. The method for preparing the composition according to claim 1, wherein The method comprises the following steps: mixing polygonatum, bletilla striata, sophora flavescens, liquorice and ethanol solution, performing reflux extraction, and collecting medicinal residues and medicinal liquid 1; mixing the medicinal residues with ethanol solution, performing reflux extraction, and collecting medicinal liquid 2; The medicinal solution 1 and the medicinal solution 2 are mixed and concentrated to obtain an extract; the extract is mixed with water for water precipitation, the precipitate is removed, and the extract is concentrated to obtain a composition.

3. The preparation method according to claim 2, characterized in that The volume fraction of the ethanol solution is 50% to 75%; the mass ratio of the total mass of polygonatum, bletilla striata, sophora flavescens and liquorice to the ethanol solution is 1:6 to 10.

4. The preparation method according to claim 2, characterized in that The reflux extraction time is 0.5h to 2h, and the reflux extraction temperature is 80°C to 100°C.

5. The preparation method according to claim 2, characterized in that The mass ratio of the medicinal residue to the ethanol solution is 1:6-10.

6. The preparation method according to claim 2, characterized in that The mass volume ratio of the extract to water is 1g:2mL~6mL; the water sinking time is 10h~15h.

7. Use of the composition according to claim 1 or the preparation method according to any one of claims 2 to 6 in preparing a product for repairing skin barrier damage.

8. Use of the composition according to claim 1 or the preparation method according to any one of claims 2 to 6 in the preparation of products with anti-inflammatory, moisturizing and / or soothing effects.

9. A skin care product with the effects of repairing skin barrier damage, anti-inflammatory, moisturizing and / or soothing, characterized in that: The skin care product is composed of the composition according to claim 1 and auxiliary materials, wherein the mass of the composition is 5% to 10% of the total mass of the skin care product.

10. The skin care product according to claim 1, characterized in that The types of skin care products include creams, lotions, essences, toners or masks.

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